Preparation and Effect of Different Adjuvants on the Immunogenic Activity of Mycobacterial Ribosomal Fraction

Abstract

Several emulsified and two nonemulsified incomplete adjuvants were examined for their adjuvant activity by use of mycobacterial ribosomal fractions as a substrate. A good adjuvant is defined as one which produces a high immunological response with the ribosomal fraction in mice to infection with virulent tubercle bacilli. Freund's incomplete adjuvant, consisting of Aquaphor and heavy mineral oil, and Arlacel A plus hexadecane were the best adjuvants tested. Aquaphor plus light mineral oil and Arlacel A plus 7- n -hexyloctadecane were not quite as effective. Peanut oil was not satisfactory when emulsified with either Aquaphor or Arlacel A. A moderate degree of immunity was produced in mice vaccinated with ribosomal fraction mixed with aluminum hydroxide gel. Sodium alginate mixed with ribosomal fraction produced a low degree of immunity only with the highest vaccinating dose. It was found that the effectiveness of the emulsified type of adjuvant depended upon the method of preparation. Careful standardization of technique to produce uniform and complete emulsification was essential for maximal adjuvant activity using minimal vaccinating doses. A rapid and practical method of preparing emulsified adjuvants is given. The mode of action of incomplete adjuvants as employed in these experiments is discussed, and it is thought that they acted primarily by protecting the ribosomes from being inactivated by host ribonuclease before they were engulfed by the macrophages.