[Correlation between CTL and Th1 cell proportion in peripheral blood of liver transplantation recipients and the success of hepatitis B vaccination].

Objective: To investigate the correlation between the proportion of CTL and Th1 cells in peripheral blood of liver transplant recipients and the success of hepatitis B vaccination. Methods: The subjects of this study were liver transplantation recipients with chronic HBV-related liver diseases in Organ transplantation institute of the third medical center of PLA general hospital. Subjects were randomly divided into two groups for prospective study. In the rapid group, one dose of 40 µg hepatitis B vaccine was administered at 0, 1, 2and 3 months, and one dose of 20 µg hepatitis B vaccine was administered at 4, 5 and 6 months. In the rapid-enhanced group, one dose of 40 µg hepatitis B vaccine was administered at 0, 1, 2 and 3 months, and one dose of 60 µg hepatitis B vaccine was administered at 4, 5 and 6months. Compare and analyze the success rate of inoculation, the titer of hepatitis B surface antibody (anti-HBs), the proportion of CTL cells in CD8(+)T cells and Th1 cells in CD4(+)T cells. Correlation analysis was performed for CTL and Th1 cells and anti-HBs, Observe the safety of vaccination. Results: The inoculation success rate, anti-HBs growth rate, CTL cell percentage increase and Th1 cell percentage increase in the rapid enhancement group were all higher than those in the rapid enhancement group, and the differences were statistically significant, they were 38.3% (23/60) vs 21.7% (13/60) (P=0.046), 91.3(72.5,124.2) vs 22.1(12.4, 31.6) (P=0.001), 1.4(0.8,1.9) vs 0.4(0.2,1.4) (P=0.001) and 7.4±2.6 vs 5.6±3.7 (P=0.001) respectively. The percentage increase of CTL cells and Th1 cells in the successful group was greater than that in the non-successful group, and the difference was statistically significant. They were 1.9(1.4,2.5) vs 0.1(0.0,1.1) (P=0.024) and 9.6±3.1 vs 2.4±2.0 (P<0.001). There was no significant correlation between anti-HBs increase (105.5±37.1) and CTL increase 1(0,3) (P=0.099), while there was significant positive correlation with Th1 increase 7(2,11) (P<0.001). No rejection reaction occurred during the study period, and there was no special abnormal change in the safety index. Conclusion: Reasonable increase of vaccine dose can up-regulate Th1 cell expression and promote the generation of anti-HBs.

[Scanning electron microscopic observation of interactions of vaginal epithelial cells with Candida albicans and Lactobacillus crispatus].

Objective: To investigate interactions of vaginal epithelial cells (VEC) with Candida albicans and the protective effect of Lactobacillus crispatus on VEC. Methods: VK2/E6E7 VEC cell line was cultured. Candida albicans and Lactobacillus crispatus suspension were prepared respectively. Ultrastructural morphological changes of VEC infected with Candida albicans and treated with Lactobacillus crispatus were observed by scanning electron microscope (SEM). Results: Candida albicans invaded into VEC mainly through endocytosis after an early infection of 6 hours. Pseudopods of VEC increased, contacted and covered the surface of pseudohyphae. After 12 hours infection, active penetration developed into the predominant invasion mechanism. Candida albicans directly penetrated through VEC, destroyed the surface structures, and even led to cell death. A reduction of pseudopods on VEC was observed. After co-incubation with Lactobacillus crispatus, SEM revealed a morphological recovery of VEC and destruction of pseudohyphae. Conclusion: s Candida albicans invade VEC mainly through endocytosis and active penetration. Lactobacillus crispatus could utilize occupying effect to exert protective effect on VEC during the infection with Candida albicans.

[Study on the production of IgG derived from vaginal epithelial cells and the effect of anti-Candida albicans].

Objective: To investigate the function of IgG secreted by vaginal epithelial cells in natural resistance to vulvovaginal candidiasis. Methods: (1)Immunohistochemical method was used to determine the expression of IgG secreted by normal vaginal epithelial cells VK2/E6E7.(2)Samples were divided into three groups by different proportions of VK2/E6E7 cells to Candida albicans ,including Candida albicans: VK2/E6E7 cells were 1∶10, 1∶1[yeast+ cells(1∶10)group and yeast+ cells(1∶1)group]and VK2/E6E7 cells as blank control group. The growth status of 3 groups were observed under inverted microscope after 24 hours. ELISA method was used to detect the production of IgG in 3 groups after 0, 3, 6, 12, 24, 48 hours. Results: (1)Immunohistochemical method showed normal vaginal epithelial cells were polygonal with pale blue nucleus and cytoplasm were distributed by brown granules, which indicated that IgG were strongly positive. While negative control group just had light blue nuclei.(2)Inverted microscope observation represented that control group had a clear outline, strong refraction and large nuclei with cobblestone-like appearance. After yeast+cells(1∶10)group co-cultured for 24 hours, Candida albicans begin to sprout and transformed to hyphae. VK2/E6E7 cells and Candida albicans were close to each other with vacuoles and small black granules in the cytoplasm. The morphology of cells were complete. Yeast+ cells(1∶1)group showed obvious invasion effect of Candida albicans to VK2/E6E7 cells with vigorous growth of hyphae, the decreased number and incomplete morphology of cells. Moreover, the connection of cells were loose. ELISA assay showed that there were statistically significant difference of IgG secretions between the 3 groups after 0, 3, 6, 12, 24, 48 hours(P<0.05). After stimulation of Candida albicans, secretion of IgG was significantly lower than that in the control group. The statistical difference of IgG secretions between yeast+ cells(1∶10)group and yeast+ cells(1∶1)group after 3, 6, 12, 24 hours were dramatic(P<0.05). The peak of IgG production showed when the ratio of Candida albicans and VK2/E6E7 was 1∶1 after 24 hours co-cultured(1.61±0.05)µg/ml. Conclusions: Candida albicans has a significant invasion role on epithelial cells. With increasing concentrations of Candida albicans, the invasion effect will be enhenced. While, after the vaginal epithelial cells co-cultured with Candida albicans, the secretion of IgG decreased significantly.