Interferon-alpha 2 produced by normal human leukocytes is predominantly interferon-alpha 2b

Area of Science:

• Immunology
• Virology
• Biochemistry

Background:

• Interferon alpha (IFN-alpha) is a crucial cytokine with antiviral properties.
• Production of biologically active IFN-alpha from human leukocytes is a complex process.

Purpose of the Study:

• To purify and characterize interferon alpha n3 produced by Sendai virus-induced peripheral blood leukocytes.
• To identify the specific subtypes of interferon alpha 2 present in the purified preparation.

Main Methods:

• Immunoaffinity and size-exclusion chromatography for IFN-alpha purification.
• Reverse-phase high-performance liquid chromatography (RP-HPLC) for protein separation.
• Amino-terminal sequencing and CNBr cleavage for protein identification and variant determination.

Main Results:

• IFN-alpha n3 was purified to >98% homogeneity.
• Interferon alpha 2 was isolated and found to exist as two closely eluting peaks (1.1 and 1.2) on RP-HPLC.
• Amino-terminal sequencing identified IFN-alpha 2b as the major component, with IFN-alpha 2a absent.

Conclusions:

• Sendai virus induction of leukocytes yields a highly purified IFN-alpha n3 preparation.
• The characterization of IFN-alpha 2 variants provides critical data for understanding leukocyte-derived interferon.
• IFN-alpha 2b is the predominant subtype in this specific IFN-alpha preparation.