anti cd28  (Bio X Cell)

Journal: bioRxiv

Article Title: Enhancement of activation-induced T cell proliferation by SIRPG in a CD47-independent manner

doi: 10.1101/2025.05.01.651731

Figure Lengend Snippet: A . PBMC from healthy donors were stained with antibodies against indicated surface markers and analyzed with FACS. The levels of SIRPG, CD28, and CCR7 of indicated populations are shown in overlay histograms in A . B-K . PBMC from two healthy donors ( B-F ) and synovial fluid cells from two patients with inflammatory arthritis ( G-K ) were stained for surface SIRPG and CD45RA and intracellular GzmK and GzmB. The stained cells were first separated into four populations based on the expression of GzmK and GzmB ( B & G ). The levels of SIRPG in the four GzmK/GzmB populations are shown in overlay histograms ( C & H ). The stained PBMC and synovial fluid cells were also separated into four populations based on the expression of SIRPG and CD45RA ( D & I ). The percentages of the SIRPG/CD45RA populations are shown in the cumulated bar graphs ( D & I ). The expression of GzmK and GzmB in each SIRPG/CD45RA population is then shown in the dot plots in E & J . The percentage of cells positive or negative for GzmB or GzmK from the SIRPG/CD45RA populations is shown in the cumulative bar graph of F and K .

Article Snippet: PBMCs were plated in 24-well plates (2-2.5 millions/1ml/well) pre-coated with anti-CD3 (2,5ug/ml, OKT3, Supplemental Table 3) and soluble anti-CD28 (2,0ug/ml, CD28.2, Supplemental Table 3) and stimulated with adalimumab (125ug/ml), etanercept (50ug/ml), certolizumab pegol (50ug/ml), tocilizumab (100ug/ml), or an isotype control anti-human IgG1 (125ug/ml, #BE0297, BioXCell, Lebanon, NH, USA) for indicated times before analyzing by flow cytometry.

Techniques: Staining, Expressing

Journal: bioRxiv

Article Title: Enhancement of activation-induced T cell proliferation by SIRPG in a CD47-independent manner

doi: 10.1101/2025.05.01.651731

Figure Lengend Snippet: A . PBMC from healthy donors were stimulated with anti-CD3/anti-CD28 and restimulated with anti-CD3 on day 6. The surface level of SIRPG in CD4+ T cells was analyzed with FACS and shown in the overlay histograms (the left panel). The MFI of SIRPG from three independent donors over the time course is shown in the right panel. Adalimumab (ada), certolizumab (cer), etanercept (eta), tocilizumab (toc), or control IgG1 was added on day 0 during the stimulation. The MFI of SIRPG from non-blasting ( B ) and blasting ( C ) T cells is shown in the overlay histograms. The cumulative results from 3 independent experiments are shown in the corresponding bar graphs. D . PBMC were stimulated in the presence or absence of ada and the surface level of SIRPG was monitored with FACS at indicated time points. Representative data from one donor is shown in the overlay histograms and cumulated results from five donors are shown in the right panel. The data points from the same donors are connected with lines. E . PBMC were stimulated and ada or control IgG was added on day 3. The level of SIRPG was analyzed on day 6. Representative overlay histograms are shown in the left panel and the cumulative results from three donors are shown in the right panel. F . PBMC were treated with ada or control IgG without anti-CD3 stimulation. The surface level of SIRPG in T and non-T cells was analyzed on day 3. Representative overlay histograms are shown in the left panel and cumulative results from three donors are shown in the right panel.

Article Snippet: PBMCs were plated in 24-well plates (2-2.5 millions/1ml/well) pre-coated with anti-CD3 (2,5ug/ml, OKT3, Supplemental Table 3) and soluble anti-CD28 (2,0ug/ml, CD28.2, Supplemental Table 3) and stimulated with adalimumab (125ug/ml), etanercept (50ug/ml), certolizumab pegol (50ug/ml), tocilizumab (100ug/ml), or an isotype control anti-human IgG1 (125ug/ml, #BE0297, BioXCell, Lebanon, NH, USA) for indicated times before analyzing by flow cytometry.

Techniques: Control