In situ staining and laser capture microdissection of lymph node residing SIV Gag-specific CD8⁺ T cells - A tool to interrogate a functional immune response ex vivo

While a plethora of data describes the essential role of systemic CD8⁺ T cells in the control of SIV replication little is known about the local in situ CD8⁺ T cell immune responses against SIV at the intact tissue level, due to technical limitations. In situ staining, using GagCM9 Qdot 655 multimers, were here combined with laser capture microdissection to detect and collect SIV Gag CM9 specific CD8⁺ T cells in lymph node tissue from SIV infected rhesus macaques. CD8⁺ T cells from SIV infected and uninfected rhesus macaques were also collected and compared to the SIV GagCM9 specific CD8⁺ T cells. Illumina bead array and transcriptional analyses were used to assess the transcriptional profiles and the three different CD8⁺ T cell populations displayed unique transcriptional patterns. This pilot study demonstrates that rapid and specific immunostaining combined with laser capture microdissection in concert with transcriptional profiling may be used to elucidate phenotypic differences between CD8⁺ T cells in SIV infection. Such technologies may be useful to determine differences in functional activities of HIV/SIV specific T cells.