In vivo activation, differentiation and effector function of antigen specific target cells by CD8+ T lymphocytes: role of Caspase1, GSDMD, RIPK3 and MLKL

• Grant number: 21/12143-7
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: March 01, 2022
• End date: September 30, 2025
• Field of knowledge: Biological Sciences - Immunology - Cellular Immunology
• Principal Investigator: João Gustavo Pessini Amarante Mendes
• Grantee: Abolaji Samson Olagunju
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

Tumor cells, as well as viruses and other intracellular pathogens, are persistently challenging the human organism. In this scenario, the activation and effector function of antigen-specific CD8+ T lymphocytes represents a major immunological response, since these cells can eliminate infected or transformed cells by inducing their demise (apoptosis). This project will investigate the possible involvement of the key regulatory and effector molecules of necroptosis (RIPK3 and MLKL) and pyroptosis (Caspase-1 and GSDMD) in the in vivo activation, differentiation and effector function of antigen-specific CD8+ T cells and immune checkpoint inhibitor- anti PD L1 on tumor growth. C57Bl/6 wild-type (WT), ripk3-/-, mlkl-/-, casp-1-/-, gsdmd-/-, gsdmd/mlkl-/- mice will be immunized with the human recombinant adenoviral vector 5 carrying the ovalbumin gene (rhAd5.OVA). The in vivo cytotoxicity assay will determine the efficiency of target cell elimination by CD8+ T lymphocytes. ELISPOT will be employed to evaluate the frequency of OVA-specific CD8+ T lymphocytes and multi-colour flow cytometry will be used to evaluate the level of activation markers, cytokines, granzymes, among others. The statistical analysis will be done using GraphPad prism v7, two-way analysis of variance (ANOVA) with the factor groups and condition (immunized, non-immunized). The univariate ANOVAs will be applied for each dependent variable and the significance level (originally 5%) will be adjusted downward accordingly to protect against an inflated type I error. (AU)

Scientific publications

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

OLAGUNJU, ABOLAJI S.; SARDINHA, ANDREW V. D.; AMARANTE-MENDES, GUSTAVO P.. Long-Lasting, Fine-Tuned Anti-Tumor Activity of Recombinant Listeria monocytogenes Vaccine Is Controlled by Pyroptosis and Necroptosis Regulatory and Effector Molecules. PATHOGENS, v. 13, n. 10, p. 11-pg., 2024-10-01. (21/07624-6, 21/13486-5, 21/12143-7, 23/02577-5, 18/25395-1)