TNF^−/− T cells do not have intrinsic defects in their proliferative capability and ability to be alloactivated.

(A) Splenocytes were incubated in quadruplicate wells for 72 hours in the presence of ConA. Proliferation over the last 20 hours was measured by H-3-thymidine incorporation and is expressed as -fold increase over controls without ConA. The mean ± SE for 3 animals per group is shown. (B) Splenocytes from preimmunized B6 wt or TNF^−/−animals were incubated for 96 hours in quadruplicate wells in the presence of allogeneic (C3HxB6) irradiated splenocytes as stimulators. Proliferation over the last 20 hours was measured by H-3-thymidine incorporation and is expressed as fold increase over incubation with irradiated syngeneic (B6) controls. The mean ± SE for one representative experiment (of 2 total) with 3 animals per group is shown. (C) C3HxB6 recipients of allogeneic (B6) TCD wt BM (5 × 10⁶ cells) and wt or TNF^−/− splenic T cells (2 × 10⁶) were killed on days 7 and 14 after BMT. Total splenocytes were counted and donor T cells were determined by flow cytometry for Ly9.1 and CD3. The mean ± SE of one representative experiment (of 3 total) with 4 animals per group is shown. (D) C3HxB6 recipients were given transplants of TCD wt B6 BM and a high dose (25 × 10⁶) of CFSE-stained wt or TNF^−/− T cells. Spleens were harvested 72 hours later and multicolor flow cytometric analysis was performed. CD25 and CD 122 expression in CD8⁺ cells is shown. Percentages of CD25 and CD122-expressing cells are indicated. One experiment representative of 2 separate experiments is shown.