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Heterogeneity and Sequence of Appearance of Rabbit Antibodies to Hog Intrinsic Factor¹

From the Gastroenterology Unit, Department of Medicine, the University of Rochester School of Medicine and Dentristry, Rochester, New York and the Department of Medicine, Center for the Health Sciences, University of California, Los Angeles, California

Abstract

The primary and secondary immune responses of rabbits to HIFC were investigated with the radioimmunoelectrophoretic method. Three anti-HIFC-Co⁶⁰B₁₂-binding antibodies were detected. When a goat anti-rabbit serum was used as a developer, M and G antibodies were found to appear simultaneously, 7 to 8 days following primary immunization. When the developing serum was not used, an illusion suggesting the later appearance of G antibody was seen. These results provide further evidence for the concurrent synthesis of M and G antibodies following primary immunization. In addition, the data suggest that early G anti-HIF antibody forms a soluble or easily dissociable complex with HIFC-Co⁶⁰B₁₂ while later G antibody forms an insoluble or less readily dissociable complex with the radioactively labeled antigen.

In two of three rabbits, a third HIFC-Co⁶⁰B₁₂-binding antibody appeared from 3 to 4 weeks after primary immunization. This antibody migrated slightly faster than G on electrophoresis in agar gel, was eluted with G-globulins from Sephadex G-200, and was resistant to 2-mercaptoethanol.

Following secondary immunization an anamnestic response was observed for the synthesis of all three antibodies.

Footnotes

This investigation was supported by Research Grant AM 08956 from the National Institutes of Health.

Presented in part at the 50th Meeting of the Federation of American Societies for Experimental biology, Atlantic City, New Jersey on April 16, 1966 (1).