Role of glycosilation in the susceptibility of "acid labile" interferon alpha to acid treatment.
Mononuclear cells from blood of healthy donors produce acid-labile interferon (IFN) alpha when stimulated with HIV-infected cells. A large proportion of this IFN appears to be glycosilated, as treatment with neuraminidase causes a shift of the isoelectric point (IP) from pH = 5.2-5.4 to pH = 5.8-6.2. To assess the role of glycosilation in determining the instability of antiviral activity after exposure to acid (pH lower than 4) peripheral blood mononuclear cells (PBMC) were induced to produce IFN with HIV-infected cells in the presence of tunicamycin, an inhibitor of glycosilation. The IFN produced under such experimental conditions (tu-IFN) was acid-stable. Tu-IFN was compared to a standard acid-labile IFN by affinity chromatography on Con A-sepharose. The elution pattern showed that tu-IFN does not bind to the gel, whereas the acid-labile IFN is eluted in two fractions, one unbound, which is stable at pH2, and one bound, which retains the initial acid-lability. These results suggest that acid-labile IFN alpha is largely glycosilated, and that the presence of glycosilated molecules contribute to render the IFN molecule unstable at acidic pH. It is to be determined whether some glycosilated molecule present in the IFN preparation, or glycosilation of the IFN molecule per se, is responsible for acid-lability of the antiviral activity.
Capobianchi
M R
MR
Institute of Virology, University La Sapienza, Roma, Italy.
Mattana
P
P
Gentile
M
M
Conciatori
G C
GC
Ankel
H
H
Dianzani
F
F
D002846:Chromatography, Affinity; D006031:Glycosylation; D006801:Humans; D006863:Hydrogen-Ion Concentration; D016898:Interferon-alpha / Q000737:chemistry*; D007526:Isoelectric Point
D016898:Interferon-alpha