The observations reported here show that the T Vi antigen is effective in stimulating antibodies when it is attached to the cells. It is also clear that the Vi antigen is readily removed in aqueous solutions, but in such organic solvents as acetone, chloroform, ether, alcohol, carbon tetrachloride and benzene it is retained by the cell. In alcohol-water mixtures the Vi antigen is progressively more soluble in those solutions where the alcohol concentration is less than 70 %. However, its solubility in water is limited, and by making progressively heavier suspensions it was found that a point is reached at which Vi antigen is retained by the cell, the suspending solution apparently being saturated. Such an aqueous suspension is able to stimulate the production of Vi antibody.
The administration of dried organisms in peanut oil resulted in the production of antibodies against both O and Vi antigens. This was in contradistinction to the results obtained when mineral oil, mineral oil + saline, or peanut oil + saline were used: in these cases the O antibody titre was lower and Vi antibodies were produced in low titre or not at all. It seems clear that the presence of water in the vaccine has the effect of removing the antigenicity of the Vi hapten; water-free, vegetable-oil vaccine is suggested by these studies as an effective agent for immunizing against all of the antigens of Salmonella typhi. The use of such a vaccine in producing active and passive immunity in animals is discussed in the paper which follows.