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HiMedia Laboratories peripheral blood mononuclear cells pbmcs
Peripheral Blood Mononuclear Cells Pbmcs, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Article Title: Upregulation of GLS1 Isoforms KGA and GAC Facilitates Mitochondrial Metabolism and Cell Proliferation in Epstein–Barr Virus Infected Cells
Article Snippet: Peripheral blood mononuclear cells (PBMC) were purchased from HiMedia laboratories, India.

Article Title: Surfactant Protein D Inhibits HIV-1 Infection of Target Cells via Interference with gp120-CD4 Interaction and Modulates Pro-Inflammatory Cytokine Production
Article Snippet: Peripheral Blood Mononuclear Cells (PBMCs) pooled from different study participants were procured from Himedia Laboratories.

Article Title: Hexokinase II inhibition by 3-bromopyruvate sensitizes myeloid leukemic cells K-562 to anti-leukemic drug, daunorubicin
Article Snippet: Whereas, peripheral blood mononuclear cells (PBMCs) were procured from HiMedia (Mumbai, India).

Article Title: Strophanthidin Attenuates MAPK, PI3K/AKT/mTOR, and Wnt/β-Catenin Signaling Pathways in Human Cancers
Article Snippet: Peripheral blood mononuclear cells (PBMCs) were purchased from HiMedia (CL003-25).

Cell Culture:

Article Title: Extract of Vernonia condensata, Inhibits Tumor Progression and Improves Survival of Tumor-allograft Bearing Mouse
Article Snippet: .. Cell lines and culture Human pre B-cell leukemic cell lines, Reh and Nalm6, human chronic myelogenous leukemic cell line, K562, T-cell leukemic cell line, Molt4 and human breast adenocarcinoma cell line, MCF7, human embryonic kidney cell line, HEK293T and peripheral blood mononuclear cells (PBMCs) were cultured in RPMI 1640 or MEM or DMEM (HiMedia, India) supplemented with 10% heat-inactivated fetal bovine serum (Gibco BRL, USA), 100 U of Penicillin-Streptomycin/ml. ..

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  • 90
    HiMedia Laboratories in vitro pbmcs
    Expression of GvpC-mBPI N fusion protein in haloarchaeal gas vesicle nanoparticles and its antibacterial and anti-inflammatory activity. ( a ) Thin-sections of Halobacterium sp. observed by transmission electron microscopy Panel A. Strain SD109 with a deletion of the gvp gene cluster and lacking GVNPs. Panel B. Strain SD109 transformed with a plasmid containing the entire gvp gene cluster producing buoyant GNPVs. Bar in Panel A indicates 0.5 um length (for both panels) ( b ) Halobacterium sp. NRC-1 (pDRK-C3-mBPI) BPI-GVNPs were produced and purified by floatation of the GVNP particles using the accelerated centrifugation and expression of GvpC-mBPINfusion protein was confirmed by Western blotting using a 1:1000 dilution of anti-His-tag primary antibody. Lane 1: <t>mBPIN-GVNPs,</t> Lane 2: WT-GVNPs. 10 6 Salmonella Typhimurium 14028 ( c ) and E. coli ( d ) were incubated with WT-GVNPs (White Bars) or mBPIN-GVNPs (Black Bars) for 2 hours at 37 °C. Cells were plated on LB agar and incubated at 37 °C overnight. Percent survival values are plotted and are means of triplicate assays. (n = 3 experiments) ( e ) 10 6 Salmonella Typhimurium 14028 were incubated with PBS (White Bars) or mBPIN-GVP-C3 fusion protein (Black Bars) for 2 hours at 37 °C. Cells were plated on LB agar and incubated at 37 °C overnight. Percent survival values are plotted and are means of triplicate assays. (n = 3 experiments) ( f ) 10 6 Salmonella Typhimurium 14028 were incubated with WT-GVNPs (Panel A) or mBPIN-GVNPs (Panel B) for 2 hours at 37 °C. Cells were fixed and observed by scanning electron microscopy. White arrow indicates membrane damage and leakage of cytosolic contents in bacteria incubated with mBPIN-GVNPs. Inserts were magnified to show cell morphology of WT-GVNP treated and mBPIN-GVNP treated bacteria. ( g ) <t>PBMCs</t> were treated with LPS (10 ng) or with LPS (10 ng) preincubated with mBPIN-GVNP. 24 h post treatment, supernatant was collected and TNFα levels were measured by ELISA. TNFαlevels were compared with un-treated control. (n = 3 experiments). Data was analyzed by students T test. Key: ***p
    In Vitro Pbmcs, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/in vitro pbmcs/product/HiMedia Laboratories
    Average 90 stars, based on 1 article reviews
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    91
    HiMedia Laboratories blood mononuclear cell pbmc culture pbmcs
    Increased shedding of DPP4 from T cells of T2DM patients . (A, B) <t>PBMC</t> culture supernatants (A) and cell lysates (B) were analyzed for DPP4 by ELISA after 48 h (control n = 9 T2DM n = 10). (C–D) Relative gene expression in CD4+ T cells for DPP4 and KLK5 in T2DM and control population (control n = 18 T2DM n = 27). (E–F) Secreted DPP4 as well as KLK5 protein were measured by ELISA (control n = 12 T2DM n = 27). (G) Linear regression analysis of secreted DPP4 protein with secreted KLK5 protein in overall population. (H–I) siKLK5 was electroporated in CD4+ T cells and gene expression and secreted DPP4 levels were measured after 72 h (n = 7). (J) Stimulated CD4+ T cells were treated with human recombinant KLK5 (150 nM) for 4 h and culture supernatant was analyzed for DPP4 by ELISA (n = 8). (K) Activated CD4+ T cells were cultured in a polylysine coated plate for 16 h, stained with DPP4 mAb (red) and visualized with fluorescence confocal microscopy. Scale bar = 10 μm. (L) Fluorescence intensity was calculated of 100 different cells across four fields by Image J software. Data are shown as the mean ± SD. Statistical analysis was performed by Mann–Whitney U test and Student's t test with Spearman correlation; *p
    Blood Mononuclear Cell Pbmc Culture Pbmcs, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/blood mononuclear cell pbmc culture pbmcs/product/HiMedia Laboratories
    Average 91 stars, based on 1 article reviews
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    90
    HiMedia Laboratories pooled peripheral blood mononuclear cell pbmc pooled peripheral blood mononuclear cells
    <t>HIV</t> transfer assay mediated by DC-SIGN . Cell surface DC-SIGN expressing HEK (DC-HEK) cells were grown in a 12-well plate to form a confluent layer. Different concentrations of proteins were added to the cells and incubated for 2 h for binding. Unbound proteins were removed and cells were challenged with 2.5 ng/mL p24 of HIV-1 (SF-162 strain) for 1 h. Unbound virus was washed off and cells were co-cultured with PHA-activated <t>PBMCs</t> for 24 h. PBMCs were separated from the DC-HEK monolayer and cultured separately for 7 days to determine viral titer of the supernatants collected on days 4 and 7. (A) C1q, ghA, ghB, ghC, and ghABC; (B) gC1qR in presence of C1q, ghA, ghB, ghC, and ghABC. Data represent mean ± SD. P
    Pooled Peripheral Blood Mononuclear Cell Pbmc Pooled Peripheral Blood Mononuclear Cells, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/pooled peripheral blood mononuclear cell pbmc pooled peripheral blood mononuclear cells/product/HiMedia Laboratories
    Average 90 stars, based on 1 article reviews
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    HiMedia Laboratories pbmc s
    Cross reactivity between Rhi o 1 and Bla g 2. (A) IgE ELISA: rBla g r was tested for its IgE binding to ten Rhi o 1 allergic sera and eight of them were IgE reactive to rBla g 2 (except patient no. 5 and 8) (B) IgE ELISA inhibition: These eight Rhi o 1 positive sera with high anti-Bla g 2 IgE titer were pooled and pre-incubated with increasing concentrations of rBla g 2 (fluid phase inhibitor). rRhi o 1 and BSA instead of rBla g 2 were used as positive (auto inhibition) and negative control (no inhibition) respectively. IgE Binding to plate bound 1 μg of rRhi o 1 was inhibited by rBla g 2 in a dose dependent manner. IC 50 for rBla g 2 is 20 ng and maximum 58% inhibition was observed with 100 ng rBla g 2. (C) A reciprocal IgE-ELISA inhibition where plate bound 1 μg of rBla g 2 in solid phase was incubated with sera mixed with rRhi o 1 or rBla g 2. In case of rRhi o 1 the IC 50 is 9 ng and maximum ~93% was achieved at 1000 ng. In case of auto inhibition with rBla g 2 full inhibition was observed. (D) Cross-stimulation experiment: <t>PBMC’s</t> sensitized with eight anti-Rhi o 1 IgE antibody were stimulated with rBla g 2 and histamine release was observed within a range 29% to 36%. No histamine release observed for healthy sera (HS1 and HS2) sensitization and BSA challenge as negative controls.
    Pbmc S, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 89/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Average 89 stars, based on 1 article reviews
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    Expression of GvpC-mBPI N fusion protein in haloarchaeal gas vesicle nanoparticles and its antibacterial and anti-inflammatory activity. ( a ) Thin-sections of Halobacterium sp. observed by transmission electron microscopy Panel A. Strain SD109 with a deletion of the gvp gene cluster and lacking GVNPs. Panel B. Strain SD109 transformed with a plasmid containing the entire gvp gene cluster producing buoyant GNPVs. Bar in Panel A indicates 0.5 um length (for both panels) ( b ) Halobacterium sp. NRC-1 (pDRK-C3-mBPI) BPI-GVNPs were produced and purified by floatation of the GVNP particles using the accelerated centrifugation and expression of GvpC-mBPINfusion protein was confirmed by Western blotting using a 1:1000 dilution of anti-His-tag primary antibody. Lane 1: mBPIN-GVNPs, Lane 2: WT-GVNPs. 10 6 Salmonella Typhimurium 14028 ( c ) and E. coli ( d ) were incubated with WT-GVNPs (White Bars) or mBPIN-GVNPs (Black Bars) for 2 hours at 37 °C. Cells were plated on LB agar and incubated at 37 °C overnight. Percent survival values are plotted and are means of triplicate assays. (n = 3 experiments) ( e ) 10 6 Salmonella Typhimurium 14028 were incubated with PBS (White Bars) or mBPIN-GVP-C3 fusion protein (Black Bars) for 2 hours at 37 °C. Cells were plated on LB agar and incubated at 37 °C overnight. Percent survival values are plotted and are means of triplicate assays. (n = 3 experiments) ( f ) 10 6 Salmonella Typhimurium 14028 were incubated with WT-GVNPs (Panel A) or mBPIN-GVNPs (Panel B) for 2 hours at 37 °C. Cells were fixed and observed by scanning electron microscopy. White arrow indicates membrane damage and leakage of cytosolic contents in bacteria incubated with mBPIN-GVNPs. Inserts were magnified to show cell morphology of WT-GVNP treated and mBPIN-GVNP treated bacteria. ( g ) PBMCs were treated with LPS (10 ng) or with LPS (10 ng) preincubated with mBPIN-GVNP. 24 h post treatment, supernatant was collected and TNFα levels were measured by ELISA. TNFαlevels were compared with un-treated control. (n = 3 experiments). Data was analyzed by students T test. Key: ***p

    Journal: Scientific Reports

    Article Title: Halobacterial nano vesicles displaying murine bactericidal permeability-increasing protein rescue mice from lethal endotoxic shock

    doi: 10.1038/srep33679

    Figure Lengend Snippet: Expression of GvpC-mBPI N fusion protein in haloarchaeal gas vesicle nanoparticles and its antibacterial and anti-inflammatory activity. ( a ) Thin-sections of Halobacterium sp. observed by transmission electron microscopy Panel A. Strain SD109 with a deletion of the gvp gene cluster and lacking GVNPs. Panel B. Strain SD109 transformed with a plasmid containing the entire gvp gene cluster producing buoyant GNPVs. Bar in Panel A indicates 0.5 um length (for both panels) ( b ) Halobacterium sp. NRC-1 (pDRK-C3-mBPI) BPI-GVNPs were produced and purified by floatation of the GVNP particles using the accelerated centrifugation and expression of GvpC-mBPINfusion protein was confirmed by Western blotting using a 1:1000 dilution of anti-His-tag primary antibody. Lane 1: mBPIN-GVNPs, Lane 2: WT-GVNPs. 10 6 Salmonella Typhimurium 14028 ( c ) and E. coli ( d ) were incubated with WT-GVNPs (White Bars) or mBPIN-GVNPs (Black Bars) for 2 hours at 37 °C. Cells were plated on LB agar and incubated at 37 °C overnight. Percent survival values are plotted and are means of triplicate assays. (n = 3 experiments) ( e ) 10 6 Salmonella Typhimurium 14028 were incubated with PBS (White Bars) or mBPIN-GVP-C3 fusion protein (Black Bars) for 2 hours at 37 °C. Cells were plated on LB agar and incubated at 37 °C overnight. Percent survival values are plotted and are means of triplicate assays. (n = 3 experiments) ( f ) 10 6 Salmonella Typhimurium 14028 were incubated with WT-GVNPs (Panel A) or mBPIN-GVNPs (Panel B) for 2 hours at 37 °C. Cells were fixed and observed by scanning electron microscopy. White arrow indicates membrane damage and leakage of cytosolic contents in bacteria incubated with mBPIN-GVNPs. Inserts were magnified to show cell morphology of WT-GVNP treated and mBPIN-GVNP treated bacteria. ( g ) PBMCs were treated with LPS (10 ng) or with LPS (10 ng) preincubated with mBPIN-GVNP. 24 h post treatment, supernatant was collected and TNFα levels were measured by ELISA. TNFαlevels were compared with un-treated control. (n = 3 experiments). Data was analyzed by students T test. Key: ***p

    Article Snippet: Anti-inflammatory activity of mBPIN-GVNPs: in vitro PBMCs were isolated from healthy human volunteers using Himedia LSM as per instructors manual.

    Techniques: Expressing, Activity Assay, Transmission Assay, Electron Microscopy, Transformation Assay, Plasmid Preparation, Produced, Purification, Centrifugation, Western Blot, Incubation, Enzyme-linked Immunosorbent Assay

    Increased shedding of DPP4 from T cells of T2DM patients . (A, B) PBMC culture supernatants (A) and cell lysates (B) were analyzed for DPP4 by ELISA after 48 h (control n = 9 T2DM n = 10). (C–D) Relative gene expression in CD4+ T cells for DPP4 and KLK5 in T2DM and control population (control n = 18 T2DM n = 27). (E–F) Secreted DPP4 as well as KLK5 protein were measured by ELISA (control n = 12 T2DM n = 27). (G) Linear regression analysis of secreted DPP4 protein with secreted KLK5 protein in overall population. (H–I) siKLK5 was electroporated in CD4+ T cells and gene expression and secreted DPP4 levels were measured after 72 h (n = 7). (J) Stimulated CD4+ T cells were treated with human recombinant KLK5 (150 nM) for 4 h and culture supernatant was analyzed for DPP4 by ELISA (n = 8). (K) Activated CD4+ T cells were cultured in a polylysine coated plate for 16 h, stained with DPP4 mAb (red) and visualized with fluorescence confocal microscopy. Scale bar = 10 μm. (L) Fluorescence intensity was calculated of 100 different cells across four fields by Image J software. Data are shown as the mean ± SD. Statistical analysis was performed by Mann–Whitney U test and Student's t test with Spearman correlation; *p

    Journal: Molecular Metabolism

    Article Title: KLK5 induces shedding of DPP4 from circulatory Th17 cells in type 2 diabetes

    doi: 10.1016/j.molmet.2017.09.004

    Figure Lengend Snippet: Increased shedding of DPP4 from T cells of T2DM patients . (A, B) PBMC culture supernatants (A) and cell lysates (B) were analyzed for DPP4 by ELISA after 48 h (control n = 9 T2DM n = 10). (C–D) Relative gene expression in CD4+ T cells for DPP4 and KLK5 in T2DM and control population (control n = 18 T2DM n = 27). (E–F) Secreted DPP4 as well as KLK5 protein were measured by ELISA (control n = 12 T2DM n = 27). (G) Linear regression analysis of secreted DPP4 protein with secreted KLK5 protein in overall population. (H–I) siKLK5 was electroporated in CD4+ T cells and gene expression and secreted DPP4 levels were measured after 72 h (n = 7). (J) Stimulated CD4+ T cells were treated with human recombinant KLK5 (150 nM) for 4 h and culture supernatant was analyzed for DPP4 by ELISA (n = 8). (K) Activated CD4+ T cells were cultured in a polylysine coated plate for 16 h, stained with DPP4 mAb (red) and visualized with fluorescence confocal microscopy. Scale bar = 10 μm. (L) Fluorescence intensity was calculated of 100 different cells across four fields by Image J software. Data are shown as the mean ± SD. Statistical analysis was performed by Mann–Whitney U test and Student's t test with Spearman correlation; *p

    Article Snippet: 2.2 Peripheral blood mononuclear cell (PBMC) culture PBMCs were isolated using HiSep™ LSM 1077 (Himedia Laboratories).

    Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Recombinant, Cell Culture, Staining, Fluorescence, Confocal Microscopy, Software, MANN-WHITNEY

    Decreased surface expression of DPP4 in Th17 cells of T2DM patients . (A, B) Comparison of relative KLK5 gene expression in sorted Th1 and Th17 cells (control, n = 10 T2DM, n = 14). (C, D) Correlations of percentage of CD26+IL17A+CD4+ T cells with plasma DPP4 activity (A; n = 26) and PBMC DPP4 activity (B; n = 16). (E, F) Correlations of percentage of CD26+IFNγ+CD4+ T cells with plasma DPP4 activity (C; n = 30) and PBMC DPP4 activity (D; n = 15). (G, H) Representative flow plots for IFNγ and CD26 staining (E) and the summary (F) of all the analyzed samples are shown (control n = 17 T2DM n = 12). (I, J) IL-17A and CD26 staining from healthy control (n = 20) and treatment naïve T2DM (n = 19) were determined by flow cytometry. Statistical analysis was performed by Mann–Whitney U test and Student's t test with Spearman correlation; *p

    Journal: Molecular Metabolism

    Article Title: KLK5 induces shedding of DPP4 from circulatory Th17 cells in type 2 diabetes

    doi: 10.1016/j.molmet.2017.09.004

    Figure Lengend Snippet: Decreased surface expression of DPP4 in Th17 cells of T2DM patients . (A, B) Comparison of relative KLK5 gene expression in sorted Th1 and Th17 cells (control, n = 10 T2DM, n = 14). (C, D) Correlations of percentage of CD26+IL17A+CD4+ T cells with plasma DPP4 activity (A; n = 26) and PBMC DPP4 activity (B; n = 16). (E, F) Correlations of percentage of CD26+IFNγ+CD4+ T cells with plasma DPP4 activity (C; n = 30) and PBMC DPP4 activity (D; n = 15). (G, H) Representative flow plots for IFNγ and CD26 staining (E) and the summary (F) of all the analyzed samples are shown (control n = 17 T2DM n = 12). (I, J) IL-17A and CD26 staining from healthy control (n = 20) and treatment naïve T2DM (n = 19) were determined by flow cytometry. Statistical analysis was performed by Mann–Whitney U test and Student's t test with Spearman correlation; *p

    Article Snippet: 2.2 Peripheral blood mononuclear cell (PBMC) culture PBMCs were isolated using HiSep™ LSM 1077 (Himedia Laboratories).

    Techniques: Expressing, Activity Assay, Flow Cytometry, Staining, Cytometry, MANN-WHITNEY

    PBMC is an important source of increased plasma DPP4 activity in T2DM patients . DPP4 activity in plasma and peripheral blood mononuclear cells (PBMC) of treatment naïve type 2 diabetes (T2DM) patients and healthy control subjects. DPP4 expression in PBMC was measured and linear regression analysis was performed with Spearman correlation. (A–C) Comparison of plasma (control n = 78 T2DM n = 135) PBMC DPP4 activity (control n = 46 T2DM n = 35) and PBMC DPP4 gene expression (control n = 49 T2DM n = 41). (D–G) Linear regression analysis of PBMC DPP4 activity with plasma DPP4 activity (D F), and PBMC DPP4 activity and gene expression (E G). (H, I) PBMCs from control (n = 22) and treatment naïve T2DM subjects (n = 28) were cultured for 48 h and supernatant (H) as well as cellular DPP4 levels (I) were analyzed by ELISA. Statistical analysis was performed by Mann–Whitney U test and Student's t test with Spearman correlation; *p

    Journal: Molecular Metabolism

    Article Title: KLK5 induces shedding of DPP4 from circulatory Th17 cells in type 2 diabetes

    doi: 10.1016/j.molmet.2017.09.004

    Figure Lengend Snippet: PBMC is an important source of increased plasma DPP4 activity in T2DM patients . DPP4 activity in plasma and peripheral blood mononuclear cells (PBMC) of treatment naïve type 2 diabetes (T2DM) patients and healthy control subjects. DPP4 expression in PBMC was measured and linear regression analysis was performed with Spearman correlation. (A–C) Comparison of plasma (control n = 78 T2DM n = 135) PBMC DPP4 activity (control n = 46 T2DM n = 35) and PBMC DPP4 gene expression (control n = 49 T2DM n = 41). (D–G) Linear regression analysis of PBMC DPP4 activity with plasma DPP4 activity (D F), and PBMC DPP4 activity and gene expression (E G). (H, I) PBMCs from control (n = 22) and treatment naïve T2DM subjects (n = 28) were cultured for 48 h and supernatant (H) as well as cellular DPP4 levels (I) were analyzed by ELISA. Statistical analysis was performed by Mann–Whitney U test and Student's t test with Spearman correlation; *p

    Article Snippet: 2.2 Peripheral blood mononuclear cell (PBMC) culture PBMCs were isolated using HiSep™ LSM 1077 (Himedia Laboratories).

    Techniques: Activity Assay, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    KLK5 is the proteolytic enzymes involved in the DPP4 shedding from PBMC . (A, B) PBMC culture supernatants were pooled (n = 8) and a human protease array profile was performed. Spots for KLK5 are marked (A). Data are represented as relative pixel density of significantly altered proteases in T2DM (B). (C) Comparison of PBMC KLK5 gene expression (control n = 17 T2DM n = 20). (D) Plasma KLK5 levels (control n = 29 T2DM n = 36). (E, F) PBMC was incubated for 24 h and 16 h with 10 μM MMP inhibitor (MMPi; control n = 6 and T2DM n = 9) and 100 μg/ml KLK inhibitor (KLKi; control n = 6 and T2DM n = 12) respectively. DPP4 release in the culture supernatants were measured by ELISA. Statistical analysis was calculated by Mann–Whitney U test and shown as box plots; p**

    Journal: Molecular Metabolism

    Article Title: KLK5 induces shedding of DPP4 from circulatory Th17 cells in type 2 diabetes

    doi: 10.1016/j.molmet.2017.09.004

    Figure Lengend Snippet: KLK5 is the proteolytic enzymes involved in the DPP4 shedding from PBMC . (A, B) PBMC culture supernatants were pooled (n = 8) and a human protease array profile was performed. Spots for KLK5 are marked (A). Data are represented as relative pixel density of significantly altered proteases in T2DM (B). (C) Comparison of PBMC KLK5 gene expression (control n = 17 T2DM n = 20). (D) Plasma KLK5 levels (control n = 29 T2DM n = 36). (E, F) PBMC was incubated for 24 h and 16 h with 10 μM MMP inhibitor (MMPi; control n = 6 and T2DM n = 9) and 100 μg/ml KLK inhibitor (KLKi; control n = 6 and T2DM n = 12) respectively. DPP4 release in the culture supernatants were measured by ELISA. Statistical analysis was calculated by Mann–Whitney U test and shown as box plots; p**

    Article Snippet: 2.2 Peripheral blood mononuclear cell (PBMC) culture PBMCs were isolated using HiSep™ LSM 1077 (Himedia Laboratories).

    Techniques: Expressing, Incubation, Enzyme-linked Immunosorbent Assay, MANN-WHITNEY

    HIV transfer assay mediated by DC-SIGN . Cell surface DC-SIGN expressing HEK (DC-HEK) cells were grown in a 12-well plate to form a confluent layer. Different concentrations of proteins were added to the cells and incubated for 2 h for binding. Unbound proteins were removed and cells were challenged with 2.5 ng/mL p24 of HIV-1 (SF-162 strain) for 1 h. Unbound virus was washed off and cells were co-cultured with PHA-activated PBMCs for 24 h. PBMCs were separated from the DC-HEK monolayer and cultured separately for 7 days to determine viral titer of the supernatants collected on days 4 and 7. (A) C1q, ghA, ghB, ghC, and ghABC; (B) gC1qR in presence of C1q, ghA, ghB, ghC, and ghABC. Data represent mean ± SD. P

    Journal: Frontiers in Immunology

    Article Title: Complement Protein C1q Interacts with DC-SIGN via Its Globular Domain and Thus May Interfere with HIV-1 Transmission

    doi: 10.3389/fimmu.2016.00600

    Figure Lengend Snippet: HIV transfer assay mediated by DC-SIGN . Cell surface DC-SIGN expressing HEK (DC-HEK) cells were grown in a 12-well plate to form a confluent layer. Different concentrations of proteins were added to the cells and incubated for 2 h for binding. Unbound proteins were removed and cells were challenged with 2.5 ng/mL p24 of HIV-1 (SF-162 strain) for 1 h. Unbound virus was washed off and cells were co-cultured with PHA-activated PBMCs for 24 h. PBMCs were separated from the DC-HEK monolayer and cultured separately for 7 days to determine viral titer of the supernatants collected on days 4 and 7. (A) C1q, ghA, ghB, ghC, and ghABC; (B) gC1qR in presence of C1q, ghA, ghB, ghC, and ghABC. Data represent mean ± SD. P

    Article Snippet: HIV-1 Transfer Assay with DC-HEK Cells and Pooled Peripheral Blood Mononuclear Cell (PBMC) Pooled peripheral blood mononuclear cells (HiMedia Laboratories, India) were cultured in RPMI 1640 medium (Sigma Aldrich) containing 10% FBS, 1% Penicillin–Streptomycin (Complete RPMI medium), and stimulated with 5 µg/mL phytohemaglutinin (PHA) and 10 U/Ml of recombinant-human IL-2 (Gibco) for 24 h. PHA/IL-2 was washed off and activated PBMCs were cultured further for 3 days in complete RPMI 1640 medium.

    Techniques: Expressing, Incubation, Binding Assay, Cell Culture

    Cross reactivity between Rhi o 1 and Bla g 2. (A) IgE ELISA: rBla g r was tested for its IgE binding to ten Rhi o 1 allergic sera and eight of them were IgE reactive to rBla g 2 (except patient no. 5 and 8) (B) IgE ELISA inhibition: These eight Rhi o 1 positive sera with high anti-Bla g 2 IgE titer were pooled and pre-incubated with increasing concentrations of rBla g 2 (fluid phase inhibitor). rRhi o 1 and BSA instead of rBla g 2 were used as positive (auto inhibition) and negative control (no inhibition) respectively. IgE Binding to plate bound 1 μg of rRhi o 1 was inhibited by rBla g 2 in a dose dependent manner. IC 50 for rBla g 2 is 20 ng and maximum 58% inhibition was observed with 100 ng rBla g 2. (C) A reciprocal IgE-ELISA inhibition where plate bound 1 μg of rBla g 2 in solid phase was incubated with sera mixed with rRhi o 1 or rBla g 2. In case of rRhi o 1 the IC 50 is 9 ng and maximum ~93% was achieved at 1000 ng. In case of auto inhibition with rBla g 2 full inhibition was observed. (D) Cross-stimulation experiment: PBMC’s sensitized with eight anti-Rhi o 1 IgE antibody were stimulated with rBla g 2 and histamine release was observed within a range 29% to 36%. No histamine release observed for healthy sera (HS1 and HS2) sensitization and BSA challenge as negative controls.

    Journal: PLoS ONE

    Article Title: Purification, Cloning and Immuno-Biochemical Characterization of a Fungal Aspartic Protease Allergen Rhi o 1 from the Airborne Mold Rhizopus oryzae

    doi: 10.1371/journal.pone.0144547

    Figure Lengend Snippet: Cross reactivity between Rhi o 1 and Bla g 2. (A) IgE ELISA: rBla g r was tested for its IgE binding to ten Rhi o 1 allergic sera and eight of them were IgE reactive to rBla g 2 (except patient no. 5 and 8) (B) IgE ELISA inhibition: These eight Rhi o 1 positive sera with high anti-Bla g 2 IgE titer were pooled and pre-incubated with increasing concentrations of rBla g 2 (fluid phase inhibitor). rRhi o 1 and BSA instead of rBla g 2 were used as positive (auto inhibition) and negative control (no inhibition) respectively. IgE Binding to plate bound 1 μg of rRhi o 1 was inhibited by rBla g 2 in a dose dependent manner. IC 50 for rBla g 2 is 20 ng and maximum 58% inhibition was observed with 100 ng rBla g 2. (C) A reciprocal IgE-ELISA inhibition where plate bound 1 μg of rBla g 2 in solid phase was incubated with sera mixed with rRhi o 1 or rBla g 2. In case of rRhi o 1 the IC 50 is 9 ng and maximum ~93% was achieved at 1000 ng. In case of auto inhibition with rBla g 2 full inhibition was observed. (D) Cross-stimulation experiment: PBMC’s sensitized with eight anti-Rhi o 1 IgE antibody were stimulated with rBla g 2 and histamine release was observed within a range 29% to 36%. No histamine release observed for healthy sera (HS1 and HS2) sensitization and BSA challenge as negative controls.

    Article Snippet: Briefly, PBMC’s (2 x 105 ) were isolated from healthy donors by HiSepTM LSM 1077 (HiMedia Laboratories, India) and bound IgE’s were stripped off by incubating in lactic acid buffer (pH 3.5) for 3 min.

    Techniques: Enzyme-linked Immunosorbent Assay, Binding Assay, Inhibition, Incubation, Negative Control