Assessing Antigen-Specific Cellular Immune Responses upon HIV/SIV Plasmid DNA Vaccination in the Nonhuman Primate Model

Hu, Xintao; Felber, Barbara K.; Valentı́n, Antonio

doi:10.1007/978-1-0716-0872-2_6

Cited by 1 publication

(3 citation statements)

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“…PBMC were isolated from EDTA-blood by Ficoll-Paque ™ Plus (GE Health #17-1440-03) gradient centrifugation. Live-frozen cells were thawed, DNaseI-treated and counted upon Acridine Orange/Ethidium Bromide dye staining, detailed in (71). The viability typically was >90%.…”

Section: Cellular Immune Responsesmentioning

confidence: 99%

“…Bronchioalveolar lavage (BAL) was collected in by perfusion with PBS, and cells were collected after centrifugation and washing, and the cells were directly assayed. In both, PBMC and BAL, antigen-specific T cell responses were measured in peptide-stimulated cells, in the presence of the protein secretion inhibitor Monensin (GolgiStop), as previously described (30,71). Briefly, 10 6 cells were seeded in 96-well plates and stimulated with different peptide pools at a final concentration of 1ug/ml for each individual peptide.…”

Section: Cellular Immune Responsesmentioning

confidence: 99%

“…For negative and positive controls, cells were cultured in medium without peptides or stimulated with a PMA-cell stimulation cocktail (eBioscience, Affymetrix Inc. San Diego, CA, USA). PMA (Phorbol 12-Myristate 13-Acetate) stimulation was performed as described (71), and the median value obtained was 36% (range 22-46) of IFN-g + T cells. After 12 hours incubation, the cells were washed and stained with antibody cocktails targeting surface proteins.…”

Section: Cellular Immune Responsesmentioning

confidence: 99%

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Comparative immunogenicity of an mRNA/LNP and a DNA vaccine targeting HIV gag conserved elements in macaques

Valentı́n

Bergamaschi²,

Rosati³

et al. 2022

Front. Immunol.

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Immunogenicity of HIV-1 mRNA vaccine regimens was analyzed in a non-human primate animal model. Rhesus macaques immunized with mRNA in lipid nanoparticle (mRNA/LNP) formulation expressing HIV-1 Gag and Gag conserved regions (CE) as immunogens developed robust, durable antibody responses but low adaptive T-cell responses. Augmentation of the dose resulted in modest increases in vaccine-induced cellular immunity, with no difference in humoral responses. The gag mRNA/lipid nanoparticle (LNP) vaccine provided suboptimal priming of T cell responses for a heterologous DNA booster vaccination regimen. In contrast, a single immunization with gag mRNA/LNP efficiently boosted both humoral and cellular responses in macaques previously primed by a gag DNA-based vaccine. These anamnestic cellular responses were mediated by activated CD8+ T cells with a phenotype of differentiated T-bet+ cytotoxic memory T lymphocytes. The heterologous prime/boost regimens combining DNA and mRNA/LNP vaccine modalities maximized vaccine-induced cellular and humoral immune responses. Analysis of cytokine responses revealed a transient systemic signature characterized by the release of type I interferon, IL-15 and IFN-related chemokines. The pro-inflammatory status induced by the mRNA/LNP vaccine was also characterized by IL-23 and IL-6, concomitant with the release of IL-17 family of cytokines. Overall, the strong boost of cellular and humoral immunity induced by the mRNA/LNP vaccine suggests that it could be useful as a prophylactic vaccine in heterologous prime/boost modality and in immune therapeutic interventions against HIV infection or other chronic human diseases.

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Section: Cellular Immune Responsesmentioning

confidence: 99%

Section: Cellular Immune Responsesmentioning

confidence: 99%

Section: Cellular Immune Responsesmentioning

confidence: 99%

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