Kaposi's sarcoma-associated herpesvirus (KSHV) is etiologically associated with AIDS and non-AIDS Kaposi's sarcoma (KS), primary effusion lymphoma, and multicentric Castleman disease. Since its discovery in 1994, the entire virus genome has been successfully sequenced; however, the biology of KSHV infection with a special emphasis to the reactivation of KSHV latency is far from being completely understood. This is due in part to the lack of a system that can monitor different aspects (lytic and latent replication cycle) of KSHV infection in Real time. Such a Real time monitoring of infection can be possible only by a non-invasive procedure. In here, we propose to examine the KSHV infection of cells by Raman tweezers. Raman tweezers involves a confocal microscopy which incorporates the use of both laser (optical) tweezers and Raman spectroscopy (LTRS). This technique can provide biochemical composition of a single living cell without chemically interfering with it and the measured vibrational energy levels can be used as fingerprints for identification of the biological cells. We propose to use a combination of Raman tweezers and biochemical studies to analyze the KSHV lytic and latent infection of target cells. The results from this proposal will serve as a basis for future studies that will be primarily focused on using Raman tweezers as a diagnostic tool to identify KSHV and other pathogens in human blood samples. Akula, Shaw, Muthukrishna PROJECT NARRATIVE Kaposi's sarcoma-associated herpesvirus (KSHV) or human herpesvirus-8 (HHV-8) causes several cancer disease conditions like Kaposi's sarcoma (KS), primary effusion lymphoma (PEL), and multicentric Castleman disease (MCD). KSHV infects people of all races around the world. For KSHV to cause the disease condition, it has to establish an infection. Upon infection, KSHV lays dormant (latent) in the cell; however, there is a small population (1-3%) of these cells that support an active replication (lytic) leading to cell death and release of new viruses. This process of terminating KSHV dormancy and proceeding to production of new viruses is not well understood. This is due in part to the lack of a system that can monitor different aspects (lytic and latent replication cycle) of KSHV infection in Real time. Such a Real time monitoring of infection can be possible only by a non-invasive procedure. In here, we propose to examine the KSHV infection of cells by Raman tweezers, an instrument based of the principles of spectroscopy. This technique can provide biochemical composition of a single living cell without chemically interfering with it and the measured vibrational energy levels can be used as fingerprints for identification of the biological cells. We propose to use a combination of Raman tweezers and biochemical studies to analyze the KSHV lytic and latent infection of target cells. Realization of this study, (1) will help us understand the biology of KSHV infection and pathogenesis; (2) will serve as a model to understand latent virus infections in general; and (3) will definitely open the door for developing novel strategies to diagnose virus infections efficiently and quickly. ? ? ?
