Molecular cloned SIVmac32H vaccine and challenge virus. NLM AIDSLINE Important note: Information in this article was accurate in 1993. The state of the art may have changed since the publication date.

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Molecular cloned SIVmac32H vaccine and challenge virus.

Symp Nonhum Primate Models AIDS. 1992 Nov 17-20;10:abstract no. 127. Unique Identifier : AIDSLINE PRIM10/93200967
Rud EW; Yon JR; Quirk J; Corbin AY; Clarke BE; Mackett M; Stott EJ; Kent KA; Corcoran T; Cook N; et al; Wellcome Research Laboratories, Dept. of Mol. Sci., Beckenham,; Kent, U.K.

Abstract: We molecularly cloned, sequenced and titrated genomic clones of the SIVmac32H vaccine challenge virus used by the British MRC AIDS Directed Programme. The env gene was subcloned out of the full length molecular clone, SIVmac32H(pJ5) and inserted into various expression vectors. The molecular clones of SIVmac32H were selected out of a lambda library made from SIVmac32H(11/88) infected cells. The resulting full length clone SIVmac32H(pJ5) was fully sequenced and transfected into C8166 cells to produce a molecularly defined vaccine challenge virus (9/90). The 9/90 pool was further passaged on macaque PBLs to produce a challenge virus free of C8166 cell components (J5m, 3/92 pool). The ability of these viruses to infect rhesus and cynomolgus macaques was assessed and the MMID50 and TCID50 was determined by inoculating serial dilutions of the cell free virus into rhesus macaques and C8166 cells, respectively. The molecular clone SIVmac32H(pJ5) contains no premature termination codons nor internal deletions in any of the known SIV open reading frames. The 9/90 pool of virus originating from SIVmac32H(pJ5) had a MMID50 of 10(6) and a TCID50 of 5 x 10(5). Both rhesus and cynomolgus macaques are infected by this virus as determined by virus recovery, PCR analysis and seroconversion. The 3/92 pool of J5m had a MMID50 of 10(4) and a TCID50 of 10(3.8). The gp160 region of pJ5 has been subcloned into vaccinia and the resulting expressed product reacts with both linear and conformational monoclonal antibodies. The gp160 sequence is essentially identical to the consensus sequence of the SIVmac32H(11/88) uncloned virus pool as determined by PCR analysis. This molecularly defined challenge virus and expressed gene products subcloned out of SIVmac32H(pJ5) are being used in a subunit vaccine trial by the British MRC AIDS directed programme. This virus will enable more precise characterization of the important immune responses required for vaccine induced protection from SIV.
Keywords: Animal Antibodies, Monoclonal Cell Line Cloning, Molecular/METHODS Codon Gene Products, env/GENETICS/IMMUNOLOGY *Genes, env Macaca fascicularis Macaca mulatta Open Reading Frames Simian Acquired Immunodeficiency Syndrome/*IMMUNOLOGY/PREVENTION & CONTROL SIV/GENETICS/*IMMUNOLOGY Transfection Vaccines, Synthetic/*GENETICS Viral Vaccines/*GENETICS ABSTRACTKWDanimalantibodies,monoclonalcelllinecloning,molecular/methodscodongeneproducts,env/genetics/immunologyKWDgenes,envmacacafascicularismacacamulattaopenreadingframessimianacquiredimmunodeficiencysyndrome/KWDimmunology/prevention&controlsiv/genetics/KWDimmunologytransfectionvaccines,synthetic/KWDgeneticsviralvaccines/KWDgeneticsabstract
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Copyright © 1993 - National Library of Medicine. Reproduced under license with the National Library of Medicine, Bethesda, MD.

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