The Virology Lab Core

Lawrence Corey, MD

The Virology Laboratory Core provides virologic, immunologic, and molecular diagnostic testing for HSV and HIV-1 in 3 laboratories: 1) The UW Virology Laboratory at Children's Hospital and Regional Medical Center ("UW-CHRMC"; Section Director: R. Ashley). This laboratory provides HSV and HIV-1 serology, HSV isolation, HSV and HIV-1 antiviral sensitivity testing, and specimen banking; 2) The UW-Molecular Virology Laboratory in the SCCA Building at Fred Hutchinson Cancer Research Center's (FHCRC) South Lake Union (SLU) campus (Section Director: L. Corey) performs HSV PCR testing and will develop a rapid HSV PCR test; 3) The UW Retrovirus Pathogenesis Laboratory in the Rosen building of the UW SLU campus provides HIV-1 testing to detect and quantify HIV-1 in tissues, cells and mucosal secretions (Section Director: S. Brodie).

These laboratories are well-established, accredited diagnostic laboratories; licensed by the State of Washington and certified by the College of American Pathologists and ACTG. The HSV Laboratories provide centralized support for defining subclinical and incident HSV infection using state-of-the-art antibody and HSV PCR testing. The Retrovirus Laboratory provides analyses for HIV-1 infection and replication in blood and mucosal lymphoid tissues and mucosal secretions using newly developed assays that quantitate the frequency and amount of HIV-1 expressed in HIV-1 infected cells. Dr. Koelle and Dr. Brodie's laboratories will collaborate to co-localize HIV-1 and HSV in mucosal and HSV skin lesions in parallel with measurements of cytokines, chemokines and hormone receptor proteins. All Core laboratories share basic protocols for specimen transport, accessioning, tracking, and resulting. Computerized database entry is standardized and coordinated with Statistical Core C for data analyses.

The Lab Core consists of 5 specific aims:

Specific Aim 1. To develop HSV serologic algorithms that accurately identify subclinical HSV infection and incident HSV infection at the U.S. and international clinical trial sites (Peru, Zambia, Zimbabwe, Cameroon, Seattle and New York City) and to provide QA/QC support for sites performing screening HSV and HIV-1 serologies assays.

Specific Aim 2. To provide centralized laboratory reference support to confirm HSV serologic status and incident HSV and HIV-1 infections.

Specific Aim 3. To develop a sensitive and specific "rapid" HSV PCR assay. To implement this assay within the laboratory support services to the Departments of Obstetrics at the University of Washington.

Specific Aim 4 . To quantitate HIV-1 DNA and RNA in lymphoid tissue, mucosal samples, plasma and purified populations of T cells in HSV-HIV-1 co-infected persons. To measure the replication rate and evolution of HIV-1 quasispecies prior to and after acyclovir. To perform in situ hybridization to co-localize HSV and HIV-1 infection in tissue.

Specific Aim 5. To perform immunocytochemisty on tissue to quantitate chemokine and hormone receptor expression in lesional biopsies

Recent Peer Reviewed Publications:

•  Posavad CM, Koelle DM, Corey L. Tipping the scales of herpes simplex virus reactivation: The important responses are local. Nature Med 1998; 4:381-82.

•  Langenberg AGM, Corey L, Ashley RL, Leong WP, Straus SE, for the Chiron HSV Vaccine Study Group. A prospective study of new infections with herpes simplex virus type 1 and herpes simplex virus type 2. N Engl J Med 1999; 341:1532-38.

•  Ashley RL, Wald A, Eagleton M. Premarket evaluation of the POCkit TM -HSV-2 type specific serologic test in culture-documented cases of genital herpes simplex virus type 2. STD 2000; 26:266-69

•  Corey L and Handsfield HH. Genital herpes and public health: addressing a global problem. J Amer Med Assoc 2000;283:791-4

•  Koelle DM, Reymond SN, Chen H, Kwok WW, McClurkan C, Gyaltsong T, Petersdorf EW, Rotkis W, Talley AR, Harrison DA. Tegument-specific, virus-reactive CD4 T cells localize to the cornea in herpes simplex virus interstitial keratitis in humans. J Virol 2000; 74:10930-38.

•  Koelle DM, Schomogyi M, Corey L. Antigen-specific t cells localize to the uterine cervix in women with genital herpes simplex virus type 2 infection. J Infect Dis 2000; 182:662-70.

•  Koelle DM, Schomogyi M, McClurkan C, Reymond SN, Chen HB. CD4 T-cell responses to herpes simplex virus type 2 major capsid protein VP5: comparison with responses to tegument and envelope glycoproteins. J Virol 2000; 74:11422-25.

•  Krone MR, Wald A, Tabet SR, Paradise M, Corey L, Celum CL. HSV-2 shedding in HIV-negative men who have sex with men: frequency, patterns, and risk factors. Clin Infect Dis 2000; 30:261-67.

•  Kwok WW, Liu AW, Novak EJ, Gebe JA, Reymond SN, Ettinger RA, Nepom GT, Koelle DM. HLA-DQ tetramers identify epitope-specific T-cells in peripheral blood of herpes simplex virus-2 (HSV-2)-infected individuals: direct detection of immunodominant antigen responsive cells. J Immunology 2000; 164:4244-49.

•  Lafferty WE, Downey L, Celum C, Wald A. HSV-1 as a cause of genital herpes: impact on surveillance and prevention. J Infect Dis 2000; 181:1454-57.

•  Mostad MB, Kreiss JK, Ryncarz AJ, Mandaliya K, Chohan B, Ndinya-Achola J, Bwayo JJ, Corey L. Cervical shedding of herpes simplex virus in human immunodeficiency virus-infected women: effects of hormonal contraception, pregnancy, and vitamin A deficiency. J Infect Dis 2000; 181:58-63

•  Pauk J, Huang ML, Brodie SJ, Wald A, Koelle DM, Schacker T, Celum C, Selke S, Corey L. Mucosal shedding sites and patterns of human herpesvirus 8 in men. NEJM 2000; 343:1369-77.

•  Posavad CM, Barcy S, Huang ML, Koelle DM, Polyak SJ, Corey L.Long-term persistence of herpes simplex virus-specific CD8+ CTL clones derived from genital lesions. J Immunol 2000;165:1146-1152.

•  Wald A, Zeh J, Selke S, Ryncarz A, Ashley R, Corey L. Reactivation of genital herpes simplex virus type-2 infection in asymptomatic seropositive persons. NEJM 2000; 342:844-50

•  Ashley RL. Sorting out the new HSV type specific antibody tests. Sex Transm Inf 2001;77:232-7.

•  Barcy S and Corey L. Herpes simplex inhibits the capacity of lymphoblastoid B cell lines to stimulate CD4+ T cells. J Immunology 2001; 166:4242-49.

•  Koelle DM, Chen H, Gavin MA, Wald A, Kwok WW, Corey L. CD8 CTL from genital herpes simplex lesions: recognition of viral tegument and immediate early proteins and lysis of infected cutaneous cells. J Immunol 2001; 166:4049-58.

•  Sucato G, Celum C, Dithmer D, Ashley R, Wald A. Demographic rather than behavioral risk factors predict herpes simplex virus type 2 infection in sexually active adolescents. Ped Infect Dis J 2001;20:422-6.

•  Vieira J, O'Hearn P, Kimball L, Chandran B, Corey L. Activation of Kaposi's sarcoma-associated herpesvirus (Human Herpesvirus 8) lytic replication by human cytomegalovirus. J Virol 2001;75:1378-86

•  Wald A, Langenberg AGM, Link K, Izu A, Ashley R, Warren T, Tyring S, Douglas JM, Corey L. Effect of condoms on reducing the transmission of herpes simplex virus type-2 from men to women. JAMA 2001; 285:3100-06

•  Wald A, Link K. Risk of human immunodeficiency virus (HIV) infection in herpes simplex virus type-2 (HSV-2) seropositive persons: a meta-analysis. J Infect Dis 2001; 185:45-52

•  Whittington WLH, Celum C, Cent A, Ashley R. Use of a glycoprotein G-based type-specific assay to detect antibodies to herpes simplex virus type 2 among persons attending sexually transmitted disease clinics. Sex Trans Dis 2001;28:99-104.