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    Product is the lyophilized powder of goat antiserum to mouse immunoglobulins IgG IgA IgM and buffer salts
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    95
    Millipore iga
    Iga, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 2032 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    IGAN Bio iga nephropathy
    Biochemical features of purified <t>IgA1</t> mAbs. (A) Detection of J chain by Western blot. (B) Analysis of monomeric (mIgA) and polymeric (pIgA) forms by Western blot after SDS-PAGE in nonreducing conditions. (C) Glycosylation of IgA1 mAbs that deposit or
    Iga Nephropathy, supplied by IGAN Bio, used in various techniques. Bioz Stars score: 92/100, based on 7694 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher iga
    RV-specific antibody responses in sera from children with acute wheeze. Frequencies and levels of a <t>IgG</t> and b <t>IgA</t> responses ( y -axes: n ) ( x -axes: red: RV-A species; green: RV-B species; blue: RV-C species). Antibody levels are color-coded and expressed as ISAC standardized units, ISU-G and ISU-A, respectively. c Median IgG levels ( y -axis: ISU-G) to VP1 peptides ( x -axis) in children grouped according to age (6–12 months: squares; 13–24 months: triangles; 25–42: circles). d Spearman’s rank correlation between the number of IgG-reactive peptides ( n , y -axis; median IgG > 15 ISU) and age ( x -axis: months). Correlation coefficient ( ρ ) and p -value are shown. e Comparison of the number of IgG-reactive VP1 peptides ( n , y -axis; median IgG > 15 ISU) in children according to age ( x -axis). Horizontal lines indicate medians. Statistically significant differences between groups are indicated (** p
    Iga, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1654 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher f ab 2 goat anti human igg iga igm secondary antibody
    RV-specific antibody responses in sera from children with acute wheeze. Frequencies and levels of a <t>IgG</t> and b <t>IgA</t> responses ( y -axes: n ) ( x -axes: red: RV-A species; green: RV-B species; blue: RV-C species). Antibody levels are color-coded and expressed as ISAC standardized units, ISU-G and ISU-A, respectively. c Median IgG levels ( y -axis: ISU-G) to VP1 peptides ( x -axis) in children grouped according to age (6–12 months: squares; 13–24 months: triangles; 25–42: circles). d Spearman’s rank correlation between the number of IgG-reactive peptides ( n , y -axis; median IgG > 15 ISU) and age ( x -axis: months). Correlation coefficient ( ρ ) and p -value are shown. e Comparison of the number of IgG-reactive VP1 peptides ( n , y -axis; median IgG > 15 ISU) in children according to age ( x -axis). Horizontal lines indicate medians. Statistically significant differences between groups are indicated (** p
    F Ab 2 Goat Anti Human Igg Iga Igm Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher peroxidase conjugated anti rabbit
    RV-specific antibody responses in sera from children with acute wheeze. Frequencies and levels of a <t>IgG</t> and b <t>IgA</t> responses ( y -axes: n ) ( x -axes: red: RV-A species; green: RV-B species; blue: RV-C species). Antibody levels are color-coded and expressed as ISAC standardized units, ISU-G and ISU-A, respectively. c Median IgG levels ( y -axis: ISU-G) to VP1 peptides ( x -axis) in children grouped according to age (6–12 months: squares; 13–24 months: triangles; 25–42: circles). d Spearman’s rank correlation between the number of IgG-reactive peptides ( n , y -axis; median IgG > 15 ISU) and age ( x -axis: months). Correlation coefficient ( ρ ) and p -value are shown. e Comparison of the number of IgG-reactive VP1 peptides ( n , y -axis; median IgG > 15 ISU) in children according to age ( x -axis). Horizontal lines indicate medians. Statistically significant differences between groups are indicated (** p
    Peroxidase Conjugated Anti Rabbit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 547 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    EUROIMMUN vca iga
    RV-specific antibody responses in sera from children with acute wheeze. Frequencies and levels of a <t>IgG</t> and b <t>IgA</t> responses ( y -axes: n ) ( x -axes: red: RV-A species; green: RV-B species; blue: RV-C species). Antibody levels are color-coded and expressed as ISAC standardized units, ISU-G and ISU-A, respectively. c Median IgG levels ( y -axis: ISU-G) to VP1 peptides ( x -axis) in children grouped according to age (6–12 months: squares; 13–24 months: triangles; 25–42: circles). d Spearman’s rank correlation between the number of IgG-reactive peptides ( n , y -axis; median IgG > 15 ISU) and age ( x -axis: months). Correlation coefficient ( ρ ) and p -value are shown. e Comparison of the number of IgG-reactive VP1 peptides ( n , y -axis; median IgG > 15 ISU) in children according to age ( x -axis). Horizontal lines indicate medians. Statistically significant differences between groups are indicated (** p
    Vca Iga, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 90/100, based on 95 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    SouthernBiotech goat anti mouse iga
    RV-specific antibody responses in sera from children with acute wheeze. Frequencies and levels of a <t>IgG</t> and b <t>IgA</t> responses ( y -axes: n ) ( x -axes: red: RV-A species; green: RV-B species; blue: RV-C species). Antibody levels are color-coded and expressed as ISAC standardized units, ISU-G and ISU-A, respectively. c Median IgG levels ( y -axis: ISU-G) to VP1 peptides ( x -axis) in children grouped according to age (6–12 months: squares; 13–24 months: triangles; 25–42: circles). d Spearman’s rank correlation between the number of IgG-reactive peptides ( n , y -axis; median IgG > 15 ISU) and age ( x -axis: months). Correlation coefficient ( ρ ) and p -value are shown. e Comparison of the number of IgG-reactive VP1 peptides ( n , y -axis; median IgG > 15 ISU) in children according to age ( x -axis). Horizontal lines indicate medians. Statistically significant differences between groups are indicated (** p
    Goat Anti Mouse Iga, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 94/100, based on 315 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    IGAN Bio iga1
    SPR analysis of lectin binding to <t>IgA1</t> myeloma proteins
    Iga1, supplied by IGAN Bio, used in various techniques. Bioz Stars score: 91/100, based on 62 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Agilent technologies iga
    <t>IgA</t> characterization in the indicated tissue of STAM and MUP-uPA mice a – e , Single splenocyte or liver cell suspensions from tumour (HCC)-bearing mice (STAM-B6) were stained with CD45, CD19, IgA, <t>CD138,</t> B220, and PD-L1, and FVD-eF780 was used to exclude dead cells. a , The gating strategies for splenocytes and liver lymphocytes: lymphocyte gate, dead cell exclusion, doublets exclusion, and CD45 + population gate. b , Flow cytometry analysis of IgA and CD19 expression of indicated strains, gated on the CD45 + population. Spleen or liver from Iga −/− mice was used to set up the gating for the populations. Tgfbr2 ΔB and Pdl1/2 −/− mice clearly showed less IgA + cells than WT mice. c – e , IgA subpopulations were gated as indicated and analysed for CD138, B220, and PD-L1 expression. IgA subpopulations were gated on the basis of CD19 expression, in the indicated strains, and the IgA + CD19 + , IgA + CD19 −/low/int , and IgA − CD19 + subpopulations were further analysed for their B220 and CD138 expression. The decisions for CD19 and CD138 levels were also based on their mean fluorescence intensity (MFI; red). The analyses showed two main populations: (1) IgA + CD19 + B220 + CD138 low/− and (2) IgA + CD19 −/int B220 − CD138 +/hi . These two populations and the IgA − CD19 + populations were further analysed for their ability to express PD-L1 (percentage and mean fluorescence intensity). f – j , Single-cell suspensions were prepared from the spleen, liver, or intestine of MUP-uPA mice kept on normal chow or HFD, and were stained with CD45, CD19, IgA, B220, CD138, CD11b, MHCII, PD-L1, and CD5, and analysed by flow cytometry. f , The B220 and CD138 expression in total IgA + cells (CD19 + and CD19 − populations) confirmed that most of the IgA + cells in spleen and liver of HFD-fed MUP-uPA mice are CD138 + cells. g , h , IgA subpopulations in liver ( g ) and spleen ( h ) were gated as indicated and analysed for CD138, B220, and MHCII expression. i , Representative dot-plots gated on IgA + cells, showing that most of the IgA + cells are not CD11b + . j , PD-L1 and CD5 expression of IgA + cells in spleen, liver, and intestine.
    Iga, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 93/100, based on 1479 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Becton Dickinson iga
    Sensitization to peanut by non-oral routes. TLR9+/+ and −/− mice were sensitized to peanut by intraperitoneal injection with alum (IP), or by repeated epicutaneous exposure (skin). A) Body temperature measured 30 min after intraperitoneal challenge with 500 g of peanut extract. B) IgE, C) <t>IgA,</t> and D) <t>IgG1</t> antibodies were measured in serum obtained 1-2 days prior to challenge. * p
    Iga, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 98/100, based on 916 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 98 stars, based on 916 article reviews
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    iga - by Bioz Stars, 2020-12
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    99
    SouthernBiotech iga antibodies
    Sensitization to peanut by non-oral routes. TLR9+/+ and −/− mice were sensitized to peanut by intraperitoneal injection with alum (IP), or by repeated epicutaneous exposure (skin). A) Body temperature measured 30 min after intraperitoneal challenge with 500 g of peanut extract. B) IgE, C) <t>IgA,</t> and D) <t>IgG1</t> antibodies were measured in serum obtained 1-2 days prior to challenge. * p
    Iga Antibodies, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 99/100, based on 215 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    SouthernBiotech goat f
    Sensitization to peanut by non-oral routes. TLR9+/+ and −/− mice were sensitized to peanut by intraperitoneal injection with alum (IP), or by repeated epicutaneous exposure (skin). A) Body temperature measured 30 min after intraperitoneal challenge with 500 g of peanut extract. B) IgE, C) <t>IgA,</t> and D) <t>IgG1</t> antibodies were measured in serum obtained 1-2 days prior to challenge. * p
    Goat F, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 96/100, based on 692 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    iga  (Bio-Rad)
    95
    Bio-Rad iga
    Sensitization to peanut by non-oral routes. TLR9+/+ and −/− mice were sensitized to peanut by intraperitoneal injection with alum (IP), or by repeated epicutaneous exposure (skin). A) Body temperature measured 30 min after intraperitoneal challenge with 500 g of peanut extract. B) IgE, C) <t>IgA,</t> and D) <t>IgG1</t> antibodies were measured in serum obtained 1-2 days prior to challenge. * p
    Iga, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 95/100, based on 552 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Glycoform iga1
    Hypothesis on the pathogenesis of IgA nephropathy. Synthesis of <t>IgA1</t> with some O -glycans deficient in galactose (autoantigen) is elevated. Gd-IgA1 is present in the circulation at increased levels (hit 1). This immunoglobulin is recognized by unique circulating
    Iga1, supplied by Glycoform, used in various techniques. Bioz Stars score: 91/100, based on 54 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Millipore human iga
    Hypothesis on the pathogenesis of IgA nephropathy. Synthesis of <t>IgA1</t> with some O -glycans deficient in galactose (autoantigen) is elevated. Gd-IgA1 is present in the circulation at increased levels (hit 1). This immunoglobulin is recognized by unique circulating
    Human Iga, supplied by Millipore, used in various techniques. Bioz Stars score: 96/100, based on 223 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    EUROIMMUN euroimmun iga
    Comparison of ten serological assays for the detection of anti-SARS CoV-2 IgG. As for in Fig 2 , the same 110 serum samples were assessed for the presence of anti-SARS CoV-2 IgG. Each sample was assayed using an in-house anti-S ELISA (shown in the graph across the top of each panel, black bars), seven colloidal gold lateral flow tests (Deep Blue, Accu-Tell, GenBody, SureScreen, Spring, Biohit and Medomics), and a commercial ELISA <t>(EUROIMMUN).</t> A chemiluminescent assay for total anti-SARS CoV-2 IgM, IgG and <t>IgA</t> (Watmind) was also included. The threshold for a positive result in the in-house ELISA is set at 4-fold above background, as indicated by the red dashed line. Results for the other tests are represented as heatmaps, with colour intensity corresponding to strength of signal for each test. For EUROIMMUN, scores of
    Euroimmun Iga, supplied by EUROIMMUN, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    iga  (Abcam)
    95
    Abcam iga
    Associations between the plasma levels of APRIL and immunoglobulin production. (A) Associations between the APRIL levels and HIV-1-specific <t>IgM</t> in HIV-1-infected individuals ( n = 59). (B) Associations between the APRIL levels and HIV-1-specific <t>IgA</t> in HIV-1-infected individuals ( n = 59). (C) Associations between the APRIL levels and neutralization titers in untreated HIV-1-infected individuals ( n = 46). (D) Associations between the APRIL levels and neutralization breadth in untreated HIV-1-infected individuals ( n = 46). LTNPs-B, long-term non-progressors at baseline; LTNPs-L, long-term non-progressors at the latest measurement; TPs, typical progressors; ART, antiretroviral therapy; ID 50 , 50% inhibitory dilution. Intergroup comparisons were performed using a Kruskal–Wallis test, followed by Dunn's post-test; p and r values were calculated by Spearman's rank correlation tests (* p
    Iga, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 185 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    IGAN Bio iga c3 ratio
    Associations between the plasma levels of APRIL and immunoglobulin production. (A) Associations between the APRIL levels and HIV-1-specific <t>IgM</t> in HIV-1-infected individuals ( n = 59). (B) Associations between the APRIL levels and HIV-1-specific <t>IgA</t> in HIV-1-infected individuals ( n = 59). (C) Associations between the APRIL levels and neutralization titers in untreated HIV-1-infected individuals ( n = 46). (D) Associations between the APRIL levels and neutralization breadth in untreated HIV-1-infected individuals ( n = 46). LTNPs-B, long-term non-progressors at baseline; LTNPs-L, long-term non-progressors at the latest measurement; TPs, typical progressors; ART, antiretroviral therapy; ID 50 , 50% inhibitory dilution. Intergroup comparisons were performed using a Kruskal–Wallis test, followed by Dunn's post-test; p and r values were calculated by Spearman's rank correlation tests (* p
    Iga C3 Ratio, supplied by IGAN Bio, used in various techniques. Bioz Stars score: 95/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Thermo Fisher goat anti mouse igg iga igm h l secondary antibody
    Associations between the plasma levels of APRIL and immunoglobulin production. (A) Associations between the APRIL levels and HIV-1-specific <t>IgM</t> in HIV-1-infected individuals ( n = 59). (B) Associations between the APRIL levels and HIV-1-specific <t>IgA</t> in HIV-1-infected individuals ( n = 59). (C) Associations between the APRIL levels and neutralization titers in untreated HIV-1-infected individuals ( n = 46). (D) Associations between the APRIL levels and neutralization breadth in untreated HIV-1-infected individuals ( n = 46). LTNPs-B, long-term non-progressors at baseline; LTNPs-L, long-term non-progressors at the latest measurement; TPs, typical progressors; ART, antiretroviral therapy; ID 50 , 50% inhibitory dilution. Intergroup comparisons were performed using a Kruskal–Wallis test, followed by Dunn's post-test; p and r values were calculated by Spearman's rank correlation tests (* p
    Goat Anti Mouse Igg Iga Igm H L Secondary Antibody, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 96/100, based on 97 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Becton Dickinson antibodies immunoglobulin a
    Temporal immune responses to OMVs derived from NTHi strain 3198-R. Shown are the median titers over time of IgM (A), <t>IgA</t> (B), and IgG1 (C) antibodies to 3198-R OMVs in sera from mice intranasally immunized with either IM-1 (solid line) or IM-2 (dashed line) as well as in sera from nonvaccinated control mice (dotted line) (n = 20 for each group). The error bars indicate the interquartile range of each data set for each time point.
    Antibodies Immunoglobulin A, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    SouthernBiotech iga
    Effect of immunization route on response. Ten mice per group were immunized three times with a 2-week interval between immunizations via intranasal (IN) or subcutaneous (SC) route or a combination of these using GLA-3M-052-PEG2000 liposomes as an adjuvant. Samples were collected 1 week after 3rd immunization. a Stool supernatants were diluted 400-fold to determine LecA-specific <t>IgA</t> titer by ELISA. b Plasma samples were diluted 100,000-fold to determine titers of IgG1 (black circles) and IgG2a (red circles) subtypes. c Splenocytes were restimulated with LecA for 72 h and production of extracellular cytokines in the culture supernatants was determined by Luminex. Cytokine levels of the unstimulated samples were at the baseline (not shown). Error bars represent standard error of the mean. For clarity, statistical significance vs. the adjuvant alone control groups is not represented but is detailed below. For the analysis of the data in plot a , Welch’s one-way ANOVA was employed with Games–Howell correction for multiple comparisons; all of the vaccine groups except for SC + SC + SC were statistically different ( p
    Iga, supplied by SouthernBiotech, used in various techniques. Bioz Stars score: 98/100, based on 2053 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    N/A
    Polyclonal Antibody to Immunoglobulin A IgA
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    N/A
    Immunoglobulin A1 Antibody is a Rabbit Polyclonal against Immunoglobulin A1
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    N/A
    Immunoglobulin A2 IgA2 Antibody is a Mouse Monoclonal antibody against Immunoglobulin A2 IgA2
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    Image Search Results


    Biochemical features of purified IgA1 mAbs. (A) Detection of J chain by Western blot. (B) Analysis of monomeric (mIgA) and polymeric (pIgA) forms by Western blot after SDS-PAGE in nonreducing conditions. (C) Glycosylation of IgA1 mAbs that deposit or

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: IgA Structure Variations Associate with Immune Stimulations and IgA Mesangial Deposition

    doi: 10.1681/ASN.2015080911

    Figure Lengend Snippet: Biochemical features of purified IgA1 mAbs. (A) Detection of J chain by Western blot. (B) Analysis of monomeric (mIgA) and polymeric (pIgA) forms by Western blot after SDS-PAGE in nonreducing conditions. (C) Glycosylation of IgA1 mAbs that deposit or

    Article Snippet: Suzuki H, Fan R, Zhang Z, Brown R, Hall S, Julian BA, Chatham WW, Suzuki Y, Wyatt RJ, Moldoveanu Z, Lee JY, Robinson J, Tomana M, Tomino Y, Mestecky J, Novak J.: Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity.

    Techniques: Purification, Western Blot, SDS Page

    Calculated pI of combined VH and V κ CDR regions from α 1KI IgA1 hybridomas. Amino acid sequences of VH and V κ CDR from hybridomas 1–10 were combined to calculate pI; α 1KI hybridomas yielding (circles) or not yielding

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: IgA Structure Variations Associate with Immune Stimulations and IgA Mesangial Deposition

    doi: 10.1681/ASN.2015080911

    Figure Lengend Snippet: Calculated pI of combined VH and V κ CDR regions from α 1KI IgA1 hybridomas. Amino acid sequences of VH and V κ CDR from hybridomas 1–10 were combined to calculate pI; α 1KI hybridomas yielding (circles) or not yielding

    Article Snippet: Suzuki H, Fan R, Zhang Z, Brown R, Hall S, Julian BA, Chatham WW, Suzuki Y, Wyatt RJ, Moldoveanu Z, Lee JY, Robinson J, Tomana M, Tomino Y, Mestecky J, Novak J.: Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity.

    Techniques:

    IgA production and deposition vary with housing conditions. Influence of environmental stimuli on (A) mesangial IgA deposition and (B) plasma IgA levels in 6-week-old and 3-month-old α 1KI mice housed in GF, SOPF, or CIS facilities. Homogeneous

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: IgA Structure Variations Associate with Immune Stimulations and IgA Mesangial Deposition

    doi: 10.1681/ASN.2015080911

    Figure Lengend Snippet: IgA production and deposition vary with housing conditions. Influence of environmental stimuli on (A) mesangial IgA deposition and (B) plasma IgA levels in 6-week-old and 3-month-old α 1KI mice housed in GF, SOPF, or CIS facilities. Homogeneous

    Article Snippet: Suzuki H, Fan R, Zhang Z, Brown R, Hall S, Julian BA, Chatham WW, Suzuki Y, Wyatt RJ, Moldoveanu Z, Lee JY, Robinson J, Tomana M, Tomino Y, Mestecky J, Novak J.: Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity.

    Techniques: Mouse Assay

    In vivo deposited IgA in RAG-2/ γ c–deficient mice after intraperitoneal grafting of α 1KI hybridomas 1, 5, 6, 9, and 10. Each hybridoma was injected in two mice, which were euthanized after tumor development. Ctrl− stands

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: IgA Structure Variations Associate with Immune Stimulations and IgA Mesangial Deposition

    doi: 10.1681/ASN.2015080911

    Figure Lengend Snippet: In vivo deposited IgA in RAG-2/ γ c–deficient mice after intraperitoneal grafting of α 1KI hybridomas 1, 5, 6, 9, and 10. Each hybridoma was injected in two mice, which were euthanized after tumor development. Ctrl− stands

    Article Snippet: Suzuki H, Fan R, Zhang Z, Brown R, Hall S, Julian BA, Chatham WW, Suzuki Y, Wyatt RJ, Moldoveanu Z, Lee JY, Robinson J, Tomana M, Tomino Y, Mestecky J, Novak J.: Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity.

    Techniques: In Vivo, Mouse Assay, Injection

    IgA glycosylation varies with housing conditions. Analysis of (A) N -acetylneuraminic acid and (B) exposed GalNac in plasma IgA from 6-week-old and 3-month-old α 1KI, α 1KI J−/−, and wild-type (WT) mice housed in GF, SOPF,

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: IgA Structure Variations Associate with Immune Stimulations and IgA Mesangial Deposition

    doi: 10.1681/ASN.2015080911

    Figure Lengend Snippet: IgA glycosylation varies with housing conditions. Analysis of (A) N -acetylneuraminic acid and (B) exposed GalNac in plasma IgA from 6-week-old and 3-month-old α 1KI, α 1KI J−/−, and wild-type (WT) mice housed in GF, SOPF,

    Article Snippet: Suzuki H, Fan R, Zhang Z, Brown R, Hall S, Julian BA, Chatham WW, Suzuki Y, Wyatt RJ, Moldoveanu Z, Lee JY, Robinson J, Tomana M, Tomino Y, Mestecky J, Novak J.: Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity.

    Techniques: Mouse Assay

    Role of mAb V regions in kidney deposition. IgA1 mAb6 or its IgG1 switched variant was injected in immunodeficient mice 2 hours before euthanasia. IgA1 and IgG1 were revealed on kidney sections with FITC-labeled (green) antisera against either (upper

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: IgA Structure Variations Associate with Immune Stimulations and IgA Mesangial Deposition

    doi: 10.1681/ASN.2015080911

    Figure Lengend Snippet: Role of mAb V regions in kidney deposition. IgA1 mAb6 or its IgG1 switched variant was injected in immunodeficient mice 2 hours before euthanasia. IgA1 and IgG1 were revealed on kidney sections with FITC-labeled (green) antisera against either (upper

    Article Snippet: Suzuki H, Fan R, Zhang Z, Brown R, Hall S, Julian BA, Chatham WW, Suzuki Y, Wyatt RJ, Moldoveanu Z, Lee JY, Robinson J, Tomana M, Tomino Y, Mestecky J, Novak J.: Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity.

    Techniques: Variant Assay, Injection, Mouse Assay, Labeling

    Influence of environmental stimuli on the monomeric-to-polymeric ratio of circulating IgA in groups of young (6–8 weeks old) and adult (3–4 months old) mice housed in GF, SOPF, or CIS facilities. (A) Plasma proteins were separated under

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: IgA Structure Variations Associate with Immune Stimulations and IgA Mesangial Deposition

    doi: 10.1681/ASN.2015080911

    Figure Lengend Snippet: Influence of environmental stimuli on the monomeric-to-polymeric ratio of circulating IgA in groups of young (6–8 weeks old) and adult (3–4 months old) mice housed in GF, SOPF, or CIS facilities. (A) Plasma proteins were separated under

    Article Snippet: Suzuki H, Fan R, Zhang Z, Brown R, Hall S, Julian BA, Chatham WW, Suzuki Y, Wyatt RJ, Moldoveanu Z, Lee JY, Robinson J, Tomana M, Tomino Y, Mestecky J, Novak J.: Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity.

    Techniques: Mouse Assay

    Kinetics of plasma IgA levels and IgA deposition in glomeruli of 1-week-old to 3-month-old α 1KI and α 1KI J−/− mice. (A) IgA was measured by ELISA in mice housed in CIS facilities (means±SEM of n =4–8 mice;

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: IgA Structure Variations Associate with Immune Stimulations and IgA Mesangial Deposition

    doi: 10.1681/ASN.2015080911

    Figure Lengend Snippet: Kinetics of plasma IgA levels and IgA deposition in glomeruli of 1-week-old to 3-month-old α 1KI and α 1KI J−/− mice. (A) IgA was measured by ELISA in mice housed in CIS facilities (means±SEM of n =4–8 mice;

    Article Snippet: Suzuki H, Fan R, Zhang Z, Brown R, Hall S, Julian BA, Chatham WW, Suzuki Y, Wyatt RJ, Moldoveanu Z, Lee JY, Robinson J, Tomana M, Tomino Y, Mestecky J, Novak J.: Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    IgA1 mesangial deposits in adult α 1KI mice housed in CIS facilities. (A) Kidney section stained with FITC–conjugated anti–human IgA (green). (B) Electron microscopy of kidney section: white arrow points to IgA deposits. (C) Electron

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: IgA Structure Variations Associate with Immune Stimulations and IgA Mesangial Deposition

    doi: 10.1681/ASN.2015080911

    Figure Lengend Snippet: IgA1 mesangial deposits in adult α 1KI mice housed in CIS facilities. (A) Kidney section stained with FITC–conjugated anti–human IgA (green). (B) Electron microscopy of kidney section: white arrow points to IgA deposits. (C) Electron

    Article Snippet: Suzuki H, Fan R, Zhang Z, Brown R, Hall S, Julian BA, Chatham WW, Suzuki Y, Wyatt RJ, Moldoveanu Z, Lee JY, Robinson J, Tomana M, Tomino Y, Mestecky J, Novak J.: Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity.

    Techniques: Mouse Assay, Staining, Electron Microscopy

    Western blot analysis of trypsin digestion products. Nine IgA1 mAbs were submitted to 10% SDS-PAGE after various digestion times (0–10 hours) by trypsin. Western blot was performed to reveal IgA determinants using HRP–linked goat anti–human

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: IgA Structure Variations Associate with Immune Stimulations and IgA Mesangial Deposition

    doi: 10.1681/ASN.2015080911

    Figure Lengend Snippet: Western blot analysis of trypsin digestion products. Nine IgA1 mAbs were submitted to 10% SDS-PAGE after various digestion times (0–10 hours) by trypsin. Western blot was performed to reveal IgA determinants using HRP–linked goat anti–human

    Article Snippet: Suzuki H, Fan R, Zhang Z, Brown R, Hall S, Julian BA, Chatham WW, Suzuki Y, Wyatt RJ, Moldoveanu Z, Lee JY, Robinson J, Tomana M, Tomino Y, Mestecky J, Novak J.: Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity.

    Techniques: Western Blot, SDS Page

    Circular dichroism analysis of IgA1 mAbs. (A) Far-ultraviolet spectra of six mAbs acquired at 20°C with a path length of 0.2 cm on a spectropolarimeter. IgA1 mAb1 and IgA1 mAb6 yield IgA deposition, and the others do not. (B) Thermal unfolding

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: IgA Structure Variations Associate with Immune Stimulations and IgA Mesangial Deposition

    doi: 10.1681/ASN.2015080911

    Figure Lengend Snippet: Circular dichroism analysis of IgA1 mAbs. (A) Far-ultraviolet spectra of six mAbs acquired at 20°C with a path length of 0.2 cm on a spectropolarimeter. IgA1 mAb1 and IgA1 mAb6 yield IgA deposition, and the others do not. (B) Thermal unfolding

    Article Snippet: Suzuki H, Fan R, Zhang Z, Brown R, Hall S, Julian BA, Chatham WW, Suzuki Y, Wyatt RJ, Moldoveanu Z, Lee JY, Robinson J, Tomana M, Tomino Y, Mestecky J, Novak J.: Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity.

    Techniques:

    Analysis of IgA deposition in the kidney of immunodeficient mice 2 hours after injection of human IgA1 mAb. Clones 1–10 were injected in two mice each. Ctrl− is a negative control after injection of saline. (A) IgA deposits in kidney sections

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: IgA Structure Variations Associate with Immune Stimulations and IgA Mesangial Deposition

    doi: 10.1681/ASN.2015080911

    Figure Lengend Snippet: Analysis of IgA deposition in the kidney of immunodeficient mice 2 hours after injection of human IgA1 mAb. Clones 1–10 were injected in two mice each. Ctrl− is a negative control after injection of saline. (A) IgA deposits in kidney sections

    Article Snippet: Suzuki H, Fan R, Zhang Z, Brown R, Hall S, Julian BA, Chatham WW, Suzuki Y, Wyatt RJ, Moldoveanu Z, Lee JY, Robinson J, Tomana M, Tomino Y, Mestecky J, Novak J.: Aberrantly glycosylated IgA1 in IgA nephropathy patients is recognized by IgG antibodies with restricted heterogeneity.

    Techniques: Mouse Assay, Injection, Clone Assay, Negative Control

    Release of CCL5, IL-8 and MCP-1. Mesangial cells (MCs) from patients with IgA nephropathy (IgAN) and controls were stimulated with IgA1 purified from blood from healthy controls (cIgA) or patients with IgAN (gd-IgA) or medium only for 24 h and the release of cytokines into the medium was investigated. Stimulation of control MCs and IgAN MCs with gd-IgA resulted in an increased release of CCL5 in both groups of cells, in contrast stimulation with cIgA did not affect either group of cells at all ( a ). Only the control MCs responded to treatment with gd-IgA with an increased release of IL-8 ( b ) and MCP-1 ( c ). Grey bars represent control MCs, black bars represent IgAN MCs * P

    Journal: BMC Nephrology

    Article Title: Mesangial cells from patients with IgA nephropathy have increased susceptibility to galactose-deficient IgA1

    doi: 10.1186/s12882-016-0251-5

    Figure Lengend Snippet: Release of CCL5, IL-8 and MCP-1. Mesangial cells (MCs) from patients with IgA nephropathy (IgAN) and controls were stimulated with IgA1 purified from blood from healthy controls (cIgA) or patients with IgAN (gd-IgA) or medium only for 24 h and the release of cytokines into the medium was investigated. Stimulation of control MCs and IgAN MCs with gd-IgA resulted in an increased release of CCL5 in both groups of cells, in contrast stimulation with cIgA did not affect either group of cells at all ( a ). Only the control MCs responded to treatment with gd-IgA with an increased release of IL-8 ( b ) and MCP-1 ( c ). Grey bars represent control MCs, black bars represent IgAN MCs * P

    Article Snippet: Background IgA nephropathy (IgAN) is the most common type of glomerular nephritis worldwide.

    Techniques: Purification

    Gene expression of matrix associated genes. One of the main findings in IgAN is expanded mesangial matrix and mesangial proliferation. In order to investigate if cells from patients with IgA-depositions in the kidney express more matrix associated genes we investigated the gene expression of selected matrix genes, namely; BGN (biglycan), COL4A1 (collagen alpha-1(IV) chain), DCN (decorin), FN1 (fibronectin), HSPG2 (perlecan) and NDST1 (heparan sulfate N-deacetylase/N-sulfotransferase 1). The expression of DCN (a small proteoglycan) was significantly higher in untreated IgAN MCs than controls. Overall we could see a trend that IgAN MCs had a higher expression of matrix genes, in the unstimulated groups, and that stimulation with cIgA and gd-IgA increased the expression for all groups. * P

    Journal: BMC Nephrology

    Article Title: Mesangial cells from patients with IgA nephropathy have increased susceptibility to galactose-deficient IgA1

    doi: 10.1186/s12882-016-0251-5

    Figure Lengend Snippet: Gene expression of matrix associated genes. One of the main findings in IgAN is expanded mesangial matrix and mesangial proliferation. In order to investigate if cells from patients with IgA-depositions in the kidney express more matrix associated genes we investigated the gene expression of selected matrix genes, namely; BGN (biglycan), COL4A1 (collagen alpha-1(IV) chain), DCN (decorin), FN1 (fibronectin), HSPG2 (perlecan) and NDST1 (heparan sulfate N-deacetylase/N-sulfotransferase 1). The expression of DCN (a small proteoglycan) was significantly higher in untreated IgAN MCs than controls. Overall we could see a trend that IgAN MCs had a higher expression of matrix genes, in the unstimulated groups, and that stimulation with cIgA and gd-IgA increased the expression for all groups. * P

    Article Snippet: Background IgA nephropathy (IgAN) is the most common type of glomerular nephritis worldwide.

    Techniques: Expressing, IF-cells, Histone Deacetylase Assay

    Mesangial cells were cultured from renal biopsies, characterized and treated with IgA. Schematic figure of harvest and culture of mesangial cells ( a ). Mesangial cells growing out from glomeruli ( b ). Immunofluorescence microscopy of smooth muscle actin labeling in mesangial cells. Smooth muscle actin is exclusively localized to the mesangial cells in the glomeruli. Mesangial cells cultured from glomeruli from renal biopsies were stained using an anti-smooth muscle actin antibody ( green ). Nuclei are stained blue with DAPI. Magnification 20× ( c ), 40× ( d ). The mesangial cells were negative for endothelial and podocyte markers, see Methods. IgA1 purified from human blood was run on HPLC to detect different sizes of IgA1 ( e ). A representative sandwich ELISA shows that IgA1 purified from patients with IgAN have a higher amount of gd-IgA than IgA1 purified from controls ( f )

    Journal: BMC Nephrology

    Article Title: Mesangial cells from patients with IgA nephropathy have increased susceptibility to galactose-deficient IgA1

    doi: 10.1186/s12882-016-0251-5

    Figure Lengend Snippet: Mesangial cells were cultured from renal biopsies, characterized and treated with IgA. Schematic figure of harvest and culture of mesangial cells ( a ). Mesangial cells growing out from glomeruli ( b ). Immunofluorescence microscopy of smooth muscle actin labeling in mesangial cells. Smooth muscle actin is exclusively localized to the mesangial cells in the glomeruli. Mesangial cells cultured from glomeruli from renal biopsies were stained using an anti-smooth muscle actin antibody ( green ). Nuclei are stained blue with DAPI. Magnification 20× ( c ), 40× ( d ). The mesangial cells were negative for endothelial and podocyte markers, see Methods. IgA1 purified from human blood was run on HPLC to detect different sizes of IgA1 ( e ). A representative sandwich ELISA shows that IgA1 purified from patients with IgAN have a higher amount of gd-IgA than IgA1 purified from controls ( f )

    Article Snippet: Background IgA nephropathy (IgAN) is the most common type of glomerular nephritis worldwide.

    Techniques: Cell Culture, Immunofluorescence, Microscopy, Labeling, Staining, Purification, High Performance Liquid Chromatography, Sandwich ELISA

    PDGF is the major growth factor for mesangial cells we therefore wanted to investigate the expression of PDGF in mesangial cells from controls or patients with IgA deposits as well as their response to stimulation with PDGF. Mesangial cells (MCs) from patients with IgA nephropathy (IgAN) and controls were stimulated with IgA1 purified from blood from healthy controls (cIgA) or patients with IgAN (gd-IgA) or medium only for 6 h and the gene expression was investigated. The gene expression of PDGFB was not affected for the control MCs when stimulated with either cIgA or gd-IgA. The gene expression of PDGFB was increased for the IgAN MCs when stimulated with either cIgA or gd-IgA and stimulation of gd-IgA on IgAN MCs had a significantly higher expression than in control MCs stimulated with gd-IgA ( a ). In contrast the gene expression of PDGFRB (gene coding for platelet-derived growth factor receptor beta) was lower in all samples from IgAN MCs compared to the control MCs, but the gene was not further affected by stimulation with either cIgA or gd-IgA ( b ). Medium was collected from IgAN MCs and control MCs after 6 and 24 h of stimulation with cIgA and gd-IgA and the release of PDGF-BB into the medium was measured. Both the IgAN MCs and control MCs released the highest amounts of PDGF-BB when stimulated with gd-IgA, but the levels was significantly higher for the IgAN MCs than for the control-MCs after 6 h ( c ) and 24 h ( d ). Mesangial cells from patients with IgAN and control were treated with 50 ng of either PDGF-AB or -BB or medium only and the relative proliferation of the cells was measured. Stimulation of the IgAN MCs with PDGF gave a significant increase of the relative proliferation compared to control MCs ( e ). Grey bars represent control MCs, black bars represent IgAN MCs * P

    Journal: BMC Nephrology

    Article Title: Mesangial cells from patients with IgA nephropathy have increased susceptibility to galactose-deficient IgA1

    doi: 10.1186/s12882-016-0251-5

    Figure Lengend Snippet: PDGF is the major growth factor for mesangial cells we therefore wanted to investigate the expression of PDGF in mesangial cells from controls or patients with IgA deposits as well as their response to stimulation with PDGF. Mesangial cells (MCs) from patients with IgA nephropathy (IgAN) and controls were stimulated with IgA1 purified from blood from healthy controls (cIgA) or patients with IgAN (gd-IgA) or medium only for 6 h and the gene expression was investigated. The gene expression of PDGFB was not affected for the control MCs when stimulated with either cIgA or gd-IgA. The gene expression of PDGFB was increased for the IgAN MCs when stimulated with either cIgA or gd-IgA and stimulation of gd-IgA on IgAN MCs had a significantly higher expression than in control MCs stimulated with gd-IgA ( a ). In contrast the gene expression of PDGFRB (gene coding for platelet-derived growth factor receptor beta) was lower in all samples from IgAN MCs compared to the control MCs, but the gene was not further affected by stimulation with either cIgA or gd-IgA ( b ). Medium was collected from IgAN MCs and control MCs after 6 and 24 h of stimulation with cIgA and gd-IgA and the release of PDGF-BB into the medium was measured. Both the IgAN MCs and control MCs released the highest amounts of PDGF-BB when stimulated with gd-IgA, but the levels was significantly higher for the IgAN MCs than for the control-MCs after 6 h ( c ) and 24 h ( d ). Mesangial cells from patients with IgAN and control were treated with 50 ng of either PDGF-AB or -BB or medium only and the relative proliferation of the cells was measured. Stimulation of the IgAN MCs with PDGF gave a significant increase of the relative proliferation compared to control MCs ( e ). Grey bars represent control MCs, black bars represent IgAN MCs * P

    Article Snippet: Background IgA nephropathy (IgAN) is the most common type of glomerular nephritis worldwide.

    Techniques: Expressing, Purification, Derivative Assay

    Gene expression of TGFB1 and TGFBR1 and release of TGFβ1. Mesangial cells (MCs) from patients with IgA nephropathy (IgAN) and controls were stimulated with IgA1 purified from blood from healthy controls (cIgA) or patients with IgAN (gd-IgA) or medium only for 6 h and the gene expression was investigated. Stimulation of control MCs and IgAN MCs with gd-IgA gave a significant increase in the gene expression of TGFB1 (gene coding for TGFβ1) ( a ). The gene expression for TGFBR1 (gene coding for TGF-beta receptor type-1) was not significantly affected by any of the treatments ( b ). IgAN MCs were stimulated with either cIgA or gd-IgA for 6 h and the release of TGFβ1into the cell culture medium was investigated. Both treatment with cIgA and gd-IgA resulted in an increased release of TGFβ1 compared to untreated cells ( c ). Grey bars represent control MCs, black bars represent IgAN MCs * P

    Journal: BMC Nephrology

    Article Title: Mesangial cells from patients with IgA nephropathy have increased susceptibility to galactose-deficient IgA1

    doi: 10.1186/s12882-016-0251-5

    Figure Lengend Snippet: Gene expression of TGFB1 and TGFBR1 and release of TGFβ1. Mesangial cells (MCs) from patients with IgA nephropathy (IgAN) and controls were stimulated with IgA1 purified from blood from healthy controls (cIgA) or patients with IgAN (gd-IgA) or medium only for 6 h and the gene expression was investigated. Stimulation of control MCs and IgAN MCs with gd-IgA gave a significant increase in the gene expression of TGFB1 (gene coding for TGFβ1) ( a ). The gene expression for TGFBR1 (gene coding for TGF-beta receptor type-1) was not significantly affected by any of the treatments ( b ). IgAN MCs were stimulated with either cIgA or gd-IgA for 6 h and the release of TGFβ1into the cell culture medium was investigated. Both treatment with cIgA and gd-IgA resulted in an increased release of TGFβ1 compared to untreated cells ( c ). Grey bars represent control MCs, black bars represent IgAN MCs * P

    Article Snippet: Background IgA nephropathy (IgAN) is the most common type of glomerular nephritis worldwide.

    Techniques: Expressing, Purification, Cell Culture

    Gene expression and release of IL-6. IL-6 expression is known to increase in mesangial cells when stimulated with gd-IgA and has been suggested as a prognostic marker for IgAN but its role is still debated. Mesangial cells (MCs) from patients with IgA nephropathy (IgAN) and controls were stimulated with IgA1 purified from blood from healthy controls (cIgA) or patients with IgAN (gd-IgA) or medium only for 6 h and the gene expression was investigated. Stimulation with IgA on both controls MCs and IgAN MCs increased the gene expression of IL-6 in a similar pattern ( a ). The release of IL-6 into the medium after 24 h of stimulation was most pronounced for the gd-IgA treatment for both groups of cells, but the IgAN MCs released significantly more IL-6 than the control MCs stimulated with gd-IgA ( b ). Grey bars represent control MCs, black bars represent IgAN MCs * P

    Journal: BMC Nephrology

    Article Title: Mesangial cells from patients with IgA nephropathy have increased susceptibility to galactose-deficient IgA1

    doi: 10.1186/s12882-016-0251-5

    Figure Lengend Snippet: Gene expression and release of IL-6. IL-6 expression is known to increase in mesangial cells when stimulated with gd-IgA and has been suggested as a prognostic marker for IgAN but its role is still debated. Mesangial cells (MCs) from patients with IgA nephropathy (IgAN) and controls were stimulated with IgA1 purified from blood from healthy controls (cIgA) or patients with IgAN (gd-IgA) or medium only for 6 h and the gene expression was investigated. Stimulation with IgA on both controls MCs and IgAN MCs increased the gene expression of IL-6 in a similar pattern ( a ). The release of IL-6 into the medium after 24 h of stimulation was most pronounced for the gd-IgA treatment for both groups of cells, but the IgAN MCs released significantly more IL-6 than the control MCs stimulated with gd-IgA ( b ). Grey bars represent control MCs, black bars represent IgAN MCs * P

    Article Snippet: Background IgA nephropathy (IgAN) is the most common type of glomerular nephritis worldwide.

    Techniques: Expressing, Marker, Purification

    RV-specific antibody responses in sera from children with acute wheeze. Frequencies and levels of a IgG and b IgA responses ( y -axes: n ) ( x -axes: red: RV-A species; green: RV-B species; blue: RV-C species). Antibody levels are color-coded and expressed as ISAC standardized units, ISU-G and ISU-A, respectively. c Median IgG levels ( y -axis: ISU-G) to VP1 peptides ( x -axis) in children grouped according to age (6–12 months: squares; 13–24 months: triangles; 25–42: circles). d Spearman’s rank correlation between the number of IgG-reactive peptides ( n , y -axis; median IgG > 15 ISU) and age ( x -axis: months). Correlation coefficient ( ρ ) and p -value are shown. e Comparison of the number of IgG-reactive VP1 peptides ( n , y -axis; median IgG > 15 ISU) in children according to age ( x -axis). Horizontal lines indicate medians. Statistically significant differences between groups are indicated (** p

    Journal: Nature Communications

    Article Title: PreDicta chip-based high resolution diagnosis of rhinovirus-induced wheeze

    doi: 10.1038/s41467-018-04591-0

    Figure Lengend Snippet: RV-specific antibody responses in sera from children with acute wheeze. Frequencies and levels of a IgG and b IgA responses ( y -axes: n ) ( x -axes: red: RV-A species; green: RV-B species; blue: RV-C species). Antibody levels are color-coded and expressed as ISAC standardized units, ISU-G and ISU-A, respectively. c Median IgG levels ( y -axis: ISU-G) to VP1 peptides ( x -axis) in children grouped according to age (6–12 months: squares; 13–24 months: triangles; 25–42: circles). d Spearman’s rank correlation between the number of IgG-reactive peptides ( n , y -axis; median IgG > 15 ISU) and age ( x -axis: months). Correlation coefficient ( ρ ) and p -value are shown. e Comparison of the number of IgG-reactive VP1 peptides ( n , y -axis; median IgG > 15 ISU) in children according to age ( x -axis). Horizontal lines indicate medians. Statistically significant differences between groups are indicated (** p

    Article Snippet: For the detection of RV-specific IgG and IgA antibodies, serum samples were diluted 1:300 and 1:20 in a sample dilution buffer (Phadia-Thermo Fisher), respectively.

    Techniques:

    SPR analysis of lectin binding to IgA1 myeloma proteins

    Journal: Biochemistry

    Article Title: Recognition of galactose-deficient O-glycans in the hinge region of IgA1 by N-acetylgalactosamine-specific snail lectins: a comparative binding study †

    doi: 10.1021/bi9019498

    Figure Lengend Snippet: SPR analysis of lectin binding to IgA1 myeloma proteins

    Article Snippet: In contrast, IgA1 from IgAN patient #1 had much lower affinity for MAA-II compared to IgA1 from IgAN patient #2 or the healthy controls ( ).

    Techniques: SPR Assay, Binding Assay

    Equilibrium binding isotherms for HAA and HPA binding to IgA1 derived from IgAN patients and healthy controls

    Journal: Biochemistry

    Article Title: Recognition of galactose-deficient O-glycans in the hinge region of IgA1 by N-acetylgalactosamine-specific snail lectins: a comparative binding study †

    doi: 10.1021/bi9019498

    Figure Lengend Snippet: Equilibrium binding isotherms for HAA and HPA binding to IgA1 derived from IgAN patients and healthy controls

    Article Snippet: In contrast, IgA1 from IgAN patient #1 had much lower affinity for MAA-II compared to IgA1 from IgAN patient #2 or the healthy controls ( ).

    Techniques: Binding Assay, Derivative Assay

    Biosensor-based analysis of lectin binding to IgA1 derived from EBV-immortalized IgA1-secreting cell lines from patients

    Journal: Biochemistry

    Article Title: Recognition of galactose-deficient O-glycans in the hinge region of IgA1 by N-acetylgalactosamine-specific snail lectins: a comparative binding study †

    doi: 10.1021/bi9019498

    Figure Lengend Snippet: Biosensor-based analysis of lectin binding to IgA1 derived from EBV-immortalized IgA1-secreting cell lines from patients

    Article Snippet: In contrast, IgA1 from IgAN patient #1 had much lower affinity for MAA-II compared to IgA1 from IgAN patient #2 or the healthy controls ( ).

    Techniques: Binding Assay, Derivative Assay

    Biosensor-based analysis of lectin binding to IgA1 derived from EBV-immortalized IgA1-secreting cell lines from patients

    Journal: Biochemistry

    Article Title: Recognition of galactose-deficient O-glycans in the hinge region of IgA1 by N-acetylgalactosamine-specific snail lectins: a comparative binding study †

    doi: 10.1021/bi9019498

    Figure Lengend Snippet: Biosensor-based analysis of lectin binding to IgA1 derived from EBV-immortalized IgA1-secreting cell lines from patients

    Article Snippet: In contrast, IgA1 from IgAN patient #1 had much lower affinity for MAA-II compared to IgA1 from IgAN patient #2 or the healthy controls ( ).

    Techniques: Binding Assay, Derivative Assay

    Equilibrium binding isotherms for HAA and HPA binding to different IgA1 myeloma proteins

    Journal: Biochemistry

    Article Title: Recognition of galactose-deficient O-glycans in the hinge region of IgA1 by N-acetylgalactosamine-specific snail lectins: a comparative binding study †

    doi: 10.1021/bi9019498

    Figure Lengend Snippet: Equilibrium binding isotherms for HAA and HPA binding to different IgA1 myeloma proteins

    Article Snippet: In contrast, IgA1 from IgAN patient #1 had much lower affinity for MAA-II compared to IgA1 from IgAN patient #2 or the healthy controls ( ).

    Techniques: Binding Assay

    O -linked glycans in the hinge region of circulatory IgA1

    Journal: Biochemistry

    Article Title: Recognition of galactose-deficient O-glycans in the hinge region of IgA1 by N-acetylgalactosamine-specific snail lectins: a comparative binding study †

    doi: 10.1021/bi9019498

    Figure Lengend Snippet: O -linked glycans in the hinge region of circulatory IgA1

    Article Snippet: In contrast, IgA1 from IgAN patient #1 had much lower affinity for MAA-II compared to IgA1 from IgAN patient #2 or the healthy controls ( ).

    Techniques:

    Model for bivalent binding of HAA or HPA to a single molecule of undergalactosylated monomeric IgA1

    Journal: Biochemistry

    Article Title: Recognition of galactose-deficient O-glycans in the hinge region of IgA1 by N-acetylgalactosamine-specific snail lectins: a comparative binding study †

    doi: 10.1021/bi9019498

    Figure Lengend Snippet: Model for bivalent binding of HAA or HPA to a single molecule of undergalactosylated monomeric IgA1

    Article Snippet: In contrast, IgA1 from IgAN patient #1 had much lower affinity for MAA-II compared to IgA1 from IgAN patient #2 or the healthy controls ( ).

    Techniques: Binding Assay

    SPR analysis of lectin binding to IgA1 myeloma proteins

    Journal: Biochemistry

    Article Title: Recognition of galactose-deficient O-glycans in the hinge region of IgA1 by N-acetylgalactosamine-specific snail lectins: a comparative binding study †

    doi: 10.1021/bi9019498

    Figure Lengend Snippet: SPR analysis of lectin binding to IgA1 myeloma proteins

    Article Snippet: In contrast, IgA1 from IgAN patient #1 had much lower affinity for MAA-II compared to IgA1 from IgAN patient #2 or the healthy controls ( ).

    Techniques: SPR Assay, Binding Assay

    Equilibrium binding isotherms for different IgA1 myeloma proteins binding to HAA ligand

    Journal: Biochemistry

    Article Title: Recognition of galactose-deficient O-glycans in the hinge region of IgA1 by N-acetylgalactosamine-specific snail lectins: a comparative binding study †

    doi: 10.1021/bi9019498

    Figure Lengend Snippet: Equilibrium binding isotherms for different IgA1 myeloma proteins binding to HAA ligand

    Article Snippet: In contrast, IgA1 from IgAN patient #1 had much lower affinity for MAA-II compared to IgA1 from IgAN patient #2 or the healthy controls ( ).

    Techniques: Binding Assay

    SPR analysis of lectin binding to IgA1 myeloma proteins

    Journal: Biochemistry

    Article Title: Recognition of galactose-deficient O-glycans in the hinge region of IgA1 by N-acetylgalactosamine-specific snail lectins: a comparative binding study †

    doi: 10.1021/bi9019498

    Figure Lengend Snippet: SPR analysis of lectin binding to IgA1 myeloma proteins

    Article Snippet: In contrast, IgA1 from IgAN patient #1 had much lower affinity for MAA-II compared to IgA1 from IgAN patient #2 or the healthy controls ( ).

    Techniques: SPR Assay, Binding Assay

    Analysis of different IgA1 proteins by lectin western blotting

    Journal: Biochemistry

    Article Title: Recognition of galactose-deficient O-glycans in the hinge region of IgA1 by N-acetylgalactosamine-specific snail lectins: a comparative binding study †

    doi: 10.1021/bi9019498

    Figure Lengend Snippet: Analysis of different IgA1 proteins by lectin western blotting

    Article Snippet: In contrast, IgA1 from IgAN patient #1 had much lower affinity for MAA-II compared to IgA1 from IgAN patient #2 or the healthy controls ( ).

    Techniques: Western Blot

    IgA characterization in the indicated tissue of STAM and MUP-uPA mice a – e , Single splenocyte or liver cell suspensions from tumour (HCC)-bearing mice (STAM-B6) were stained with CD45, CD19, IgA, CD138, B220, and PD-L1, and FVD-eF780 was used to exclude dead cells. a , The gating strategies for splenocytes and liver lymphocytes: lymphocyte gate, dead cell exclusion, doublets exclusion, and CD45 + population gate. b , Flow cytometry analysis of IgA and CD19 expression of indicated strains, gated on the CD45 + population. Spleen or liver from Iga −/− mice was used to set up the gating for the populations. Tgfbr2 ΔB and Pdl1/2 −/− mice clearly showed less IgA + cells than WT mice. c – e , IgA subpopulations were gated as indicated and analysed for CD138, B220, and PD-L1 expression. IgA subpopulations were gated on the basis of CD19 expression, in the indicated strains, and the IgA + CD19 + , IgA + CD19 −/low/int , and IgA − CD19 + subpopulations were further analysed for their B220 and CD138 expression. The decisions for CD19 and CD138 levels were also based on their mean fluorescence intensity (MFI; red). The analyses showed two main populations: (1) IgA + CD19 + B220 + CD138 low/− and (2) IgA + CD19 −/int B220 − CD138 +/hi . These two populations and the IgA − CD19 + populations were further analysed for their ability to express PD-L1 (percentage and mean fluorescence intensity). f – j , Single-cell suspensions were prepared from the spleen, liver, or intestine of MUP-uPA mice kept on normal chow or HFD, and were stained with CD45, CD19, IgA, B220, CD138, CD11b, MHCII, PD-L1, and CD5, and analysed by flow cytometry. f , The B220 and CD138 expression in total IgA + cells (CD19 + and CD19 − populations) confirmed that most of the IgA + cells in spleen and liver of HFD-fed MUP-uPA mice are CD138 + cells. g , h , IgA subpopulations in liver ( g ) and spleen ( h ) were gated as indicated and analysed for CD138, B220, and MHCII expression. i , Representative dot-plots gated on IgA + cells, showing that most of the IgA + cells are not CD11b + . j , PD-L1 and CD5 expression of IgA + cells in spleen, liver, and intestine.

    Journal: Nature

    Article Title: Inflammation-induced IgA+ cells dismantle anti-liver cancer immunity

    doi: 10.1038/nature24302

    Figure Lengend Snippet: IgA characterization in the indicated tissue of STAM and MUP-uPA mice a – e , Single splenocyte or liver cell suspensions from tumour (HCC)-bearing mice (STAM-B6) were stained with CD45, CD19, IgA, CD138, B220, and PD-L1, and FVD-eF780 was used to exclude dead cells. a , The gating strategies for splenocytes and liver lymphocytes: lymphocyte gate, dead cell exclusion, doublets exclusion, and CD45 + population gate. b , Flow cytometry analysis of IgA and CD19 expression of indicated strains, gated on the CD45 + population. Spleen or liver from Iga −/− mice was used to set up the gating for the populations. Tgfbr2 ΔB and Pdl1/2 −/− mice clearly showed less IgA + cells than WT mice. c – e , IgA subpopulations were gated as indicated and analysed for CD138, B220, and PD-L1 expression. IgA subpopulations were gated on the basis of CD19 expression, in the indicated strains, and the IgA + CD19 + , IgA + CD19 −/low/int , and IgA − CD19 + subpopulations were further analysed for their B220 and CD138 expression. The decisions for CD19 and CD138 levels were also based on their mean fluorescence intensity (MFI; red). The analyses showed two main populations: (1) IgA + CD19 + B220 + CD138 low/− and (2) IgA + CD19 −/int B220 − CD138 +/hi . These two populations and the IgA − CD19 + populations were further analysed for their ability to express PD-L1 (percentage and mean fluorescence intensity). f – j , Single-cell suspensions were prepared from the spleen, liver, or intestine of MUP-uPA mice kept on normal chow or HFD, and were stained with CD45, CD19, IgA, B220, CD138, CD11b, MHCII, PD-L1, and CD5, and analysed by flow cytometry. f , The B220 and CD138 expression in total IgA + cells (CD19 + and CD19 − populations) confirmed that most of the IgA + cells in spleen and liver of HFD-fed MUP-uPA mice are CD138 + cells. g , h , IgA subpopulations in liver ( g ) and spleen ( h ) were gated as indicated and analysed for CD138, B220, and MHCII expression. i , Representative dot-plots gated on IgA + cells, showing that most of the IgA + cells are not CD11b + . j , PD-L1 and CD5 expression of IgA + cells in spleen, liver, and intestine.

    Article Snippet: Antibodies specific for the following antigens were used (anti-mouse antigens were abbreviated as ‘m’, and anti-human as ‘h’): Tim-3 (mB8.2C12- AlexaF647; hF38-2E2-PerCP-eF710); CD138 (m281-2-PE/APC; hDL-101- APC); IgA (mA-6E1-PE/FITC; m11-44-2-Biotin; mRMA-1; hIS11-8E10- FITC; DAKO hA0262); CD8α (m53-6.7-PerCp-Cy5.5/PE-Cy7; hRPA-T8-PE; hC8/144B); CD45 (m30-F11-V500; hHI30-eF450); CD20 (mAISB12-PE); CD44 (mIM7-PE/FITC/eF450); CD4 (mRM4-5-eF450; mGk1.5-FITC/PE/PE-Cy7; hRPA-T4-APC); B220 (mRA3-6B2-eF450/FITC); CD19 (meBio1D3-eF660/PE-Cy7; hHIB19-PE); IgM (mII/41-PE-Cy7; hSA-DA4-eF450); IgD (m11- 26c.2a-Pacific-Blue; m11-26c-PerCP-eF710); TNF (mMP6-XT22-FITC/PE); IFNγ (mXMG1.2-eF660); GrzB (mNGZB-eF660); CD107a (meBio1D4B-PE); PD-1 (mJ43-eF450/FITC; mRMP1-30-APC; hMIH4-FITC); PD-L1 (mMIH5- PE-Cy7/PE; hMIH1-PE-Cy7); FAS-L1 (mMFL3-FITC); Ki-67 (mSolA15-eF450); IgG2a (m2a-15F8-FITC/APC; mRMG2a-62-PE); IgG1 (mM1-14D12-eF450; mRMG1-1-FITC); IL-10 (mJES5-16E3-PE); CD11c (mN418-eF450/PE); CD11b (mM1/70-eF450/eF660); MHCII (mM5/114.15.2-FITC/PE); Gr-1 (m1A8-Ly6G-PerCP- eF710); IL-17 (meBio17B7-PE-Cy7); Foxp3 (mFJK-16s-eF450); Perforin (meBioOMAK-D-FITC); CXCR5 (mSPRCL5-PE/APC/Biotin); CD62L (mMEL- 14-PE/APC); SIINFEKL/H-2Kb (meBio25-D1.16-APC); F4/80 (mBM8-PE/FITC); anti-Biotin (BK-1/39-PE); CD5 (m53-7.3-FITC/eF450) and NK1.1 (mPK136- APC); CD31 (m390-eF450) (most from eBioscience and Biolegend); pAb rabbit to CD3 (DAKO, IS503); and alpha-SMA (m-ab5694; DAKO, h1A4).

    Techniques: Mouse Assay, Staining, Flow Cytometry, Cytometry, Expressing, Fluorescence

    IgA + plasmocytes regulate tumour killing by CD8 + T cells a , Dih10 and dihXY HCC cells were transfected with an inducible ovalbumin (Ova) expression vector, and Ova expression and presentation were confirmed by flow cytometry, using an antibody that recognized the SIINFEKL peptide on the MHCI molecule H-2Kb. b – h , Ova-expressing dih cells or controls (dih–RFP) were starved overnight (5% cell death), after which their medium was changed and B cells from WT, Iga −/− , Pdl1/2 −/− , or SW-HEL mice were added in the presence of TGFβ (5 ng ml −1 ) and CTGF (3 ng ml −1 ), for an additional 24 h. Thereafter, the medium was replaced and CFSE-labelled OT-I T cells were added to the cultures that either contained or did not contain the B cells described above. After 4–6 days, the co-cultured cells were analysed by flow cytometry, while the secretory IgA was analysed by ELISA ( n = 2–4 wells per group per day). a – j , Experiments were repeated with two different Ova-expressing HCC and one prostate cancer cell lines. Shown are the representative flow cytometry histograms or plots depicting ( b ) OT-I CD8 + T-cell proliferation, ( c ) PD-L1 and SIINFEKL/H-2Kb expression on cancer cells, ( d ) PD-L1 expression on B cells, ( e ) cancer cell death. f , Relative dih-Ova–RFP killing by OT-I CD8 + T cells in the presence or absence of the indicated B cells. g , Total secretory IgA and anti-OVA-IgA antibody amounts in culture supernatants. h , Percentages of OT-I CD8 + cells in each culture, as indicated. i , j , TRC2-Ova–RFP cells or its control cell line (TRC2–RFP) were co-cultured with OT-I cells and splenic B cells (WT and Il10 −/− ), as described for a – h. i , Proliferation of OT-I cells was analysed using CFSE ( n = 3, 7, 5, 4). j , The amounts of secretory IgA were analysed using ELISA as indicated ( n = 3 per group). k – o , Liver cells from indicated 3-month-old mice were stained and analysed by flow cytometry. Experiments were repeated at least two or three times. Each dot represents one mouse. Shown are the percentage of CD8 + T cells among CD45 + cells ( n = 6, 5, 3, 7, 4) ( k ), absolute CD8 + T-cell number per gram of liver ( n = 3, 3, 7, 4) ( l ), the percentage of CD8 + CD44 + Ki-67 + T cells with representative scatter plots ( n = 3 or 4 per group) ( m , n ), and the representative scatter plots of perforin and GrzB among CD8 + CD44 + Ki-67 + T cells ( o ). p , q , Liver cell suspensions from the indicated mice were stained as shown and analysed by flow cytometry to determine the absolute CD8 + T-cell number in both STAM-BL6 and STAM-FVB mice ( n = 8, 4, 5, 3, 5, 5, 6, 7) ( p ), and the percentage of T EM cells using CD8, CD44, and CD62L ( n = 4, 4, 6, 9) ( q ). r , Liver cells from indicated 3-, 6-, and 11-month-old mice kept on HFD ( n = 4–10) were stained and analysed by flow cytometry. Shown are the percentage of CD8 + IFNγ + CD107a + T cells. Detailed n values are shown in . s – y , Liver cell suspensions from the indicated mice were stained as shown and analysed by flow cytometry to determine the percentage of Th17 cells using CD4 and IL-17a ( n = 4, 7, 9, 10, 4, 9, 11) ( s ), the percentage of regulatory T cells using CD4 and Foxp3 ( n = 4, 3, 8, 11, 7, 13) ( t ), the percentage of Tfh-like cells using CXCR5, PD1 and CD4 (3, 5, 6, 5) ( u ), the percentage of B220 + CD19 + B cells ( n = 4, 7, 7, 4, 5, 3, 9, 3, 11) ( v ), absolute B220 + CD19 + B-cell number per gram of liver ( n = 5, 7, 12, 27, 15, 7) ( w ), the percentage of IgG + cells ( n = 4, 3, 14, 11) ( x ), and CD138 + plasma cells ( n = 4, 7, 4, 7, 7, 8, 5, 8, 6, 6, 13, 27, 17) ( y ). Two-sided t -test (means ± s.e.m.; f – m ) and Mann–Whitney test (median; p – y ) were used to determine significance. * P

    Journal: Nature

    Article Title: Inflammation-induced IgA+ cells dismantle anti-liver cancer immunity

    doi: 10.1038/nature24302

    Figure Lengend Snippet: IgA + plasmocytes regulate tumour killing by CD8 + T cells a , Dih10 and dihXY HCC cells were transfected with an inducible ovalbumin (Ova) expression vector, and Ova expression and presentation were confirmed by flow cytometry, using an antibody that recognized the SIINFEKL peptide on the MHCI molecule H-2Kb. b – h , Ova-expressing dih cells or controls (dih–RFP) were starved overnight (5% cell death), after which their medium was changed and B cells from WT, Iga −/− , Pdl1/2 −/− , or SW-HEL mice were added in the presence of TGFβ (5 ng ml −1 ) and CTGF (3 ng ml −1 ), for an additional 24 h. Thereafter, the medium was replaced and CFSE-labelled OT-I T cells were added to the cultures that either contained or did not contain the B cells described above. After 4–6 days, the co-cultured cells were analysed by flow cytometry, while the secretory IgA was analysed by ELISA ( n = 2–4 wells per group per day). a – j , Experiments were repeated with two different Ova-expressing HCC and one prostate cancer cell lines. Shown are the representative flow cytometry histograms or plots depicting ( b ) OT-I CD8 + T-cell proliferation, ( c ) PD-L1 and SIINFEKL/H-2Kb expression on cancer cells, ( d ) PD-L1 expression on B cells, ( e ) cancer cell death. f , Relative dih-Ova–RFP killing by OT-I CD8 + T cells in the presence or absence of the indicated B cells. g , Total secretory IgA and anti-OVA-IgA antibody amounts in culture supernatants. h , Percentages of OT-I CD8 + cells in each culture, as indicated. i , j , TRC2-Ova–RFP cells or its control cell line (TRC2–RFP) were co-cultured with OT-I cells and splenic B cells (WT and Il10 −/− ), as described for a – h. i , Proliferation of OT-I cells was analysed using CFSE ( n = 3, 7, 5, 4). j , The amounts of secretory IgA were analysed using ELISA as indicated ( n = 3 per group). k – o , Liver cells from indicated 3-month-old mice were stained and analysed by flow cytometry. Experiments were repeated at least two or three times. Each dot represents one mouse. Shown are the percentage of CD8 + T cells among CD45 + cells ( n = 6, 5, 3, 7, 4) ( k ), absolute CD8 + T-cell number per gram of liver ( n = 3, 3, 7, 4) ( l ), the percentage of CD8 + CD44 + Ki-67 + T cells with representative scatter plots ( n = 3 or 4 per group) ( m , n ), and the representative scatter plots of perforin and GrzB among CD8 + CD44 + Ki-67 + T cells ( o ). p , q , Liver cell suspensions from the indicated mice were stained as shown and analysed by flow cytometry to determine the absolute CD8 + T-cell number in both STAM-BL6 and STAM-FVB mice ( n = 8, 4, 5, 3, 5, 5, 6, 7) ( p ), and the percentage of T EM cells using CD8, CD44, and CD62L ( n = 4, 4, 6, 9) ( q ). r , Liver cells from indicated 3-, 6-, and 11-month-old mice kept on HFD ( n = 4–10) were stained and analysed by flow cytometry. Shown are the percentage of CD8 + IFNγ + CD107a + T cells. Detailed n values are shown in . s – y , Liver cell suspensions from the indicated mice were stained as shown and analysed by flow cytometry to determine the percentage of Th17 cells using CD4 and IL-17a ( n = 4, 7, 9, 10, 4, 9, 11) ( s ), the percentage of regulatory T cells using CD4 and Foxp3 ( n = 4, 3, 8, 11, 7, 13) ( t ), the percentage of Tfh-like cells using CXCR5, PD1 and CD4 (3, 5, 6, 5) ( u ), the percentage of B220 + CD19 + B cells ( n = 4, 7, 7, 4, 5, 3, 9, 3, 11) ( v ), absolute B220 + CD19 + B-cell number per gram of liver ( n = 5, 7, 12, 27, 15, 7) ( w ), the percentage of IgG + cells ( n = 4, 3, 14, 11) ( x ), and CD138 + plasma cells ( n = 4, 7, 4, 7, 7, 8, 5, 8, 6, 6, 13, 27, 17) ( y ). Two-sided t -test (means ± s.e.m.; f – m ) and Mann–Whitney test (median; p – y ) were used to determine significance. * P

    Article Snippet: Antibodies specific for the following antigens were used (anti-mouse antigens were abbreviated as ‘m’, and anti-human as ‘h’): Tim-3 (mB8.2C12- AlexaF647; hF38-2E2-PerCP-eF710); CD138 (m281-2-PE/APC; hDL-101- APC); IgA (mA-6E1-PE/FITC; m11-44-2-Biotin; mRMA-1; hIS11-8E10- FITC; DAKO hA0262); CD8α (m53-6.7-PerCp-Cy5.5/PE-Cy7; hRPA-T8-PE; hC8/144B); CD45 (m30-F11-V500; hHI30-eF450); CD20 (mAISB12-PE); CD44 (mIM7-PE/FITC/eF450); CD4 (mRM4-5-eF450; mGk1.5-FITC/PE/PE-Cy7; hRPA-T4-APC); B220 (mRA3-6B2-eF450/FITC); CD19 (meBio1D3-eF660/PE-Cy7; hHIB19-PE); IgM (mII/41-PE-Cy7; hSA-DA4-eF450); IgD (m11- 26c.2a-Pacific-Blue; m11-26c-PerCP-eF710); TNF (mMP6-XT22-FITC/PE); IFNγ (mXMG1.2-eF660); GrzB (mNGZB-eF660); CD107a (meBio1D4B-PE); PD-1 (mJ43-eF450/FITC; mRMP1-30-APC; hMIH4-FITC); PD-L1 (mMIH5- PE-Cy7/PE; hMIH1-PE-Cy7); FAS-L1 (mMFL3-FITC); Ki-67 (mSolA15-eF450); IgG2a (m2a-15F8-FITC/APC; mRMG2a-62-PE); IgG1 (mM1-14D12-eF450; mRMG1-1-FITC); IL-10 (mJES5-16E3-PE); CD11c (mN418-eF450/PE); CD11b (mM1/70-eF450/eF660); MHCII (mM5/114.15.2-FITC/PE); Gr-1 (m1A8-Ly6G-PerCP- eF710); IL-17 (meBio17B7-PE-Cy7); Foxp3 (mFJK-16s-eF450); Perforin (meBioOMAK-D-FITC); CXCR5 (mSPRCL5-PE/APC/Biotin); CD62L (mMEL- 14-PE/APC); SIINFEKL/H-2Kb (meBio25-D1.16-APC); F4/80 (mBM8-PE/FITC); anti-Biotin (BK-1/39-PE); CD5 (m53-7.3-FITC/eF450) and NK1.1 (mPK136- APC); CD31 (m390-eF450) (most from eBioscience and Biolegend); pAb rabbit to CD3 (DAKO, IS503); and alpha-SMA (m-ab5694; DAKO, h1A4).

    Techniques: Transfection, Expressing, Plasmid Preparation, Flow Cytometry, Cytometry, Mouse Assay, Cell Culture, Enzyme-linked Immunosorbent Assay, Staining, MANN-WHITNEY

    IgA + cells in NASH livers express PD-L1 a , Single-cell suspensions were prepared from the livers of tumour-bearing mice (STAM-B6) as indicated. Shown are the PD-L1 and IgA staining in the indicated strains. Spleen or liver from Iga −/− and Pdl1/2 −/− mice were used as control for IgA and PD-L1 staining. b , Liver single-cell suspensions of MUP-uPA mice and indicated strains were stained for CD45, IgA, and PD-L1. Percentages of liver PD-L1 + cells in CD45 + IgA − (labelled: CD45 + ) or IgA + cell populations from the indicated strains are shown ( n = 8, 6, 6, 7, 6, 6, 18, 16, 11, 12). c , Percentages of liver PD-L1 + CD45 + cells are shown in the indicated strains ( n = 5, 6, 6, 16, 10). The data were validated in at least three different experiments. d , e , CD138 + cells were divided into IgA + and IgA − populations and analysed for PD-L1 expression. d , Shown is the mean fluorescence intensity of PD-L1 expression for CD138 + cells from spleen (gated on either CD138 + IgA − or CD138 + IgA + cells). e , Mean fluorescence intensity of PD-L1 expression is shown for CD138 + plasmocytes from livers of 19 mice (gated on either CD138 + IgA − or CD138 + IgA + plasma cells). f , Percentages of intestinal IgA + cells in MUP-uPA mice kept on normal chow or HFD ( n = 6, 13; left). Percentages of PD-L1 + IgA + cells in intestine and liver are included for comparison ( n = 7, 8; right). g , Percentages of liver IgA + cells are shown in the indicated strains kept on HFD at the age of 3 months ( n = 9, 7). h , Percentages of PD-L1 + cells are shown gated on CD45 + CD19 − IgA − cells in the indicated strains ( n = 3, 4, 5). The data were validated at least in two or three experiments. Two-sided Mann– Whitney tests (median; b , c , e–h ) were used to determine significance. * P

    Journal: Nature

    Article Title: Inflammation-induced IgA+ cells dismantle anti-liver cancer immunity

    doi: 10.1038/nature24302

    Figure Lengend Snippet: IgA + cells in NASH livers express PD-L1 a , Single-cell suspensions were prepared from the livers of tumour-bearing mice (STAM-B6) as indicated. Shown are the PD-L1 and IgA staining in the indicated strains. Spleen or liver from Iga −/− and Pdl1/2 −/− mice were used as control for IgA and PD-L1 staining. b , Liver single-cell suspensions of MUP-uPA mice and indicated strains were stained for CD45, IgA, and PD-L1. Percentages of liver PD-L1 + cells in CD45 + IgA − (labelled: CD45 + ) or IgA + cell populations from the indicated strains are shown ( n = 8, 6, 6, 7, 6, 6, 18, 16, 11, 12). c , Percentages of liver PD-L1 + CD45 + cells are shown in the indicated strains ( n = 5, 6, 6, 16, 10). The data were validated in at least three different experiments. d , e , CD138 + cells were divided into IgA + and IgA − populations and analysed for PD-L1 expression. d , Shown is the mean fluorescence intensity of PD-L1 expression for CD138 + cells from spleen (gated on either CD138 + IgA − or CD138 + IgA + cells). e , Mean fluorescence intensity of PD-L1 expression is shown for CD138 + plasmocytes from livers of 19 mice (gated on either CD138 + IgA − or CD138 + IgA + plasma cells). f , Percentages of intestinal IgA + cells in MUP-uPA mice kept on normal chow or HFD ( n = 6, 13; left). Percentages of PD-L1 + IgA + cells in intestine and liver are included for comparison ( n = 7, 8; right). g , Percentages of liver IgA + cells are shown in the indicated strains kept on HFD at the age of 3 months ( n = 9, 7). h , Percentages of PD-L1 + cells are shown gated on CD45 + CD19 − IgA − cells in the indicated strains ( n = 3, 4, 5). The data were validated at least in two or three experiments. Two-sided Mann– Whitney tests (median; b , c , e–h ) were used to determine significance. * P

    Article Snippet: Antibodies specific for the following antigens were used (anti-mouse antigens were abbreviated as ‘m’, and anti-human as ‘h’): Tim-3 (mB8.2C12- AlexaF647; hF38-2E2-PerCP-eF710); CD138 (m281-2-PE/APC; hDL-101- APC); IgA (mA-6E1-PE/FITC; m11-44-2-Biotin; mRMA-1; hIS11-8E10- FITC; DAKO hA0262); CD8α (m53-6.7-PerCp-Cy5.5/PE-Cy7; hRPA-T8-PE; hC8/144B); CD45 (m30-F11-V500; hHI30-eF450); CD20 (mAISB12-PE); CD44 (mIM7-PE/FITC/eF450); CD4 (mRM4-5-eF450; mGk1.5-FITC/PE/PE-Cy7; hRPA-T4-APC); B220 (mRA3-6B2-eF450/FITC); CD19 (meBio1D3-eF660/PE-Cy7; hHIB19-PE); IgM (mII/41-PE-Cy7; hSA-DA4-eF450); IgD (m11- 26c.2a-Pacific-Blue; m11-26c-PerCP-eF710); TNF (mMP6-XT22-FITC/PE); IFNγ (mXMG1.2-eF660); GrzB (mNGZB-eF660); CD107a (meBio1D4B-PE); PD-1 (mJ43-eF450/FITC; mRMP1-30-APC; hMIH4-FITC); PD-L1 (mMIH5- PE-Cy7/PE; hMIH1-PE-Cy7); FAS-L1 (mMFL3-FITC); Ki-67 (mSolA15-eF450); IgG2a (m2a-15F8-FITC/APC; mRMG2a-62-PE); IgG1 (mM1-14D12-eF450; mRMG1-1-FITC); IL-10 (mJES5-16E3-PE); CD11c (mN418-eF450/PE); CD11b (mM1/70-eF450/eF660); MHCII (mM5/114.15.2-FITC/PE); Gr-1 (m1A8-Ly6G-PerCP- eF710); IL-17 (meBio17B7-PE-Cy7); Foxp3 (mFJK-16s-eF450); Perforin (meBioOMAK-D-FITC); CXCR5 (mSPRCL5-PE/APC/Biotin); CD62L (mMEL- 14-PE/APC); SIINFEKL/H-2Kb (meBio25-D1.16-APC); F4/80 (mBM8-PE/FITC); anti-Biotin (BK-1/39-PE); CD5 (m53-7.3-FITC/eF450) and NK1.1 (mPK136- APC); CD31 (m390-eF450) (most from eBioscience and Biolegend); pAb rabbit to CD3 (DAKO, IS503); and alpha-SMA (m-ab5694; DAKO, h1A4).

    Techniques: Mouse Assay, Staining, Expressing, Fluorescence, MANN-WHITNEY

    Liver-IgA + cells are oligoclonal, and effects of HFD on control WT, Iga −/− , and Cd8a −/− mice a , b , B cells and plasmocytes (CD19 + , CD138 + , IgA + cells) were sorted from spleens ( n = 3) and livers ( n = 3) of HCC-bearing MUP-uPA mice and their μ (IgM) and α (IgA) locus genetic diversities were determined by BCR sequencing. c , Circulating ALT in HFD-fed WT, Iga −/− , and Cd8a −/− mice at 6 months of age ( n = 5, 4, 8). d – f , Paraffin-embedded and frozen liver sections from the above mice were stained with haematoxylin and eosin, Oil Red O, or Sirius Red, as indicated. The data were validated at least twice. Scale bars: haematoxylin and eosin, 100 μm; Sirius Red, 100 μm; Oil Red O, 50 μm. The Sirius Red ( n = 3, 4, 8) ( d ) and Oil Red O ( n = 3, 3, 6) ( e ) stained areas were quantitated. g – i , Serum cholesterol ( g ), serum triglycerides ( h ), and liver triglycerides ( i ) were measured ( n = 5, 4, 3). Two-sided t -test ( a , d – i ) and Mann–Whitney test ( c ) were used to determine significance. * P

    Journal: Nature

    Article Title: Inflammation-induced IgA+ cells dismantle anti-liver cancer immunity

    doi: 10.1038/nature24302

    Figure Lengend Snippet: Liver-IgA + cells are oligoclonal, and effects of HFD on control WT, Iga −/− , and Cd8a −/− mice a , b , B cells and plasmocytes (CD19 + , CD138 + , IgA + cells) were sorted from spleens ( n = 3) and livers ( n = 3) of HCC-bearing MUP-uPA mice and their μ (IgM) and α (IgA) locus genetic diversities were determined by BCR sequencing. c , Circulating ALT in HFD-fed WT, Iga −/− , and Cd8a −/− mice at 6 months of age ( n = 5, 4, 8). d – f , Paraffin-embedded and frozen liver sections from the above mice were stained with haematoxylin and eosin, Oil Red O, or Sirius Red, as indicated. The data were validated at least twice. Scale bars: haematoxylin and eosin, 100 μm; Sirius Red, 100 μm; Oil Red O, 50 μm. The Sirius Red ( n = 3, 4, 8) ( d ) and Oil Red O ( n = 3, 3, 6) ( e ) stained areas were quantitated. g – i , Serum cholesterol ( g ), serum triglycerides ( h ), and liver triglycerides ( i ) were measured ( n = 5, 4, 3). Two-sided t -test ( a , d – i ) and Mann–Whitney test ( c ) were used to determine significance. * P

    Article Snippet: Antibodies specific for the following antigens were used (anti-mouse antigens were abbreviated as ‘m’, and anti-human as ‘h’): Tim-3 (mB8.2C12- AlexaF647; hF38-2E2-PerCP-eF710); CD138 (m281-2-PE/APC; hDL-101- APC); IgA (mA-6E1-PE/FITC; m11-44-2-Biotin; mRMA-1; hIS11-8E10- FITC; DAKO hA0262); CD8α (m53-6.7-PerCp-Cy5.5/PE-Cy7; hRPA-T8-PE; hC8/144B); CD45 (m30-F11-V500; hHI30-eF450); CD20 (mAISB12-PE); CD44 (mIM7-PE/FITC/eF450); CD4 (mRM4-5-eF450; mGk1.5-FITC/PE/PE-Cy7; hRPA-T4-APC); B220 (mRA3-6B2-eF450/FITC); CD19 (meBio1D3-eF660/PE-Cy7; hHIB19-PE); IgM (mII/41-PE-Cy7; hSA-DA4-eF450); IgD (m11- 26c.2a-Pacific-Blue; m11-26c-PerCP-eF710); TNF (mMP6-XT22-FITC/PE); IFNγ (mXMG1.2-eF660); GrzB (mNGZB-eF660); CD107a (meBio1D4B-PE); PD-1 (mJ43-eF450/FITC; mRMP1-30-APC; hMIH4-FITC); PD-L1 (mMIH5- PE-Cy7/PE; hMIH1-PE-Cy7); FAS-L1 (mMFL3-FITC); Ki-67 (mSolA15-eF450); IgG2a (m2a-15F8-FITC/APC; mRMG2a-62-PE); IgG1 (mM1-14D12-eF450; mRMG1-1-FITC); IL-10 (mJES5-16E3-PE); CD11c (mN418-eF450/PE); CD11b (mM1/70-eF450/eF660); MHCII (mM5/114.15.2-FITC/PE); Gr-1 (m1A8-Ly6G-PerCP- eF710); IL-17 (meBio17B7-PE-Cy7); Foxp3 (mFJK-16s-eF450); Perforin (meBioOMAK-D-FITC); CXCR5 (mSPRCL5-PE/APC/Biotin); CD62L (mMEL- 14-PE/APC); SIINFEKL/H-2Kb (meBio25-D1.16-APC); F4/80 (mBM8-PE/FITC); anti-Biotin (BK-1/39-PE); CD5 (m53-7.3-FITC/eF450) and NK1.1 (mPK136- APC); CD31 (m390-eF450) (most from eBioscience and Biolegend); pAb rabbit to CD3 (DAKO, IS503); and alpha-SMA (m-ab5694; DAKO, h1A4).

    Techniques: Mouse Assay, Sequencing, Staining, MANN-WHITNEY

    Sensitization to peanut by non-oral routes. TLR9+/+ and −/− mice were sensitized to peanut by intraperitoneal injection with alum (IP), or by repeated epicutaneous exposure (skin). A) Body temperature measured 30 min after intraperitoneal challenge with 500 g of peanut extract. B) IgE, C) IgA, and D) IgG1 antibodies were measured in serum obtained 1-2 days prior to challenge. * p

    Journal: Mucosal immunology

    Article Title: Reduced severity of peanut-induced anaphylaxis in TLR9-deficient mice is associated with selective defects in humoral immunity

    doi: 10.1038/mi.2012.55

    Figure Lengend Snippet: Sensitization to peanut by non-oral routes. TLR9+/+ and −/− mice were sensitized to peanut by intraperitoneal injection with alum (IP), or by repeated epicutaneous exposure (skin). A) Body temperature measured 30 min after intraperitoneal challenge with 500 g of peanut extract. B) IgE, C) IgA, and D) IgG1 antibodies were measured in serum obtained 1-2 days prior to challenge. * p

    Article Snippet: Peanut-specific IgG1 and IgA were detected by applying serum dilutions to peanut-coated plates and detecting with biotinylated anti-IgG1 and IgA, respectively (antibodies from BD Pharmingen, San Diego, CA).

    Techniques: Mouse Assay, Injection

    Impact of TLR9 deficiency on peanut-induced anaphylaxis and antibody and cytokine responses. TLR9 +/+ or −/− mice were orally sensitized to peanut with cholera toxin (PN/CT), followed by intraperitoneal challenge with crude peanut extract. (A) Body temperature measured 30 min after peanut challenge. Peanut (PN)-specific IgE (B), IgG1 (C) and IgA (D) as measured by ELISA in serum obtained 1-2 days prior to challenge. Cytokine secretion (E) measured after re-stimulation of spleen cells with peanut extract. Spleen cells were harvested immediately after allergen challenge. A is combined data from 2 independent experiments. B-E are from one representative experiment of 2, with n = 10 (+/+) and 8 (−/−). *** p

    Journal: Mucosal immunology

    Article Title: Reduced severity of peanut-induced anaphylaxis in TLR9-deficient mice is associated with selective defects in humoral immunity

    doi: 10.1038/mi.2012.55

    Figure Lengend Snippet: Impact of TLR9 deficiency on peanut-induced anaphylaxis and antibody and cytokine responses. TLR9 +/+ or −/− mice were orally sensitized to peanut with cholera toxin (PN/CT), followed by intraperitoneal challenge with crude peanut extract. (A) Body temperature measured 30 min after peanut challenge. Peanut (PN)-specific IgE (B), IgG1 (C) and IgA (D) as measured by ELISA in serum obtained 1-2 days prior to challenge. Cytokine secretion (E) measured after re-stimulation of spleen cells with peanut extract. Spleen cells were harvested immediately after allergen challenge. A is combined data from 2 independent experiments. B-E are from one representative experiment of 2, with n = 10 (+/+) and 8 (−/−). *** p

    Article Snippet: Peanut-specific IgG1 and IgA were detected by applying serum dilutions to peanut-coated plates and detecting with biotinylated anti-IgG1 and IgA, respectively (antibodies from BD Pharmingen, San Diego, CA).

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Hypothesis on the pathogenesis of IgA nephropathy. Synthesis of IgA1 with some O -glycans deficient in galactose (autoantigen) is elevated. Gd-IgA1 is present in the circulation at increased levels (hit 1). This immunoglobulin is recognized by unique circulating

    Journal: Kidney Diseases

    Article Title: New Insights into the Pathogenesis of IgA Nephropathy

    doi: 10.1159/000382134

    Figure Lengend Snippet: Hypothesis on the pathogenesis of IgA nephropathy. Synthesis of IgA1 with some O -glycans deficient in galactose (autoantigen) is elevated. Gd-IgA1 is present in the circulation at increased levels (hit 1). This immunoglobulin is recognized by unique circulating

    Article Snippet: These techniques also revealed significant heterogeneity of O -glycans of IgA1 in the circulation of healthy controls (table ), but it remains to be determined precisely which IgA1 glycoform(s) in patients with IgA nephropathy is associated with disease expression and/or prognosis.

    Techniques:

    Structure and O -glycosylation of human circulatory IgA1: monomeric IgA1 and hinge-region amino acid sequence with the sites of O -glycan attachment (top and middle panels). There are nine threonine and serine amino acids in the hinge region of monomeric

    Journal: Kidney Diseases

    Article Title: New Insights into the Pathogenesis of IgA Nephropathy

    doi: 10.1159/000382134

    Figure Lengend Snippet: Structure and O -glycosylation of human circulatory IgA1: monomeric IgA1 and hinge-region amino acid sequence with the sites of O -glycan attachment (top and middle panels). There are nine threonine and serine amino acids in the hinge region of monomeric

    Article Snippet: These techniques also revealed significant heterogeneity of O -glycans of IgA1 in the circulation of healthy controls (table ), but it remains to be determined precisely which IgA1 glycoform(s) in patients with IgA nephropathy is associated with disease expression and/or prognosis.

    Techniques: Sequencing

    Structure of IgA1 and Synthesis of IgA1 O-Glycans

    Journal: Kidney Diseases

    Article Title: New Insights into the Pathogenesis of IgA Nephropathy

    doi: 10.1159/000382134

    Figure Lengend Snippet: Structure of IgA1 and Synthesis of IgA1 O-Glycans

    Article Snippet: These techniques also revealed significant heterogeneity of O -glycans of IgA1 in the circulation of healthy controls (table ), but it remains to be determined precisely which IgA1 glycoform(s) in patients with IgA nephropathy is associated with disease expression and/or prognosis.

    Techniques:

    Comparison of ten serological assays for the detection of anti-SARS CoV-2 IgG. As for in Fig 2 , the same 110 serum samples were assessed for the presence of anti-SARS CoV-2 IgG. Each sample was assayed using an in-house anti-S ELISA (shown in the graph across the top of each panel, black bars), seven colloidal gold lateral flow tests (Deep Blue, Accu-Tell, GenBody, SureScreen, Spring, Biohit and Medomics), and a commercial ELISA (EUROIMMUN). A chemiluminescent assay for total anti-SARS CoV-2 IgM, IgG and IgA (Watmind) was also included. The threshold for a positive result in the in-house ELISA is set at 4-fold above background, as indicated by the red dashed line. Results for the other tests are represented as heatmaps, with colour intensity corresponding to strength of signal for each test. For EUROIMMUN, scores of

    Journal: PLoS Pathogens

    Article Title: Comparative assessment of multiple COVID-19 serological technologies supports continued evaluation of point-of-care lateral flow assays in hospital and community healthcare settings

    doi: 10.1371/journal.ppat.1008817

    Figure Lengend Snippet: Comparison of ten serological assays for the detection of anti-SARS CoV-2 IgG. As for in Fig 2 , the same 110 serum samples were assessed for the presence of anti-SARS CoV-2 IgG. Each sample was assayed using an in-house anti-S ELISA (shown in the graph across the top of each panel, black bars), seven colloidal gold lateral flow tests (Deep Blue, Accu-Tell, GenBody, SureScreen, Spring, Biohit and Medomics), and a commercial ELISA (EUROIMMUN). A chemiluminescent assay for total anti-SARS CoV-2 IgM, IgG and IgA (Watmind) was also included. The threshold for a positive result in the in-house ELISA is set at 4-fold above background, as indicated by the red dashed line. Results for the other tests are represented as heatmaps, with colour intensity corresponding to strength of signal for each test. For EUROIMMUN, scores of

    Article Snippet: Interestingly, the in-house ELISA IgM and EUROIMMUN IgA results showed particularly good agreement (94.5%), although the EUROIMMUN detected IgA more frequently in early samples compared with the IgM detected by in-house ELISA ( ).

    Techniques: Enzyme-linked Immunosorbent Assay

    Comparison of nine serological assays for the detection of anti-SARS CoV-2 IgM and IgA. 110 serum samples from 87 individuals with confirmed SARS-CoV-2 infection (RNA+ by RT-PCR) were assayed for anti-SARS CoV-2 IgM using an in-house anti-S ELISA (shown in the graph across the top of each panel, black bars), seven colloidal gold lateral flow tests (Deep Blue, Accu-Tell, GenBody, SureScreen, Spring, Biohit and Medomics), and for anti-S1 IgA using a commercial ELISA (EUROIMMUN). The threshold for a positive result in the in-house ELISA is set at 4-fold above background, as indicated by the red dashed line. Results for the other tests are represented as heatmaps, with colour intensity corresponding to strength of signal for each test. For EUROIMMUN, scores of

    Journal: PLoS Pathogens

    Article Title: Comparative assessment of multiple COVID-19 serological technologies supports continued evaluation of point-of-care lateral flow assays in hospital and community healthcare settings

    doi: 10.1371/journal.ppat.1008817

    Figure Lengend Snippet: Comparison of nine serological assays for the detection of anti-SARS CoV-2 IgM and IgA. 110 serum samples from 87 individuals with confirmed SARS-CoV-2 infection (RNA+ by RT-PCR) were assayed for anti-SARS CoV-2 IgM using an in-house anti-S ELISA (shown in the graph across the top of each panel, black bars), seven colloidal gold lateral flow tests (Deep Blue, Accu-Tell, GenBody, SureScreen, Spring, Biohit and Medomics), and for anti-S1 IgA using a commercial ELISA (EUROIMMUN). The threshold for a positive result in the in-house ELISA is set at 4-fold above background, as indicated by the red dashed line. Results for the other tests are represented as heatmaps, with colour intensity corresponding to strength of signal for each test. For EUROIMMUN, scores of

    Article Snippet: Interestingly, the in-house ELISA IgM and EUROIMMUN IgA results showed particularly good agreement (94.5%), although the EUROIMMUN detected IgA more frequently in early samples compared with the IgM detected by in-house ELISA ( ).

    Techniques: Infection, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

    Sensitivity and specificity comparison of serological assays. ( A ) Specificity was determined for each serological assay using a panel of 50 pre-pandemic serum samples the St Thomas’ emergency admissions cohort (STH Healthy, March 2019). Overall sensitivity was determined for each serological assay, based on results for 110 serum samples from known SARS-CoV-2-positive individuals (shown in Figs 3 and 4 ). For the lateral flow assays, a positive result for IgM, IgG or both is considered positive. For the EUROIMMUN and in-house ELISAs, sensitivities for IgA, IgM and IgG are calculated separately. ( B ) Sensitivity and 95% confidence intervals (vertical lines) were determined for each serological assay at increasing days POS. 95% sensitivity is indicated by the horizontal dashed line. Results for each test were categorised according to whether the serum sample was from

    Journal: PLoS Pathogens

    Article Title: Comparative assessment of multiple COVID-19 serological technologies supports continued evaluation of point-of-care lateral flow assays in hospital and community healthcare settings

    doi: 10.1371/journal.ppat.1008817

    Figure Lengend Snippet: Sensitivity and specificity comparison of serological assays. ( A ) Specificity was determined for each serological assay using a panel of 50 pre-pandemic serum samples the St Thomas’ emergency admissions cohort (STH Healthy, March 2019). Overall sensitivity was determined for each serological assay, based on results for 110 serum samples from known SARS-CoV-2-positive individuals (shown in Figs 3 and 4 ). For the lateral flow assays, a positive result for IgM, IgG or both is considered positive. For the EUROIMMUN and in-house ELISAs, sensitivities for IgA, IgM and IgG are calculated separately. ( B ) Sensitivity and 95% confidence intervals (vertical lines) were determined for each serological assay at increasing days POS. 95% sensitivity is indicated by the horizontal dashed line. Results for each test were categorised according to whether the serum sample was from

    Article Snippet: Interestingly, the in-house ELISA IgM and EUROIMMUN IgA results showed particularly good agreement (94.5%), although the EUROIMMUN detected IgA more frequently in early samples compared with the IgM detected by in-house ELISA ( ).

    Techniques: Serologic Assay

    Associations between the plasma levels of APRIL and immunoglobulin production. (A) Associations between the APRIL levels and HIV-1-specific IgM in HIV-1-infected individuals ( n = 59). (B) Associations between the APRIL levels and HIV-1-specific IgA in HIV-1-infected individuals ( n = 59). (C) Associations between the APRIL levels and neutralization titers in untreated HIV-1-infected individuals ( n = 46). (D) Associations between the APRIL levels and neutralization breadth in untreated HIV-1-infected individuals ( n = 46). LTNPs-B, long-term non-progressors at baseline; LTNPs-L, long-term non-progressors at the latest measurement; TPs, typical progressors; ART, antiretroviral therapy; ID 50 , 50% inhibitory dilution. Intergroup comparisons were performed using a Kruskal–Wallis test, followed by Dunn's post-test; p and r values were calculated by Spearman's rank correlation tests (* p

    Journal: Frontiers in Medicine

    Article Title: High APRIL Levels Are Associated With Slow Disease Progression and Low Immune Activation in Chronic HIV-1-Infected Patients

    doi: 10.3389/fmed.2020.00299

    Figure Lengend Snippet: Associations between the plasma levels of APRIL and immunoglobulin production. (A) Associations between the APRIL levels and HIV-1-specific IgM in HIV-1-infected individuals ( n = 59). (B) Associations between the APRIL levels and HIV-1-specific IgA in HIV-1-infected individuals ( n = 59). (C) Associations between the APRIL levels and neutralization titers in untreated HIV-1-infected individuals ( n = 46). (D) Associations between the APRIL levels and neutralization breadth in untreated HIV-1-infected individuals ( n = 46). LTNPs-B, long-term non-progressors at baseline; LTNPs-L, long-term non-progressors at the latest measurement; TPs, typical progressors; ART, antiretroviral therapy; ID 50 , 50% inhibitory dilution. Intergroup comparisons were performed using a Kruskal–Wallis test, followed by Dunn's post-test; p and r values were calculated by Spearman's rank correlation tests (* p

    Article Snippet: The bound antibodies were detected with horseradish peroxidase (HRP)-conjugated goat anti-human IgG, IgM, and IgA (1:10,000, Abcam) and mouse anti-human IgG1, IgG2 (1:1,000, Abcam), IgG3 (1:1,000, Thermo Fisher, Waltham, Massachusetts, USA), and IgG4 (1:4,000, Abcam).

    Techniques: Infection, Neutralization

    Temporal immune responses to OMVs derived from NTHi strain 3198-R. Shown are the median titers over time of IgM (A), IgA (B), and IgG1 (C) antibodies to 3198-R OMVs in sera from mice intranasally immunized with either IM-1 (solid line) or IM-2 (dashed line) as well as in sera from nonvaccinated control mice (dotted line) (n = 20 for each group). The error bars indicate the interquartile range of each data set for each time point.

    Journal: PLoS ONE

    Article Title: Intranasal Immunization with Nontypeable Haemophilus influenzae Outer Membrane Vesicles Induces Cross-Protective Immunity in Mice

    doi: 10.1371/journal.pone.0042664

    Figure Lengend Snippet: Temporal immune responses to OMVs derived from NTHi strain 3198-R. Shown are the median titers over time of IgM (A), IgA (B), and IgG1 (C) antibodies to 3198-R OMVs in sera from mice intranasally immunized with either IM-1 (solid line) or IM-2 (dashed line) as well as in sera from nonvaccinated control mice (dotted line) (n = 20 for each group). The error bars indicate the interquartile range of each data set for each time point.

    Article Snippet: Quantitation of antibodies Immunoglobulin A (IgA), IgG1, and IgM antibodies as well as half-maximum total Ig titers (IgA, IgG, and IgM) to OMVs were quantified by indirect enzyme-linked immunosorbent assay (ELISA) using 96-well ELISA microplates (BD Falcon) essentially as described previously , .

    Techniques: Derivative Assay, Mouse Assay

    Characterization of the immune response from mice intraperitoneally immunized with NTHi OMVs. (A) Shown are the median IgG1and IgA titers to OMVs derived from NTHi strain 2019-R in sera from mice intraperitoneally immunized with IM-1 collected at day 0 and 39 (n = 7). (B) Shown are the median half-maximum total Ig titers to OMVs derived from NTHi strains 2019-R in sera collected at day 39 from mice intraperitoneally immunized with IM-1 (i.p. IM-1) as well as from nonvaccinated control mice (co) (n = 6 for the co group and n = 7 for the i.p. IM-1 group). The error bars indicate the interquartile range of each data set for each time point. Significant differences between the data sets are marked by asterisks ( P

    Journal: PLoS ONE

    Article Title: Intranasal Immunization with Nontypeable Haemophilus influenzae Outer Membrane Vesicles Induces Cross-Protective Immunity in Mice

    doi: 10.1371/journal.pone.0042664

    Figure Lengend Snippet: Characterization of the immune response from mice intraperitoneally immunized with NTHi OMVs. (A) Shown are the median IgG1and IgA titers to OMVs derived from NTHi strain 2019-R in sera from mice intraperitoneally immunized with IM-1 collected at day 0 and 39 (n = 7). (B) Shown are the median half-maximum total Ig titers to OMVs derived from NTHi strains 2019-R in sera collected at day 39 from mice intraperitoneally immunized with IM-1 (i.p. IM-1) as well as from nonvaccinated control mice (co) (n = 6 for the co group and n = 7 for the i.p. IM-1 group). The error bars indicate the interquartile range of each data set for each time point. Significant differences between the data sets are marked by asterisks ( P

    Article Snippet: Quantitation of antibodies Immunoglobulin A (IgA), IgG1, and IgM antibodies as well as half-maximum total Ig titers (IgA, IgG, and IgM) to OMVs were quantified by indirect enzyme-linked immunosorbent assay (ELISA) using 96-well ELISA microplates (BD Falcon) essentially as described previously , .

    Techniques: Mouse Assay, Derivative Assay

    Mucosal immune responses to OMVs derived from NTHi strain 2019-R and 3198-R. Shown are the median IgA titers to OMVs derived from NTHi strain 2019-R (A) and 3198-R (B) extracted from fecal pellets collected at day 39 from mice intranasally immunized with either IM-1 or IM-2 as well as from nonvaccinated control mice (co) (n = 10 for each group). The error bars indicate the interquartile range of each data set. Significant differences between the data sets are marked by asterisks ( P

    Journal: PLoS ONE

    Article Title: Intranasal Immunization with Nontypeable Haemophilus influenzae Outer Membrane Vesicles Induces Cross-Protective Immunity in Mice

    doi: 10.1371/journal.pone.0042664

    Figure Lengend Snippet: Mucosal immune responses to OMVs derived from NTHi strain 2019-R and 3198-R. Shown are the median IgA titers to OMVs derived from NTHi strain 2019-R (A) and 3198-R (B) extracted from fecal pellets collected at day 39 from mice intranasally immunized with either IM-1 or IM-2 as well as from nonvaccinated control mice (co) (n = 10 for each group). The error bars indicate the interquartile range of each data set. Significant differences between the data sets are marked by asterisks ( P

    Article Snippet: Quantitation of antibodies Immunoglobulin A (IgA), IgG1, and IgM antibodies as well as half-maximum total Ig titers (IgA, IgG, and IgM) to OMVs were quantified by indirect enzyme-linked immunosorbent assay (ELISA) using 96-well ELISA microplates (BD Falcon) essentially as described previously , .

    Techniques: Derivative Assay, Mouse Assay

    Temporal immune responses to OMVs derived from NTHi strain 2019-R. Shown are the median titers over time of IgM (A), IgA (B), and IgG1 (C) antibodies to 2019-R OMVs in sera from mice intranasally immunized with either IM-1 (solid line) or IM-2 (dashed line) as well as in sera from nonvaccinated control mice (dotted line) (n = 20 for each group). The error bars indicate the interquartile range of each data set for each time point.

    Journal: PLoS ONE

    Article Title: Intranasal Immunization with Nontypeable Haemophilus influenzae Outer Membrane Vesicles Induces Cross-Protective Immunity in Mice

    doi: 10.1371/journal.pone.0042664

    Figure Lengend Snippet: Temporal immune responses to OMVs derived from NTHi strain 2019-R. Shown are the median titers over time of IgM (A), IgA (B), and IgG1 (C) antibodies to 2019-R OMVs in sera from mice intranasally immunized with either IM-1 (solid line) or IM-2 (dashed line) as well as in sera from nonvaccinated control mice (dotted line) (n = 20 for each group). The error bars indicate the interquartile range of each data set for each time point.

    Article Snippet: Quantitation of antibodies Immunoglobulin A (IgA), IgG1, and IgM antibodies as well as half-maximum total Ig titers (IgA, IgG, and IgM) to OMVs were quantified by indirect enzyme-linked immunosorbent assay (ELISA) using 96-well ELISA microplates (BD Falcon) essentially as described previously , .

    Techniques: Derivative Assay, Mouse Assay

    Effect of immunization route on response. Ten mice per group were immunized three times with a 2-week interval between immunizations via intranasal (IN) or subcutaneous (SC) route or a combination of these using GLA-3M-052-PEG2000 liposomes as an adjuvant. Samples were collected 1 week after 3rd immunization. a Stool supernatants were diluted 400-fold to determine LecA-specific IgA titer by ELISA. b Plasma samples were diluted 100,000-fold to determine titers of IgG1 (black circles) and IgG2a (red circles) subtypes. c Splenocytes were restimulated with LecA for 72 h and production of extracellular cytokines in the culture supernatants was determined by Luminex. Cytokine levels of the unstimulated samples were at the baseline (not shown). Error bars represent standard error of the mean. For clarity, statistical significance vs. the adjuvant alone control groups is not represented but is detailed below. For the analysis of the data in plot a , Welch’s one-way ANOVA was employed with Games–Howell correction for multiple comparisons; all of the vaccine groups except for SC + SC + SC were statistically different ( p

    Journal: NPJ Vaccines

    Article Title: Adjuvant composition and delivery route shape immune response quality and protective efficacy of a recombinant vaccine for Entamoeba histolytica

    doi: 10.1038/s41541-018-0060-x

    Figure Lengend Snippet: Effect of immunization route on response. Ten mice per group were immunized three times with a 2-week interval between immunizations via intranasal (IN) or subcutaneous (SC) route or a combination of these using GLA-3M-052-PEG2000 liposomes as an adjuvant. Samples were collected 1 week after 3rd immunization. a Stool supernatants were diluted 400-fold to determine LecA-specific IgA titer by ELISA. b Plasma samples were diluted 100,000-fold to determine titers of IgG1 (black circles) and IgG2a (red circles) subtypes. c Splenocytes were restimulated with LecA for 72 h and production of extracellular cytokines in the culture supernatants was determined by Luminex. Cytokine levels of the unstimulated samples were at the baseline (not shown). Error bars represent standard error of the mean. For clarity, statistical significance vs. the adjuvant alone control groups is not represented but is detailed below. For the analysis of the data in plot a , Welch’s one-way ANOVA was employed with Games–Howell correction for multiple comparisons; all of the vaccine groups except for SC + SC + SC were statistically different ( p

    Article Snippet: Horseradish peroxidase-conjugated secondary antibodies specific for IgA (catalog #1040-05) and IgG (catalog #1071-05 and 1081-05) subtypes were purchased from Southern Biotechnology and diluted as per the manufacturer’s instructions.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay, Luminex

    Biological activity of adjuvant formulation: importance of PEG length and complementary roles for GLA and 3M-052. Five mice per group were immunized three times with a 2-week interval between immunizations using a mixed intranasal/subcutaneous regimen. Mice were euthanized a week after third immunization and samples collected. a Stool supernatants were diluted 250-fold and anti-LecA IgA titer was determined by ELISA. b Plasma samples were diluted 100,000-fold and titers of anti-LecA IgG1 (black circles) and IgG2a (red circles) subtypes were determined by ELISA. c Intracellular IFN-γ levels were measured using flow cytometry as described (negative values obtained after subtracting values from unstimulated cells were considered zero; the outlier test described below was conducted prior to this data transformation). Error bars represent standard error of the mean. For the analysis of the data in plot a , data were analyzed by one-way ANOVA with Sidak’s correction for selected comparisons. For the analysis of the data in plot b , data were log-transformed with a small offset if necessary and Sidak’s correction for selected comparisons was employed. All formulations in plot b elicited statistically increased ( p

    Journal: NPJ Vaccines

    Article Title: Adjuvant composition and delivery route shape immune response quality and protective efficacy of a recombinant vaccine for Entamoeba histolytica

    doi: 10.1038/s41541-018-0060-x

    Figure Lengend Snippet: Biological activity of adjuvant formulation: importance of PEG length and complementary roles for GLA and 3M-052. Five mice per group were immunized three times with a 2-week interval between immunizations using a mixed intranasal/subcutaneous regimen. Mice were euthanized a week after third immunization and samples collected. a Stool supernatants were diluted 250-fold and anti-LecA IgA titer was determined by ELISA. b Plasma samples were diluted 100,000-fold and titers of anti-LecA IgG1 (black circles) and IgG2a (red circles) subtypes were determined by ELISA. c Intracellular IFN-γ levels were measured using flow cytometry as described (negative values obtained after subtracting values from unstimulated cells were considered zero; the outlier test described below was conducted prior to this data transformation). Error bars represent standard error of the mean. For the analysis of the data in plot a , data were analyzed by one-way ANOVA with Sidak’s correction for selected comparisons. For the analysis of the data in plot b , data were log-transformed with a small offset if necessary and Sidak’s correction for selected comparisons was employed. All formulations in plot b elicited statistically increased ( p

    Article Snippet: Horseradish peroxidase-conjugated secondary antibodies specific for IgA (catalog #1040-05) and IgG (catalog #1071-05 and 1081-05) subtypes were purchased from Southern Biotechnology and diluted as per the manufacturer’s instructions.

    Techniques: Activity Assay, Mouse Assay, Enzyme-linked Immunosorbent Assay, Flow Cytometry, Cytometry, Transformation Assay