<it>In situ </it>detection of Gag-specific CD8<sup>+ </sup>cells in the GI tract of SIV infected Rhesus macaques
<p>Abstract</p> <p>Background</p> <p>SIV and HIV predominantly replicate in lymphoid tissue, but the study of virus specific CD8<sup>+ </sup>T cells in intact lymphoid tissue is difficult, as traditional <it>in situ </it>tetramer staining require...
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2010-02-01
|
| Series: | Retrovirology |
| Online Access: | http://www.retrovirology.com/content/7/1/12 |
| _version_ | 1811251238632685568 |
|---|---|
| author | McElrath M Juliana Ohlen Claes Cao Jianhong Vazquez Julio McDonald David Diem Kurt Laing Kerry Zhu Jia Tjernlund Annelie Picker Louis J Corey Lawrence |
| author_facet | McElrath M Juliana Ohlen Claes Cao Jianhong Vazquez Julio McDonald David Diem Kurt Laing Kerry Zhu Jia Tjernlund Annelie Picker Louis J Corey Lawrence |
| author_sort | McElrath M Juliana |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>SIV and HIV predominantly replicate in lymphoid tissue, but the study of virus specific CD8<sup>+ </sup>T cells in intact lymphoid tissue is difficult, as traditional <it>in situ </it>tetramer staining requires fresh tissue.</p> <p>Results</p> <p>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<sup>+ </sup>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.</p> <p>Conclusions</p> <p>The use of Qdot multimers provide an anatomic and quantitative evaluation of SIV specific CD8<sup>+ </sup>T cell responses in SIV pathogenesis, and may prove useful to studies of SIV specific CD8<sup>+ </sup>T cell responses elicited by vaccines and other immunotherapies in the non-human primate model.</p> |
| first_indexed | 2024-04-12T16:16:49Z |
| format | Article |
| id | doaj.art-c402d79685f042f28b2434ced9e94bc9 |
| institution | Directory Open Access Journal |
| issn | 1742-4690 |
| language | English |
| last_indexed | 2024-04-12T16:16:49Z |
| publishDate | 2010-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Retrovirology |
| spelling | doaj.art-c402d79685f042f28b2434ced9e94bc92022-12-22T03:25:42ZengBMCRetrovirology1742-46902010-02-01711210.1186/1742-4690-7-12<it>In situ </it>detection of Gag-specific CD8<sup>+ </sup>cells in the GI tract of SIV infected Rhesus macaquesMcElrath M JulianaOhlen ClaesCao JianhongVazquez JulioMcDonald DavidDiem KurtLaing KerryZhu JiaTjernlund AnneliePicker Louis JCorey Lawrence<p>Abstract</p> <p>Background</p> <p>SIV and HIV predominantly replicate in lymphoid tissue, but the study of virus specific CD8<sup>+ </sup>T cells in intact lymphoid tissue is difficult, as traditional <it>in situ </it>tetramer staining requires fresh tissue.</p> <p>Results</p> <p>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<sup>+ </sup>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.</p> <p>Conclusions</p> <p>The use of Qdot multimers provide an anatomic and quantitative evaluation of SIV specific CD8<sup>+ </sup>T cell responses in SIV pathogenesis, and may prove useful to studies of SIV specific CD8<sup>+ </sup>T cell responses elicited by vaccines and other immunotherapies in the non-human primate model.</p>http://www.retrovirology.com/content/7/1/12 |
| spellingShingle | McElrath M Juliana Ohlen Claes Cao Jianhong Vazquez Julio McDonald David Diem Kurt Laing Kerry Zhu Jia Tjernlund Annelie Picker Louis J Corey Lawrence <it>In situ </it>detection of Gag-specific CD8<sup>+ </sup>cells in the GI tract of SIV infected Rhesus macaques Retrovirology |
| title | <it>In situ </it>detection of Gag-specific CD8<sup>+ </sup>cells in the GI tract of SIV infected Rhesus macaques |
| title_full | <it>In situ </it>detection of Gag-specific CD8<sup>+ </sup>cells in the GI tract of SIV infected Rhesus macaques |
| title_fullStr | <it>In situ </it>detection of Gag-specific CD8<sup>+ </sup>cells in the GI tract of SIV infected Rhesus macaques |
| title_full_unstemmed | <it>In situ </it>detection of Gag-specific CD8<sup>+ </sup>cells in the GI tract of SIV infected Rhesus macaques |
| title_short | <it>In situ </it>detection of Gag-specific CD8<sup>+ </sup>cells in the GI tract of SIV infected Rhesus macaques |
| title_sort | it in situ it detection of gag specific cd8 sup sup cells in the gi tract of siv infected rhesus macaques |
| url | http://www.retrovirology.com/content/7/1/12 |
| work_keys_str_mv | AT mcelrathmjuliana itinsituitdetectionofgagspecificcd8supsupcellsinthegitractofsivinfectedrhesusmacaques AT ohlenclaes itinsituitdetectionofgagspecificcd8supsupcellsinthegitractofsivinfectedrhesusmacaques AT caojianhong itinsituitdetectionofgagspecificcd8supsupcellsinthegitractofsivinfectedrhesusmacaques AT vazquezjulio itinsituitdetectionofgagspecificcd8supsupcellsinthegitractofsivinfectedrhesusmacaques AT mcdonalddavid itinsituitdetectionofgagspecificcd8supsupcellsinthegitractofsivinfectedrhesusmacaques AT diemkurt itinsituitdetectionofgagspecificcd8supsupcellsinthegitractofsivinfectedrhesusmacaques AT laingkerry itinsituitdetectionofgagspecificcd8supsupcellsinthegitractofsivinfectedrhesusmacaques AT zhujia itinsituitdetectionofgagspecificcd8supsupcellsinthegitractofsivinfectedrhesusmacaques AT tjernlundannelie itinsituitdetectionofgagspecificcd8supsupcellsinthegitractofsivinfectedrhesusmacaques AT pickerlouisj itinsituitdetectionofgagspecificcd8supsupcellsinthegitractofsivinfectedrhesusmacaques AT coreylawrence itinsituitdetectionofgagspecificcd8supsupcellsinthegitractofsivinfectedrhesusmacaques |