This core will provide support to the overall program by providing centralized viral and immunologic assays, as well as providing scientific guidance to each project. This will streamline, standardize, and integrate experimental methodology between Projects. There are two Aims:
Aim 1. To perform immunologic and virologic assays in support of the interventions in the Projects, and develop or optimize novel assays as needed. ? Flow cytometric analyses to quantify reconstitution in the huBLT mouse model ? Flow cytometric identification of transduced cells expressing surface C46 ? Phenotyping of transduced T and B cells ? Evaluation of antigen-specific T cell proliferation by flow cytometry ? Detection of bNAb expressing B cells by flow cytometry ? Quantification of soluble bNAb (Design of quantitative ELISA) ? Assessment of bNAb-directed anti-idiotype humoral immunity (Design of an anti-idiotype ELISA) ? Monitoring for cellular immunity against transduced gene products ? Monitoring for humoral immunity against cell surface transduced gene products C46 and/or huEGFRt (Design of a flow cytometric assay) ? Measurement of plasma viremia in the huBLT mouse model ? Antiviral assays for cellular immunity in the huBLT mouse model ? Antiviral assays for bNAb produced in vivo ? Sequencing of HIV-1 env for potential escape mutations ? Quantitation of the latent reservoir in the huBLT mouse model ? Testing functionality of CMV-specific CAR constructs ? Measurement of pro-inflammatory cytokines ? Clonal tracking of transduced cells by vector integration site deep sequencing Aim 2. To provide scientific and technical consultation to the Projects. The Core investigators will be available for scientific and technical consultation for each of the scientific projects.
