Anti mouse cd 3 antibody

For survival experiments, 5–6-month-old female BALB/c mice were used because we observed that young mice treated with anti-CD3 antibodies underwent severe weight loss but did not consistently die, even at very high doses of the anti-CD3 antibody. MTR was freshly dissolved in PBS and injected intraperitoneally at 60 mg kg⁻¹ per day for three days before injection of anti-CD3 antibodies. Various doses of anti-CD3 antibody were tested, and it was found that 125 μg per mouse resulted in the death of about half the mice; this was the dose chosen for further experiments. Thirty minutes before the intraperitoneal injection of the anti-mouse CD3 antibody (BioXcell, 145–2C11), MTR was intraperitoneally injected at 60 mg kg⁻¹ into the contralateral side. A single additional dose of 30 mg kg⁻¹ MTR was injected intraperitoneally on the following day. Control groups were injected with PBS at the same times. For experiments with conditional TH-knockout mice, 4–6-week-old LysM^creTh^fl/fl (Th^ΔLysM) mice with C57Bl/6 background were used and LysM^creTh^+/+ mice of the same age were used as control. In these experiments, 200 μg per mouse anti-mouse CD3 antibody was injected intraperitoneally.