anti igg1 igg4 antibodies Search Results


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  • 99
    Thermo Fisher igg2 antibodies
    Viremic HIV controllers exhibited higher <t>IgG</t> antibodies to HIV-1 p24 associated with greater isotype diversification
    Igg2 Antibodies, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore igg2
    Western blot analysis of the serum <t>IgG</t> reactivities of 15 sera from kala-azar patients in our longitudinal study, with LAg being incorporated into liposomes following separation by SDS-PAGE. (A) Reactivity with acute-phase sera; (B) corresponding reactivity after SAG treatment; (C) cross-reactivity with serum from patients with malaria (lanes 1 to 3), typhoid (lanes 4 to 6), tuberculosis (lanes 7 to 9), and leprosy (lanes 10 to 12) and healthy controls (lanes 13 to 15). The major bands (in kilodaltons) are marked on the left.
    Igg2, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 639 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Nordic-Mubio igg2b
    Induction of anti-nucleolar autoantibodies (ANolA) of <t>IgG1</t> and <t>IgG2a</t> isotypes in outbred Institute of Cancer Research (ICR), Naval Medical Research Institute (NMRI) and Black Swiss mice treated with silver nitrate (AgNO 3 ). Groups of female ICR (solid and open circles), NMRI (solid and open squares) and Black Swiss (solid and open hexagons) outbred mice were injected repeatedly subcutaneously (s.c.) with AgNO 3 (solid symbols) or NaCl (open symbols) for 4 weeks. At the end of each experiment the mice were bled and killed. The sera were tested for the presence of immunoglobulin (Ig)G1 (a) and IgG2a (b) ANolA by using an indirect immunofluorescence (IIF) method. Each symbol represents a single animal. The mean values ± standard error are presented as thin vertical lines. Values in parentheses represent the proportions of silver-treated animals, which produce ANolA.
    Igg2b, supplied by Nordic-Mubio, used in various techniques. Bioz Stars score: 85/100, based on 24 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Boehringer Ingelheim anti igg1 igg4 antibodies
    Induction of anti-nucleolar autoantibodies (ANolA) of <t>IgG1</t> and <t>IgG2a</t> isotypes in outbred Institute of Cancer Research (ICR), Naval Medical Research Institute (NMRI) and Black Swiss mice treated with silver nitrate (AgNO 3 ). Groups of female ICR (solid and open circles), NMRI (solid and open squares) and Black Swiss (solid and open hexagons) outbred mice were injected repeatedly subcutaneously (s.c.) with AgNO 3 (solid symbols) or NaCl (open symbols) for 4 weeks. At the end of each experiment the mice were bled and killed. The sera were tested for the presence of immunoglobulin (Ig)G1 (a) and IgG2a (b) ANolA by using an indirect immunofluorescence (IIF) method. Each symbol represents a single animal. The mean values ± standard error are presented as thin vertical lines. Values in parentheses represent the proportions of silver-treated animals, which produce ANolA.
    Anti Igg1 Igg4 Antibodies, supplied by Boehringer Ingelheim, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam anti igg1 igg2a igg3 antibody m204 3
    Induction of anti-nucleolar autoantibodies (ANolA) of <t>IgG1</t> and <t>IgG2a</t> isotypes in outbred Institute of Cancer Research (ICR), Naval Medical Research Institute (NMRI) and Black Swiss mice treated with silver nitrate (AgNO 3 ). Groups of female ICR (solid and open circles), NMRI (solid and open squares) and Black Swiss (solid and open hexagons) outbred mice were injected repeatedly subcutaneously (s.c.) with AgNO 3 (solid symbols) or NaCl (open symbols) for 4 weeks. At the end of each experiment the mice were bled and killed. The sera were tested for the presence of immunoglobulin (Ig)G1 (a) and IgG2a (b) ANolA by using an indirect immunofluorescence (IIF) method. Each symbol represents a single animal. The mean values ± standard error are presented as thin vertical lines. Values in parentheses represent the proportions of silver-treated animals, which produce ANolA.
    Anti Igg1 Igg2a Igg3 Antibody M204 3, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Thermo Fisher mouse anti human igg1 igg2 igg3 igg4 antibodies
    Induction of anti-nucleolar autoantibodies (ANolA) of <t>IgG1</t> and <t>IgG2a</t> isotypes in outbred Institute of Cancer Research (ICR), Naval Medical Research Institute (NMRI) and Black Swiss mice treated with silver nitrate (AgNO 3 ). Groups of female ICR (solid and open circles), NMRI (solid and open squares) and Black Swiss (solid and open hexagons) outbred mice were injected repeatedly subcutaneously (s.c.) with AgNO 3 (solid symbols) or NaCl (open symbols) for 4 weeks. At the end of each experiment the mice were bled and killed. The sera were tested for the presence of immunoglobulin (Ig)G1 (a) and IgG2a (b) ANolA by using an indirect immunofluorescence (IIF) method. Each symbol represents a single animal. The mean values ± standard error are presented as thin vertical lines. Values in parentheses represent the proportions of silver-treated animals, which produce ANolA.
    Mouse Anti Human Igg1 Igg2 Igg3 Igg4 Antibodies, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    89
    Sanquin igg4 antibody depletion igg1 anti igg4 antibody
    <t>IgG</t> 4 /IgE ratios to peanut and peanut allergens in children with PA (n = 65) and PS children (n = 27). Values are presented as medians and interquartile ranges. P values refer to the comparison between children with PA and PS children using the Mann-Whitney U test.
    Igg4 Antibody Depletion Igg1 Anti Igg4 Antibody, supplied by Sanquin, used in various techniques. Bioz Stars score: 89/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Bio-Rad igg4
    Anti-cE5 IgG1 and <t>IgG4</t> responses. (A) Box plots of anti-cE5 IgG1 and IgG4 levels among the 207 individuals of the Aug 1994 survey. Boxes, whiskers, dots and P value as in Figure 1 . IgG1 and IgG4 levels are expressed in ng/ml. (B) Scatter plot reporting the IgG1 (red) and IgG4 (black) antibody response to cE5 as function of age (n = 207). The best-fit lines (solid) and 95% CI (broken lines) are shown. Note that three data points are outside the axis limits.
    Igg4, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 95/100, based on 98 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    95
    Abcam anti igg1
    Effects of TLR ‐7 agonist R848 in transgenic mice. A , Detection of GFP in bone marrow–derived macrophages ( BMM s) from mTLR ‐7−/− (knockout [ KO ]) mice and transgenic (Tg) mice by flow cytometric analysis, after staining with anti‐ CD 11b antibody. At least 2 independent experiments were performed. B , Interleukin‐6 ( IL ‐6) and IL ‐12p40 levels in BMM s from knockout mice and transgenic mice left unstimulated or stimulated with R848. After 24 hours of incubation, IL ‐6 and IL ‐12p40 were detected in culture medium by enzyme‐linked immunosorbent assay ( ELISA ). Results are from triplicate wells. At least 2 independent experiments were performed. C , Weight of SMG s, kidneys, lungs, and liver in 8‐week‐old knockout, wild‐type ( WT ), and transgenic mice treated with topical R848 for 4 weeks (n = 8 per group). D and F , H E‐stained ( D ) and Masson's trichrome–stained ( F ) sections of SMG s, pancreas, kidneys, lungs, and liver from representative 8‐week‐old R848‐treated knockout, WT , and transgenic mice. Bars = 100 μm. E and G , Focus score ( E ) and fibrosis score ( G ) for each organ in 8‐week‐old knockout, WT , and transgenic mice left untreated or treated with R848 (n = 8 per group). The fibrosis score was calculated from Masson's trichrome staining as described in Patients and Methods. HPF = high‐power field. H , Serial sections of SMG s from a representative 8‐week‐old R848‐treated transgenic mouse, stained with H E and for <t>IgG1,</t> TLR ‐7, CD 206, CD 317, and IL ‐33. Mayer's hematoxylin (blue) counterstained; bars = 100 μm. I , Serum IgG, IgG1, and IgG2a levels in knockout, WT , and transgenic mice (n = 10 per group) before and after R848 treatment, as determined by ELISA . Symbols represent individual mice; horizontal lines show the mean. J , Serum IL ‐33 levels, determined by ELISA , in knockout, WT , and transgenic mice left untreated or treated with R848 (n = 10 per group). In B , C , E , G , and J , bars show the mean ± SD . * = P
    Anti Igg1, supplied by Abcam, used in various techniques. Bioz Stars score: 95/100, based on 55 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    igg1  (Abcam)
    99
    Abcam igg1
    Serum levels of total IgG (A), <t>IgG1</t> (B), IgG2a (C) and IgG2b (D) antibodies (OD 450 ± SD) in immunized and control groups of mice, at week 0, 2, 4, and 6. Significant differences between control and immunized groups are marked with (*), where * P
    Igg1, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 316 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Thermo Fisher anti igg
    IFNαR 1 Is Required for Dysregulated Spt-AFC and Spt-GC Formation in B6. Sle1b Mice (A) Flow cytometric analysis of surface expression of IFNαR 1 on B220 + Fas low GL-7 low non-GC B cells (red), B220 + Fas h GL-7 hi GC B cells (blue), CD4 + PD-1 low CXCR5 low non-Tfh (orange), and CD4 + PD-1 hi CXCR5 hi Tfh (green) in female B6 and B6. Sle1b mice. Error bars represent mean ± SEM. (B and C) The percentages of GC B cells (B) and Tfh cells (C) in splenocytes from 3- and 6-month-old female mice. (D) Spleen sections from 6-month-old B6, B6.IFNαR 1 −/− , B6. Sle1b , and B6. Sle1b .IFNαR 1 −/− female mice (5 mice per group) were stained with anti-CD4 Ab (red) GL-7 (green) and anti-IgD Ab (blue), and GC areas were measured for 10 GCs per spleen section (right panels). The scale bars represent 50 μm. (E and F) Each symbol represents a measured GC. IgM + (E) and <t>IgG</t> + (F) AFCs in the spleens of 6-month-old B6, B6.IFNαR 1 −/− , B6. Sle1b , and B6. Sle1b .IFNαR 1 −/− female mice are shown. (G-J) Analysis of serum titers of IgM (G), IgG (H), IgG1 (I), and IgG2c (J) antibodies in 6-month-old mice by <t>ELISA.</t> Each symbol represents a mouse, and these data represent at least two-three independent experiments. Statistical significance was determined by one-way ANOVA with a follow-up Tukey multiple-comparison test (NS, not significant, *p
    Anti Igg, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 98/100, based on 446 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Beckman Coulter pe conjugated mouse anti human igg1 igg2 igg3 igg4 antibody
    Complement-dependent cytotoxicity (CDC) of anti-FLRT2 antibody . CDC activities using two healthy control IgG and two anti-FLRT2 positive IgG, B11-8 and X10-48, at a concentration of 1.28 mg/ml with 1:3 diluted complement  (A) , 1.28 mg/ml of IgG, and 1:3 diluted complement with recombinant FLRT2 at the indicated concentrations  (B) , and various IgG concentrations with 1:6 diluted complement  (C)  against HUVECs were measured with the WST-1 assay.  (D)  CDC activities against mock transfected HEK293T cells (empty, left) and FLRT2-expressing HEK293T cells (FLRT2, right) by using 1.28 mg/ml of IgG and 1:3 diluted complement were measured with the WST-1 assay. HEK293T cells negative or positive for FLRT2 expression were stained with anti-FLRT2 antibody followed by secondary antibody against human IgG1, IgG2, IgG3, IgG4  (E) , and IgM  (F) , and analyzed with flow cytometry.  (G)  ADCC activities using control IgG, B11-8, and X10-48, at a concentration of 1.28 mg/ml with an E:T ratio of 25:1 were determined with the lactate dehydrogenase detection method. Error bars indicate SD. * P
    Pe Conjugated Mouse Anti Human Igg1 Igg2 Igg3 Igg4 Antibody, supplied by Beckman Coulter, used in various techniques. Bioz Stars score: 85/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abbexa igg antibodies
    Complement-dependent cytotoxicity (CDC) of anti-FLRT2 antibody . CDC activities using two healthy control IgG and two anti-FLRT2 positive IgG, B11-8 and X10-48, at a concentration of 1.28 mg/ml with 1:3 diluted complement  (A) , 1.28 mg/ml of IgG, and 1:3 diluted complement with recombinant FLRT2 at the indicated concentrations  (B) , and various IgG concentrations with 1:6 diluted complement  (C)  against HUVECs were measured with the WST-1 assay.  (D)  CDC activities against mock transfected HEK293T cells (empty, left) and FLRT2-expressing HEK293T cells (FLRT2, right) by using 1.28 mg/ml of IgG and 1:3 diluted complement were measured with the WST-1 assay. HEK293T cells negative or positive for FLRT2 expression were stained with anti-FLRT2 antibody followed by secondary antibody against human IgG1, IgG2, IgG3, IgG4  (E) , and IgM  (F) , and analyzed with flow cytometry.  (G)  ADCC activities using control IgG, B11-8, and X10-48, at a concentration of 1.28 mg/ml with an E:T ratio of 25:1 were determined with the lactate dehydrogenase detection method. Error bars indicate SD. * P
    Igg Antibodies, supplied by Abbexa, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BioLegend anti mouse igg1
    Ranitidine significantly enhanced levels of <t>IgG2a</t> capable of binding to E0771-GFP cancer cells. Serum was diluted 1:10 and added to cultured E0771-GFP cells and anti-isotype antibodies for IgG2a (A) or kappa light chain (B) were added. Relative fluorescence intensity (RFI) was used as an indicator for antibody binding to E0771-GFP cells relative to the average of control MFI. Box plot represents mean and range of data from 20 to 24 individual mice. * p
    Anti Mouse Igg1, supplied by BioLegend, used in various techniques. Bioz Stars score: 95/100, based on 83 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Bio-Rad anti igg1
    dAd5GNC evoked anti-GNC antibodies in mice. ( a ) Total anti-GNC <t>IgG</t> antibody titers over time. BALB/c mice ( n = 20) were vaccinated intramuscularly with 4 µg dAd5GNC at 0, 3, and 6 week. Antibody titers were assessed by ELISA against BSA
    Anti Igg1, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 91/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    BioLegend igg1 pe
    dAd5GNC evoked anti-GNC antibodies in mice. ( a ) Total anti-GNC <t>IgG</t> antibody titers over time. BALB/c mice ( n = 20) were vaccinated intramuscularly with 4 µg dAd5GNC at 0, 3, and 6 week. Antibody titers were assessed by ELISA against BSA
    Igg1 Pe, supplied by BioLegend, used in various techniques. Bioz Stars score: 96/100, based on 111 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    VMRD Inc igg1
    dAd5GNC evoked anti-GNC antibodies in mice. ( a ) Total anti-GNC <t>IgG</t> antibody titers over time. BALB/c mice ( n = 20) were vaccinated intramuscularly with 4 µg dAd5GNC at 0, 3, and 6 week. Antibody titers were assessed by ELISA against BSA
    Igg1, supplied by VMRD Inc, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Becton Dickinson igg2
    Anti- S. aureus IgG1 and <t>IgG2</t> titers in paired maternal and cord serum samples from the case and control groups. (a) Anti- S. aureus IgG1 titers and (d) anti- S. aureus IgG2 titers in the maternal serum and umbilical cord serum samples from the case (n=48) and control group (n=98). In (b) and (c) , correlation coefficients between paired maternal and umbilical cord serum of anti- Sa IgG1 in the case and control group, respectively. In (e) and (f) , correlation coefficients between the paired maternal and umbilical cord serum of anti- Sa IgG2 in the case and control group, respectively. In (a) and (d) , the box represents the 25 th –75 th percentiles, and the median is represented by the line within the box. The whiskers represent the 5 th –95 th percentiles.
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    Image Search Results


    Viremic HIV controllers exhibited higher IgG antibodies to HIV-1 p24 associated with greater isotype diversification

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Viremic HIV controllers exhibit high plasmacytoid dendritic cell\reactive opsonophagocytic IgG antibody responses against HIV-1 p24 associated with greater antibody isotype diversification

    doi: 10.4049/jimmunol.1402918

    Figure Lengend Snippet: Viremic HIV controllers exhibited higher IgG antibodies to HIV-1 p24 associated with greater isotype diversification

    Article Snippet: The extent of IgG1 and IgG2 depletion was assessed by ELISA using a similar protocol to that used for detection of HIV p24- and HIV gp140-specific IgG1 and IgG2 antibodies, but plates were coated with 1μg/mL recombinant protein G (Pierce® ) instead of antigens and antibodies were detected with either biotinylated anti-human IgG1 (Invitrogen) diluted 1:2000 or biotinylated anti-human IgG2 (Southern Biotech) diluted 1:4000.

    Techniques:

    pDC reactive opsonophagocytic antibody responses against HIV-1 p24 are mediated via FcγRIIa and not decreased by depletion of IgG2 from immunoglobulin preparations

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Viremic HIV controllers exhibit high plasmacytoid dendritic cell\reactive opsonophagocytic IgG antibody responses against HIV-1 p24 associated with greater antibody isotype diversification

    doi: 10.4049/jimmunol.1402918

    Figure Lengend Snippet: pDC reactive opsonophagocytic antibody responses against HIV-1 p24 are mediated via FcγRIIa and not decreased by depletion of IgG2 from immunoglobulin preparations

    Article Snippet: The extent of IgG1 and IgG2 depletion was assessed by ELISA using a similar protocol to that used for detection of HIV p24- and HIV gp140-specific IgG1 and IgG2 antibodies, but plates were coated with 1μg/mL recombinant protein G (Pierce® ) instead of antigens and antibodies were detected with either biotinylated anti-human IgG1 (Invitrogen) diluted 1:2000 or biotinylated anti-human IgG2 (Southern Biotech) diluted 1:4000.

    Techniques:

    IgG antibody responses against HIV-1 p24 correlated inversely with plasma HIV viral load in all viremic patients

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    Article Title: Viremic HIV controllers exhibit high plasmacytoid dendritic cell\reactive opsonophagocytic IgG antibody responses against HIV-1 p24 associated with greater antibody isotype diversification

    doi: 10.4049/jimmunol.1402918

    Figure Lengend Snippet: IgG antibody responses against HIV-1 p24 correlated inversely with plasma HIV viral load in all viremic patients

    Article Snippet: The extent of IgG1 and IgG2 depletion was assessed by ELISA using a similar protocol to that used for detection of HIV p24- and HIV gp140-specific IgG1 and IgG2 antibodies, but plates were coated with 1μg/mL recombinant protein G (Pierce® ) instead of antigens and antibodies were detected with either biotinylated anti-human IgG1 (Invitrogen) diluted 1:2000 or biotinylated anti-human IgG2 (Southern Biotech) diluted 1:4000.

    Techniques:

    Western blot analysis of the serum IgG reactivities of 15 sera from kala-azar patients in our longitudinal study, with LAg being incorporated into liposomes following separation by SDS-PAGE. (A) Reactivity with acute-phase sera; (B) corresponding reactivity after SAG treatment; (C) cross-reactivity with serum from patients with malaria (lanes 1 to 3), typhoid (lanes 4 to 6), tuberculosis (lanes 7 to 9), and leprosy (lanes 10 to 12) and healthy controls (lanes 13 to 15). The major bands (in kilodaltons) are marked on the left.

    Journal: Infection and Immunity

    Article Title: Characterization of Immunoglobulin G and Its Subclass Response to Indian Kala-Azar Infection before and after Chemotherapy

    doi: 10.1128/IAI.72.2.863-870.2004

    Figure Lengend Snippet: Western blot analysis of the serum IgG reactivities of 15 sera from kala-azar patients in our longitudinal study, with LAg being incorporated into liposomes following separation by SDS-PAGE. (A) Reactivity with acute-phase sera; (B) corresponding reactivity after SAG treatment; (C) cross-reactivity with serum from patients with malaria (lanes 1 to 3), typhoid (lanes 4 to 6), tuberculosis (lanes 7 to 9), and leprosy (lanes 10 to 12) and healthy controls (lanes 13 to 15). The major bands (in kilodaltons) are marked on the left.

    Article Snippet: For IgG subclass reactivity, the incubation of the membranes with human sera at a 1:100 dilution was followed by a 1-h reaction with anti-human IgG1, IgG2, IgG3, and IgG4 monoclonal antibodies (1:300 dilution; Sigma Immunochemicals).

    Techniques: Western Blot, SDS Page

    Western blot analysis of kala-azar serum IgG reactivity with SDS-PAGE-separated (6-μg) membrane proteins of LAg before (A) and after (B) SAG treatment. Each lane represents one VL patient, and longitudinal samples have identical lane numbers. The apparent molecular masses of prominent bands in kilodaltons are marked on the left.

    Journal: Infection and Immunity

    Article Title: Characterization of Immunoglobulin G and Its Subclass Response to Indian Kala-Azar Infection before and after Chemotherapy

    doi: 10.1128/IAI.72.2.863-870.2004

    Figure Lengend Snippet: Western blot analysis of kala-azar serum IgG reactivity with SDS-PAGE-separated (6-μg) membrane proteins of LAg before (A) and after (B) SAG treatment. Each lane represents one VL patient, and longitudinal samples have identical lane numbers. The apparent molecular masses of prominent bands in kilodaltons are marked on the left.

    Article Snippet: For IgG subclass reactivity, the incubation of the membranes with human sera at a 1:100 dilution was followed by a 1-h reaction with anti-human IgG1, IgG2, IgG3, and IgG4 monoclonal antibodies (1:300 dilution; Sigma Immunochemicals).

    Techniques: Western Blot, SDS Page

    Immunoblot profiles of LAg-specific IgG subclass reactivities of four (A to D) kala-azar patients before and after SAG treatment. Lanes 1, pretreatment sera; lanes 2, posttreatment sera. The positions of prominent bands at apparent molecular masses in kilodaltons are shown on the left.

    Journal: Infection and Immunity

    Article Title: Characterization of Immunoglobulin G and Its Subclass Response to Indian Kala-Azar Infection before and after Chemotherapy

    doi: 10.1128/IAI.72.2.863-870.2004

    Figure Lengend Snippet: Immunoblot profiles of LAg-specific IgG subclass reactivities of four (A to D) kala-azar patients before and after SAG treatment. Lanes 1, pretreatment sera; lanes 2, posttreatment sera. The positions of prominent bands at apparent molecular masses in kilodaltons are shown on the left.

    Article Snippet: For IgG subclass reactivity, the incubation of the membranes with human sera at a 1:100 dilution was followed by a 1-h reaction with anti-human IgG1, IgG2, IgG3, and IgG4 monoclonal antibodies (1:300 dilution; Sigma Immunochemicals).

    Techniques:

    Induction of anti-nucleolar autoantibodies (ANolA) of IgG1 and IgG2a isotypes in outbred Institute of Cancer Research (ICR), Naval Medical Research Institute (NMRI) and Black Swiss mice treated with silver nitrate (AgNO 3 ). Groups of female ICR (solid and open circles), NMRI (solid and open squares) and Black Swiss (solid and open hexagons) outbred mice were injected repeatedly subcutaneously (s.c.) with AgNO 3 (solid symbols) or NaCl (open symbols) for 4 weeks. At the end of each experiment the mice were bled and killed. The sera were tested for the presence of immunoglobulin (Ig)G1 (a) and IgG2a (b) ANolA by using an indirect immunofluorescence (IIF) method. Each symbol represents a single animal. The mean values ± standard error are presented as thin vertical lines. Values in parentheses represent the proportions of silver-treated animals, which produce ANolA.

    Journal: Clinical and Experimental Immunology

    Article Title: Mercury and silver induce B cell activation and anti-nucleolar autoantibody production in outbred mouse stocks: are environmental factors more important than the susceptibility genes in connection with autoimmunity?

    doi: 10.1111/j.1365-2249.2008.03801.x

    Figure Lengend Snippet: Induction of anti-nucleolar autoantibodies (ANolA) of IgG1 and IgG2a isotypes in outbred Institute of Cancer Research (ICR), Naval Medical Research Institute (NMRI) and Black Swiss mice treated with silver nitrate (AgNO 3 ). Groups of female ICR (solid and open circles), NMRI (solid and open squares) and Black Swiss (solid and open hexagons) outbred mice were injected repeatedly subcutaneously (s.c.) with AgNO 3 (solid symbols) or NaCl (open symbols) for 4 weeks. At the end of each experiment the mice were bled and killed. The sera were tested for the presence of immunoglobulin (Ig)G1 (a) and IgG2a (b) ANolA by using an indirect immunofluorescence (IIF) method. Each symbol represents a single animal. The mean values ± standard error are presented as thin vertical lines. Values in parentheses represent the proportions of silver-treated animals, which produce ANolA.

    Article Snippet: The numbers of splenic cells secreting antibodies belonging to different Ig classes and subclasses were quantified utilizing the protein A plaque assay described by Gronowicz et al. [ ], employing rabbit anti-mouse IgM, IgG1, IgG3 (Organon Teknika, Durham, NC, USA) and IgG2b (Nordic Immunological Laboratories, Tillburg, the Netherlands) as the developing reagents.

    Techniques: Mouse Assay, Injection, Immunofluorescence

    Induction of anti-nucleolar autoantibodies (ANolA) of IgG1 and IgG2a isotypes in outbred Institute of Cancer Research (ICR), Naval Medical Research Institute (NMRI) and Black Swiss mice treated with mercuric chloride (HgCl 2 ). Groups of female ICR (solid and open circles), NMRI (solid and open squares) and Black Swiss (solid and open hexagons) outbred mice were injected repeatedly subcutaneously (s.c.) with HgCl 2 (solid symbols) or NaCl (open symbols) for 4 weeks. At the end of each experiment the mice were bled and killed. The sera were tested for the presence of IgG1 (a) and IgG2a (b) ANolA by using an indirect immunofluorescence (IIF) method. Each symbol represents a single animal. The mean values ± standard error are presented as thin vertical lines. Values in parentheses represent the proportions of mercury-treated animals, which produce ANolA.

    Journal: Clinical and Experimental Immunology

    Article Title: Mercury and silver induce B cell activation and anti-nucleolar autoantibody production in outbred mouse stocks: are environmental factors more important than the susceptibility genes in connection with autoimmunity?

    doi: 10.1111/j.1365-2249.2008.03801.x

    Figure Lengend Snippet: Induction of anti-nucleolar autoantibodies (ANolA) of IgG1 and IgG2a isotypes in outbred Institute of Cancer Research (ICR), Naval Medical Research Institute (NMRI) and Black Swiss mice treated with mercuric chloride (HgCl 2 ). Groups of female ICR (solid and open circles), NMRI (solid and open squares) and Black Swiss (solid and open hexagons) outbred mice were injected repeatedly subcutaneously (s.c.) with HgCl 2 (solid symbols) or NaCl (open symbols) for 4 weeks. At the end of each experiment the mice were bled and killed. The sera were tested for the presence of IgG1 (a) and IgG2a (b) ANolA by using an indirect immunofluorescence (IIF) method. Each symbol represents a single animal. The mean values ± standard error are presented as thin vertical lines. Values in parentheses represent the proportions of mercury-treated animals, which produce ANolA.

    Article Snippet: The numbers of splenic cells secreting antibodies belonging to different Ig classes and subclasses were quantified utilizing the protein A plaque assay described by Gronowicz et al. [ ], employing rabbit anti-mouse IgM, IgG1, IgG3 (Organon Teknika, Durham, NC, USA) and IgG2b (Nordic Immunological Laboratories, Tillburg, the Netherlands) as the developing reagents.

    Techniques: Mouse Assay, Injection, Immunofluorescence

    IgG 4 /IgE ratios to peanut and peanut allergens in children with PA (n = 65) and PS children (n = 27). Values are presented as medians and interquartile ranges. P values refer to the comparison between children with PA and PS children using the Mann-Whitney U test.

    Journal: The Journal of Allergy and Clinical Immunology

    Article Title: IgG4 inhibits peanut-induced basophil and mast cell activation in peanut-tolerant children sensitized to peanut major allergens

    doi: 10.1016/j.jaci.2015.01.012

    Figure Lengend Snippet: IgG 4 /IgE ratios to peanut and peanut allergens in children with PA (n = 65) and PS children (n = 27). Values are presented as medians and interquartile ranges. P values refer to the comparison between children with PA and PS children using the Mann-Whitney U test.

    Article Snippet: IgG4 antibody depletion IgG1 anti-IgG4 antibody (clone MH164-4; Sanquin, Amsterdam, The Netherlands) was coupled to cyanogen bromide–activated Sepharose (GE Healthcare, Hertfordshire, United Kingdom) during an overnight incubation at 4°C.

    Techniques: MANN-WHITNEY

    Inhibition of peanut-induced activation of mast cells sensitized with plasma from a patient with PA in the presence of mock- or IgG 4 -depleted plasma samples from peanut-sensitized tolerant patients (median inhibition, 75% vs 30%, respectively; P = .007, n = 12). % Inhibition = (% CD63 + cells sensitized with plasma from a patient with PA − % CD63 + cells sensitized with plasma from a patient with PA in the presence of test plasma)/% CD63 + cells sensitized with plasma from a patient with PA. The P value refers to the comparison between IgG 4 - and mock-depleted paired samples by using the Wilcoxon signed-rank test. ** P

    Journal: The Journal of Allergy and Clinical Immunology

    Article Title: IgG4 inhibits peanut-induced basophil and mast cell activation in peanut-tolerant children sensitized to peanut major allergens

    doi: 10.1016/j.jaci.2015.01.012

    Figure Lengend Snippet: Inhibition of peanut-induced activation of mast cells sensitized with plasma from a patient with PA in the presence of mock- or IgG 4 -depleted plasma samples from peanut-sensitized tolerant patients (median inhibition, 75% vs 30%, respectively; P = .007, n = 12). % Inhibition = (% CD63 + cells sensitized with plasma from a patient with PA − % CD63 + cells sensitized with plasma from a patient with PA in the presence of test plasma)/% CD63 + cells sensitized with plasma from a patient with PA. The P value refers to the comparison between IgG 4 - and mock-depleted paired samples by using the Wilcoxon signed-rank test. ** P

    Article Snippet: IgG4 antibody depletion IgG1 anti-IgG4 antibody (clone MH164-4; Sanquin, Amsterdam, The Netherlands) was coupled to cyanogen bromide–activated Sepharose (GE Healthcare, Hertfordshire, United Kingdom) during an overnight incubation at 4°C.

    Techniques: Inhibition, Activation Assay

    Anti-cE5 IgG1 and IgG4 responses. (A) Box plots of anti-cE5 IgG1 and IgG4 levels among the 207 individuals of the Aug 1994 survey. Boxes, whiskers, dots and P value as in Figure 1 . IgG1 and IgG4 levels are expressed in ng/ml. (B) Scatter plot reporting the IgG1 (red) and IgG4 (black) antibody response to cE5 as function of age (n = 207). The best-fit lines (solid) and 95% CI (broken lines) are shown. Note that three data points are outside the axis limits.

    Journal: Parasites & Vectors

    Article Title: Differential antibody response to the Anopheles gambiae gSG6 and cE5 salivary proteins in individuals naturally exposed to bites of malaria vectors

    doi: 10.1186/s13071-014-0549-8

    Figure Lengend Snippet: Anti-cE5 IgG1 and IgG4 responses. (A) Box plots of anti-cE5 IgG1 and IgG4 levels among the 207 individuals of the Aug 1994 survey. Boxes, whiskers, dots and P value as in Figure 1 . IgG1 and IgG4 levels are expressed in ng/ml. (B) Scatter plot reporting the IgG1 (red) and IgG4 (black) antibody response to cE5 as function of age (n = 207). The best-fit lines (solid) and 95% CI (broken lines) are shown. Note that three data points are outside the axis limits.

    Article Snippet: After washing, blocking and washing again as above wells were incubated overnight at 4°C with serial dilutions, from 1 μg/ml to 0.0078 μg/ml (1 → 0.5 → 0.25 → 0.125 → 0.0625 → 0.03125 → 0.0156 → 0.0078), of purified native human IgG1 or IgG4 (ABD Serotec, Kidlington, Oxford, UK) in 50 μl of blocking reagent.

    Techniques:

    Seasonal pattern of the anti-cE5 IgG response. IgG response to the cE5 protein in the three surveys: Aug 1994 (n = 117, average age ±95% CI = 17.2 ± 3.0), Oct 1994 (n = 121, average age ±95% CI = 16.8 ± 3.1) and Mar 1995 (n = 121, average age ±95% CI = 16.0 ± 2.9). (A) Box plots of OD values among responders from the different surveys as indicated. Boxes, whiskers and dots as in Figure 1 . IgG levels are expressed as OD 492 . (B) Seroprevalence of anti-cE5 IgG antibodies. Error bars as in Figure 1 .

    Journal: Parasites & Vectors

    Article Title: Differential antibody response to the Anopheles gambiae gSG6 and cE5 salivary proteins in individuals naturally exposed to bites of malaria vectors

    doi: 10.1186/s13071-014-0549-8

    Figure Lengend Snippet: Seasonal pattern of the anti-cE5 IgG response. IgG response to the cE5 protein in the three surveys: Aug 1994 (n = 117, average age ±95% CI = 17.2 ± 3.0), Oct 1994 (n = 121, average age ±95% CI = 16.8 ± 3.1) and Mar 1995 (n = 121, average age ±95% CI = 16.0 ± 2.9). (A) Box plots of OD values among responders from the different surveys as indicated. Boxes, whiskers and dots as in Figure 1 . IgG levels are expressed as OD 492 . (B) Seroprevalence of anti-cE5 IgG antibodies. Error bars as in Figure 1 .

    Article Snippet: After washing, blocking and washing again as above wells were incubated overnight at 4°C with serial dilutions, from 1 μg/ml to 0.0078 μg/ml (1 → 0.5 → 0.25 → 0.125 → 0.0625 → 0.03125 → 0.0156 → 0.0078), of purified native human IgG1 or IgG4 (ABD Serotec, Kidlington, Oxford, UK) in 50 μl of blocking reagent.

    Techniques:

    IgG antibody response to the An. gambiae cE5 and gSG6 proteins. Comparison of the anti-cE5 and anti-gSG6 IgG responses among exposed individuals during the malaria transmission season (survey Aug 1994, n = 207, average age ±95% CI = 22.7 ± 16.9). (A) Box plots of OD values among responders to both cE5 and gSG6 antigens (n = 156). Boxes display median values, 25th and 75th percentiles. Whiskers represent 5–95 percentiles and dots the outliers. The IgG response to the cE5 protein in non-exposed individuals (n = 59, average age ±95% CI = 26.3 ± 16.1) is shown as control (C). IgG levels are expressed as OD 492 . P value determined by the Wilcoxon matched-pairs signed rank test (****, p

    Journal: Parasites & Vectors

    Article Title: Differential antibody response to the Anopheles gambiae gSG6 and cE5 salivary proteins in individuals naturally exposed to bites of malaria vectors

    doi: 10.1186/s13071-014-0549-8

    Figure Lengend Snippet: IgG antibody response to the An. gambiae cE5 and gSG6 proteins. Comparison of the anti-cE5 and anti-gSG6 IgG responses among exposed individuals during the malaria transmission season (survey Aug 1994, n = 207, average age ±95% CI = 22.7 ± 16.9). (A) Box plots of OD values among responders to both cE5 and gSG6 antigens (n = 156). Boxes display median values, 25th and 75th percentiles. Whiskers represent 5–95 percentiles and dots the outliers. The IgG response to the cE5 protein in non-exposed individuals (n = 59, average age ±95% CI = 26.3 ± 16.1) is shown as control (C). IgG levels are expressed as OD 492 . P value determined by the Wilcoxon matched-pairs signed rank test (****, p

    Article Snippet: After washing, blocking and washing again as above wells were incubated overnight at 4°C with serial dilutions, from 1 μg/ml to 0.0078 μg/ml (1 → 0.5 → 0.25 → 0.125 → 0.0625 → 0.03125 → 0.0156 → 0.0078), of purified native human IgG1 or IgG4 (ABD Serotec, Kidlington, Oxford, UK) in 50 μl of blocking reagent.

    Techniques: Transmission Assay

    Comparison of the IgG response to cE5 and gSG6 in different age groups. (A) Box plots of OD values among responders to cE5 (1–15 n = 94; 16–30 n = 50; > 30 n = 53) and to gSG6 (1–15 n = 84; 16–30 n = 40; > 30 n = 38) in the three age groups as indicated. Boxes, whiskers and dots as in Figure 1 . IgG levels are expressed as OD 492 . P values determined by the Kruskal-Wallis (cE5 P = 0.001; gSG6 ns) and the Mann–Whitney tests (***, p

    Journal: Parasites & Vectors

    Article Title: Differential antibody response to the Anopheles gambiae gSG6 and cE5 salivary proteins in individuals naturally exposed to bites of malaria vectors

    doi: 10.1186/s13071-014-0549-8

    Figure Lengend Snippet: Comparison of the IgG response to cE5 and gSG6 in different age groups. (A) Box plots of OD values among responders to cE5 (1–15 n = 94; 16–30 n = 50; > 30 n = 53) and to gSG6 (1–15 n = 84; 16–30 n = 40; > 30 n = 38) in the three age groups as indicated. Boxes, whiskers and dots as in Figure 1 . IgG levels are expressed as OD 492 . P values determined by the Kruskal-Wallis (cE5 P = 0.001; gSG6 ns) and the Mann–Whitney tests (***, p

    Article Snippet: After washing, blocking and washing again as above wells were incubated overnight at 4°C with serial dilutions, from 1 μg/ml to 0.0078 μg/ml (1 → 0.5 → 0.25 → 0.125 → 0.0625 → 0.03125 → 0.0156 → 0.0078), of purified native human IgG1 or IgG4 (ABD Serotec, Kidlington, Oxford, UK) in 50 μl of blocking reagent.

    Techniques: MANN-WHITNEY

    Effects of TLR ‐7 agonist R848 in transgenic mice. A , Detection of GFP in bone marrow–derived macrophages ( BMM s) from mTLR ‐7−/− (knockout [ KO ]) mice and transgenic (Tg) mice by flow cytometric analysis, after staining with anti‐ CD 11b antibody. At least 2 independent experiments were performed. B , Interleukin‐6 ( IL ‐6) and IL ‐12p40 levels in BMM s from knockout mice and transgenic mice left unstimulated or stimulated with R848. After 24 hours of incubation, IL ‐6 and IL ‐12p40 were detected in culture medium by enzyme‐linked immunosorbent assay ( ELISA ). Results are from triplicate wells. At least 2 independent experiments were performed. C , Weight of SMG s, kidneys, lungs, and liver in 8‐week‐old knockout, wild‐type ( WT ), and transgenic mice treated with topical R848 for 4 weeks (n = 8 per group). D and F , H E‐stained ( D ) and Masson's trichrome–stained ( F ) sections of SMG s, pancreas, kidneys, lungs, and liver from representative 8‐week‐old R848‐treated knockout, WT , and transgenic mice. Bars = 100 μm. E and G , Focus score ( E ) and fibrosis score ( G ) for each organ in 8‐week‐old knockout, WT , and transgenic mice left untreated or treated with R848 (n = 8 per group). The fibrosis score was calculated from Masson's trichrome staining as described in Patients and Methods. HPF = high‐power field. H , Serial sections of SMG s from a representative 8‐week‐old R848‐treated transgenic mouse, stained with H E and for IgG1, TLR ‐7, CD 206, CD 317, and IL ‐33. Mayer's hematoxylin (blue) counterstained; bars = 100 μm. I , Serum IgG, IgG1, and IgG2a levels in knockout, WT , and transgenic mice (n = 10 per group) before and after R848 treatment, as determined by ELISA . Symbols represent individual mice; horizontal lines show the mean. J , Serum IL ‐33 levels, determined by ELISA , in knockout, WT , and transgenic mice left untreated or treated with R848 (n = 10 per group). In B , C , E , G , and J , bars show the mean ± SD . * = P

    Journal: Arthritis & Rheumatology (Hoboken, N.j.)

    Article Title: Activated M2 Macrophages Contribute to the Pathogenesis of IgG4‐Related Disease via Toll‐like Receptor 7/Interleukin‐33 Signaling

    doi: 10.1002/art.41052

    Figure Lengend Snippet: Effects of TLR ‐7 agonist R848 in transgenic mice. A , Detection of GFP in bone marrow–derived macrophages ( BMM s) from mTLR ‐7−/− (knockout [ KO ]) mice and transgenic (Tg) mice by flow cytometric analysis, after staining with anti‐ CD 11b antibody. At least 2 independent experiments were performed. B , Interleukin‐6 ( IL ‐6) and IL ‐12p40 levels in BMM s from knockout mice and transgenic mice left unstimulated or stimulated with R848. After 24 hours of incubation, IL ‐6 and IL ‐12p40 were detected in culture medium by enzyme‐linked immunosorbent assay ( ELISA ). Results are from triplicate wells. At least 2 independent experiments were performed. C , Weight of SMG s, kidneys, lungs, and liver in 8‐week‐old knockout, wild‐type ( WT ), and transgenic mice treated with topical R848 for 4 weeks (n = 8 per group). D and F , H E‐stained ( D ) and Masson's trichrome–stained ( F ) sections of SMG s, pancreas, kidneys, lungs, and liver from representative 8‐week‐old R848‐treated knockout, WT , and transgenic mice. Bars = 100 μm. E and G , Focus score ( E ) and fibrosis score ( G ) for each organ in 8‐week‐old knockout, WT , and transgenic mice left untreated or treated with R848 (n = 8 per group). The fibrosis score was calculated from Masson's trichrome staining as described in Patients and Methods. HPF = high‐power field. H , Serial sections of SMG s from a representative 8‐week‐old R848‐treated transgenic mouse, stained with H E and for IgG1, TLR ‐7, CD 206, CD 317, and IL ‐33. Mayer's hematoxylin (blue) counterstained; bars = 100 μm. I , Serum IgG, IgG1, and IgG2a levels in knockout, WT , and transgenic mice (n = 10 per group) before and after R848 treatment, as determined by ELISA . Symbols represent individual mice; horizontal lines show the mean. J , Serum IL ‐33 levels, determined by ELISA , in knockout, WT , and transgenic mice left untreated or treated with R848 (n = 10 per group). In B , C , E , G , and J , bars show the mean ± SD . * = P

    Article Snippet: Antibodies used included anti‐CD68 (catalog no. ab955; Abcam), anti‐CD163 (catalog no. NCL‐CD163; Leica Biosystems), anti‐CD123 (catalog no. NCL‐L‐CD123; Leica Biosystems), anti–TLR‐8 (catalog no. 85859; Abcam), anti–TLR‐9 (catalog no. 134368; Abcam), and anti–IL‐33 (catalog no. 54385; Abcam) mouse monoclonal antibodies; anti‐CD11c (catalog no. 52632; Abcam), and anti–TLR‐7 (catalog no. 124928; Abcam) rabbit monoclonal antibodies; anti‐CD317 (catalog no. DDX0390p‐100; Novus) rat monoclonal antibodies; and anti–TLR‐10 (catalog no. 53631; Abcam), anti‐CD206 (catalog no. AF2534; R & D Systems), and anti‐IgG1 (catalog no. 97236; Abcam) goat polyclonal antibodies.

    Techniques: Transgenic Assay, Mouse Assay, Derivative Assay, Knock-Out, Staining, Incubation, Enzyme-linked Immunosorbent Assay

    Schematic model of the role of Toll‐like receptor 7 ( TLR ‐7)–positive M2 macrophages in the initiation of Ig4‐related disease. TLR ‐7 expressed on M2 macrophages recognize some RNA viruses or self RNA from apoptotic cells. Activated M2 macrophages secrete interleukin‐33 ( IL ‐33) and promote production of Th2 cytokines that lead to IgG4 class‐switching and fibrosis. TGF β = transforming growth factor β.

    Journal: Arthritis & Rheumatology (Hoboken, N.j.)

    Article Title: Activated M2 Macrophages Contribute to the Pathogenesis of IgG4‐Related Disease via Toll‐like Receptor 7/Interleukin‐33 Signaling

    doi: 10.1002/art.41052

    Figure Lengend Snippet: Schematic model of the role of Toll‐like receptor 7 ( TLR ‐7)–positive M2 macrophages in the initiation of Ig4‐related disease. TLR ‐7 expressed on M2 macrophages recognize some RNA viruses or self RNA from apoptotic cells. Activated M2 macrophages secrete interleukin‐33 ( IL ‐33) and promote production of Th2 cytokines that lead to IgG4 class‐switching and fibrosis. TGF β = transforming growth factor β.

    Article Snippet: Antibodies used included anti‐CD68 (catalog no. ab955; Abcam), anti‐CD163 (catalog no. NCL‐CD163; Leica Biosystems), anti‐CD123 (catalog no. NCL‐L‐CD123; Leica Biosystems), anti–TLR‐8 (catalog no. 85859; Abcam), anti–TLR‐9 (catalog no. 134368; Abcam), and anti–IL‐33 (catalog no. 54385; Abcam) mouse monoclonal antibodies; anti‐CD11c (catalog no. 52632; Abcam), and anti–TLR‐7 (catalog no. 124928; Abcam) rabbit monoclonal antibodies; anti‐CD317 (catalog no. DDX0390p‐100; Novus) rat monoclonal antibodies; and anti–TLR‐10 (catalog no. 53631; Abcam), anti‐CD206 (catalog no. AF2534; R & D Systems), and anti‐IgG1 (catalog no. 97236; Abcam) goat polyclonal antibodies.

    Techniques:

    Pathologic and serologic findings in human TLR ‐7–transgenic/mouse TLR ‐7–deficient (hu TLR ‐7–transgenic/ mTLR ‐7 −/− ) mice on a C57 BL /6 background. A , A wild‐type ( WT ) C57 BL /6 mouse and an hu TLR ‐7–transgenic/ mTLR ‐7−/− (transgenic [Tg]) mouse at 4 weeks old. B , Weight of SMG s, kidneys, lungs, and liver in 4‐week‐old WT mice (n = 5) and transgenic mice (n = 5). Bars show the mean ± SD . C , H E‐stained sections of SMG s, pancreas, kidneys, lungs, and liver from representative WT and transgenic mice. Bars = 100 μm. D , Focus score for each organ in WT mice (n = 5) and transgenic mice (n = 5). The focus score was estimated as described in Patients and Methods. Bars show the mean ± SD . HPF = high‐power field. E , Masson's trichrome–stained sections of SMG s, pancreas, kidneys, lungs, and liver from representative WT and transgenic mice. Masson's trichrome was used to stain nuclei (purple), cytoplasm (red), and collagen (connective or fibrotic tissue; blue) Bars = 100 μm. F , Fibrosis score for each organ in WT mice (n = 5) and transgenic mice (n = 5). The fibrosis score was calculated from Masson's trichrome staining as described in Patients and Methods. Bars show the mean ± SD . G , Serum IgG, IgG1, and IgG2a levels in WT mice (n = 10) and transgenic mice (n = 10), as determined by enzyme‐linked immunosorbent assay. Symbols represent individual mice; horizontal lines show the mean. * = P

    Journal: Arthritis & Rheumatology (Hoboken, N.j.)

    Article Title: Activated M2 Macrophages Contribute to the Pathogenesis of IgG4‐Related Disease via Toll‐like Receptor 7/Interleukin‐33 Signaling

    doi: 10.1002/art.41052

    Figure Lengend Snippet: Pathologic and serologic findings in human TLR ‐7–transgenic/mouse TLR ‐7–deficient (hu TLR ‐7–transgenic/ mTLR ‐7 −/− ) mice on a C57 BL /6 background. A , A wild‐type ( WT ) C57 BL /6 mouse and an hu TLR ‐7–transgenic/ mTLR ‐7−/− (transgenic [Tg]) mouse at 4 weeks old. B , Weight of SMG s, kidneys, lungs, and liver in 4‐week‐old WT mice (n = 5) and transgenic mice (n = 5). Bars show the mean ± SD . C , H E‐stained sections of SMG s, pancreas, kidneys, lungs, and liver from representative WT and transgenic mice. Bars = 100 μm. D , Focus score for each organ in WT mice (n = 5) and transgenic mice (n = 5). The focus score was estimated as described in Patients and Methods. Bars show the mean ± SD . HPF = high‐power field. E , Masson's trichrome–stained sections of SMG s, pancreas, kidneys, lungs, and liver from representative WT and transgenic mice. Masson's trichrome was used to stain nuclei (purple), cytoplasm (red), and collagen (connective or fibrotic tissue; blue) Bars = 100 μm. F , Fibrosis score for each organ in WT mice (n = 5) and transgenic mice (n = 5). The fibrosis score was calculated from Masson's trichrome staining as described in Patients and Methods. Bars show the mean ± SD . G , Serum IgG, IgG1, and IgG2a levels in WT mice (n = 10) and transgenic mice (n = 10), as determined by enzyme‐linked immunosorbent assay. Symbols represent individual mice; horizontal lines show the mean. * = P

    Article Snippet: Antibodies used included anti‐CD68 (catalog no. ab955; Abcam), anti‐CD163 (catalog no. NCL‐CD163; Leica Biosystems), anti‐CD123 (catalog no. NCL‐L‐CD123; Leica Biosystems), anti–TLR‐8 (catalog no. 85859; Abcam), anti–TLR‐9 (catalog no. 134368; Abcam), and anti–IL‐33 (catalog no. 54385; Abcam) mouse monoclonal antibodies; anti‐CD11c (catalog no. 52632; Abcam), and anti–TLR‐7 (catalog no. 124928; Abcam) rabbit monoclonal antibodies; anti‐CD317 (catalog no. DDX0390p‐100; Novus) rat monoclonal antibodies; and anti–TLR‐10 (catalog no. 53631; Abcam), anti‐CD206 (catalog no. AF2534; R & D Systems), and anti‐IgG1 (catalog no. 97236; Abcam) goat polyclonal antibodies.

    Techniques: Transgenic Assay, Mouse Assay, Staining, Enzyme-linked Immunosorbent Assay

    Hematoxylin and eosin staining in IMT and IgG4-related IPT. (A) Original alveolar structures were totally destroyed by intersecting fascicles of spindle tumor cells in IMTs; (B) tumor cells were admixed with numerous lymphocytes, plasma cells and lymphoid follicles (red arrows) in IgG4-related IPTs; (C) a large number of inflammatory cells were infiltrative and involves the small bronchi (green arrow) in IPTs; (D) inflammatory infiltration of vessel walls or obstructive phlebitis (yellow arrow) were only observed in the tumors diagnosed as IgG4-related IPT.

    Journal: Journal of Thoracic Disease

    Article Title: Pulmonary inflammatory myofibroblastic tumor versus IgG4-related inflammatory pseudotumor: differential diagnosis based on a case series

    doi: 10.21037/jtd.2017.02.89

    Figure Lengend Snippet: Hematoxylin and eosin staining in IMT and IgG4-related IPT. (A) Original alveolar structures were totally destroyed by intersecting fascicles of spindle tumor cells in IMTs; (B) tumor cells were admixed with numerous lymphocytes, plasma cells and lymphoid follicles (red arrows) in IgG4-related IPTs; (C) a large number of inflammatory cells were infiltrative and involves the small bronchi (green arrow) in IPTs; (D) inflammatory infiltration of vessel walls or obstructive phlebitis (yellow arrow) were only observed in the tumors diagnosed as IgG4-related IPT.

    Article Snippet: Rabbit anti-IgG4 and anti-IgG antibodies were obtained from Abcam Trading (Shanghai) Company Ltd., Shanghai, China.

    Techniques: Staining

    The results of statistical analysis based on the clinicopathological and immunohistochemical (IHC) findings. (A) Overall survival (OS) between the IMT and IgG4-related IPT group were found no statistically significant difference (P=0.197); (B) a borderline value was found in disease-free survival (DFS) between the two group (P=0.056); (C) Pearson correlation analysis showed that the lymphocytes + plasmocytes level had a possible linear positive correlation with the number of IgG4-positive plasma cells (r=0.914, P

    Journal: Journal of Thoracic Disease

    Article Title: Pulmonary inflammatory myofibroblastic tumor versus IgG4-related inflammatory pseudotumor: differential diagnosis based on a case series

    doi: 10.21037/jtd.2017.02.89

    Figure Lengend Snippet: The results of statistical analysis based on the clinicopathological and immunohistochemical (IHC) findings. (A) Overall survival (OS) between the IMT and IgG4-related IPT group were found no statistically significant difference (P=0.197); (B) a borderline value was found in disease-free survival (DFS) between the two group (P=0.056); (C) Pearson correlation analysis showed that the lymphocytes + plasmocytes level had a possible linear positive correlation with the number of IgG4-positive plasma cells (r=0.914, P

    Article Snippet: Rabbit anti-IgG4 and anti-IgG antibodies were obtained from Abcam Trading (Shanghai) Company Ltd., Shanghai, China.

    Techniques: Immunohistochemistry

    Design, biochemical characterization, and gene silencing activity of I4R-9r PwSN. (A) Schematic representation of the Peptide-Woven SiRNA Nanocomplex (PwSN), with siRNA noncovalently bound to I4R-9r tandem peptides composed of a tumor targeting peptide (I4R, green) and Cell penetrating peptide (9R, blue) separated by a 2-glycine spacer. (B) siRNA encapsulation by I4R-9r tandem peptides was monitored by gel retardation assay with molar ratios of 1:5 to 1:50 (siRNA-to-peptide). (C) Representative confocal microscopy images of HeLa cells treated with siRNA-FITC carried I4R-9r PwSN vs . free siRNA-FITC. (D) Representative histograms from flow cytometry for cellular uptake of I4R-9r PwSN (blue), 9r PwSN (grey), and free siRNA (black). (E) Representative histograms from flow cytometry for cellular uptake of I4R-9r PwSN (blue) vs. free siRNA (black) in the presence of indicated concentrations of anti-IL4R antibody or an IgG control. (F) HeLa cells were transfected with nanocomplexes carrying siRNA against GAPDH. The GAPDH protein expression is monitored by Western blot and presented with mean of relative immunoblot intensities. Lipofectamine was used as a positive control. The error bars represent SD from cumulative data of six independent experiments.

    Journal: PLoS ONE

    Article Title: Design of a Multicomponent Peptide-Woven Nanocomplex for Delivery of siRNA

    doi: 10.1371/journal.pone.0118310

    Figure Lengend Snippet: Design, biochemical characterization, and gene silencing activity of I4R-9r PwSN. (A) Schematic representation of the Peptide-Woven SiRNA Nanocomplex (PwSN), with siRNA noncovalently bound to I4R-9r tandem peptides composed of a tumor targeting peptide (I4R, green) and Cell penetrating peptide (9R, blue) separated by a 2-glycine spacer. (B) siRNA encapsulation by I4R-9r tandem peptides was monitored by gel retardation assay with molar ratios of 1:5 to 1:50 (siRNA-to-peptide). (C) Representative confocal microscopy images of HeLa cells treated with siRNA-FITC carried I4R-9r PwSN vs . free siRNA-FITC. (D) Representative histograms from flow cytometry for cellular uptake of I4R-9r PwSN (blue), 9r PwSN (grey), and free siRNA (black). (E) Representative histograms from flow cytometry for cellular uptake of I4R-9r PwSN (blue) vs. free siRNA (black) in the presence of indicated concentrations of anti-IL4R antibody or an IgG control. (F) HeLa cells were transfected with nanocomplexes carrying siRNA against GAPDH. The GAPDH protein expression is monitored by Western blot and presented with mean of relative immunoblot intensities. Lipofectamine was used as a positive control. The error bars represent SD from cumulative data of six independent experiments.

    Article Snippet: The binding specificity of I4R-9r/FITC-siRNA nanocomplexes was assessed by blocking experiments in which HeLa cells were preincubated with antibody to human IL-4R (ab50018, abcam) and IgG control antibody (ab91361, abcam) at 4°C for 1 hr.

    Techniques: Activity Assay, Electrophoretic Mobility Shift Assay, Confocal Microscopy, Flow Cytometry, Cytometry, Transfection, Expressing, Western Blot, Positive Control

    Visualization of LRP/LR on the surface of early-stage (SW-480 and HT-29) and late-stage (DLD-1) colorectal cancer and poorly invasive (MCF-7) tumorigenic cells. (A) Cells labeled with primary antibody IgG1-iS18 and FITC-coupled secondary antibody specific

    Journal: Molecular Medicine

    Article Title: Anti-LRP/LR–Specific Antibody IgG1-iS18 Significantly Impedes Adhesion and Invasion in Early- and Late-Stage Colorectal Carcinoma Cells

    doi: 10.2119/molmed.2016.00169

    Figure Lengend Snippet: Visualization of LRP/LR on the surface of early-stage (SW-480 and HT-29) and late-stage (DLD-1) colorectal cancer and poorly invasive (MCF-7) tumorigenic cells. (A) Cells labeled with primary antibody IgG1-iS18 and FITC-coupled secondary antibody specific

    Article Snippet: The membrane was then probed with the primary anti-LRP/LR–specific antibody IgG1-iS18 (1:10000) for 1 h. Thereafter, the membrane was washed three times with 1 × PBS-Tween before incubating with goat antihuman horseradish peroxidase (HRP) secondary antibody (1:5000) (Abcam).

    Techniques: Labeling

    (A) Detection of total 37 kDa LRP levels in protein lysates of MCF-7, SW-480, HT-29 and DLD-1 cells. IgG1-iS18 was used as the primary antibody, with HRP-coupled secondary antibody; 42kDa β-actin was used as a loading control. (B) Densitometric

    Journal: Molecular Medicine

    Article Title: Anti-LRP/LR–Specific Antibody IgG1-iS18 Significantly Impedes Adhesion and Invasion in Early- and Late-Stage Colorectal Carcinoma Cells

    doi: 10.2119/molmed.2016.00169

    Figure Lengend Snippet: (A) Detection of total 37 kDa LRP levels in protein lysates of MCF-7, SW-480, HT-29 and DLD-1 cells. IgG1-iS18 was used as the primary antibody, with HRP-coupled secondary antibody; 42kDa β-actin was used as a loading control. (B) Densitometric

    Article Snippet: The membrane was then probed with the primary anti-LRP/LR–specific antibody IgG1-iS18 (1:10000) for 1 h. Thereafter, the membrane was washed three times with 1 × PBS-Tween before incubating with goat antihuman horseradish peroxidase (HRP) secondary antibody (1:5000) (Abcam).

    Techniques:

    Quantification of cell surface LRP/LR levels on tumorigenic cell lines. Cells were labeled with IgG1-iS18 primary antibody and antihuman phycoerythrin–coupled secondary antibody. Thereafter, 20,000 cells from each tumorigenic cell line were analyzed

    Journal: Molecular Medicine

    Article Title: Anti-LRP/LR–Specific Antibody IgG1-iS18 Significantly Impedes Adhesion and Invasion in Early- and Late-Stage Colorectal Carcinoma Cells

    doi: 10.2119/molmed.2016.00169

    Figure Lengend Snippet: Quantification of cell surface LRP/LR levels on tumorigenic cell lines. Cells were labeled with IgG1-iS18 primary antibody and antihuman phycoerythrin–coupled secondary antibody. Thereafter, 20,000 cells from each tumorigenic cell line were analyzed

    Article Snippet: The membrane was then probed with the primary anti-LRP/LR–specific antibody IgG1-iS18 (1:10000) for 1 h. Thereafter, the membrane was washed three times with 1 × PBS-Tween before incubating with goat antihuman horseradish peroxidase (HRP) secondary antibody (1:5000) (Abcam).

    Techniques: Labeling

    Detection of 37 kDa/67 kDa LRP/LR levels on the surfaces of tumorigenic cell lines. Cells were treated with IgG1-iS18 and IgG1-HD37 (control antibody). (A and E) represent MCF-7, (B and F) represent SW-480, (C and G) represent HT-29, and (D and H) represent

    Journal: Molecular Medicine

    Article Title: Anti-LRP/LR–Specific Antibody IgG1-iS18 Significantly Impedes Adhesion and Invasion in Early- and Late-Stage Colorectal Carcinoma Cells

    doi: 10.2119/molmed.2016.00169

    Figure Lengend Snippet: Detection of 37 kDa/67 kDa LRP/LR levels on the surfaces of tumorigenic cell lines. Cells were treated with IgG1-iS18 and IgG1-HD37 (control antibody). (A and E) represent MCF-7, (B and F) represent SW-480, (C and G) represent HT-29, and (D and H) represent

    Article Snippet: The membrane was then probed with the primary anti-LRP/LR–specific antibody IgG1-iS18 (1:10000) for 1 h. Thereafter, the membrane was washed three times with 1 × PBS-Tween before incubating with goat antihuman horseradish peroxidase (HRP) secondary antibody (1:5000) (Abcam).

    Techniques:

    Anti-M2e antibody response in sera. BALB/c mice (n = 6/group) were immunized i.n. with 10 μg/0.02 ml of Flg-HA2-2-4M2e, Flg-HA2-1-4M2e, or Flg-4M2e on days 0, 14, 28. Mice in the control groups were administered with Flg-his (10 μg/0.02 ml) or PBS. Two weeks post second boost, (A) M2e-specific IgG (M2eh and M2es) and (B) IgA response in sera were evaluated by ELISA. ( C ) Anti-M2e IgG subclasses, tested against M2e peptide in sera, were determined by ELISA. Statistical significance was determined using the Mann-Whitney U-test. The P values between immunized and control group are indicated. **—significant difference from control groups, p

    Journal: PLoS ONE

    Article Title: Combination of M2e peptide with stalk HA epitopes of influenza A virus enhances protective properties of recombinant vaccine

    doi: 10.1371/journal.pone.0201429

    Figure Lengend Snippet: Anti-M2e antibody response in sera. BALB/c mice (n = 6/group) were immunized i.n. with 10 μg/0.02 ml of Flg-HA2-2-4M2e, Flg-HA2-1-4M2e, or Flg-4M2e on days 0, 14, 28. Mice in the control groups were administered with Flg-his (10 μg/0.02 ml) or PBS. Two weeks post second boost, (A) M2e-specific IgG (M2eh and M2es) and (B) IgA response in sera were evaluated by ELISA. ( C ) Anti-M2e IgG subclasses, tested against M2e peptide in sera, were determined by ELISA. Statistical significance was determined using the Mann-Whitney U-test. The P values between immunized and control group are indicated. **—significant difference from control groups, p

    Article Snippet: Polyclonal HRP-labeled goat anti-mouse IgG, IgG1, IgG2a, IgG2b, or IgG3 antibodies (Abcam, UK) were used.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    Measuring IgG1 (a) and IgG2a (b) and IgG1/IgG2a (c) antibodies in the vaccinated mice using ELISA method three weeks after immunization. Significant differences were designated as ∗ P

    Journal: Journal of Immunology Research

    Article Title: Evolution of the Immune Response against Recombinant Proteins (TcpA, TcpB, and FlaA) as a Candidate Subunit Cholera Vaccine

    doi: 10.1155/2017/2412747

    Figure Lengend Snippet: Measuring IgG1 (a) and IgG2a (b) and IgG1/IgG2a (c) antibodies in the vaccinated mice using ELISA method three weeks after immunization. Significant differences were designated as ∗ P

    Article Snippet: The plates were kept at 37°C for 2 h. After washing, 50 μ L of 1/10000 diluted antibodies against IgG1 and IgG2a conjugated classes was added to HRP (Abcam, UK) and they were kept at 37°C for 2 h. Finally, light absorption at wavelength of 490 nm was determined by adding 100 μ L of o-phenylenediamine (OPD) substrate solution to each well and incubation for 10 min.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Serum levels of total IgG (A), IgG1 (B), IgG2a (C) and IgG2b (D) antibodies (OD 450 ± SD) in immunized and control groups of mice, at week 0, 2, 4, and 6. Significant differences between control and immunized groups are marked with (*), where * P

    Journal: Iranian Journal of Parasitology

    Article Title: Immunogenicity of Multiepitope Vaccine Candidate against Toxoplasma gondii Infection in BALB/c Mice

    doi:

    Figure Lengend Snippet: Serum levels of total IgG (A), IgG1 (B), IgG2a (C) and IgG2b (D) antibodies (OD 450 ± SD) in immunized and control groups of mice, at week 0, 2, 4, and 6. Significant differences between control and immunized groups are marked with (*), where * P

    Article Snippet: The wells were again washed prior to the addition of 100 μl of HRP conjugated anti-mouse IgG, IgG1, IgG2a and IgG2b antibodies (Abcam, USA) at a 1:10000 dilution in PBS and the plate was incubated at 37 °C for 30 min. Another three rounds of washes were carried out and the color reaction was allowed to develop by adding 100 μl of TMB substrate followed by 15 min incubation.

    Techniques: Mouse Assay

    IFNαR 1 Is Required for Dysregulated Spt-AFC and Spt-GC Formation in B6. Sle1b Mice (A) Flow cytometric analysis of surface expression of IFNαR 1 on B220 + Fas low GL-7 low non-GC B cells (red), B220 + Fas h GL-7 hi GC B cells (blue), CD4 + PD-1 low CXCR5 low non-Tfh (orange), and CD4 + PD-1 hi CXCR5 hi Tfh (green) in female B6 and B6. Sle1b mice. Error bars represent mean ± SEM. (B and C) The percentages of GC B cells (B) and Tfh cells (C) in splenocytes from 3- and 6-month-old female mice. (D) Spleen sections from 6-month-old B6, B6.IFNαR 1 −/− , B6. Sle1b , and B6. Sle1b .IFNαR 1 −/− female mice (5 mice per group) were stained with anti-CD4 Ab (red) GL-7 (green) and anti-IgD Ab (blue), and GC areas were measured for 10 GCs per spleen section (right panels). The scale bars represent 50 μm. (E and F) Each symbol represents a measured GC. IgM + (E) and IgG + (F) AFCs in the spleens of 6-month-old B6, B6.IFNαR 1 −/− , B6. Sle1b , and B6. Sle1b .IFNαR 1 −/− female mice are shown. (G-J) Analysis of serum titers of IgM (G), IgG (H), IgG1 (I), and IgG2c (J) antibodies in 6-month-old mice by ELISA. Each symbol represents a mouse, and these data represent at least two-three independent experiments. Statistical significance was determined by one-way ANOVA with a follow-up Tukey multiple-comparison test (NS, not significant, *p

    Journal: Cell reports

    Article Title: B-Cell-Intrinsic Type 1 Interferon Signaling Is Crucial for Loss of Tolerance and the Development of Autoreactive B Cells

    doi: 10.1016/j.celrep.2018.06.046

    Figure Lengend Snippet: IFNαR 1 Is Required for Dysregulated Spt-AFC and Spt-GC Formation in B6. Sle1b Mice (A) Flow cytometric analysis of surface expression of IFNαR 1 on B220 + Fas low GL-7 low non-GC B cells (red), B220 + Fas h GL-7 hi GC B cells (blue), CD4 + PD-1 low CXCR5 low non-Tfh (orange), and CD4 + PD-1 hi CXCR5 hi Tfh (green) in female B6 and B6. Sle1b mice. Error bars represent mean ± SEM. (B and C) The percentages of GC B cells (B) and Tfh cells (C) in splenocytes from 3- and 6-month-old female mice. (D) Spleen sections from 6-month-old B6, B6.IFNαR 1 −/− , B6. Sle1b , and B6. Sle1b .IFNαR 1 −/− female mice (5 mice per group) were stained with anti-CD4 Ab (red) GL-7 (green) and anti-IgD Ab (blue), and GC areas were measured for 10 GCs per spleen section (right panels). The scale bars represent 50 μm. (E and F) Each symbol represents a measured GC. IgM + (E) and IgG + (F) AFCs in the spleens of 6-month-old B6, B6.IFNαR 1 −/− , B6. Sle1b , and B6. Sle1b .IFNαR 1 −/− female mice are shown. (G-J) Analysis of serum titers of IgM (G), IgG (H), IgG1 (I), and IgG2c (J) antibodies in 6-month-old mice by ELISA. Each symbol represents a mouse, and these data represent at least two-three independent experiments. Statistical significance was determined by one-way ANOVA with a follow-up Tukey multiple-comparison test (NS, not significant, *p

    Article Snippet: Briefly, for IgG or IgM detection, ELISA plates (Thermo Fisher Scientific, Waltham, MA) were coated with anti-IgM or anti-IgG (Invitrogen, Grand Island, NY) capture antibodies and detected using biotinylated antimouse IgM (Jackson ImmunoResearch, West Grove, PA) or alkaline-phosphatase conjugated anti-mouse IgG (Molecular Probes, Grand Island, NY).

    Techniques: Single-particle Tracking, Mouse Assay, Flow Cytometry, Expressing, Staining, Enzyme-linked Immunosorbent Assay

    Complement-dependent cytotoxicity (CDC) of anti-FLRT2 antibody . CDC activities using two healthy control IgG and two anti-FLRT2 positive IgG, B11-8 and X10-48, at a concentration of 1.28 mg/ml with 1:3 diluted complement  (A) , 1.28 mg/ml of IgG, and 1:3 diluted complement with recombinant FLRT2 at the indicated concentrations  (B) , and various IgG concentrations with 1:6 diluted complement  (C)  against HUVECs were measured with the WST-1 assay.  (D)  CDC activities against mock transfected HEK293T cells (empty, left) and FLRT2-expressing HEK293T cells (FLRT2, right) by using 1.28 mg/ml of IgG and 1:3 diluted complement were measured with the WST-1 assay. HEK293T cells negative or positive for FLRT2 expression were stained with anti-FLRT2 antibody followed by secondary antibody against human IgG1, IgG2, IgG3, IgG4  (E) , and IgM  (F) , and analyzed with flow cytometry.  (G)  ADCC activities using control IgG, B11-8, and X10-48, at a concentration of 1.28 mg/ml with an E:T ratio of 25:1 were determined with the lactate dehydrogenase detection method. Error bars indicate SD. * P

    Journal: Arthritis Research & Therapy

    Article Title: A novel autoantibody against fibronectin leucine-rich transmembrane protein 2 expressed on the endothelial cell surface identified by retroviral vector system in systemic lupus erythematosus

    doi: 10.1186/ar3897

    Figure Lengend Snippet: Complement-dependent cytotoxicity (CDC) of anti-FLRT2 antibody . CDC activities using two healthy control IgG and two anti-FLRT2 positive IgG, B11-8 and X10-48, at a concentration of 1.28 mg/ml with 1:3 diluted complement (A) , 1.28 mg/ml of IgG, and 1:3 diluted complement with recombinant FLRT2 at the indicated concentrations (B) , and various IgG concentrations with 1:6 diluted complement (C) against HUVECs were measured with the WST-1 assay. (D) CDC activities against mock transfected HEK293T cells (empty, left) and FLRT2-expressing HEK293T cells (FLRT2, right) by using 1.28 mg/ml of IgG and 1:3 diluted complement were measured with the WST-1 assay. HEK293T cells negative or positive for FLRT2 expression were stained with anti-FLRT2 antibody followed by secondary antibody against human IgG1, IgG2, IgG3, IgG4 (E) , and IgM (F) , and analyzed with flow cytometry. (G) ADCC activities using control IgG, B11-8, and X10-48, at a concentration of 1.28 mg/ml with an E:T ratio of 25:1 were determined with the lactate dehydrogenase detection method. Error bars indicate SD. * P

    Article Snippet: Secondary antibodies included 1:50 diluted fluorescein isothiocyanate (FITC) or phycoerythrin (PE)-conjugated goat anti-human IgG (Abcam, Cambridge, UK), PE-conjugated donkey anti-goat IgG (Abcam), PE-conjugated mouse anti-human IgG1/IgG2/IgG3/IgG4 antibody (Beckman Coulter, Fullerton, CA, USA), and DyLight 650-conjugated anti human IgM antibody (Abcam).

    Techniques: Concentration Assay, Recombinant, WST-1 Assay, Transfection, Expressing, Staining, Flow Cytometry, Cytometry

    Ranitidine significantly enhanced levels of IgG2a capable of binding to E0771-GFP cancer cells. Serum was diluted 1:10 and added to cultured E0771-GFP cells and anti-isotype antibodies for IgG2a (A) or kappa light chain (B) were added. Relative fluorescence intensity (RFI) was used as an indicator for antibody binding to E0771-GFP cells relative to the average of control MFI. Box plot represents mean and range of data from 20 to 24 individual mice. * p

    Journal: Frontiers in Immunology

    Article Title: Ranitidine Inhibition of Breast Tumor Growth Is B Cell Dependent and Associated With an Enhanced Antitumor Antibody Response

    doi: 10.3389/fimmu.2018.01894

    Figure Lengend Snippet: Ranitidine significantly enhanced levels of IgG2a capable of binding to E0771-GFP cancer cells. Serum was diluted 1:10 and added to cultured E0771-GFP cells and anti-isotype antibodies for IgG2a (A) or kappa light chain (B) were added. Relative fluorescence intensity (RFI) was used as an indicator for antibody binding to E0771-GFP cells relative to the average of control MFI. Box plot represents mean and range of data from 20 to 24 individual mice. * p

    Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) Enzyme-linked immunosorbent assay plates were coated with anti-mouse IgG1 (BioLegend) or anti-mouse IgG2a (BioLegend) at a concentration of 2.5 µg/mL in borate buffer (2.5 µg/mL, pH 8.3) and placed at 4°C overnight.

    Techniques: Binding Assay, Cell Culture, Fluorescence, Mouse Assay

    Ranitidine enhances increased IgG2a antibody production and binding which significantly correlates with tumor weight. Tumor weights from non-treated (A,C) or ranitidine-treated mice (B,D) were plotted relative to the amount of IgG2a antibody that is specific to GFP (A,B) or to E0771-GFP tumor cell-binding IgG2a (C,D) . Each dot represents individual mice and lines represent the correlation line. Correlation analyses were performed to obtain r and p -value.

    Journal: Frontiers in Immunology

    Article Title: Ranitidine Inhibition of Breast Tumor Growth Is B Cell Dependent and Associated With an Enhanced Antitumor Antibody Response

    doi: 10.3389/fimmu.2018.01894

    Figure Lengend Snippet: Ranitidine enhances increased IgG2a antibody production and binding which significantly correlates with tumor weight. Tumor weights from non-treated (A,C) or ranitidine-treated mice (B,D) were plotted relative to the amount of IgG2a antibody that is specific to GFP (A,B) or to E0771-GFP tumor cell-binding IgG2a (C,D) . Each dot represents individual mice and lines represent the correlation line. Correlation analyses were performed to obtain r and p -value.

    Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) Enzyme-linked immunosorbent assay plates were coated with anti-mouse IgG1 (BioLegend) or anti-mouse IgG2a (BioLegend) at a concentration of 2.5 µg/mL in borate buffer (2.5 µg/mL, pH 8.3) and placed at 4°C overnight.

    Techniques: Binding Assay, Mouse Assay

    Ranitidine enhanced anti-OVA IgG2a and IgG1 antibody levels in OVA-Alum immunized naïve C57BL/6 mice but not in B16-OVA tumor-bearing mice. C57BL/6 mice were immunized and boosted with OVA with alum as an adjuvant (A,B) . C57BL/6 mice (ranitidine-treated or non-treated) were implanted with OVA-expressing B16 tumor cells (C,D) . After 18 days, the plasma from the mice were serially diluted and enzyme-linked immunosorbent assay was used to detect OVA-specific IgG1 (A,C) and IgG2a (B,D ). Data points represent mean ± SEM of 12 mice. Significance was assessed using two-way ANOVA followed by uncorrected Fisher’s LSD multiple comparisons. * p

    Journal: Frontiers in Immunology

    Article Title: Ranitidine Inhibition of Breast Tumor Growth Is B Cell Dependent and Associated With an Enhanced Antitumor Antibody Response

    doi: 10.3389/fimmu.2018.01894

    Figure Lengend Snippet: Ranitidine enhanced anti-OVA IgG2a and IgG1 antibody levels in OVA-Alum immunized naïve C57BL/6 mice but not in B16-OVA tumor-bearing mice. C57BL/6 mice were immunized and boosted with OVA with alum as an adjuvant (A,B) . C57BL/6 mice (ranitidine-treated or non-treated) were implanted with OVA-expressing B16 tumor cells (C,D) . After 18 days, the plasma from the mice were serially diluted and enzyme-linked immunosorbent assay was used to detect OVA-specific IgG1 (A,C) and IgG2a (B,D ). Data points represent mean ± SEM of 12 mice. Significance was assessed using two-way ANOVA followed by uncorrected Fisher’s LSD multiple comparisons. * p

    Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) Enzyme-linked immunosorbent assay plates were coated with anti-mouse IgG1 (BioLegend) or anti-mouse IgG2a (BioLegend) at a concentration of 2.5 µg/mL in borate buffer (2.5 µg/mL, pH 8.3) and placed at 4°C overnight.

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

    Alterations in anti-GFP IgG1 and IgG2a antibody in ranitidine-treated E0771-GFP tumor-bearing mice. Plasma from C57BL/6 and CCR2 −/− tumor-bearing mice (treated and non-treated) were serially diluted and an enzyme-linked immunosorbent assay was used to detect GFP-specific IgG1 (A) or IgG2a (B) . Data points represent mean ± SEM of 10–34 mice. Significance was assessed using two-way ANOVA followed by uncorrected Fisher’s LSD multiple comparisons. * p

    Journal: Frontiers in Immunology

    Article Title: Ranitidine Inhibition of Breast Tumor Growth Is B Cell Dependent and Associated With an Enhanced Antitumor Antibody Response

    doi: 10.3389/fimmu.2018.01894

    Figure Lengend Snippet: Alterations in anti-GFP IgG1 and IgG2a antibody in ranitidine-treated E0771-GFP tumor-bearing mice. Plasma from C57BL/6 and CCR2 −/− tumor-bearing mice (treated and non-treated) were serially diluted and an enzyme-linked immunosorbent assay was used to detect GFP-specific IgG1 (A) or IgG2a (B) . Data points represent mean ± SEM of 10–34 mice. Significance was assessed using two-way ANOVA followed by uncorrected Fisher’s LSD multiple comparisons. * p

    Article Snippet: Enzyme-Linked Immunosorbent Assay (ELISA) Enzyme-linked immunosorbent assay plates were coated with anti-mouse IgG1 (BioLegend) or anti-mouse IgG2a (BioLegend) at a concentration of 2.5 µg/mL in borate buffer (2.5 µg/mL, pH 8.3) and placed at 4°C overnight.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    dAd5GNC evoked anti-GNC antibodies in mice. ( a ) Total anti-GNC IgG antibody titers over time. BALB/c mice ( n = 20) were vaccinated intramuscularly with 4 µg dAd5GNC at 0, 3, and 6 week. Antibody titers were assessed by ELISA against BSA

    Journal: Molecular Therapy

    Article Title: Cocaine Analog Coupled to Disrupted Adenovirus: A Vaccine Strategy to Evoke High-titer Immunity Against Addictive Drugs

    doi: 10.1038/mt.2010.280

    Figure Lengend Snippet: dAd5GNC evoked anti-GNC antibodies in mice. ( a ) Total anti-GNC IgG antibody titers over time. BALB/c mice ( n = 20) were vaccinated intramuscularly with 4 µg dAd5GNC at 0, 3, and 6 week. Antibody titers were assessed by ELISA against BSA

    Article Snippet: For the rabbit anti-mouse isotype-specific antibody, 100 µl of anti-IgG1, IgG2a, IgG2b, or IgM (BioRad) was added to each independent well and incubated in 1% dry milk in PBS for 90 minutes, 23 °C.

    Techniques: Mouse Assay, Enzyme-linked Immunosorbent Assay

    Anti- S. aureus IgG1 and IgG2 titers in paired maternal and cord serum samples from the case and control groups. (a) Anti- S. aureus IgG1 titers and (d) anti- S. aureus IgG2 titers in the maternal serum and umbilical cord serum samples from the case (n=48) and control group (n=98). In (b) and (c) , correlation coefficients between paired maternal and umbilical cord serum of anti- Sa IgG1 in the case and control group, respectively. In (e) and (f) , correlation coefficients between the paired maternal and umbilical cord serum of anti- Sa IgG2 in the case and control group, respectively. In (a) and (d) , the box represents the 25 th –75 th percentiles, and the median is represented by the line within the box. The whiskers represent the 5 th –95 th percentiles.

    Journal: Clinics

    Article Title: Passive acquisition of anti-Staphylococcus aureus antibodies by newborns via transplacental transfer and breastfeeding, regardless of maternal colonization

    doi: 10.6061/clinics/2016(12)02

    Figure Lengend Snippet: Anti- S. aureus IgG1 and IgG2 titers in paired maternal and cord serum samples from the case and control groups. (a) Anti- S. aureus IgG1 titers and (d) anti- S. aureus IgG2 titers in the maternal serum and umbilical cord serum samples from the case (n=48) and control group (n=98). In (b) and (c) , correlation coefficients between paired maternal and umbilical cord serum of anti- Sa IgG1 in the case and control group, respectively. In (e) and (f) , correlation coefficients between the paired maternal and umbilical cord serum of anti- Sa IgG2 in the case and control group, respectively. In (a) and (d) , the box represents the 25 th –75 th percentiles, and the median is represented by the line within the box. The whiskers represent the 5 th –95 th percentiles.

    Article Snippet: Biotinylated anti-human IgG1 (555869; BD Pharmingen, San Diego, CA, USA) or IgG2 (555874; BD Pharmingen, San Diego, CA, USA) was used as the secondary antibody, both at dilutions of 1:500, and incubated for 90 minutes.

    Techniques:

    Placental transfer ratios (%) of anti- S. aureus IgG1 and IgG2 from case and control groups. Comparison of the data is presented as box and whisker plots. The box represents the 25 th –75 th percentiles, and the median is represented by the line within the box. The whiskers represent the 5 th –95 th percentiles.

    Journal: Clinics

    Article Title: Passive acquisition of anti-Staphylococcus aureus antibodies by newborns via transplacental transfer and breastfeeding, regardless of maternal colonization

    doi: 10.6061/clinics/2016(12)02

    Figure Lengend Snippet: Placental transfer ratios (%) of anti- S. aureus IgG1 and IgG2 from case and control groups. Comparison of the data is presented as box and whisker plots. The box represents the 25 th –75 th percentiles, and the median is represented by the line within the box. The whiskers represent the 5 th –95 th percentiles.

    Article Snippet: Biotinylated anti-human IgG1 (555869; BD Pharmingen, San Diego, CA, USA) or IgG2 (555874; BD Pharmingen, San Diego, CA, USA) was used as the secondary antibody, both at dilutions of 1:500, and incubated for 90 minutes.

    Techniques: Whisker Assay