关键词: 疱疹病毒2型，人, 树突细胞, 单纯疱疹病毒疫苗

Abstract: Fan Jianyong, Wang Ying, Yang Huilan, Liang Jie, Li Cuihua. Department of Dermatology, Guangzhou General Hospital of Guangzhou Military Command, Guangzhou 510010, China Corresponding author: Fan Jianyong, Email: fanjianyong@hotmail.com 【Abstract】 Objective To evaluate the specific immune response induced by a dendritic cell-based adenovirus-mediated vaccine carrying the herpes simplex virus type 2 glycoprotein D gene （pAdeno-HSV-2 gD-DC） in BALB/c mice. Methods Forty BALB/c mice were equally divided into four groups: blank control group receiving no treatment, pAdeno-DC group immunized with pAdeno-DC, pAdeno-HSV-2 gD-DC group immunized with the previously constructed vaccine pAdeno-HSV-2 gD-DC, DC group immunized with DCs only. Totally, three rounds of vaccination were conducted at a 7-day interval. Ten days after the last vaccination, serum samples were collected and spleen cells were isolated from these mice. Enzyme-linked immunosorbent assay （ELISA） was performed to measure the level of IgG antibody against HSV-2 gD in the serum samples. Some spleen cells were stimulated with HSV-2 gD protein （10 mg/L） for 72 hours; then, ELISA was carried out to determine the levels of interferon （IFN）-γ and interleukin （IL）-4 in the supernatant, and 3-（4,5-dimethylthiazol-2-yl）-2,5 diphenyl tetrazolium bromide （MTT） assay to estimate the proliferative activity of these cells. The cytotoxicity of spleen cells was also evaluated based on the measurement of lactate dehydrogenase （LDH） release. Results The serum level of IgG antibody against HSV-2 gD （given in the absorbance value at 450 nm） was 0.313 ± 0.034 in the pAdeno-HSV-2 gD-DC group, significantly higher than that in the pAdeno-DC group, DC group and blank control group （0.034 ± 0.009, 0.028 ± 0.009 and 0.026 ± 0.010 respectively, all P < 0.05）. Increased proliferative activity and cytotoxicity were observed in spleen cells from the pAdeno-HSV-2 gD-DC group compared with those from the pAdeno-DC group, DC group and blank control group （cell stimulation index: 1.600 ± 0.215 vs. 1.063 ± 0.070, 1.056 ± 0.063 and 1.020 ± 0.051, all P < 0.05; percentage of cytotoxicity: 37.1% vs. 16.0%, 14.9% and 15.7%, all P < 0.05）. The levels of IFN-γ and IL-4 （both given in the absorbance value at 450 nm） were 0.568 ± 0.031 and 0.544 ± 0.043 respectively in the supernatant of spleen cells from the pAdeno-HSV-2 gD-DC group, compared to 0.266 ± 0.021 and 0.278 ± 0.037 respectively in the pAdeno-DC group （both P < 0.05）, 0.271 ± 0.023 and 0.275 ± 0.044 respectively in the DC group （both P < 0.05）, and 0.252 ± 0.012 and 0.245 ± 0.051 respectively in the blank control group （both P < 0.05）. Conclusion The vaccine pAdeno-HSV-2 gD-DC could induce a specific and strong immune response in BALB/c mice.

Key words: Herpesvirus 2, human, Dendritic cells, Herpes simplex virus vaccines