RAPID DIAGNOSIS OF HIV CELLULAR INFECTION

CLINICAL LABORATORY TESTS CURRENTLY EMPLOYED FOR HUMAN IMMUNODEFICIENCY VIRUS (HIV) ANTIBODY DETECTION MERELY INDICATE THE POSSIBILITY OF EXPOSURE TO THAT VIRUS. IN CONTRAST, THE PRECISE DIAGNOSIS OF HIV INFECTION RELIES UPON THE DEMONSTRATION OF ACTIVE, ISOLATABLE VIRUS OR THE PRESENCE OF SPECIFIC GENE SEQUENCES OR VIRAL ANTIGENS. ALTHOUGH SIGNIFICANT INFORMATION EXISTS REGARDING THE DISEASE-RELATED OCCURRENCE OF ANTIGENEMIA AND HIV GENE EXPRESSION, INSIGNIFICANT ATTENTION HAS BEEN GIVEN TO THE EXAMINATION OF ANTIGEN-EXPRESSION BY INFECTED CELLS. THE LONG-TERM GOAL OF THIS APPLICATION IS TO PROVIDE TECHNOLOGY FOR THE RAPID DIAGNOSIS OF HIV INFECTION AS BASED UPON THE DEMONSTRATION OF CELL-BOUND VIRAL ANTIGEN. THE SPECIFIC AIMS OF THE PRESENT WORK ARE TO SYSTEMATICALLY INVESTIGATE THOSE PARAMETERS ASSOCIATED WITH THE CONSTRUCTION OF SENSITIVE AND PRECISE IMMUNOCYTOCHEMICAL TESTS FOR HIV DETECTION. MURINE MONOCLONAL ANTIBODIES WHICHREACT WITH VARIOUS HIV CORE AND ENVELOPE COMPONENTS WILL BE USED AS PROBES. PARAMETERS TO BE STUDIED SHALL INCLUDE: (1) FIXATION EFFECTS; (2) PRE-ENRICHMENT OF ANTIGEN-POSITIVECELLS; (3) CYTOCHEMICAL AMPLIFICATION OF IMMUNOSTAINING; AND(4) THE USE OF MONOCLONAL ANTIBODIES SINGLY AND IN COMBINATIONS. RESULTS WILL BE COMPARED TO THOSE OBTAINED USING IN SITU HYBRIDIZATION WITH RNA PROBES, A TECHNIQUE PREVIOUSLY DEMONSTRATED TO DETECT HIV-INFECTED CELLS. THE PRE-CLINICAL UTILITY OF IMMUNOCYTOCHEMICAL TESTS WILL BEINVESTIGATED VERSUS A PANEL OF BLOOD CELL SPECIMENS OBTAINEDFROM PATIENTS INFECTED WITH HIV. THE POSSIBLE RELATIONSHIP BETWEEN CELLULAR ANTIGEN EXPRESSION AND DISEASE STAGE, LEVEL OF SERUM ANTIBODY, PRESENCE OF HIV GENESEQUENCES AND VIRAL ISOLATION DATA WILL BE EXAMINED.