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Open Access Highly Accessed Research

In situ detection of Gag-specific CD8+ cells in the GI tract of SIV infected Rhesus macaques

Annelie Tjernlund1, Jia Zhu1, Kerry Laing1, Kurt Diem2, David McDonald3, Julio Vazquez3, Jianhong Cao4, Claes Ohlen5, M Juliana McElrath12, Louis J Picker6789 and Lawrence Corey12*

Author Affiliations

1 Vaccine & Infectious Disease Institute, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

2 Departments of Medicine and Laboratory Medicine, University of Washington, Seattle, WA, USA

3 Scientific Imaging, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

4 Immune monitoring lab, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

5 AIDS Vaccine Program, SAIC-Frederick/NCI-Frederick, Frederick, Maryland, USA

6 Vaccine and Gene Therapy Institute, Department of Pathology, Oregon Health and Science University, Beaverton, OR, USA

7 Vaccine and Gene Therapy Institute, Department of Molecular Microbiology, Oregon Health and Science University, Beaverton, OR, USA

8 Vaccine and Gene Therapy Institute, Department of Immunology, Oregon Health and Science University, Beaverton, OR, USA

9 Vaccine and Gene Therapy Institute, Oregon National Primate Research Center, Oregon Health and Science University, Beaverton, OR, USA

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Retrovirology 2010, 7:12  doi:10.1186/1742-4690-7-12

Published: 16 February 2010

Abstract

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.