Journal: BMC Immunology

Article Title: Activation of Toll-like receptor 2 enhances peripheral and tumor-infiltrating CD8 + T cell cytotoxicity in patients with gastric cancer

doi: 10.1186/s12865-021-00459-z

Figure Lengend Snippet: The influence of TLR2 activation on inhibitory molecules expression in peripheral and tumor-infiltrating CD8 + T cells from NC and GC patients. CD8 + T cells were purified from the same subjects as in Fig. , including peripheral bloods of NC ( n = 9) and GC patients ( n = 11), as well as from tumor residency of GC patients ( n = 11), and were cultured for 12 h in the presence of anti-CD3/CD28 with or without TLR2 agonist Pam3Csk4. Lymphocyte activation gene-3 (LAG-3) and CD279 (programmed death-1, PD-1) expression in CD8 + T cells was measured by flow cytometry. a The representative flow dots of LAG-3 and PD-1 positive cells in peripheral CD8 + T cells from NC and GC patients, as well as in tumor-infiltrating CD8 + T cells from GC patents with or without Pam3Csk4 stimulation. The percentage of b LAG-3 + and c PD-1 + cells within CD8 + T cells was compared. The columns indicated means, and the bars indicated standard deviation. Significances were determined by LSD- t test or paired t test

Article Snippet: Cells were stained with anti-CD3-PerCP Cy5.5 (eBioscience), anti-CD8-APC (BD Bioscience, San Jose, CA), anti-TLR2-PE (eBioscience), anti-TLR7-FITC (eBioscience), anti-lymphocyte activation gene-3 (LAG-3)-PerCP (BD Bioscience), and anti-CD279 (programmed death-1, PD-1)-FITC (BD Bioscience) following manufacturer’s instructions.

Techniques: Activation Assay, Expressing, Purification, Cell Culture, Flow Cytometry, Standard Deviation

Journal: Frontiers in Immunology

Article Title: Lack of Association Between Sex Hormones, MDSCs, LDGs and pDCs in Males and Females With Systemic Lupus Erythematosus

doi: 10.3389/fimmu.2022.888501

Figure Lengend Snippet: Plasmacytoid dendritic cells are under expressed, while LDGs and G-MDSCs are overexpressed in SLE. (A) Representative gating strategy for pDC identification. PBMCs were first gated as Lin - , and subsequently gated as CD123+, HLA-DR+. (B) pDC expression was decreased among males and females with active SLE. (C) Representative gating strategy for LDGs and G-MDSCs. LDGs were identified as CD15+CD14-SSC low , while G-MDSCs were identified as CD15+CD14-SSC high . (D) G-MDSCs and (E) LDGs were significantly increased among males and females with SLE. (F) Representative gating strategy for M-MDSCs. M-MDSCs were first identified as HLA-DR-, CD33+, and then gated for CD11b+, before gating as CD15-, CD14+. (G) M-MDSCs frequency was consistent among SLE patients and healthy controls. Comparisons were made using Kruskal-Wallis Test with Dunn’s test for multiple comparisons. Bars represent median and interquartile range. aSLE F (N = 15), iSLE F (N = 50), HC F (N = 14), aSLE M (N = 5), iSLE M (N = 6), HC M (N = 5). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: The following antibodies were used for staining: PE CF594-conjugated anti-CD15; APCH7-conjugated anti-CD14; PE-conjugated anti-CD33; APC-conjugated anti- HLA-DR, FITC-conjugated anti-Lineage; PE/Cy7-conjugated anti-CD123; PE-conjugated anti-TLR7 (all from BD Biosciences, San Jose, CA); A700-conjugated anti-CD11b (Life technologies, Carlsbad, CA); FITC-conjugated anti-TLR7 and FITC-conjugated anti-Rabbit IgG isotype control (both from Invitrogen, Waltham, MA); biotinylated-anti TLR9 (Abcam, Cambridge, UK); unconjugated anti-human FC block, PerCP/Cy5.5-conjugated Streptavidin; biotinylated anti-Mouse IgG2a and PE-conjugated anti-Mouse IgG1κ (all from eBioscience inc, Santa Clara, CA).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: Lack of Association Between Sex Hormones, MDSCs, LDGs and pDCs in Males and Females With Systemic Lupus Erythematosus

doi: 10.3389/fimmu.2022.888501

Figure Lengend Snippet: Plasmacytoid dendritic cells are under expressed, while LDGs and G-MDSCs are overexpressed in SLE. (A) Representative gating strategy for pDC identification. PBMCs were first gated as Lin - , and subsequently gated as CD123+, HLA-DR+. (B) pDC expression was decreased among males and females with active SLE. (C) Representative gating strategy for LDGs and G-MDSCs. LDGs were identified as CD15+CD14-SSC low , while G-MDSCs were identified as CD15+CD14-SSC high . (D) G-MDSCs and (E) LDGs were significantly increased among males and females with SLE. (F) Representative gating strategy for M-MDSCs. M-MDSCs were first identified as HLA-DR-, CD33+, and then gated for CD11b+, before gating as CD15-, CD14+. (G) M-MDSCs frequency was consistent among SLE patients and healthy controls. Comparisons were made using Kruskal-Wallis Test with Dunn’s test for multiple comparisons. Bars represent median and interquartile range. aSLE F (N = 15), iSLE F (N = 50), HC F (N = 14), aSLE M (N = 5), iSLE M (N = 6), HC M (N = 5). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001.

Article Snippet: The following antibodies were used for staining: PE CF594-conjugated anti-CD15; APCH7-conjugated anti-CD14; PE-conjugated anti-CD33; APC-conjugated anti- HLA-DR, FITC-conjugated anti-Lineage; PE/Cy7-conjugated anti-CD123; PE-conjugated anti-TLR7 (all from BD Biosciences, San Jose, CA); A700-conjugated anti-CD11b (Life technologies, Carlsbad, CA); FITC-conjugated anti-TLR7 and FITC-conjugated anti-Rabbit IgG isotype control (both from Invitrogen, Waltham, MA); biotinylated-anti TLR9 (Abcam, Cambridge, UK); unconjugated anti-human FC block, PerCP/Cy5.5-conjugated Streptavidin; biotinylated anti-Mouse IgG2a and PE-conjugated anti-Mouse IgG1κ (all from eBioscience inc, Santa Clara, CA).

Techniques: Expressing