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Received Oct 7; Accepted Mar Abstract SIVsmmPBj14 is a highly pathogenic lentivirus which causes acute diarrhea, rash, massive lymphocyte proliferation predominantly in the gastrointestinal tract, and death within 7 to 14 days.
In cell culture, the virus has mitogenic effects on resting macaque T lymphocytes. In a previous study, replacement of amino acid residues 17 and 18 of the Nef protein of SIVmac with the corresponding amino acid residues of the Nef protein of SIVsmmPBj14 yielded a PBj-like virus that caused extensive activation of resting T lymphocytes in cultures and acute PBj-like disease when inoculated into pig-tailed macaques.
This study suggested that nef played a major role in both processes. However, when inoculated into macaques, the virus failed to replicate productively or cause disease. Infection by each virus is initiated by binding of the viral glycoprotein to specific receptors on target cells; this is followed by internalization of the virus, reverse transcription of the viral genome, and integration of viral DNA into the host cell DNA However, the productive phase of replication is dependent on the ability of the virus or other immunological factors to cause activation of the latently infected cell.
The mechanisms of the activation process are not known. In macaques inoculated with SIVmac or SHIVKU-1, activation of infected cells develops predictably within 1 week and is reflected by the onset of viremia, extravasation of infected cells from the blood to other tissues such as the brain, and the ability of cultured peripheral blood mononuclear cells PBMC to produce virus without treatment of the cells with a mitogen 1113 Therefore, the mechanisms causing activation of the cells in vivo and in cell cultures seem distinct.
Under culture conditions, this virus has a mitogenic effect as indicated by incorporation of [3H]thymidine, proliferation of the cells, and production of virus.
Protracted diarrhea develops, and this results in the death of the animals within 2 weeks 7 — 9. Site-directed mutagenesis of SIVmac nef endowed this virus with the novel ability to cause activation of the cells in vitro and also the hyperacute PBj-like disease in the animals 3.
We had passaged SHIV-4 sequentially in animals and derived a highly virulent virus stock 1213 The virus was highly mitogenic in resting PBMC, and infected cells produced virus in culture. The new virus was eliminated more rapidly by the two inoculated macaques than was the SHIVPPc, which was inoculated into two other animals.
These data show that the mechanisms by which SHIV causes activation and productive infection in resting PBMC are different from those that cause the same end result in vivo. Whether the dichotomy of biological effects of this virus in cell culture versus in vivo is due to its HIV-1 envelope is not known, but the phenomenon posed the question of whether the pathogenic mechanisms of SIV are different from those of HIV The C cell line was used as the source of indicator cells with which to measure the infectivity and cytopathicity of the viruses used in this study.
Virus suspensions were assayed for infectivity by the use of the fusion cytopathicity technique in C cells, and fresh stocks were prepared by inoculating the same cells at a multiplicity of infection MOI of 0. Serum was obtained from a macaque that had been inoculated with SHIVKU-1 and had developed neutralizing antibodies to the virus.
This serum had antibody titers of less than 1: Normal macaque serum did not neutralize any of the viruses. The resulting amplified fragment contained an NcoI site at the beginning of the nef gene in addition to an internal NcoI site.This file contains additional information, probably added from the digital camera or scanner used to create or digitize it.
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