ABSTRACT
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.
ABSTRACT
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.