Different induction patterns of mRNA for IFN-alpha and -beta in human mononuclear leukocytes after in vitro stimulation with herpes simplex virus-infected fibroblasts and Sendai virus.
Human peripheral blood mononuclear leukocytes (PBL) were stimulated in vitro by HSV type 1-infected glutaraldehyde-fixed fibroblasts, or Sendai virus (SV). The PBL containing mRNA for IFN-alpha 2 or -beta 1 were clearly identified by RNA-RNA in situ hybridization by using 35S-labeled alpha 2- and beta 1-probes. Although the two inducers gave similar levels of IFN in the culture medium (about 20 U/10(4) PBL), the patterns of expression of mRNA at the cellular level differed. The HSV induced only IFN-alpha mRNA in the PBL, with a lag of 1 to 2 h, and with a peak frequency of about 10 labeled cells/10(4) PBL at 6 h. Grain counts were high, the majority of cells having more than 50 grains. They were morphologically medium to large lymphocytes. The HSV-infected glutaraldehyde-fixed fibroblasts therefore induce IFN-alpha 2 mRNA in infrequent but highly efficient PBL, each cell capable of producing as much as 2 antiviral units of IFN-alpha. In contrast, SV induced both IFN-alpha 2 and -beta 1 mRNA in PBL, and without clear lags. IFN-beta 1 mRNA-positive PBL peaked somewhat earlier (4 h) than cells containing IFN-alpha 2 mRNA (6 h), and their mean frequencies were approximately 80 and 60/10(4) PBL, respectively, in a panel of PBL from six blood donors. Grain counts were lower than with the HSV inducer, the majority of cells having less than 50 grains, and most labeled cells were morphologically monocytes. The frequency of labeled PBL rapidly decreased with increasing culture time with both the SV and HSV inducers, was low at 12 h and almost absent at 24 h.
Gobl
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Interferon Laboratory, Uppsala University, Sweden.
Funa
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Alm
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GV
D002472:Cell Transformation, Viral; D005347:Fibroblasts / Q000378:metabolism*; D006801:Humans; D007369:Interferon Inducers / Q000502:physiology; D007370:Interferon Type I / Q000096:biosynthesis* / Q000235:genetics; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D009693:Nucleic Acid Hybridization; D010222:Parainfluenza Virus 1, Human / Q000502:physiology*; D012333:RNA, Messenger / Q000096:biosynthesis*; D018139:Simplexvirus / Q000502:physiology*
D007369:Interferon Inducers; D007370:Interferon Type I; D012333:RNA, Messenger