Study Of T Lymphocyte Subsets In PBMC Of Different State Patients With Hepatitis B

Objective We use a new technique, the Biotin-Streptavidin(BSA). to detect the levels of T lymphocyte subsets (CD₃⁺,CD₄^-,CD₈^-, CD₂₅⁺) in PBMC of 188 cases with hepatitis B, so as to, we can study more about its role in the pathogenesis of hepatitis B and the useful of BSA in detection of T lymphocyte subsets. Methods Using the ficoll separate the PBMC from fresh and heparinized blood by centrifugal.then suspend it in RPMI-1640 liquid culture medium with 10% calf serum, and adjust its consistency about 2×10⁶/ml.Then, taking 10μ1. determine the percent of T lymphocyte subsets (CD₃⁺,CD₄⁺,CD₈^-,CD₂₅⁺) by BSA.In additional, take 0.5ml and put it into culture plate, and use PHA as stimulative factor, after 48 hours culturing, determine the percent of CD₂₅⁺ by BSA. Moreover, we test the infection rates of HBV DNA in PBMC.Results In the patients with hepatitis B, the levels of CD₃⁺. CD₄⁺, and CD₄⁺/CD₈^- were lower, but CD₈⁺was higher than those in normal controls (P<0.01); and the total expression levels of CD25 in PBMC before and after being stimulated with PHA were also lower than those in normal controls (P<0.01); of the patients with hepatitis B, the levels of T lymphocyte subsets and CD25 in PBMC with HBV-DNA (+) were lower than those with HBV-DNA (—), which were significantly different (P0.01); in addition, the levels of mIL-2R in PBMC were lower in silence than those in inducting with PHA (P<0.01).Conclusion the cellular immune functions are obviously disorder in patients with hepatitis B. the levels of T lymphocyte subsets decreased, and its propotion was unbalance, which might closely associate with the chronicity of the patients with hepatitis B; the BSA was a useful technique in detection of T lymphocyte subsets.