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Prolonged Immunization

II. Effect on Complement-Fixing, Incomplete and Combining Antibodies in Rabbit Anti-Mouse Erythrocyte Serum¹

From the Departments of Pathology, The Chicago Medical School and Mount Sinai Hospital, Chicago, Illinois

Abstract

1. Rabbit anti-mouse erythrocyte sera from each of 7 courses of immunization were studied with 9 immunologic techniques, namely, agglutinin titrations with antiglobulin serum, in serum-albumin medium, with papain treated cells, and in saline medium; hemolysin and complement fixation titrations; determination of combining ability with fluorescein and I¹²⁵ labeled sera; and production of anemia in mice.

2. Formalinized erythrocytes can be used successfully to study certain immunologic reactions.

3. In vitro reactions, except for complement fixation and coating with fluorescent antibodies, did come near their peak levels in 2-course sera; after 4 or more courses, titers of all in vitro reactions were essentially parallel to the base line. Whether these tests are demonstrating different reactions of the same antibody or reactions of different antibodies requires further investigation.

4. On the other hand, the anemia producing ability showed marked decrease in sera collected after more than 3 courses of immunization.

5. Our studies permit the conclusion that no in vitro reaction has yet been found to serve as a reliable indicator of the in vivo potency of a rabbit anti-mouse erythrocyte serum.

Footnotes

¹ Supported in part by Grant AM-03243 from the National Institute of Arthritis and Metabolic Diseases, United States Public Health Service, and the Leukemia Research Foundation, Inc., Chicago, Illinois.