Research And Development Of Adjuvants In The New Crown Vaccine And HIV Vaccine

Objectives:Muramyl dipeptide(MDP)is an immunostimulating peptide composed of N-acetyltetracytic acid and L-alanine-D-isoglutamine,which is a peptide molecule.MDP and its derivatives as adjuvants have been extensively studied to effectively enhance the immune responses of vaccines.In this study,we invesigated the effect of MDP in improving the efficacy of SARS-CoV-2 protein subunit vaccine.Methods:The animal strains we use are Bal B/C mice,females,6 to 8 weeks old,and then 30 mice are randomly divided into 6 each groups,a total of 5 groups.They are 50μg control group,"20μg MDP+10μg vaccine" group,"20 μg MDP+50 μg vaccine"group,"100μg MDP+10 μg vaccine" group,"100μg MDP+50μg vaccine" group.S protein was used as the antigen of SARS-CoV-2 protein subunit vaccine.Mice in each group received 2 vaccinations at week 0 and week 3 by intramuscular injection.Blood samples were collected after 2 weeks of the last immunization.The humoral immune response was diluted 100,1000,10000 times by the serum of mice,and the specific IgG antibody was detected by ELISA.Results:① When serum of mice was diluted at 1:100,"compared with the 50μg vaccine group,the 20μg MDP+50μg vaccine group with 20μg MDP at the same dose of vaccine induced significantly enhanced IgG antibody response in mice(P<0.05)"."Compared with the 50μg vaccine group,the 100μg MDP+10μg vaccine group with 100μg MDP at reduced vaccine dose induced a significantly enhanced IgG antibody response in mice(P<0.05)." "Compared with the 50μg vaccine group,the 100μg MDP+50μg vaccine group with 100μg MDP at the same dose of vaccine induced a significantly enhanced IgG antibody response in mice(P<0.05)." "Compared with the 50μg vaccine group,the 20μg MDP+1 0μg vaccine group with 20μg MDP at reduced vaccine dose did not induce a significantly enhanced IgG antibody response in mice(P>0.05)."② When serum of mice was diluted at 1:1000,"Compared with the 50μg vaccine group,the 100μg MDP+50μg vaccine group with 100μg MDP at the same dose of vaccine induced significantly enhanced IgG antibody response in mice(P<0.05)"."Compared with the 50μg vaccine group,the other three groups did not induce significantly enhanced IgG antibody response in vivo(P>0.05)."③ Compared with 50μg vaccine group,there was no significant difference in IgG antibody of 4 groups after 10000 times dilution of serum of mice(P>0.05).Conclusions:As an adjuvant,MDP can effectively enhance the humoral immune response of the SARS-CoV-2 vaccines in mice.Objectives: Negative charged biomolecules(genes)and drugs can be effectively delivered by layered double hydroxide(LDH)in vitro and in vivo.Through modifying amantadine(Ada)on the surface of NiZnAl,the immunological effects of layered double hydroxides NiZnAl-Ada and Ada in improving HIV DNA vaccines were investigated.Methods: 24 Bal B/C mice were randomly divided into four groups: blank group,HIV DNA vaccine group,“HIV DNA vaccine + Ada” group “HIV DNA vaccine 十NiZnAl-Ada,5group,(n = 6).HIV Gag DNA vaccine and HIV Env DNA vaccine were cultured in E.coli.and combined immunization with nano adjuvant NiZnAl-Ada and Ada.Each group received three vaccinations via intradermal injection at an interval of two weeks.Two weeks later,after the final injection,mouse spleens and blood are harvested for the next humoral and cellular detection.The humoral immune response was detected at the 500,1000,5000 times dilution of mouse serum,and the GAG and gp 140-specific IgG antibodies were detected by ELISA.Elispot detected IFN-y cytokine secretion at the single cell level.The secretion of CD 107 a,IFN-y,TNF-α,IL-2,IL-4 in CD4+ T and CD8+ T cells was detected by flow cytometrc assay.Secretion of B cell surface molecules CD40,CD69 and dendritic cell surface molecules CD80,CD86.Results:ELISA data showed that:①"Compared with the blank control group,HIV DNA vaccine group induced significantly enhanced Gag specific IgG antibody response in mice after 500,1000 and 5000 fold dilution of mouse serum(P<0.05)"."Ada+HIV DNA vaccine group induced significantly enhanced Gag specific IgG antibody response in mice compared with blank control group(P<0.05)"."Compared with the blank control group,the NiZnAl-Ada+HIV DNA vaccine group induced significantly enhanced Gag specific IgG antibody response in mice(P<0.05)n.nAda+HIV DNA vaccine group did not induce a significantly enhanced Gag specific IgG antibody response in mice compared to the HIV DNA vaccine group(P>0.05)"."The NiZnAl-Ada+HIV DNA vaccine group did not induce significantly enhanced GAG specific IgG antibody response in mice compared with the HIV DNA vaccine group(P>0.05)".?" Compared with the blank control group,the HIV DNA vaccine group induced significantly enhanced gpl40 specific IgG antibody response in mice(P<0.05)” when the serum of mice was diluted 500,1000,and 5000 times."Ada+HIV DNA vaccine group induced a significantly enhanced gpl40 specific IgG antibody response in mice compared to the blank control group(P<0.05)’V’NiZnAl-Ada+HIV DNA vaccine group induced significantly enhanced gpl40 specific IgG antibody response in mice compared with blank control group(P<0.05)n.nAda+HIV DNA vaccine group did not induce a significantly enhanced gpl40 specific IgG antibody response in mice compared to the HIV DNA vaccine group(P>0.05)n." NiZnAl-Ada+HIV DNA vaccine group did not induce significantly enhanced gpl40 specific IgG antibody response in mice compared with the HIV DNA vaccine group(P>0.05)’’.Elispot experiment results:?MCompared with the blank control group,the HIV DNA vaccine group induced significantly enhanced WN-y expression in mice(P <0.05)’’."Ada+HIV DNA vaccine group induced significantly increased expression of IFN-y in mice compared with blank control group(P < 0.05)." Compared with the blank control group,the Niznal-Ada+HIV DNA vaccine group induced significantly enhanced expression of IFN-y in mice(P<0.05)”.②”Compared with the HIV DNA vaccine group,Ada+HIV DNA vaccine group did not induce significantly enhanced IFN-y expression in mice(P > 0.05)M.nNiZnAl-Ada +HIV DNA vaccine group did not induce significantly enhanced IFN-y expression in mice compared to the HIV DNA vaccine group(P > 0.05)n.Flow cytometry results: ② "Compared with the blank control group,in the HIV DNA vaccine group,the Ada+HTV DNA vaccine group and the NiZxiAl-Ada +HIV DNA vaccine group?The secretion of CD 107a5 IFN-y,TNF-a,IL-2 and IL-4 in CD4+T cells was not statistically significant(P > 0.05)" /’Compared with HTV DNA vaccine group,CD107 a,IFN-y,TNF-a,IL-2 and IL-4 in CD4+ T cells of Ada+HIV DNA vaccine group and NiZnAl-Ada+HIV DNA vaccine group were not statistically significant(P>0.05)".②’’Compared with the blank control group,in the HIV DNA vaccine group,Ada+HIV DNA vaccine group and NiZnAl_Ada+HIV DNA vaccine group,The secretion of CD107a5 IFN-y,TNF-a,IL-2 and IL-4 in CD8+ T cells was not statistically significant(P>0.05)M."Compared with the HIV DNA vaccine group,CD 107 a,IFN-y,TNF-a,IL-2 and IL-4 levels in CD8+ T cells were not statistically significant in Ada+HIV DNA vaccine group and NiZnAl-Ada+HIV DNA vaccine group(P>0.05)M.?"Compared with the blank control group,CD80 secretion of dendritic cells in the NiZnAl-Ada+HIV DNA vaccine group was statistically significant(P<0.05)"."Compared with the blank control group,the secretion of CD80 in the HIV DNA vaccine group and the Ada+HIV DNA vaccine group was not statistically significant(P>0.05)"."Compared with the HIV DNA vaccine group,the secretion of CD80 in Ada+HIV DNA vaccine group and NiZnAl-Ada+HIV DNA vaccine group was not statistically significant(P> 0.05)”."Compared with the blank control group,the secretion of CD86 in the HIV DNA vaccine group,Ada+HIV DNA vaccine group and NiZnAl-Ada+HIV DNA vaccine group was not statistically significant(P>0.05)"."Compared with the HIV DNA vaccine group,the secretion of CD86 in Ada+HIV DNA vaccine group and NiZnAl-Ada+HIV DNA vaccine group was not statistically significant(P> 0.05)"."Compared with the blank control group,there was no significant difference in the secretion of CD40 and CD69 in the HIV DNA vaccine group,Ada+HIV DNA vaccine group and NiZnAl-Ada +HIV DNA vaccine group(P>0.05)"."Compared with the HIV DNA vaccine group,there was no significant difference in the secretion of CD40 and CD69 in the Ada+HIV DNA vaccine group and the NiZnAl-Ada +HIV DNA vaccine group(P > 0.05).Conclusions: 1.There was no significant difference in the humoral immune response induced by HIV DNA vaccine as an adjuvant of NiZnAl-Ada.2.There was no significant difference in the immunoresponse of IFN-y cells induced by HIV DNA vaccine with the adjuvant of NiZnAl-Ada.3.There was no significant difference in T cell immune response induced by HIV DNA vaccine as an adjuvant of NiZnAl-Ada.