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Skin-Sensitizing Properties of Guinea Pig Antibodies to Keyhole Limpet Hemocyanin¹

From the Division of Chemistry and Chemical Engineering, California Institute of Technology, Pasadena, California 91109

Abstract

Guinea pigs were immunized with keyhole limpet hemocyanin (KLH) emulsified in complete Freund's adjuvant. A crude guinea pig anti-KLH -globulin pool was prepared and three different antibody preparations were obtained as follows: A) by acid dissociation of specific KLH anti-KLH precipitates and separation of the KLH from the antibodies by ultracentrifugation; B) by acid dissociation of the immune precipitates and isolation of the antibodies by gel filtration through Sephadex G 200; C) by chromatography on DEAE-cellulose.

The preparations were absorbed with a rabbit anti-guinea pig 7 S 1 serum and were subsequently found free of 7 S 1 when tested by radioimmunoelectrophoresis. Antigenically pure 7 S 2 anti-KLH antibodies as well as unabsorbed controls were tested by passive cutaneous anaphylaxis in the guinea pig. The results indicated that antibodies associated with the 7 S 2 class elicited positive PCA reactions in the guinea pig.

Footnotes

This investigation was supported by the National Institute of Allergy and Infectious Diseases, Grants A1-01339 and 5T1 AI-127. (Contribution no. 3636.)