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Development of Primary M, G, and A Anti-TNP Plaque-Forming Cell Responses in Vitro¹

From The Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115

Abstract

Trinitrophenylated bacteriophage X174 (TNP-X) is a potent in vitro immunogen capable of stimulating specific anti-TNP plaque-forming cell (PFC) responses in cultures of mouse spleen cells. Under optimal conditions, 10 x 10⁶ spleen cells and 3 x 10⁹ TNP-X particles in 1 ml of culture medium, TNP-specific PFC responses representing the major mouse immunoglobulin classes, M, ₁, ₂, and A were detected. A study of kinetics of appearance of these immunoglobulin class-specific PFC revealed that an increase in M PFC was detected within 24 hr of initiation of the cultures; the maximum M PFC responses were on days 4 and 5. ₁ and ₂ PFC were not detectable until day 3 of culture and reached maxima on days 5 and 6. The anti-TNP PFC response to TNP-X was almost completely suppressed by the addition of 100 µg TNP guinea pig albumin; the addition of free X at a 10-fold excess over the TNP-X immunogen suppressed the anti-TNP response about 70%.

Footnotes

¹ This investigation was supported by United States Public Health Service Research Grants AI-09897 and AI-09920 from the National Institute of Allergy and Infectious Diseases.

² Present address: Department of Medicine, University Hospital, University of California, San Diego, La Jolla, California 92037.

³ Recipient of United States Public Health Service Research Career Development Award 1K4-AI-70, 173 from the National Institute of Allergy and Infectious Diseases.

⁴ Address correspondence and reprint requests to Carl W. Pierce, Department of Pathology, Harvard Medical School, Boston, Massachusetts 02115.