Antisense oligodeoxynucleotides as probes of T-lymphocyte gene function.
Conventional and thiophosphonate-derivatized oligonucleotides were employed to specifically regulate functional gene expression in murine T-cell hybridomas. For example, induction of apoptotic cell death following activation of T-cell hybridomas was examined using antisense oligonucleotides corresponding to several protooncogenes. We found that antisense oligodeoxynucleotides corresponding to c-myc inhibited both the characteristic DNA fragmentation and the loss of cell viability following activation without affecting production of lymphokines. Functional antisense oligonucleotides corresponding to c-fos had no effect in this system. These results demonstrate the use of antisense oligonucleotides to regulate function in T-cell hybridomas and provide valuable insights into the molecular bases of this biological phenomenon. Antisense oligonucleotides were also used to study another problem, the relation of T-cell-derived antigen-specific immunoregulatory factors to the T-cell receptor (TCR). Because the translation start of each TCR gene usually varies from one T cell to another, antisense oligonucleotides corresponding to the TCR V alpha or V beta of different cells were shown to act in a cell-specific manner. Furthermore, this method was used to demonstrate that a soluble antigen-specific regulatory activity produced by one of the T-cell lines depends on expression of the specific TCRa, an observation that has since been confirmed by gene transfer experiments. Expression of the CD3-TCR complex on the cell surface was also blocked by antisense oligonucleotides corresponding to CD3 gamma and CD3 zeta; however, neither these nor TCR V beta antisense oligonucleotides had any effect on production of the soluble regulatory activity.
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La Jolla Institute for Allergy and Immunology, California 92037.
Zheng
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Shi
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D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D001483:Base Sequence; D005786:Gene Expression Regulation; D016259:Genes, myc / Q000235:genetics; D006825:Hybridomas; D051379:Mice; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000494:pharmacology*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D011519:Proto-Oncogenes / Q000235:genetics; D011948:Receptors, Antigen, T-Cell / Q000187:drug effects*; D013601:T-Lymphocytes / Q000187:drug effects*; D013873:Thionucleotides / Q000737:chemistry
D015345:Oligonucleotide Probes; D016376:Oligonucleotides, Antisense; D011948:Receptors, Antigen, T-Cell; D013873:Thionucleotides