Clone jes5 2a5

Functional grade-blocking anti-VCAM-1 (clone 429, eBioscience, 16-1061), ICAM-1 (clone YN1, eBioscience, 160541), VLA4/α4 integrin (clone R1-2, eBioscience, 16-0492), high+low-affinity LFA-1/CD11a (clone M17/4, eBioscience, 16-0111) and low-affinity CD11a (clone 2D7, BD, 553118), Rat IgG2a (clone eBR2a, eBioscience, 16-4321) and Rat IgG2b (clone eB149/10H5, eBioscience, 16-4031) were used to treat endothelial layers or T cells at 4 °C for 30 min at 10 μg ml⁻¹. In some experiments, low-endotoxin, no-azide (LEAF) grade anti-IL-10 (clone JES5-2A5, Biolegend, 504903), LEAF grade anti-TGF-β1 (clone 19D8, Biolegend, 521703) or LEAF Grade Rat IgG1k (RTK2071, Biolegend, 400413) were incubated with T cells at 10 μg ml⁻¹ for 30 min at 4 °C before transmigration. T cells were washed 2 × before use except in the case of anti-TGF-β1, anti-IL-10 and control Rat IgG1k, while VCAM-1 and ICAM-1 blockades of endothelial cells were also maintained during assays.

Following a protocol we applied previously [6 (link)], mice received intraperitoneal injections of either a neutralizing anti-il-10 antibody (clone JES5-2A5; Biolegend #504909) or an isotype matched irrelevant IgG₁ (Biolegend #400432) at day-3 (50 μg), day-7 (100 μg), and day-9 (50 μg) post-infection with H. diminuta for a total of 200 μg of antibody. DNBS was administered at 8 days post-infection and mice were necropsied 3 days later.