TY - JOUR
T1 - In situ detection of Gag-specific CD8+ cells in the GI tract of SIV infected Rhesus macaques
AU - Tjernlund, Annelie
AU - Zhu, Jia
AU - Laing, Kerry
AU - Diem, Kurt
AU - McDonald, David
AU - Vazquez, Julio
AU - Cao, Jianhong
AU - Ohlen, Claes
AU - McElrath, M. Juliana
AU - Picker, Louis J.
AU - Corey, Lawrence
N1 - Funding Information:
We thank Rachel Tompa for editorial assistance, Renee Rosemary Hukkanen, Andy Sylwester and Leo Stamatatos for providing samples. This work was supported by NIH grant R37AI042528, NIH grant RR00166, the Collaboration for AIDS Vaccine Discovery grant #38645 and Immune Correlates grant #41185 from the Bill & Melinda Gates Foundation. AT was supported through the Swedish Research Council and the Swedish Society for Medical Research.
PY - 2010/2/16
Y1 - 2010/2/16
N2 - Background: SIV and HIV predominantly replicate in lymphoid tissue, but the study of virus specific CD8+ T cells in intact lymphoid tissue is difficult, as traditional in situ tetramer staining requires fresh tissue.Results: In this report, we demonstrate a novel technique using Qdot 655-conjugated peptide-MHC multimers to directly visualize SIV specific cells in cryopreserved tissue biopsies from chronically SIVmac239 infected Rhesus macaques. Qdot 655 multimers showed similar sensitivity and specificity to APC-conjugated tetramers by flow cytometry analysis, but yielded ten-fold higher signal intensity when imaged by fluorescence microscopy. Using this technique, we detected CD8+ T cells which recognize an immunodominant epitope (Gag CM9) in the spleen, lymph nodes, ileum and colon. In all these tissues, the Gag CM9 positive cells were mainly located in the extra follicular T cell zone. In the ileum and colon, we found Gag CM9 positive cells concentrated in Peyer's patches and solitary lymphoid follicles, a pattern of localization not previously described.Conclusions: The use of Qdot multimers provide an anatomic and quantitative evaluation of SIV specific CD8+ T cell responses in SIV pathogenesis, and may prove useful to studies of SIV specific CD8+ T cell responses elicited by vaccines and other immunotherapies in the non-human primate model.
AB - Background: SIV and HIV predominantly replicate in lymphoid tissue, but the study of virus specific CD8+ T cells in intact lymphoid tissue is difficult, as traditional in situ tetramer staining requires fresh tissue.Results: In this report, we demonstrate a novel technique using Qdot 655-conjugated peptide-MHC multimers to directly visualize SIV specific cells in cryopreserved tissue biopsies from chronically SIVmac239 infected Rhesus macaques. Qdot 655 multimers showed similar sensitivity and specificity to APC-conjugated tetramers by flow cytometry analysis, but yielded ten-fold higher signal intensity when imaged by fluorescence microscopy. Using this technique, we detected CD8+ T cells which recognize an immunodominant epitope (Gag CM9) in the spleen, lymph nodes, ileum and colon. In all these tissues, the Gag CM9 positive cells were mainly located in the extra follicular T cell zone. In the ileum and colon, we found Gag CM9 positive cells concentrated in Peyer's patches and solitary lymphoid follicles, a pattern of localization not previously described.Conclusions: The use of Qdot multimers provide an anatomic and quantitative evaluation of SIV specific CD8+ T cell responses in SIV pathogenesis, and may prove useful to studies of SIV specific CD8+ T cell responses elicited by vaccines and other immunotherapies in the non-human primate model.
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U2 - 10.1186/1742-4690-7-12
DO - 10.1186/1742-4690-7-12
M3 - Article
C2 - 20158906
AN - SCOPUS:77949444381
SN - 1742-4690
VL - 7
JO - Retrovirology
JF - Retrovirology
M1 - 12
ER -