FIG. 6.
T-cell responses in vitro and in vivo. (A) Normal survival of peripheral CD4+ T cells upon stimulation with CD3 MAb (aCD3), ConA, etoposide, anti-Fas plus cycloheximide (aFas/CHX), or dexamethasone (Dexa). One representative out of two experiments is shown. (B) Activation-induced cell death (AICD). CD4+ T cells were stimulated in CD3 MAb-coated plates, grown in IL-2, and restimulated overnight with CD3 antibody. Apoptosis was measured by annexin V and propidium iodide staining. The data are expressed as induction of apoptosis relative to control samples that were not restimulated. (C) Normal T-cell proliferation in TRIM-deficient mice. Purified CD4+ splenic T cells were stimulated in CD3 MAb-coated 96-well plates or with ConA, SEB, or PMA plus ionomycin (Iono) for 72 h, pulsed with [H3]thymidine, and processed for standard scintillation counting. One out of five independent experiments is shown. Three mice of each genotype were included in each experiment. (D) Migration assay. Freshly isolated lymph node cells were added to the insert of a transwell plate, with the lower chamber containing medium alone or medium supplemented with SDF1α. After 3 h, cells that had migrated into the lower wells were collected and counted by flow cytometry. The data shown are representative for three independently performed experiments. The migration is expressed as induction relative to that of control samples incubated with medium alone. (E) Serum immunoglobulin titers. The concentration of serum immunoglobulins was measured by ELISA. Bars represent mean values. (F and G) Humoral immune response. Mice were immunized with the T-dependent antigen DNP-KLH, and DNP-specific immunoglobulins were analyzed at day 10 (F), or they were reimmunized at day 14 with the same antigen and DNP-specific immunoglobulins were measured at day 28 (G) as described in Materials and Methods.