Jackson, Ronald J.Boyle, David B.Ranasinghe, Charani2026-01-012026-01-011064-3745PubMed:24715282https://hdl.handle.net/1885/733799597Although recombinant DNA and recombinant viral vectors expressing HIV antigens have yielded positive outcomes in animal models, these vaccines have not been effectively translated to humans. Despite this, there is still a high level of optimism that poxviral-based vaccine strategies could offer the best hope for developing an effective vaccine against not only HIV-1 but also other chronic diseases where good-quality T and B cell immunity is needed for protection. In this chapter we discuss step by step (1) how recombinant poxviral vectors co-expressing HIV antigens and promising mucosal/systemic adjuvants (e.g., IL-13Rα2) are constructed, (2) how these vectors can be used in alternative heterologous prime-boost immunization strategies, (3) how systemic and mucosal samples are prepared for analysis, followed by (4) two immunological assays: multicolor intracellular cytokine staining and tetramer/homing maker analysis that are used to evaluate effective systemic and mucosal T cell immunity.30enPublisher Copyright: © Springer Science+Business Media New York 2014.DNA vaccinesFowlpox virusHoming marker analysisIntracellular cytokine stainingMucosal/systemic cell preparationMucosal/systemic vaccinationPoxvirus vectorsPrime-boost immunizationT cell immunityTetramer analysisVaccinia virus201410.1007/978-1-4939-0410-5_584907536541