Propagation of a Human Herpesvirus from AIDS-Associated Kaposi's Sarcoma
Kimberly E. Foreman, Ph.D., Jacques Friborg, Ph.D., Wing-pui Kong, Ph.D., Clive Woffendin, Ph.D., Peter J. Polverini, D.D.S., Brian J. Nickoloff, M.D., Ph.D., and Gary J. Nabel, M.D., Ph.D.
Background Although unique DNA sequences related to gammaherpesviruseshave been found in Kaposi's sarcoma lesions, it is uncertainwhether this DNA encodes a virus that is able to reproduce.
Methods We isolated and propagated a filterable agent whoseDNA sequences were found to be identical to those of the Kaposi'ssarcomaassociated herpesvirus (KSHV). We obtained early-passagespindle cells from skin lesions of patients with the acquiredimmunodeficiency syndrome (AIDS) who had Kaposi's sarcoma andcultured them with cells of the human embryonal-kidney epithelial-cellline 293. We characterized the virus according to its effectson cellular morphology and viral replication and its appearanceon electron microscopy.
Results KSHV was cytotoxic to 293 cells and was detected bythe polymerase chain reaction (PCR) in infected cells but notuninfected ones. Cytotoxicity and positive PCR signals wereconsistently maintained with viral titers of 1 million per milliliteror higher for about 20 serial infections of 293 cells. The viralcopy number was relatively low (1 to 10 copies per cell). Viralreplication was confirmed by Southern blot analysis of DNA isolatedfrom the enriched nuclear fraction of infected cells and bya semiquantitative PCR using dilutions of the lysates of infectedcells to detect the 233-bp viral DNA fragment originally describedin association with Kaposi's lesions. Electron microscopy revealedherpesvirus-like particles in about 1 percent of cells frominfected cultures, as compared with none in cells from uninfectedcultures.
Conclusions A herpesvirus with DNA sequences identical to thoseof KSHV can be propagated from skin lesions of patients withAIDS-associated Kaposi's sarcoma. (N Engl J Med 1997;336:163-71.)
Source Information
From the Department of Pathology, Skin Disease Research Laboratories, Cardinal Bernardin Cancer Center, Loyola University Medical Center, Maywood, Ill. (K.E.F., B.J.N.); and the Departments of Internal Medicine and Biological Chemistry (J.F., W.K., C.W., G.J.N.) and Oral Pathology (P.J.P.), Howard Hughes Medical Institute, University of Michigan Medical Center, Ann Arbor.
Address reprint requests to Dr. Nabel at University of Michigan Medical Center, MSRBI, Rm. 4520, 1150 W. Medical Center Dr., Ann Arbor, MI 48109-0650.
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