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Production of monoclonal antibodies to human IFN‐α and their use for analysis of the antigenic composition of various natural interferons

We have established three hybridoma cell lines (designated EBI‐1, ‐2, and ‐3) secreting immunoglobulin G (IgG) antibodies specific for human alpha‐interferon (HuIFN‐α) by fusing P3X63Ag8.6.5.3 myeloma cells with spleen cells of mice immunized with human “lymphoblastoid” (NC‐37) interferon (IFN). The antibodies were used to study the antigenic diversity of a number of IFNs derived from blood leukocytes and lymphoid cell lines. When incubated with 10 U of IFN, EBI‐1 antibodies completely neutralized the antiviral activity of HulFN‐αrA produced in E coli as well as of IFN produced spontaneously in several lymphoblastoid cell lines. In contrast, only 60‐70% of alpha‐interferon (IFN‐α) induced in leukocytes, lymphoma, or lymphoblastoid cell lines by Sendai virus or poly (l):(C) could be neutralized by the antibody; furthermore, IFN‐α produced spontaneously in bromodeoxyuridine‐treated lymphoma cells was also only partially neutralized. EBI‐2 and EBI‐3 antibodies were tested against a smaller set of samples and found to react similarly. Our result indicate that human IFN‐α subtypes can be grouped into at least two antigenically distinguishable subsets, one of which being represented by IFN‐αA, and that these subsets can be expressed differently under different conditions in a given cell type.