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The Journal of Immunology, 1948, 60: 383-398.
Copyright © 1948 by The American Association of Immunologists, Inc.

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Antibody Formation and Sensitization with the Aid of Adjuvants*

Jules Freund, K. Jefferson Thomson, H. B. Hough, H. E. Sommer and T. M. Pisani

From the Division of Applied Immunology, The Public Health Research Institute of The City of New York, Inc.

Abstract

1. Antibody formation was enhanced and sustained when the antigen (typhoid bacillus) was incorporated into a water-in-oil emulsion prepared with paraffin oil and Falba or anhydrous lanolin or Arlacel A (mannide monooleate) or myricin combined with cholesterol.
2. Oil-in-water emulsion prepared with Tween 80 (polyoxyethylene sorbitan monooleate) and paraffin oil was ineffective. When oleic acid combined with myricin was used as an emulsifying agent antibody formation was neither enhanced nor sustained.
3. Emulsions prepared with "lecithin" and paraffin oil are unstable mixed water-in-oil and oil-in-water emulsions. Antibody formation with such emulsions was highly accelerated but not as well sustained as with Falba and paraffin oil.
4. Antibody formation is scant when peanut oil is substituted for paraffin oil.
5. The addition of killed mycobacteria, namely M. butyricum or M. tuberculosis or M. phlei to water-in-oil emulsions further augments antibody formation.
6. Lipid extract from M. tuberculosis has a similar augmenting effect.
7. Cholesterol also increases the adjuvant effect of water-in-oil emulsion.
8. A large amount of cerotic acid added to water-in-oil emulsions retards antibody formation.
9. In experiments with a small number of guinea pigs sensitization to picryl chloride was produced only by emulsion containing killed tubercle bacilli. Other substances which augment antibody formation such as lipid extract from M. tuberculosis and cholesterol did not produce this effect.
10. The enhancing effect on antibody formation and the alteration of sensitization may occur concomitantly but they appear to be distinct phenomena.

Footnotes

* This study was aided by a grant from the John and Mary R. Markle Foundation.




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