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TEIXEIRA, F. M. E. Evaluation of response to the DNA vaccine LAMP1/p55Gag of HIV-1 and generation of follicular T cells in the neonatal mice immunization 2018. 131p. Master Thesis (Immunology) Instituto de Ciências Biomédicas. Universidade de São Paulo, São Paulo, 2018 The number of young people infected with HIV has been increasing in recent decades, which highlights the need for vaccine strategies that are immunogenic at early phase of life able of inducing long-term response. The chimeric LAMP1/p55Gag vaccine, associates the gene encoding LAMP-1 (lysosomal associated membrane protein) and the HIV-1 gag gene, directs the traffic of viral protein to MIIC compartments, leading to presentation of the viral peptides through class II of the Major Histocompatibility Complex (MHC II). This chimeric vaccine is immunogenic in adult and neonatal BALB/c mice and crucial to induce long-term T and B responses, producing high levels of antibodies. However, the immunological mechanisms involved in the induction of the humoral response of the LAMP/Gag vaccine, such as the generation of T follicular helper cells (TFH), are unknown in the neonatal period. The objective of this study was to evaluate the generation of TFH, and follicular cytotoxic T cells and B cells in neonates submitted to immunization with the LAMP/Gag (LG) and Gag (G) vaccines. The results show that neonatal immunization at seven daysold in C57BL/6 mice strain with the LG vaccine is able to increasing the frequency of IFN-γ-secreting cells to the immunodominant peptides of the HIV-1 gag region and of CD8+IFN-γ+ T cells compared to the Gag vaccine. Neonatal immunization with the LG vaccine led to the production of high titers of anti -p24 IgG1 antibodies and the increased percentage of Gag-specific IgG1 secreting cells. Neonatal priming with LG is able to promote long-lasting anti-Gag humoral and cellular response. Moreover, neonatal (ip) immunization with LG was able to induce TFH cells (CD4+CXCR5+PD-1+Bcl-6 +), follicular CD8 + T cells (TFC) (CD8+CXCR5+) and germinal center (GC) formation in the mesenteric lymph nodes, whereas both vaccines induced follicular B cells (CD19+CXCR5+). Despite the lower frequency of TFH in neonates in relation to adult counterpartin the immunization with LG, a similar percentage of TFC cells was observed. Conventional immunization by intradermal immunization induced an increased number of TFH cells in inguinal lymph nodes on the third day after booster vaccination, despite the similar frequency of follicular TFH, TFC and B cells. In this period the LG vaccine also induced generation of CG B cells. Another peculiarity of the LG neonatal vaccine was the increase in the gene expression of the enzyme activation-induced cytidine deaminase (AID).The results show that the LAMP/Gag vaccine is immunogenic in the neonatal phase of C57BL/6 mice for the generation of antigen-specific humoral and cellular response and long-term response, similar to BALB/c mice. The findings showed effective induction of follicular T cells, maturation of lymphoid tissues with formation of GCs and up-regulation oftranscripts for AID. Taken together, our findings demonstrate that the LAMP/Gag chimeric vaccine strategy is effective at this time in life, and has an important adjuvant role in the maturation of the humoral response.

Avaliação da resposta à vacina de DNA &gt;i&lt;LAMP-1/p55Gag&gt;/i&lt; do HIV-1 e da geração de células T foliculares na imunização de camundongos neonatos