The long term goal of this project is to develop further a novel technique involving immunofluorescence in conjunction with flow cytometry. This technique will be validated by comparing data from the flow cytometric technique with that derived from the quantitative polymerase chain reaction (PCR) and infectivity assays. Validation of this novel flow cytometric technique will provide a relatively inexpensive and rapid method for detecting and quantitating the number of HIV-infected peripheral blood mononuclear cells (PBMC) in HIV- seropositive individuals. This technique will be used to monitor disease progression in HIV-infected individuals and monitor the effects of anti- retroviral chemotherapies on virus burden in these individuals.
The specific aims of this grant proposal are: 1. Validate the flow cytometric technique by comparing data obtained from flow cytometry with that derived from PCR and infectivity assays. 2. Use the flow cytometric technique to monitor patients who are receiving anti-retroviral chemotherapy such as ZDV, ddc and ddi. 3. To use the flow cytometric technique to identify the cells in the peripheral blood that are infected with HIV. 4. To use the flow cytometric to identify the cells in the peripheral blood that are infected with cytomegalovirus (CMV). Detection and quantitation of the PBMC infected with CMV may help elucidate the role that this opportunistic infection plays in the pathogenesis of AIDS. Successful completion of these studies will validate a technique that will make monitoring of the number of HIV-infected PBMC rapid and relatively inexpensive.
| McSharry, J J (2000) Analysis of virus-infected cells by flow cytometry. Methods 21:249-57 |
| McSharry, J J; Lurain, N S; Drusano, G L et al. (1998) Rapid ganciclovir susceptibility assay using flow cytometry for human cytomegalovirus clinical isolates. Antimicrob Agents Chemother 42:2326-31 |
| McSharry, J M; Lurain, N S; Drusano, G L et al. (1998) Flow cytometric determination of ganciclovir susceptibilities of human cytomegalovirus clinical isolates. J Clin Microbiol 36:958-64 |
| McSharry, J J (1994) Uses of flow cytometry in virology. Clin Microbiol Rev 7:576-604 |
