Anti cd3 mab clone 17a2

For the depletion of CD4+ T cells and CD8+ T cells in vivo, 200 µg of anti-CD8 mAb (clone 2.43; BioXcell) and anti-CD4 mAb (clone GK1.5; BioXcell) were injected intraperitoneally into mice four times at days 1, 3, 5, and 7 before challenge. Non-depleted control mice were given isotype control rat IgG2b antibodies (clone LTF-2, BioXcell). Blood and lungs were taken from the mice 24 h after the last antibody injection and were subjected to flow cytometry to confirm the depletion. For flow cytometric analysis, we used anti-CD8 mAb (clone 53-6.7; Biolegend) and anti-CD4 mAb (clone RM4-5; Biolegend) directed against different epitope of CD4 and CD8 molecules to that of the depleting antibodies. For the depletion of NK cells, 20 µl of anti-asialo GM1 antiserum (Wako Pure Chemical Industries) were injected intraperitoneally into mice four times at days 1, 3, 5, and 7 before challenge. Non-depleted control mice were given normal rabbit serum (Wako Pure Chemical Industries). The spleens were taken from the mice 24 h after the last antibody injection to confirm the depletion by flow cytometry. For flow cytometric analysis, we used anti-CD3 mAb (clone 17A2; Biolegend) and anti-CD49b mAb (clone DX5; Biolegend).