Abstract

The main goal of this proposal is to investigate the role of HCMV in the pathogenesis of KS associated with AIDS. Our approach is to determine if HCMV serves as a """"""""putative"""""""" cofactor in the induction of KS. Using subfragment probes of the transforming Towne XbaI-E fragment identified by us we propose to: 1) analyze KS tissue DNA for the presence, state, and expression of HCMV transforming DNA and myc-related sequences. The presence of these sequences will be determined by Southern blot hybridization and the expression by Northern blot hybridization. Molecular hybridization studies involving HCMV Towne XbaI-E will employ viral DNA probes lacking homology to cell DNA as well as probes of the myc-containing BamHI-J&M subfragments of the transforming Towne XbaI-E fragment. 2) Determine transforming activity in normal diploid SHE cells and NIH 3T3 cells of Towne Xbal-E and myc-related sequences (identified in #1 above). Transfection studies will be carried out employing KS tissue DNA in order to identify and isolate the transforming gene(s) and to determine any homology to the Towne XbaI-E transformed fragment. 3) Examine HCMV strainsisolated for altered or amplified transforming fragments and myc-related sequences. Restriction enzyme analysis will be used to assay structural alterations in DNA. Transfection assays in NIH 3T3 and SHE cells will be carried out to determine transforming activity. 4) Quantify HCMV genomes (copy number) in peripheral blood lymphocytes from patients groups (healthy homosexuals, AIDS patients without KS, and AIDS patients with KS). Since peripheral blood lymphocytes are an accessible tissue and have been shown to be a reservoir of virus in active HCMV infection, we propose to analyze these cells as useful source of the proposed HCMV cofactor for KS. Peripheral blood lymphocytes will be initially screened for the presence of HCMV sequences by spot hybridization using viral-specific DNA probes. The number of copies of viral genomes will be quatitated and the physical state (free or integrated) of viral genomes in peripheral blood lymphocytes will be investigated by Southern blot hybridization.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project--Cooperative Agreements (U01)
Project #
5U01CA037259-02
Application #
3548366
Study Section
(SSS)
Project Start
1984-04-01
Project End
1987-03-31
Budget Start
1985-04-01
Budget End
1986-03-31
Support Year
2
Fiscal Year
1985
Total Cost
Indirect Cost

  Institution

Name
Georgetown University
Department
Type
School of Medicine & Dentistry
DUNS #
049515844
City
Washington
State
DC
Country
United States
Zip Code
20057

  Publications

Muralidhar, S; Doniger, J; Mendelson, E et al. (1996) Human cytomegalovirus mtrII oncoprotein binds to p53 and down-regulates p53-activated transcription. J Virol 70:8691-700
Thompson, J; Choudhury, S; Kashanchi, F et al. (1994) A transforming fragment within the direct repeat region of human herpesvirus type 6 that transactivates HIV-1. Oncogene 9:1167-75
Thompson, J; Doniger, J; Rosenthal, L J (1994) A 79 amino acid oncogene is responsible for human cytomegalovirus mtrII induced malignant transformation. Arch Virol 136:161-72
Inamdar, A; Thompson, J; Kashanchi, F et al. (1992) Identification of two promoters within human cytomegalovirus morphologic transforming region II. Intervirology 34:146-53
Choudhury, S; Woodworth, C D; Inamdar, A et al. (1992) Differences in retention and expression of transfected human cytomegalovirus Towne XbaI-E transforming fragment in human cervical and NIH 3T3 lines. Intervirology 33:187-96
Jahan, N; Razzaque, A; Greenspan, J et al. (1989) Analysis of human KS biopsies and cloned cell lines for cytomegalovirus, HIV-1, and other selected DNA virus sequences. AIDS Res Hum Retroviruses 5:225-31
Jahan, N; Razzaque, A; Brady, J et al. (1989) The human cytomegalovirus mtrII colinear region in strain Tanaka is transformation defective. J Virol 63:2866-9
Jariwalla, R J; Razzaque, A; Lawson, S et al. (1989) Tumor progression mediated by two cooperating DNA segments of human cytomegalovirus. J Virol 63:425-8
Razzaque, A; Jahan, N; McWeeney, D et al. (1988) Localization and DNA sequence analysis of the transforming domain (mtrII) of human cytomegalovirus. Proc Natl Acad Sci U S A 85:5709-13
Sedarati, F; Rosenthal, L J (1988) Isolation and partial characterization of nucleocapsid forms from cells infected with human cytomegalovirus strains AD169 and Towne. Intervirology 29:86-100

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