Construction of hybridomas from nude mice with phenotypes of precursors of regulatory T cells. I. A model for study of development of T-cell function in immune systems.

Functional helper and suppressor hybridomas were constructed from fractionated Type II NFR/nude mouse splenocytes which resemble precursors of T cells. After carrier priming with rabbit erythrocytes (RE), splenocytes from Type II NFR/nude mice were fractionated on nylon wool columns. Cells from two distinct fractions (Pool 1 and Pool 2) were fused with BW5147 lymphoma cells. Hybrids were selected in HAT medium and subsequently cloned. Three hybridomas constructed using Pool 1 splenocytes were capable of complementing nude mouse PFC responses to the trinitrophenyl group (TNP) of TNP-RE. Four hybridomas constructed using Pool 2 splenocytes were observed to suppress anti-TNP PFC responses. These hybridomas express the T-cell surface markers TLa, Thy 1, Lyt 1, Lyt 2 in patterns which suggest that they represent expanded clones of T cells from precursors that are arrested in their development. They appear to be blocked short of development of carrier specificity in their functional phenotypes as regulatory T cells.