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目的分析表达HIV-1 gag蛋白的重组乳酸乳球菌通过口服、滴鼻、皮下注射以及上述3种方式联合免疫小鼠诱导的体液免疫反应。方法将50只BALB/c小鼠随机分成5组,每组10只。表达gag的重组乳酸乳球菌分别通过口服、滴鼻、皮下注射以及上述3种方式联合免疫小鼠,对照组口服含PMG36e空载体的乳酸乳球菌,接种菌量均为109菌落形成单位(CFU),连续免疫3次,每次间隔2周,其中口服和滴鼻每次连续免疫3 d。采集免疫前和每次免疫后2周小鼠的血清,ELISA检测gag特异性的IgG水平。结果表达gag的重组乳酸乳球菌经口服、滴鼻、皮下注射以及3种方式联合免疫均诱导小鼠产生了gag特异性IgG,且联合免疫组gag IgG水平显著高于其他各实验组(P<0.01)。初次免疫后6周,口服和皮下注射免疫组gag IgG水平显著高于滴鼻免疫组(P<0.01)。免疫3次后各组血清中gag抗体的阳性率:口服免疫组分别为40%、40%、90%;滴鼻免疫组分别为10%、20%、20%;皮下免疫组分别为10%、60%、90%;联合免疫组的阳性率在初次免疫后2周已达到100%。结论表达HIV-1 gag的重组乳酸乳球菌载体疫苗经口服、滴鼻、皮下注射及3种方式联合免疫均能诱导小鼠产生特异性的体液免疫反应,多种途径联合免疫可增强乳酸乳球菌载体疫苗的免疫效果。
OBJECTIVE: To analyze the humoral immune responses induced by recombinant Lactococcus lactis expressing HIV-1 gag protein by oral, intranasal, subcutaneous injection and co-immunization of the above three methods. Methods Fifty BALB / c mice were randomly divided into 5 groups with 10 mice in each group. Recombinant Lactococcus lactis expressing gag were co-immunized by oral administration, intranasal injection, subcutaneous injection and the above three methods respectively. The control group was orally administered with Lactococcus lactis containing PMG36e empty vector and the inoculum volume was 109 colony forming units (CFU) , Continuous immunization 3 times, each interval of 2 weeks, of which oral and nasal continuous immunization 3 d each time. Serum was collected before immunization and 2 weeks after each immunization, and gag-specific IgG levels were detected by ELISA. Results Recombinant Lactococcus lactis expressing gag could induce gag-specific IgG in mice by oral administration, intranasal injection, subcutaneous injection and combined immunization. The gag IgG level in combined immunized group was significantly higher than that in other experimental groups (P < 0.01). At 6 weeks after the first immunization, gag IgG levels in oral and subcutaneous immunized groups were significantly higher than those in intranasal immunized groups (P <0.01). After three times of immunization, the positive rate of gag antibody in each group was 40%, 40% and 90% respectively in the oral immunized group, 10%, 20% and 20% in the intranasal immunized group, and 10% in the subcutaneous immunized group respectively , 60% and 90%, respectively. The positive rate of the combined immunization group reached 100% two weeks after the first immunization. Conclusion Recombinant Lactococcus lactis expressing HIV-1 gag vaccine can induce specific humoral immune response in mice by oral administration, intranasal injection, subcutaneous injection and combination of three ways. Combined immunization with various pathways can enhance the growth of Lactococcus lactis The immune effect of vector vaccine.