The mode of action of mineral-oil emulsion adjuvants on antibody production in mice.

Mice given a single subcutaneous inoculation of highly-purified ovalbumin in water-in-oil emulsion yielded peak antibody titres which were about 500 times those stimulated by the same dose of ovalbumin without adjuvant, and which remained at the peak level for at least a year. Neither the oil, nor the emulsifier, nor the emulsion itself when injected at a separate site from the antigen, stimulated a response of this type. The response was not affected by a simultaneous subcutaneous injection of anti-ovalbumin, or by the later inoculation of a different antigen in water-in-oil emulsion, or by further inoculations of ovalbumin in saline.

A study of factors influencing the sensitivity to antibody of tanned, ovalbumin-coated, formalinized sheep red cells.

Comparative tests were done to find the best conditions under which formalinized sheep red cells can be tanned and then coated with chromatographically purified ovalbumin. Variation in the time taken for tanning or coating, the amount of antigen used, the temperature during coating, the use of different batches of tannic acid, ovalbumin or of cells formalinized by different methods had little effect on sensitivity to agglutination by antibody. A slight increase in sensitivity was obtained by coating at a low pH, by increasing the quantity of tannic acid employed and by heat denaturation of the antigen.

Ovalbumin and conalbumin in the tanned red cell agglutination test.

The studies reported here indicate that tanned red cells coated with a mixture of chromatographically purified ovalbumin and conalbumin give agglutination titres which are directly related to the amount of antibody (as measured by quantitative precipitation) present against either of these antigens in mixed sera. With whole (unabsorbed) mixed sera, these coated cells record the titre of the major antibody present, whether this is anti-ovalbumin or anti-conalbumin. There was no evidence of greater sensitivity to anti-conalbumin.