Interferons (IFN) have had increasing clinical usage in the treatment of a variety of disorders, at times being used in combination with chemo-or radiotherapy. However, interferons may have inhibitory effects on hematopoietic stem cell proliferation and the effects of this cytokine's use on long-term hematologic function have not been studied. We performed the competitive repopulation assay in the murine system, using cells exposed to irradiation or single-dose chemotherapy with or without concomitant IFN-␥ use. IFN-␥ alone had no deleterious effects on hematopoietic stem cell productivity. We measured the repopulating ability of exhaustible multilineage precursors that were present at early stages of marrow repopulation after competitive repopulation (30 days). These progenitors were minimally impacted by cyclophosphamide (CTX) with or without IFN-␥. Irradiation (XRT) and CTX alone produced significant repopulating defects in the most primitive hematopoietic stem cell, PHSC. Addition of IFN to either treatment regimen resulted in protection of PHSC, with improved repopulating ability, although the levels of donor marrow reached control levels only when CTX and IFN were used together. The results of multiple use of IFN with chemotherapy must be studied further, but IFN may offer hematologic radio-and chemoprotection, in addition to its antitumor properties in clinical protocols for treatment of cancers. Am.