Evaluation Of Immunological Effects Of HPV Bivalent Vaccine Via Nasal Mucosal Immunity

Objective The human papilloma virus(HPV)vaccine was administered to the mouse via the nasal mucosal immunization route,and the immune effect of the nasal mucosal immune pathway was evaluated by comparison with the subcutaneous vaccine group.Methods Fortyfive female BALB/c mice(18-20g)aged 6-8 weeks were randomly divided into three groups:nasal saline control group(A),nasal vaccine experimental group(B)and subcutaneous vaccine experimental group(C).The mice in the above three groups were immunized at 0 weeks,2weeks,and 4 weeks,and the mice were sacrificed in the sixth week and blood samples were collected by eyeballs.The IgG content of specific antibodies in serum was detected by ELISA;The spleen cell suspension was prepared from the spleen,the spleen lymphocytes were isolated and cultured,and the specific proliferation of mouse spleen lymphocytes was detected by MTT assay.Finally,the release of cytokine IFN-γ and IL-4 in the supernatant of mouse spleen lymphocytes stimulated by HPV16 L1 protein,HPV18 L1 protein and HPV16/18 L1 protein were detected by ELISA.In addition,lung lavage fluid,nasal lavage fluid and vaginal lavage fluid were collected,and the specific sIgA antibody content in the lavage fluid was detected by ELISA.Results The results of ELISA showed that BALB/c mice were immunized with bivalent HPV vaccine,and the mucosal immune response was effectively induced and enhanced after three immunizations.The specific sIgA antibody against HPV16 L1 protein and HPV18 L1 protein in the nasal lavage fluid of mice was obvious.The sIgA antibody was significantly increased in the nasal vaccine group compared with the control group and the subcutaneous vaccine group(P<0.05).The specific sIgA antibody against the HPV16 L1 protein was significantly increased in the mouse lung lavage fluid.There was significant difference between the vaccine group and the control group(P<0.01).There was significant difference between the nasal vaccine group and the subcutaneous vaccine group(P<0.05).The specific sIgA antibody against HPV18 L1 protein in the lung lavage fluid of mice was significantly increased.The difference between the nasal vaccine group and the control group was statistically significant(P<0.01).There were statistical differences between the nasal vaccine group and the subcutaneous vaccine group(P<0.001).The specific sIgA antibody against HPV16 L1 protein in mouse vaginal lavage fluid increased significantly,the differencebetween the nasal vaccine group and the control group was statistically significant(P<0.05),the specific sIgA antibody against HPV18 L1 protein in the vaginal lavage fluid was significantly increased,and the difference was statistically significant between the nasal vaccine group and the control group(P<0.01).Compared with the nasal saline group,the specific antibody IgG against HPV16 L1 protein in the serum of the nasal vaccine group was significantly increased,and the difference were statistically significant(P<0.05),the specific IgG antibody against HPV18 L1 protein in the serum of the nasal vaccine group was significantly increased,and the difference was statistically significant(P<0.01).MTT assay showed that the stimulation index of nasal vaccine group and subcutaneous vaccine group increased significantly after the stimulation of HPV16 L1 protein,HPV18 L1 protein,HPV16/18 L1 protein,and the proliferation of spleen lymphocytes was greatly improved.After stimulation by HPV16 L1 protein,HPV18 L1 protein,HPV16/18 L1 protein,the release of IFN-γ were significantly increased in the spleen cell culture supernatant of the nasal vaccine group and the subcutaneous vaccine group.Conclusion The bivalent HPV vaccine can produce specific IgG antibodies in serum after nasal mucosal immunization,which effectively induces humoral immune response.In addition,it can also induce local mucosal immune responses in the nose,lungs,vagina,etc.,and produce specific mucosal antibody sIgA.Therefore,the nasal mucosal immune pathway can be used as one of the mucosal vaccine inoculation methods in the near future.