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Murine delayed hypersensitivity (DTH) was induced by the cooperation of two subsets of T cells. One of these subsets (Tmd) produces the antigen-specific DTH augmentation factor (DAF). The other (Tm) receives DAF which is needed for the induction of DTH by antigen stimulation. These two types of T cells require the presence of the thymus only for a short term in their differentiation. In this paper, bone marrow cells were transferred to irradiated syngeneic mice and the rapid recovery of the functions of Tprod and Tm were investigated suggesting a low degree of thymus-dependency in their development. When the reconstituted mice were immunized 2 weeks after bone marrow transplantation, the degree of delayed footpad reaction elicited 1 week later recovered to the same level as that in immunized normal mice. The functions of both Td and Tm also recovered to the normal levels within such a period. The recovery of Tm, appeared to occur about 1 week earlier than that of Tpmd. Furthermore, DAF was fractionated from the sera ofmice immunized with chicken erythrocytes (CRBC) by DEAE chromatography and tested for binding to solubilized CRBC antigens. The DAF molecule had a molecular weight of 150-170 K as shown by blotting analysis. Delayed hypersensitivity (DTH) is positively regulated by helper T cells when T cells recognize protein antigens in association with class II MHC determinants on antigen-presenting cells. These helper T cells express Lytl and L3T4 antigens but not Lyt2 antigen. Recently ‘Present address: Department of Bacteriology, School of Medicine, Niigata University, 757 Ichibancho, Asahimachidouri, Niigata 951, Japan Abbreviations.‘ DAF, delayed hypersensitivity augmentation factor; DTH, delayed hypersensitivity; Tpmd, DAF producer T cell; Tm, DAF acceptor T cell; CRBC, chicken erythrocytes; HPLC, high performance liquid chromatography several papers have shown that murine helper T cells can be devided into two subsets which are distinguished by their patterns of lymphokine secretion (17, 2, 3) or some functional criteria (28, 27, 7, 16, 11). In previous studies, we have demonstrated that helper T cells are ontogenically heterologous and there are differences in thymus-dependency in their development or differentiation (25, 24, 8-10, 20). In fact, DTH to Xenogeneic erythrocytes and protection against an infection of Listeria monocytogenes were induced at normal levels in mice thymectomized neonatally within 1 day after birth, while antigen-specific helper, suppressor and killer activities or tuberculin-

EARLY DEVELOPMENT OF TWO HELPER T CELL SUBSETS FOR DELAYED HYPERSENSITIVITY AFTER BONE MARROW TRANSPLANTATION