Objective　To establish the method of alloantigen-specific T lymphocyte clones. Methods　PBMC of the recipient was stimulated by donor splenocytes for twice and the stimulated T lymphocytes were cloned by limited dilution. Results　Three clones of T lymphocytes were obtained. By analysis of immunohistochemistry, two of the three clones were CD3+CD4-CD8+, another one was CD3+CD4-CD8-. The latter was analyzed by flowcytometry and CD3+ was 98.87%, CD4- 97.05%, CD8- 97.14%. Antigen specificity of the cloned T lymphocytes was determined by stimulation of donor splenocytes and non-related individual splenocytes. When stimulated by donor splenocytes, the cloned T lymphocytes proliferated and did not show proliferative role when stimulated by non-related individual splenocytes. Conclusion　Our method of cloning alloantigen-specific T lymphocytes is successful.

Objective To investigate the changes of silver-stainng protein of nuclear organizer regions(AgNORs)expression of T lymphocytes and helper T (Th) cell subsets in peripheral mononuclear cells (PBMC) from cancer patients and to analyze the relationship between them.Methods AgNORs expression in T cells activated by a multiple clonal activator was examined using a special image system. After activation of PBMC with ionomycin plus PMA for 5 h, both interferon-? (IFN ?) and interleukin-4 (IL-4)producing cell frequencies were detected by enzyme-linked immunospot assay (ELISPOT). The stimulated with ionomycin plus PMA for 24 h, the IFN ?-and IL-4-production were assayed by ELISA.Resuts AgNORs expression of activated T cells in PBMC from cancer patients (n=86) and healthy persons (n=44) was (4.90?1.20) I.S% and (7.82?1.28) I.S% respectively. Th2 frequency was significantly higher and the ratio of Th1 to Th2 frequency was more lower in cancer patient group. Moreover, the IFN ? and IL-4 production in cancer patient group were changed in the way similar to Th subset frequencies. In lung cancer patients the AgNORs expression, Th1 frequency, Th1/Th2 ratio and IFN ? production were significantly decreased in metastasis or in recurrence status,AgNORs expression in cancer patients correlated with their Th1 frequencies, Th1/Th2 ratio, IFN ? production levels and Karnofsky′s estimators.Conclusions In analyzed cancer patient cases, the expression of AgNORs and the function of Th1 cell were significantly decreased and correlated.