The Journal of Experimental Medicine
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Journal of Experimental Medicine, Vol 149, 1227-1237, Copyright © 1979 by Rockefeller University Press

ARTICLES

Synthesis of two classes of antibody, gammaM and gammaG or gammaM and gammaA, by identical cells. Amplification of the antibody response to pneumococcal polysaccharide type III

VJ Pasanen, R Asofsky and PJ Baker

Class-specific plaque-forming cell (PFC) (gammaM, gamma1, gamma2, and gammaA) responses to type III pneumococcal polysaccharide (SSS-III) were studied in BALB/c x C57BL/6F1 (CBF1) mice with and without induction of an allogeneic effect. Gamma1, gamma2, and gammaA PFC were detected in two ways: (a) With the sequential development of the assay slides, first for direct (gammaM)PFC followed by incubation with class- specific antiimmunoglobulin and complement for the development of additional gamma1, gamma2, and gammaA PFC (gammaM-independent gamma1, gamma2, and gammaA PFC); and (b) by blocking gammaM PFC with goat anti- gammaM and simultaneously developing gamma1, gamma2, and gammaA PFC (total gamma1-, gamma2-, and gammaA-secreting PFC). The results showed that whereas gammaM PFC arose on the 3rd d after immunization, gamma1-, gamma2-, and gammaA-secreting PFC arose on the 4th to 5th d after immunization. They appeared in association with gammaM-secreting PFC because they were detected with the gammaM blocking method but not with the sequential method. By the 7th d most gamma1, gamma2, and gammaA PFC were detected by the sequential method as well, indicating that those antibodies were secreted independently of cells secreting gammaM. When the numbers of double-class-secreting PFC were evaluated on the 5th d, the following results were obtained: 83% of gammaM PFC were secreting either gamma1 (25%), gamma2 (55%), or gammaA (2%). We interpret these data as evidence for an antigen-driven class differentiation from gammaM to gammaA and from gammaM to gammaG in the majority of anti-SSS- III-secreting clones without T-cell help. When an allogeneic effect was provided by inoculation of parental BALB/c spleen cells together with antigen, the numbers of all classes of PFC were increased. Furthermore, the frequency of gammaM-gammaG (108%) or gammaM-gammaA (9%) double- class secretors was increased, and gammaM-independent gammaG and gammaA secretors were detected earlier, indicating an overall maturation- promoting effect. In addition, prolonged appearance of gammaA PFC was dependent on the allogeneic effect.
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