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| Status |
Public on Sep 06, 2011 |
| Title |
Colorectal tissue gene expression in SIV negative and SIV positive Rhesus macaques and sooty mangabeys |
| Platform organism |
Macaca mulatta |
| Sample organisms |
Cercocebus atys; Macaca mulatta |
| Experiment type |
Expression profiling by array
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| Summary |
In SIV/HIV infection, the gastrointestinal tissue dominates as an important site due to the impact of massive mucosal CD4 depletion and immune activation-induced tissue pathology. Unlike AIDS-susceptible rhesus macaques, natural hosts do not progress to AIDS and resolve immune activation earlier. Here, we examine the role of dendritic cells in mediating immune activation and disease progression. We demonstrate that plasmacytoid dendritic cells (pDC) in the blood upregulate β7-integrin and are rapidly recruited to the colorectum following a pathogenic SIV infection in rhesus macaques. These pDC were capable of producing proinflammatory cytokines and primed a Tc1 response in vitro. Consistent with the upregulation of β7-integrin on pDC, in vivo blockade of α4β7-integrin dampened pDC recruitment to the colorectum and resulted in reduced immune activation. The upregulation of β7-integrin expression on pDC in the blood was also observed in HIV-infected humans but not in chronically SIV-infected sooty mangabeys that show low levels of immune activation. Our results uncover a new mechanism by which pDC influence immune activation in colorectal tissue following pathogenic immunodeficiency virus infections.
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| Overall design |
SIV negative controls (n=4) and week 12 post SIV infected (n=4) groups of Rhesus macaques and SIV negative controls (n=4) and week 55 post SIV infected (n=4) groups of Sooty mangabeys colorectal tissue biopsies were collected in to RNA later reagent (Qiagen) and were homogenized with syringe and needle method. RNA was extracted with Rneasy mini kit (Qiagen) and was used for microarray experiments. Rhesus GeneChip assays were performed in the Yerkes Microarray Core Facility (www.microarray.emory.edu) , one of the Affymetrix Microarray Core Labs.The 0.5µg of total RNA sample was analyzed on Rhesus Macaque Genome GeneChip that consists of over 52,000 probe sets (Affymetrix, Santa Clara, CA). Target RNA labeling, hybridization and post-hybridization processing were performed following the Affymetrix GeneChip Expression Analysis standard protocols. In brief, The 5 μg of RNA sample was first reverse-transcribed using T7-Oligo(dT) Promoter Primer and SuperScript II in the first-strand cDNAs synthesis reaction. Following RNase H-mediated second-stranded cDNA synthesis, the double-stranded cDNAs were purified by use of a GeneChip sample clean-up module and served as templates in the generation of biotinylated complementary RNAs (cRNAs) in the presence of T7 RNA Polymerase and a biotinylated nucleotide analog/ribonucleotide mix by in vitro transcription (IVT) reaction. The biotinylated cRNAs were cleaned up, fragmented, and hybridized to the rhesus macaque expression arrays at 45°C for 16 h with constant rotation at 60 rpm. The gene chips were then washed and stained with Affymetrix fluidics stations 450 and scanned on Affymetrix scanner 3000. The images are processed to collect raw data with GeneChip Operating Software (GCOS) 1.4.
Tissue: Colorectal tissue
Time after SIV infection: 12 weeks for SIV infected Rhesus macaques, 55 weeks for Sooty mangabeys
Infection: SIVmac251 infection for Rhesus macaques, SIVsm infection for Sooty mangabeys
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| Contributor(s) |
Dyavar-Shetty R, Amara R |
| Citation(s) |
21693759 |
| Submission date |
Jun 15, 2011 |
| Last update date |
Jul 18, 2012 |
| Contact name |
RAVI DYAVAR SHETTY |
| E-mail(s) |
rdyavar@emory.edu
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| Phone |
404-727-9233
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| Organization name |
emory university
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| Department |
Microbiology and Immunology
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| Lab |
Rama Amara
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| Street address |
954 Gatewood RD NE
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| City |
Atlanta |
| State/province |
Georgia |
| ZIP/Postal code |
30329 |
| Country |
USA |
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| Platforms (1) |
| GPL3535 |
[Rhesus] Affymetrix Rhesus Macaque Genome Array |
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| Samples (16)
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| Relations |
| BioProject |
PRJNA140767 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE29980_RAW.tar |
76.6 Mb |
(http)(custom) |
TAR (of CEL) |
| Processed data included within Sample table |
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