title: Biphasic in vitro regulation of retroviral replication by CD8+ cells from nonhuman primates.
abstract: CD8+ T cells from naturally infected disease-resistant sooty mangabeys (Cercocebus atys) secrete a soluble factor which inhibits the in vitro replication of the simian immunodeficiency virus (SIV). To gain further insight on the mechanism(s) involved, CD8+ effector T cells and target cells from sooty mangabeys were immortalized and cloned. The target cells were then stably transfected with an SIV-LTR-CAT construct or with the parental CAT plasmid as a control. A quantitative RT-PCR method, providing the necessary sensitivity, was developed to monitor the influence of the cloned CD8+ T cells on the CATmRNA contained in the target cells. It could be demonstrated that a soluble factor was secreted by the cloned CD8+ T cells from sooty mangabeys, which appeared to regulate CATmRNA activity in a dose-dependent and reversible manner. Kinetic experiments showed that the CATmRNA transcriptional activity was initially augmented at 30 min postcoculture and was followed by a marked decrease in transcriptional activity after a few hours. This immediate early response could be mitigated utilizing H7, Calmodulin, or PDTC (a pyrrolidone derivative of dithiocarbamate), suggesting that the pathway was protein kinase-dependent and that the NF-kappa B site may be involved. The inhibitory effect could also be overcome using a protein synthesis inhibitor, suggesting that protein synthesis was needed to negatively regulate CATmRNA activity and hence SIV promoter activity.
authors.affiliation: Department of Pathology, Emory University School of Medicine, Atlanta, GA.
authors.forename: M
authors.initials: M
authors.lastname: Knuchel
authors.forename: D P
authors.initials: DP
authors.lastname: Bednarik
authors.forename: N
authors.initials: N
authors.lastname: Chikkala
authors.forename: A A
authors.initials: AA
authors.lastname: Ansari
mesh_terms: D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D000199:Actins / Q000235:genetics; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001483:Base Sequence; D016827:CD8 Antigens / Q000276:immunology*; D002147:Calmodulin / Q000494:pharmacology; D002460:Cell Line; D002461:Cell Line, Transformed; D016665:Cercocebus atys; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D002999:Clone Cells; D017931:DNA Primers / Q000737:chemistry; D007546:Isoquinolines / Q000494:pharmacology; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000737:chemistry; D010879:Piperazines / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D047428:Protein Kinase Inhibitors; D011759:Pyrrolidines / Q000494:pharmacology; D012333:RNA, Messenger / Q000737:chemistry; D015302:Simian Immunodeficiency Virus / Q000502:physiology*; D050378:T-Lymphocytes, Regulatory / Q000187:drug effects / Q000276:immunology / Q000382:microbiology*; D013859:Thiocarbamates / Q000494:pharmacology; D014158:Transcription, Genetic; D014162:Transfection; D014779:Virus Replication / Q000276:immunology*
chemical_list: D000199:Actins; D000970:Antineoplastic Agents; D016827:CD8 Antigens; D002147:Calmodulin; D017931:DNA Primers; D007546:Isoquinolines; D015345:Oligonucleotide Probes; D010879:Piperazines; D047428:Protein Kinase Inhibitors; D011759:Pyrrolidines; D012333:RNA, Messenger; D013859:Thiocarbamates; C020972:pyrrolidine dithiocarbamic acid; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D015500:Chloramphenicol O-Acetyltransferase