AEGiS-11IAC: The infectious nature of the novel herpesvirus-like DNA sequences detected in Kaposi's sarcoma.

11th International AIDS Conference

Vancouver, British Columbia — July 7-12, 1996

The infectious nature of the novel herpesvirus-like DNA sequences detected in Kaposi's sarcoma.

Int Conf AIDS 1996 Jul 7-12; 11:215 (abstract no. Th.A.273)
Blackbourn DJ, Ambroziak JA, Lennette ET, Nelson J, Northfelt DW, Gullett J, Glogau RG, Levy JA; Cancer Research Institute, University of California, San Francisco, CA, USA. Fax: (415) 476-8365.

OBJECTIVE: To determine whether the Kaposi's sarcoma (KS)-associated novel gammaherpesvirus-like DNA sequences are associated with an infectious agent.

METHODS: The novel herpesvirus DNA sequences were detected by the polymerase chain reaction (PCR) according to Chang et al. (Science 266: 1865; 1994). The putative virus represented by these DNA sequences is referred to as human herpesvirus 8 (HHV8). HHV8 PCR positive CD19+ cells from HIV-seropositive KS patient peripheral blood mononuclear cells (PBMC) were activated in culture. KS tumor samples were obtained by punch biopsy and cocultured with either PBMC, CD19+ cells or umbilical cord blood MC (UBMC). Filtered (0.45 micromolar) fluids from these cultures were inoculated onto target cells which were either HHV8-uninfected adult CD19+ cells, UBMC or the HPV-transformed endothelial cell line, BB19. After three to nine days of culture, these target cells were tested by diagnostic HHV8 PCR.

RESULTS: Inoculation of filtered culture fluids from activated CD19+ cells from each of three KS patients onto either adult CD19+ or UBMC target cells resulted in an HHV8 PCR positive signal. Furthermore, inoculation of either adult CD19+ or UBMC target cells with pooled coculture fluids from KS biopsies from two patients also yielded an HHV8 PCR positive signal. When pooled coculture fluid from the biopsies of one patient was diluted 100-fold and inoculated onto adult CD19+ cells, an HHV8 PCR positive signal could still be detected. In addition, the HPV-transformed human endothelial cell line BB19 was found to be susceptible to HHV8 infection by inoculation with filtered cell culture fluids derived from either a cocultured KS biopsy or activated CD19+ cells from a patient.

CONCLUSIONS: These data provide biological evidence for the infectious nature of HHV8 and for the susceptibility of an endothelial cell line to HHV8 infection.

Keywords: AEGIS, Sarcoma, Kaposi, Herpesvirus 8, Human, Polymerase Chain Reaction, Simplexvirus, Papillomavirus, Human, Coculture, Cell Line, Transformed, Biopsy, Adult, Human, ICA11

960707
ThA273