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  • Title: Oligoclonal and polyclonal CD4 and CD8 lymphocytes in aplastic anemia and paroxysmal nocturnal hemoglobinuria measured by V beta CDR3 spectratyping and flow cytometry.
    Author: Risitano AM, Kook H, Zeng W, Chen G, Young NS, Maciejewski JP.
    Journal: Blood; 2002 Jul 01; 100(1):178-83. PubMed ID: 12070025.
    Abstract:
    We have hypothesized that in aplastic anemia (AA) the presence of antigen-specific T cells is reflected by their contribution to the expansion of a particular variable beta chain (V beta) subfamily and also by clonal CDR3 skewing. To determine the role of disease-specific "signature" T-cell clones in AA, we studied preferential V beta usage by flow cytometry and analyzed V beta-CDR3 regions for the presence of oligoclonality. We first established the contribution of each V beta family to the total CD4(+) and CD8(+) lymphocyte pool; in AA and paroxysmal nocturnal hemoglobinuria, a seemingly random overrepresentation of different V beta families was observed. On average, we found expansion in 3 (of 22 examined) V beta families per patient. When the contribution of individual V beta families to the effector pool was examined, more striking V beta skewing was found. V beta-CDR3 size distribution was analyzed for the expanded V beta families in isolated CD4(+) and CD8(+) populations; underrepresented V beta families displayed more pronounced CDR3 skewing. Expanded CD4(+)V beta subfamilies showed mostly a polyclonal CDR3 size distribution with only 38% of skewing in expanded V beta families. In contrast, within overrepresented CD8(+)V beta types, marked CDR3 skewing (82%) was seen, consistent with nonrandom expansion of specific CD8(+) T-cell clones. No preferential expansion of particular V beta families was observed, in relation to HLA-type. In patients examined after immunosuppressive therapy, an abnormal V beta-distribution pattern was retained, but the degree of expansion of individual V beta was lower. As V beta skewing may correlate with relative V beta size, oligoclonality in combination with numerical V beta expansion can be applied to recognition of disease-specific T-cell receptors.
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