Immune Phenotyping Core

SUMMARY The goal of the Immune Phenotyping Core (Core C) is to provide tools and reagents to the different projects that will be used to characterize the immune responses induced by vaccinations and infections with coronaviruses, influenza viruses and dengue viruses. The Core will leverage existing state-of-the-art serological techniques established in the Krammer laboratory, as well as multiplex analysis of cytokine and chemokine plasma profile and Cytek Aurora Spectral Flow Cytometry profiling of PBMCs and human tonsillar histocultures (HC) currently used and optimized in the Fernandez-Sesma laboratory. The following aims re proposed: Aim 1: Characterization of antibody responses to coronavirus, influenza virus and dengue virus vaccination and infection. The Core will provide assays, reagents and protocols to measure binding and functional antibody responses against SARS-CoV-2 for Project 1 and influenza viruses for Project 2. Additionally, the secretion of IgM, IgG and IgA in the supernatant of human tonsil histocultures (HC) treated with the different SARS-CoV-2 vaccines types (Project 1), influenza virus vaccines and viruses (Project 2) will be assessed. Aim 2: Characterization of cytokine/chemokine responses to coronavirus, influenza virus and dengue virus vaccination and infection. The Core will analyze the levels of cytokines and chemokines in the plasma of vaccinated/infected individuals and the supernatant of human tonsil histocultures treated with different vaccines for Projects 1, 2 and 3. Aim 3: Characterization of cellular responses to coronavirus, influenza virus and dengue virus vaccination and infection. Analysis of the cellular immune profiles of PBMCs from vaccinated/infected individuals over time, using Spectral Flow cytometry. Human tonsillar HC will be also analyzed by Cytek Aurora Spectral Flow Cytometry in order to capture early immune signatures and changes in cell populations corresponding to adaptive immune responses in those HC after treatment with different vaccines. We will obtain high-resolution data at the single-cell level to resolve the most challenging cell populations including cells expressing viral antigens. PBMCs will also be subjected to a complementary transcriptomics analysis by RNAseq conducted by the Genomics Core. These tools will serve to generate immune signatures representative of the longitudinal immune responses to vaccination and/or infection in study participants enrolled in observational non-interventional cohort studies in coordination with the Data Management and Analysis Core (Core E). Data obtained using these immunological techniques will be analyzed by the Data management and Analysis Core comparing them across the different systems used in the projects as well as in combination with the genomic data obtained in the Genomics Core (Core D) from the same samples. All data generated by the VIVA Projects and Cores, including the Immune Phenotyping Core, will be deposited by the Data Analysis Core into ImmPort.