接种纯化人用狂犬病疫苗对Th1／Th2类细胞因子谱的影响

Th1/Th2 cytokines response to purified rabies vaccine for human use

目的研究人用狂犬病纯化疫苗（RV）对人体抗原特异性Th1／Th2细胞因子谱的影响。方法对20例暴露者进行RV的全程接种，在开始接种RV的第0天、14天、45天时分别采血并分离人外周血单个核细胞（PBMC）与RV进行培养，采用ELISA法检测血清抗狂犬病病毒抗体，体外培养检测淋巴细胞转化增殖能力（MTT法），流式微球分析技术（CBA）法检测细胞培养上清液中Th1类细胞因子（IFN-γ、TNF、IL-2）及Th2类细胞因子（IL--4、IL-5、IL-10）的水平。结果暴露组全程注射RV后，19例于第45天、1例于第60天检测血清抗狂犬病病毒抗体阳性，阳性率100％。RV刺激后，暴露组第14天、第45天淋巴细胞增殖能力显著增高，第45天淋巴细胞增殖能力明显高于第0天（P〈0．05）；当RV刺激后，暴露组第14天、第45天产生IFN-γ、IL-2、IL-4、IL-5的含量高于未刺激组及暴露组第0天水平（P〈0．05）；TNF、IL-10的含量各组间比较差异无统计学意义（P〉0．05）。结论RV在刺激机体产生体液免疫的同时，可以诱导机体产生特异性细胞免疫，Th1的免疫应答尤为显著，提示细胞免疫在预防和控制狂犬病病毒感染中发挥重要的协同作用。

Objective To study the effects of purified rabies vaccine for human use （RV） on specific Th1/Th2 cytokines in human. Methods Twenty cases were injected intramuscularly with 5 full doses of RV. PBMCs were isolated from the blood sample collected at day 0, 14, 45 after the RV inoculation. Neutralizing antibody was determined by ELISA, and the proliferation of lymphocyte by in vitro test. The levels of RV specific IFN-γ, TNF, IL-2,IL-4, IL-5, IL-10 in the culture supernatants were detected by cytometric bead array （CBA）. Results The neutralizing antibody was tested positive in 19 cases 45 days after inoculation and 1 case after 60 days, with the positive rate reaching 100%. After stimulation with RV, the lymphocyte transformation index at day 14, 45 in cases were significantly higher than those day of 0 （P 〈 0.05）, and similar results were confirmed with IFN-γ, IL-2, IL-4, IL-5 tested by CBA （P 〈 0.05）. Conclusion The RV could induce humoral and antigen-specific cellular immune responses in human, tested by showing good protective effect on rabies virus.