Major Areas of Interest
- Development and application of novel imaging methods for analysis of tissue cells
- T cell dynamics in secondary lymphoid organs in natural infection (HIV/SIV) and after vaccination
- Generating “tissue imaging signatures” in cancer and cancer/HIV
The mission of the Tissue Analysis Core is the development, optimization and application of cutting-edge imaging assays for the analysis of human and nonhuman primate (NHP) tissues in support of VRC goals.
Imaging methodologies are a prerequisite for the understanding of:
- the spatial analysis of tissue immune cells and with respect to tissue structure
- the local interactions between immune cells, tissue stromal cells and tissue-expressed antigens (pathogens, cancer, self-antigens)
- the role of local inflammatory mediators for the dynamics of tissue immune cells
A comprehensive tissue analysis requires the application of complementary technologies allowing for the characterization of phenotype, localization, function and molecular signatures of cells under investigation. Merging cutting-edge technologies like multidimensional imaging, multiparameter flow cytometry and deep sequencing allows for the generation of “imaging signatures” that could provide information about viral persistence in specific tissue areas-sanctuaries and assist in the discovery of biomarkers for disease progression and interventional strategies with specific immunotherapies and vaccines.
Current technologies employed in the Tissue Analysis Core include:
- multispectral confocal imaging (A),Credit: NIAID
- RNAscope methodology that allows for the simultaneous detection of mRNA and several protein markers (B),Credit: NIAID
- live imaging and v) TIRF microscopy (C).Credit: NIAID
Several analytical tools (Imaris, Voloom, Histo-cytometry platform and in house developed algorithms) are used for the quantitative analysis of the obtained imaging objects.
Dr. Petrovas has been working in the field of HIV/SIV pathogenesis for more than 15 years. He obtained his Ph.D. in immunology studying anti-phospholipid antibodies in autoimmunity and HIV at the School of Medicine, National University of Athens, Greece. He pursued post-doctoral work at the Department of Biochemistry, Boston University and Department of Microbiology and Immunology, Drexel University before he joined the Immunology Laboratory at VRC in 2005. His previous work focused on the mechanisms mediating the “exhaustion” of SIV/HIV- specific CD8 T cells and he was the first to describe PD-1 as a mediator of SIV/HIV-specific CD8 T cell survival. More recently, his research has been focused on the dynamics of CD4 and CD8 T cells at tissue level and particularly in the lymph node follicles in HIV and SIV infection. His work was the first to describe the phenotype, localization, molecular profile and dynamics of Tfh CD4 T cells in acute and chronic SIV infection. Currently, he is characterizing the dynamics of tissue immune cells in HIV/SIV and cancer and developing tools to understand the interplay between inflammation, adaptive immunity and tissue antigens.
Constantinos Petrovas, Sara Ferrando-Martinez, Michael Y Gerner, Joseph P Casazza, Amarendra Pegu, Arik Cooper, Jason Hataye, Sarah Andrews, David Ambrozak, Perla M Del Río Estrada, Eli Boritz, Robert Paris, Eirini Moysi, Kristin L Boswell, Ezequiel Ruiz-Mateos, Ilias Vagios, Manuel Leal, Yuria Ablanedo-Terrazas, Amaranta Rivero, Luz Alicia Gonzalez-Hernandez, Adrian B McDermott, Gustavo Reyes-Teran, Fernando Docobo, Giuseppe Pantaleo, Daniel C Douek, Michael R Betts, Ronald N Germain, John R Mascola and Richard A Koup. Follicular CD8 T cells accumulate in HIV and kill infected cells via bispecific antibodies. Science Translational Medicine (in press).
Yamamoto T, Lynch RM, Gautam R, Matus-Nicodemos R, Schmidt SD, Boswell KL, Darko S, Wong P, Sheng Z, Petrovas C, McDermott AB, Seder RA, Keele BF, Shapiro L, Douek DC, Nishimura Y, Mascola JR, Martin MA, Koup RA. Quality and quantity of TFH cells are critical for broad antibody development in SHIVAD8 infection, Science Translational Medicine, 2015 Jul 29;7(298)
Boswell KL, Paris R, Boritz E, Ambrozak D, Yamamoto T, Darko S, Wloka K, Wheatley A, Narpala S, McDermott A, Roederer M, Haubrich R, Connors M, Ake J, Douek DC, Kim J, Petrovas C, Koup RA. Loss of circulating CD4 T cells with B cell helper function during chronic HIV infection, Plos Pathogen, 2014 Jan 30;10(1).
Cooper A, García M, Petrovas C, Yamamoto T, Koup RA, Nabel GJ. HIV-1 Causes CD4 Cell Death through DNA-Dependent Protein Kinase during Viral Integration, Nature, 2013 Jun 20;498(7454):376-9.
Petrovas C, Yamamoto T, Gerner MY, Boswell KL, Wloka K, Smith EC, Ambrozak DR, Sandler NG, Timmer KJ, Sun X, Pan L, Poholek A, Rao SS, Brenchley JM, Alam SM, Tomaras GD, Roederer M, Douek DC, Seder RA, Germain RN, Haddad EK, Koup RA. CD4 T follicular helper dynamics during SIV infection, J Clin Investigation, 2012 Sep 4; 122(9): 3281-94.
Petrovas C, Casazza JP, Brenchley JM, Price DA, Gostick E, Adams WC, Precopio ML, Schacker T, Roederer M, Douek DC, Koup RA. PD-1 is a regulator of virus-specific CD8+ T cell survival in HIV infection, Journal of Experimental Medicine, 2006 Oct 2; 203(10): 2281-92.