The early (18-hr) human mixed lymphocyte reaction: identification and isolation of activated T-cell clones.

This study provides direct evidence that de novo expression of the activation antigens B1 49.9 (49.9) (interleukin-2 receptor) and 4F2 enables identification of alloactivated cells within 18 hr of initiation of human mixed lymphocyte reactions (MLR). Using a dual-parameter flow cytometer (simultaneous assessment of immunofluorescence and DNA content on the same cell), it was demonstrated that these activation antigens emerge before activated cells enter into S/G2/M phase of the cell cycle. Family studies illustrate that early activation antigen appearance occurs in response to a mismatch at chromosome 6, and invariably heralds the proliferative outcome at 6 days of MLR. In order to directly study the small alloactivated T-cell population, 49.9-positive cells were isolated using a cell sorter after 18 hr of MLR and cloned by limiting dilution using purified recombinant interleukin-2 (rIL-2). Antigen-specific T4-positive T-cell clones were isolated. Analysis of these clones demonstrates that antigen-specific reactivity is acquired within 18 hr in the MLR. These methods should permit a dissection of the early events of alloactivation.