- 25-7108-42 - Invitrogen Antibodies IL10 antibody

Submitted references Early loss of T lymphocyte 4-1BB receptor expression is associated with higher short-term mortality in alcoholic hepatitis.

Eriksen LL, Nielsen MA, Laursen TL, Deleuran B, Vilstrup H, Støy S
PloS one 2021;16(8):e0255574

Altered T-Cell Subsets are Associated with Dysregulated Cytokine Secretion of CD4(+) T Cells During HIV Infection.

Wang D, Jiang Y, Song Y, Zeng Y, Li C, Wang X, Liu Y, Xiao J, Kong Y, Zhao H
Journal of inflammation research 2021;14:5149-5163

Cysticercus cellulosae Regulates T-Cell Responses and Interacts With the Host Immune System by Excreting and Secreting Antigens.

Fan X, Zhang Y, Ouyang R, Luo B, Li L, He W, Liu M, Jiang N, Yang F, Wang L, Zhou B
Frontiers in cellular and infection microbiology 2021;11:728222

CAIX Regulates GBM Motility and TAM Adhesion and Polarization through EGFR/STAT3 under Hypoxic Conditions.

Huang BR, Liu YS, Lai SW, Lin HJ, Shen CK, Yang LY, Lu DY
International journal of molecular sciences 2020 Aug 14;21(16)

Multiple Sclerosis-Associated Changes in the Composition and Immune Functions of Spore-Forming Bacteria.

Cekanaviciute E, Pröbstel AK, Thomann A, Runia TF, Casaccia P, Katz Sand I, Crabtree E, Singh S, Morrissey J, Barba P, Gomez R, Knight R, Mazmanian S, Graves J, Cree BAC, Zamvil SS, Baranzini SE
mSystems 2018 Nov-Dec;3(6)

Chloroquine modulates antitumor immune response by resetting tumor-associated macrophages toward M1 phenotype.

Chen D, Xie J, Fiskesund R, Dong W, Liang X, Lv J, Jin X, Liu J, Mo S, Zhang T, Cheng F, Zhou Y, Zhang H, Tang K, Ma J, Liu Y, Huang B
Nature communications 2018 Feb 28;9(1):873

Pre-existing neutralizing antibody mitigates B cell dysregulation and enhances the Env-specific antibody response in SHIV-infected rhesus macaques.

Jaworski JP, Bryk P, Brower Z, Zheng B, Hessell AJ, Rosenberg AF, Wu TT, Sanz I, Keefer MC, Haigwood NL, Kobie JJ
PloS one 2017;12(2):e0172524

Heterologous Immunity between Adenoviruses and Hepatitis C Virus: A New Paradigm in HCV Immunity and Vaccines.

Singh S, Vedi S, Samrat SK, Li W, Kumar R, Agrawal B
PloS one 2016;11(1):e0146404

Follicular regulatory T cells impair follicular T helper cells in HIV and SIV infection.

Miles B, Miller SM, Folkvord JM, Kimball A, Chamanian M, Meditz AL, Arends T, McCarter MD, Levy DN, Rakasz EG, Skinner PJ, Connick E
Nature communications 2015 Oct 20;6:8608

Daclizumab reverses intrathecal immune cell abnormalities in multiple sclerosis.

Lin YC, Winokur P, Blake A, Wu T, Romm E, Bielekova B
Annals of clinical and translational neurology 2015 May;2(5):445-55

Granzyme B production distinguishes recently activated CD8(+) memory cells from resting memory cells.

Nowacki TM, Kuerten S, Zhang W, Shive CL, Kreher CR, Boehm BO, Lehmann PV, Tary-Lehmann M
Cellular immunology 2007 May;247(1):36-48

Mycobacterium bovis BCG attenuates surface expression of mature class II molecules through IL-10-dependent inhibition of cathepsin S.

Sendide K, Deghmane AE, Pechkovsky D, Av-Gay Y, Talal A, Hmama Z
Journal of immunology (Baltimore, Md. : 1950) 2005 Oct 15;175(8):5324-32

Mycobacterium bovis BCG attenuates surface expression of mature class II molecules through IL-10-dependent inhibition of cathepsin S.

Sendide K, Deghmane AE, Pechkovsky D, Av-Gay Y, Talal A, Hmama Z
Journal of immunology (Baltimore, Md. : 1950) 2005 Oct 15;175(8):5324-32

Role of the phosphatidylinositol 3 kinase-Akt pathway in the regulation of IL-10 and IL-12 by Porphyromonas gingivalis lipopolysaccharide.

Martin M, Schifferle RE, Cuesta N, Vogel SN, Katz J, Michalek SM
Journal of immunology (Baltimore, Md. : 1950) 2003 Jul 15;171(2):717-25

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Normal human peripheral blood mononuclear cells were cultured for 6 days under Th2-polarizing conditions. The cells were unstimulated (left) or restimulated (right) with the Cell Stimulation Cocktail (plus protein transport inhibitors) (Product # 00-4975-03) then intracellularly stained with Anti-Human CD4 PE (Product # 12-0049-42) and Anti-Human IL-10 PE-Cyanine7 (Product # 25-7108-42) using the Intracellular Fixation & Permeabilization Buffer Set (Product # 88-8824-00) and protocol. Total viable cells, as determined by Fixable Viability Dye eFluor® 450 (Product # 65-0863-14), were used for analysis.

Supportive validation

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Experimental details: Fig 4 Cross-reactive CD4 + and CD8 + T cells obtained from Ad vector immunized mice produce cytokines upon ex vivo stimulation with various HCV proteins. Splenocytes obtained from Ad vector immunized mice were cultured with HCV core, NS3, NS4 or NS5 antigens at 5 mug/ml, and analyzed after 5 days for intracellular IFN-gamma and IL-10 expression profile of CD4 + and CD8 + T cells by flow cytometry. Data are obtained from a pool (n = 5) of spleen cells and are representative of two independent experiments.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Fig 5 Cross-reactive CD4 + and CD8 + T cells obtained from Ad vector immunized mice produce cytokines upon ex vivo stimulation with HCV peptides. Splenocytes obtained from Ad vector immunized mice were cultured with representative peptides derived from HCV core, NS3, NS4 or NS5 at 5 mug/ml each, and analyzed after 5 days for intracellular IFN-gamma and IL-10 expression profile of CD4 + and CD8 + T cells by flow cytometry. Data are obtained from a pool (n = 5) of spleen cells and are representative of two independent experiments.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 T FR exhibit an enhanced regulatory phenotype in ex vivo HIV infection. Tonsil cells were mock-, X4-, or R5-spinoculated and cultured under experimental conditions as indicated. T FR were then analysed for expression of regulatory receptors and cytokine production by intracellular cytokine staining. ( a ) Percentage of total (surface and intracellular) T FR CTLA-4 expression ( n =15). ( b ) Percentage of surface T FR LAG-3 expression ( n =8). ( c ) Production of IL-10 by T FR ( n =7). ( d ) Production of TGF-beta-1 (measured as LAP) by T FR ( n =5). The horizontal bars of each graph indicate the median value and are listed where appropriate for clarity. Statistical analyses were performed by Friedman ( a , b ) or Mann-Whitney tests ( c , d ) and significance is denoted by asterisks where * P

Supportive validation

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Experimental details: FIG 2 Spore-forming bacteria from MS patients inhibit IL-10 + Treg differentiation in vitro . (A and B) Representative flow cytometry plots (A) and quantification (B) of CD4 + FoxP3 + Tregs within CD3 + lymphocytes differentiated in response to spore-forming bacteria isolated from controls or untreated MS patients. n = 7 PBMC donors; each dot represents an average response from PBMC donor to isolates from 6 control or MS bacterial donors. * * , P < 0.01, two-tailed repeated measures t test. (C and D) Representative flow cytometry plots (C) and quantification (D) of IL-10 + lymphocyte population within CD3 + CD4 + FoxP3 + Tregs differentiated in response to spore-forming bacteria isolated from controls or untreated MS patients. n = 6 bacterial donors per group. * , P < 0.05, two-tailed t test. Error bars, mean +- SEM. The experiment was repeated with nonoverlapping PBMC and bacterial donors and gave the same results. (E) Quantification of T effector cell proliferation in response to Tregs differentiated in the presence of spore-forming bacteria from MS patients or controls. n = 3 bacterial donors per group, each representing an average of 3 technical replicates. (F) Linear correlation between IL-10 + population within CD3 + CD4 + FoxP3 + Tregs and Clostridia-Bacilli relative abundances. R 2 = 0.214, P = 0.0459. Black dots, MS patients. Light gray dots, controls.

Supportive validation

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Experimental details: Figure 4 The polarizations of monocyte and GBM progression in a co-cultured model under hypoxic conditions. ( A ) THP-1 monocytes were co-cultured with U87-green fluorescent protein (GFP) or U251-GFP under hypoxic conditions for 48 h. The co-cultured cells were plotted on a side scatter versus FITC. The THP-1 monocytes (GFP-negative cells) were analyzed to assess the levels of cluster of differentiation (CD) 206, arginase 1 (Arg1), interleukin (IL)-10, interferon (IFN)-gamma, and tumor necrosis factor (TNF)-alpha by flow cytometry. ( B ) The median fluorescence intensity (MFI) of CD206, Arg1, IL-10, IFN-gamma, and TNF-alpha in THP-1 monocytes in the co-cultured model under hypoxic conditions for 48 h. * p < 0.05 compared with the THP-1 monocyte group. Quantitative data are presented as the mean +- standard error (representative of n = 3). ( C ) U87-GFP and U251-GFP were treated with U104 (CAIX inhibitor) and subsequently co-cultured with THP-1 monocytes under hypoxic conditions for 48 h. THP-1 monocytes (GFP-negative cells) were analyzed to assess the levels of CD206, Arg1, and IL-10 by flow cytometry. ( D ) U87-GFP or U251-GFP was treated with U104 and subsequently co-cultured with THP-1 monocytes under hypoxic conditions for 48 h. The co-cultured cells were plotted on a side scatter versus FITC. The GFP-positive gated GBM was analyzed to assess the levels of programmed death ligand 1 (PD-L1) by flow cytometry. ( E ) The levels of CAIX, PD-L1, tumor growth factor-beta, and c

Supportive validation

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Experimental details: 10.1371/journal.pone.0255574.g002 Fig 2 Cytokine production in 4-1BB + and 4-1BB - CD4 + T lymphocytes. Flow cytometry of peripheral blood mononuclear cells stimulated for 48 hours with anti-CD3 and anti-CD28 given PMA and brefaldin-A for the last 4 hours. (A) Typical flow cytometry plot of 4-1BB + CD4 + T lymphocytes from a patient with alcoholic hepatitis showing interferon-gamma (IFNy) on x-axis and interleukin-10 (IL-10) on y-axis. Frequencies of interferon-gamma + (B, n = 6) and IL-10 + (C, n = 3)) 4-1BB + and 4-1BB - CD4 + T lymphocytes in patients with alcoholic hepatitis. Differences between groups compared using paried T test. Graphs shown as median with interquartile range.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 5 IL-10 + lymphocyte assay. The production of IL-10 + lymphocyte subsets by flow cytometry. (A) Both Cysticercus cellulosae ESA and CA induced the production of IL-10 + lymphocyte. (B) CD4 + T lymphocyte was the main source Cysticercus cellulosae ESA and CA-induced IL-10 expression. (C) In the presence or absence of LPS, Cysticercus cellulosae ESA and CA significantly inhibited the expression frequency of CD8 + IL-10 + lymphocyte. (D) Cysticercus cellulosae ESA could be induced the expression of CD4 - CD8 - IL-10 + cell under the presence or absence of LPS. While, CA only decreased the expression in the presence of LPS. (E) Similar to (C) , the production of IL-10 in CD4 + CD8 + lymphocyte was inhibited by Cysticercus cellulosae ESA. All dates were shown as the means +- SD, * P < 0.05, ** P < 0.01, *** P < 0.001.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 The production of Foxp3 + IL-10 + lymphocyte. The co-expressing Foxp3 and IL-10 cell were detected via tricolor flow cytometry, which shown that the production of Foxp3 + IL-10 + lymphocyte was inhibited by Cysticercus cellulosae ESA. Besides, CA had significantly decreased the production only under the presence of LPS. Results were expressed as the mean+- SD, * P < 0.05.

25-7108-42

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