The Application Of Peptide Nanomaterials, NME And 2F, In HIV Vaccine Adjuvant And Detection

The human immunodeficiency virus (HIV) can attack the body's immune system,especially CD4 T lympHocytes, resulting in collapse of immune system and deficiency of resistance to bacteria invasion. Therefore, the human body is susceptible to various diseases.In order to improve the efficiency of prevention and treatment of Acquired Immune Deficiency Syndrome (AIDS), development of high-potency vaccines and high-sensitivity early diagnostic reagents are urgently need to be studied.The DNA vaccine is an important type of HIV vaccine, which plays an important role in early immunotherapy of AIDS. The ideal DNA vaccine should be able to overcome its low immunogenicity and the high variability of the HIV virus during replication. Therefore,in this study, NME (NapGFFYNMe) was chosen as the main research object, and its effects on humoral immunity and cellular immunity, including the immunogenicity of the NME HIV DNA vaccine against the adjuvant and polyfunctional T cell immune response, the broad IgG subclass response and the V1/V2-specific IgG response, were studied. The results showed that the T cell immune response to the random combination of 5 cytokines(IL-2, IL-4, TNF-?, IFN-? and CD107a) were 1.05%,0.93%,1.37%, the the T cell immune response to the random combination of 4 cytokines (IL-2, IL-4, TNF-?, IFN-? and CD107a)were 2.39%, 2.61%, 4.86%, the T cell immune response to the random combination of 3 cvtykines (IL-2, IL-4, TNF-a, IFN-y and CD107a) were 3.31%, 3.22%, 2.7%. These results confirmed that NME vaccine adjuvant could effectively enhance the effect of HIV DNA vaccine on the immune response of the whole body.On the other hand, in order to improve the sensitivity of early diagnosis of HIV, a short peptide of 2F (NapFFKYp) was designed in this study. We got a small molecule polypeptide NapFFKYp was self-assembled into super molecular hydrogel under the action of alkaline pHospHatase. This material has ? -sheet structure. Since the fluorescence of congo red could be enhanced and form a cascade signal amplification when it was in a ?-sheet environment, we establish a 2F-Congo red assay for the detection of HIV-1 p24. The results showed that the limit of the 2F-Congo red detection reached 2.44 pg/mL, which was lower than that of clinical ELISA (5 pg/mL). Besides, the inter-batch variability, intra-batch variability and precision of 2F-Congo red for HIV detection were (5.4%, 2.3% and 2.9%,respectively), which were similar to those of clinical ELISA (4.1%,2.7% and 3.2%,respectively). The variability and precision of 2F-Congo red was consistent to that of clinical ELISA for quantifying HIV-1 p24. 2F-Congo red thus shows a better performance for the detection of HIV infection at the early stages, as detection by clinical ELISA is difficult at the early stages of infection.