Naloxone/alum mixture a potent adjuvant for HIV-1 vaccine: induction of cellular and poly-isotypic humoral immune responses

Abstract

In the present study we used a fusion peptide from HIV-1 p24 and Nef as vaccine model and adjuvant activity of Naloxone/alum mixture was evaluated in a peptide vaccine model. HIV-1 p24-Nef fusion peptide was synthesized. Female BALB/c mice were divided into five groups. The first group immunized subcutaneously with the p24-Nef fusion peptide adjuvanted with Naloxone/alum mixture and boosted with same protocol. The second was immunized with fusion peptide adjuvanted in alum. The control groups were injected with NLX (Group 3), Alum (Group 4), or PBS (Groups 5) under the same conditions. To determine the type of induced immune response, sera and splenocytes were analyzed by commercial ELISA method for total IgG and isotypes and cytokine secretion (IL-4 & IFN-γ), respectively. We have also used the ELISPOT assay to monitor changes in the frequency of IFN-γ-producing T cells. The proliferation of T cells was assessed using Brdu method and T-cell cytotoxicity was assessed with CFSE method. Immunization of mice with HIV-1 p24-Nef fusion peptide formulated in Naloxone/ alum mixture significantly increased lymphocyte proliferation and shifted cytokine responses toward Th1 profile compared to all other groups. Analysis of humoral immune responses revealed that administration of HIV-1 p24-Nef fusion peptide with Naloxone/alum mixture significantly increased specific IgG responses and also increased IgG1,IgG2a, IgG2b, IgG3, and IgM vs. alum-adjuvanted vaccine groups. Naloxone/alum mixture as an adjuvant could improve cellular and humoral immune response for HIV vaccine model and this adjuvant maybe useful for HIV vaccine model in human clinical trial.

Formulation of a recombinant HIV‐1 polytope candidate vaccine with naloxone/alum mixture: induction of multi‐cytokine responses with a higher regulatory mechanism

“…Assessment of the IFN-c cytokine showed that mice immunized with the NLX/alumadjuvanted vaccine showed a significant increase in the IFN-c cytokine as compared with those immunized with alum-adjuvanted vaccine. The synergistic effect of NLX and alum in the induction of IFN-c and subsequent Th1 responses were consistent with the previous studies [17,26].…”

“…In the present study, mice immunized with the NLX/alumadjuvanted vaccine exhibited a significant increase in the proliferation responses of lymphocytes when compared with those immunized with the alumand NLX-adjuvanted vaccines. This result showed the synergistic immunostimulating effect of NLX/ alum mixture on the lymphoproliferative response in vaccinated mice, as confirmed by other studies [15,17,26].…”

“…In this study, more immunologic responses and also immunoregulatory activity of NLX/alum mixture adjuvant are determined in an HIV-1 polytope vaccine model. Herein, we analyzed a wide variety of cytokines than our previous study [26] to find a cytokine network of the NLX/alum mixture as an adjuvant for the formulation in an HIV-1 vaccine. Surprisingly, we found a novel potential function of NLX/alum adjuvant on the immunopathology effect of vaccine.…”

“…Previous study shows the adjuvant activity of NLX and NLX/alum in vaccine model [26]. But other immunologic patterns inducing by this adjuvant and its immunoregulatory effect have not been clarified.…”

Leishmaniasis in humans: drug or vaccine therapy?

“…The results of the studies demonstrated that the TSA gene was the most protective vaccine with the later including a CD8 + response. 132 , 142 – 146 DNA vaccine studies were performed in clinical trial Phases I and II and animals that were tested included dogs and mice. However, these promising results should be carefully considered and further confirmed by subsequent studies.…”