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J Virol, February 1998, p. 1431-1437, Vol. 72, No. 2
0022-538X/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Identification of Highly Attenuated Mutants of Simian Immunodeficiency Virus

Ronald C. Desrosiers,1,* Jeffrey D. Lifson,2 James S. Gibbs,1 Susan C. Czajak,1 Anita Y. M. Howe,1 Larry O. Arthur,2 and R. Paul Johnson1,3

New England Regional Primate Research Center, Harvard Medical School, Southborough, Massachusetts 017721; NCI-Frederick Cancer Research and Development Center, Frederick, Maryland 217012; and Infectious Disease Unit and Partners AIDS Research Center, Massachusetts General Hospital, Boston, Massachusetts 021153

Received 11 August 1997/Accepted 22 October 1997

Deletion mutants of the pathogenic clone of simian immunodeficiency virus isolate 239 (SIVmac239) were derived that are missing nef, vpr, and upstream sequences (US) in the U3 region of the LTR (SIVmac239Delta 3), nef, vpx, and US (SIVmac239Delta 3x), and nef, vpr, vpx, and US (SIVmac239Delta 4). These multiply deleted derivatives replicated well in the continuously growing CEMx174 cell line and were infectious for rhesus monkeys. However, on the basis of virus load measurements, strength of antibody responses, and lack of disease progression, these mutants were highly attenuated. Measurements of cell-associated viral load agreed well with assays of plasma viral RNA load and with the strengths of the antibody responses; thus, these measurements likely reflected the extent of viral replication in vivo. A derivative of SIVmac239 lacking vif sequences (SIVmac239Delta vif) could be consistently grown only in a vif-complementing cell line. This Delta vif virus appeared to be very weakly infectious for rhesus monkeys on the basis of sensitive antibody tests only. The weak antibody responses elicited by SIVmac239Delta vif were apparently in response to low levels of replicating virus since they were not elicited by heat-inactivated virus and the anti-SIV antibody responses persisted for greater than 1 year. These results, and the results of previous studies, allow a rank ordering of the relative virulence of nine mutant strains of SIVmac according to the following order: Delta vpr > Delta vpx > Delta vprDelta vpx congruent  Delta nef > Delta 3 > Delta 3x >=  Delta 4 > Delta vif > Delta 5. The results also demonstrate that almost any desired level of attenuation can be achieved, ranging from still pathogenic in a significant proportion of animals (Delta vpr and Delta vpx) to not detectably infectious (Delta 5), simply by varying the number and location of deletions in these five loci.

* Corresponding author. Mailing address: New England Regional Primate Research Center, Harvard Medical School, One Pine Hill Dr., Box 9102, Southborough, MA 01772-9102. Phone: (508) 624-8042. Fax: (508) 624-8190. E-mail: rdesrosi{at}warren.med.harvard.edu.




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