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User name	BÃ©atrice Jacquelin
Email	beatrice.jacquelin@pasteur.fr
Phone / Fax number	0145688792 /
Location	Institut Pasteur (UnitÃ© de RÃ©gulations des Infections RÃ©trovirales) - 25-28, rue du Docteur Roux - Paris, France

Scientific description

SIVagm-infected African green monkeys (AGM) do not develop chronic T cell activation and AIDS, despite similar viral loads as HIV-1 infected humans and SIVmac-infected rhesus macaques (RM). To understand the underlying molecular basis, we longitudinally assessed the gene expression profiles of blood- and lymph node (LN)-derived CD4+ cells from AGMs and RMs in response to SIVagm and SIVmac infection, respectively. Gene expression was analyzed using human long oligonucleotide arrays which return a robust signal with CD4+ cell RNA from AGM (>13,500 genes) and RM (>14,500 genes). The molecular signature of acute infection was characterized by strong up-regulation of type I Interferon-Stimulated Genes (ISG). ISG expression returned to basal level after day 28 post-infection in AGMs but was sustained in RMs, especially in LN. ISG are a surrogate marker for IFN-alpha activity. Our results suggest that IFN-alpha levels and/or the associated responses are rapidly controlled in AGM. This control might be involved in the weak inflammatory response that characterizes primary SIVagm infection.

Technical description

In this serie, there are six AGM female (Chlorocebus sabaeus from Barbados), 3 years old, weighing 2.8â€“3.4 kg. AGM were intravenously infected with 250 TCID50 of SIVagm_sab92018 on day 0. 10ml of whole blood was collected from those animals under anaesthesia in heparinized tubes at 4 time points before infection (days -90, -70, -40 and -8) to get a robust baseline, 4 time points during the acute phase of the infection (days 1, 6, 14, 28) and 3 time points during the chronic phase (days 41, 65 and 115). PBMC were isolated freshly from blood samples by Ficoll procedures. Biopsies of peripheral Lymph nodes were performed by excision at 2 time points before infection (days -90, -8) and 6 time points post-infection for AGMs (days 1, 6, 14, 28, 65, 590). After careful removal of adhering connective and fat tissues, the LN fragments were mechanically disrupted on a sterile stainless steel mesh and filtered to collect LN mononuclear cells (LNMC). The recovered PBMCs and LNMCs were then washed, counted, and the the CD4+ cell subset was further isolated from those tissues using the MACS magnetic labeling system (Miltenyi Biotec, Germany) according to instructions of the manufacturer. Isolated cells consists of >95% of CD4+ T lymphocytes (CD3+ CD4+ cells). The total RNA was extracted and analyzed on the ABI Human Whole Genome Arrays v2.0. With the ABI array protocol 600ng of total RNA were used to generate sufficient hybridization signal.

References

Jacquelin B, Mayau V, Targat B, Liovat AS, Kunkel D, Petitjean G, Dillies MA, Roques P, Butor C, Silvestri G, Giavedoni LD, Lebon P, BarrÃ©-Sinoussi F, Benecke A, Muller-Trutwin M (2009) Strong but Rapidly Controlled Interferon Type I Response in Nonpathogenic SIV Infection of African Green Monkeys. J. Clinical Invest. doi:10.1172/JCI40093. in the press.

-- see also published comment: Manches O, Bhardwaj N (2009) Resolution of immune activation defines nonpathogenic SIV infection. J. Clinical Invest. doi:10.1172/JCI41509.

Pubmed : http://www.ncbi.nlm.nih.gov/pubmed/19959873

Jacquelin B, Mayau V, Targat B, Liovat AS, Kunkel D, Petitjean G, Dillies MA, Roques P, Butor C, Giavedoni LD, Lebon P, BarrÃ©-Sinoussi F, Benecke A, Muller-Trutwin M (2009) Inflammatory control in AIDS-resistant non human primates. Retrovirology. 6:S2:123.