Type I interferon subtypes produced by human peripheral mononuclear cells from one normal donor stimulated by viral and non-viral inducing factors.
Through the activation of Toll-like receptors (TLRs) or cytosolic RNA helicases, a large number of pathogenic or synthetic components can induce the transcription of genes coding for type I interferons (IFNs). This family of related cytokines includes notably, a single IFN-beta protein and 13 different IFN-alpha subtypes, whose biological activities are probably not the same. The aim of this study was to characterize the type I IFN subtypes produced in vitro by human peripheral blood mononuclear cells (PBMCs) in response to specific inducers. Thus, PBMCs obtained from a single donor, were exposed to various agents including Sendai virus, Herpes simplex virus-1 (HSV-1), poliovirus-IgG complexes and serum from a patient with systemic lupus erythematosus (SLE). Six hours later, mRNA was extracted and amplified by RT-PCR using primers which recognize IFN-B mRNA and the different IFN-A mRNA subtypes. IFN-A subtypes were identified by cloning and sequencing the amplification product. Antiviral activity was assayed in supernatant at 18 hours. Human PBMCs were found to express constitutively type I IFNs mRNA. Antiviral activity and expression of IFN-A and IFN-B mRNA increased with each inducing agent. Although almost all the IFN-A subtypes were detected, their relative abundance appeared to be dependent upon the inducing agent. Incubation of PBMCs with a neutralizing monoclonal antibody directed against the type I IFN receptor (IFNAR) did not affect the level of antiviral activity in the supernatant of induced PBMCs. Our results suggest that the level of IFN-alpha expressed by PBMCs cells is independent of IFNAR feedback signalling and that the nature of the inducing agent modifies the pattern of IFN-A subtypes preferentially expressed by these cells.
Palmer
Pierre
P
Université Paris Descartes - Faculté de Médecine - Laboratory of Virology, Cochin-Saint-Vincent-de-Paul Hospital, Assistance Publique-Hôpitaux de Paris (AP-HP), 82, avenue Denfert-Rochereau, 75674 Paris Cedex 14, France.
Tovey
Michael G
MG
Raschilas
Franck
F
Brassart
Lilia
L
Meritet
Jean-François
JF
Porcher
Raphaël
R
Lebon
Pierre
P
D000998:Antiviral Agents / Q000378:metabolism; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003600:Cytosol / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000378:metabolism; D007370:Interferon Type I / Q000378:metabolism*; D016898:Interferon-alpha / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000378:metabolism*; D008180:Lupus Erythematosus, Systemic / Q000097:blood*; D017955:Poliovirus / Q000378:metabolism; D020365:RNA Helicases / Q000378:metabolism; D053633:Receptor, Interferon alpha-beta / Q000378:metabolism; D029082:Sendai virus / Q000378:metabolism
D000998:Antiviral Agents; D007074:Immunoglobulin G; D007370:Interferon Type I; D016898:Interferon-alpha; D053633:Receptor, Interferon alpha-beta; D020365:RNA Helicases