The role of purinergic receptor signaling in the establishment of HIV latency

• Applicant: Dr. Ilona Tóth
• Subject Area: Virology ; Immunology
• Term: from 2015 to 2017
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 279339224

This application proposes a project to examine the role of purinergic receptor signaling in HIV infection. In particular, the role of ATP and adenosine signaling in the HIV replicative cycle and in the establishment of viral latency will be analyzed. To investigate the role of purinergic receptors (P1 for adenosine, P2X and P2Y for ATP) in HIV entry, replication and latency, in vitro infection assays of healthy, human peripheral blood mononuclear cells with wildtype HIV (JRFL) and reporter HIV virions (R7GEmC, pMM310) are planned in combination with specific purinergic receptor agonists and antagonists. (Provided by Prof. Jacobson, Molecular Recognition Section, NIH). To investigate the role of purinergic receptor signaling in the clinical course of HIV infection,whole transcriptome analysis using next generation sequencing will be carried out on single sorted, HIV-infected CD4+ T cells from HIV-infected people at different stages of disease, both on and off combination anti-retroviral therapy (cART). A droplet-based microfluidic cell sorting procedure for the identification and isolation of HIV-infected cells is already established in the Douek Laboratory. Samples of peripheral blood, lymph node and gut mucosa will be obtained through Dr. Doueks ongoing collaborations with investigators at the University of California San Francisco (Dr. Steven Deeks), Case Western Reserve University (Dr. Michael Lederman), The University of Minnesota (Dr. Timothy Schacker) and through the NIH clinic. Currently, longitudinal samples are available from over 100 HIV-infected individuals. The experimental work will be conducted at the National Institute of Allergy and Infectious Diseases (NIAID) at the campus of the National Institutes of Health (Bethesda, Maryland) in the Human Immunology Section of the Vaccine Research Center (VRC), headed by Daniel Douek M.D., MRCP, Ph.D. Adequate S3 facilities and technical equipment for handling and single-sorting of HIV infected materials and whole transcriptome analysis are accessible in the Genome Analysis Core of the VRC, headed by Daniel Douek.

• DFG Programme: Research Fellowships
• International Connection: USA
• Host: Dr. Daniel Douek