Generation and characterization of a monoclonal antibody that inhibits stromal cell-dependent B lymphopoiesis

We generated a mAb, R4, against a stromal cell line, ST2, which is one of several stromal cell lines that can support in vitro B lymphopoiesis. Flow cytometry studies using R4 revealed that R4 reacted with several stromal, fibroblast, and some B cell lines, but not with freshly isolated bone marrow cells, splenocytes, thymocytes, or T cell lines. R4 could immunoprecipitate a M(r) = 24-kDa protein (p24) and a M(r) = 55-kDa protein (p55) from a stromal cell line and a B cell line, respectively. R4 inhibited the development of B cell progenitors from bone marrow cells in cultures supported by stromal cell lines, ST2, or op/op fibroblast line, but did not inhibit myelopoiesis supported by ST2. R4 also inhibited the growth of a stromal cell-dependent pre-B cell line, DW8, but not of two stromal cell-independent B cell lines, 70Z/3 and M12, despite the fact that R4 reacted all three of these lines. Because R4 inhibited B lymphopoiesis in the transwell culture where stromal cells are not allowed to contact pre-B cells, R4 Ag may not directly affect cellular contact. In vivo treatment of pregnant mice with R4 caused a reduction in the number of B cell progenitors in the bone marrows of some of the newborn offspring.