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Blood. 2002 Aug 15;100(4):1185-91.

Intracellular interferon-gamma in circulating and marrow T cells detected by flow cytometry and the response to immunosuppressive therapy in patients with aplastic anemia.

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  • 1Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892-1652, USA.


Immunosuppressive therapy leads to meaningful hematologic improvement in most patients with aplastic anemia (AA). Failure to respond and a later relapse could be due to deficient numbers of hematopoietic stem cells, inadequate treatment of the immune process, or a nonimmunologic etiology. Interferon-gamma (IFN-gamma) has been implicated in the pathophysiology of hematopoietic failure in AA. On the basis of previous findings showing overexpression of IFN-gamma in bone marrow (BM) and peripheral blood (PB) in this disease, we hypothesized that quantitation of IFN-gamma might be applied to predict and monitor responses to immunosuppressive therapy. We measured expression of IFN-gamma in lymphocytes obtained from 123 AA patients, using intracellular 2-color fluorescent staining and flow cytometry. Of 70 patients with severe AA, 36 (51%) demonstrated increased IFN-gamma in circulating T cells. IFN-gamma was detected in only 4 of 53 patients who had recovered from AA. IFN-gamma was not found in PB lymphocytes of patients with other hematologic diseases and heavy transfusion burdens or in healthy volunteers. Among 62 AA patients who were assessed before first treatment with immunosuppressive drugs, 27 of 28 (96%) with circulating IFN-gamma-containing T cells subsequently responded to therapy; in contrast, only 11 of 34 (32%) patients whose PB lacked IFN-gamma lymphocytes improved to transfusion independence. IFN-gamma-containing lymphocytes declined following treatment in all cases. Of 17 patients assessed during relapse, IFN-gamma was present in T cells prior to the blood count decline in 13, and 12 responded to reinstitution of immunosuppressive drugs. Of 30 BMs tested prior to first treatment, 20, all in responding patients, were positive for IFN-gamma, whereas the negative tests were obtained in 10 nonresponding patients. IFN-gamma is increased in the PB lymphocytes of many patients with AA, and these cells decline with therapy. The presence of intracellular IFN-gamma may predict response to immunosuppressive treatment and also the onset of relapse.

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