Adjuvant Properties of Aluminum and Calcium Compounds

Mice were immunized with either poly(Glu, Arg, Ala) or poly(Glu, Lys, Phe) contained in two different adjuvant preparations, alum (A1K(SO4)2) or complete Freund's adjuvant (CFA), and in vitro antigen-driven proliferative responses of lymph node cells were assayed 4-16 days later. After immunization with antigens on alum, endogenous interleukin 1 (IL-1) as well as interleukin 4 (IL-4) production was required for the proliferation of the elicited T cells as inclusion of either polyclonal goat anti-mouse IL-1 alpha or monoclonal anti-mouse IL-4 (11B11) in the cultures inhibited proliferative responses to antigen. In contrast, proliferative responses of cells elicited by antigen in CFA were not inhibited by either anti-IL-1 or anti-IL-4. Monoclonal antimouse CD4 (GK 1.5) inhibited proliferative responses regardless of which adjuvant was used to elicit antigen-reactive cells. These data indicated that phenotypically different subpopulations of CD4+ cells were elicited by the same antigen administered in different adjuvant preparations, Th2-like cells after immunization with polymers on alum and Th1-like cells after immunization with antigens in CFA. An examination of the isotypes of polymer-specific antibodies present in the sera of immunized mice also supported this conclusion.

Antibodies against two physicochemically purified haemagglutinins (HAs) of Bordetella pertussis (filamentous HA and leucocytosis-promoting-factor HA) protect laboratory animals from pertussis. A vaccine containing these two HAs was prepared and tested in trials involving about 5000 children. Culture supernatant of Bordetella pertussis, phase I, was treated with ammonium sulphate, and a crude extract of the HAs was extracted from the precipitate by the use of concentrated sodium chloride. This crude extract was fractionated by sucrose density gradient centrifugation to obtain an HA preparation practically free of endotoxin. The HA preparation was treated with formalin to destroy its ability to induce leucocytosis and to cause histamine sensitisation. Aluminium hydroxide was added to the preparation as an adjuvant. The component vaccine is not only potent as judged by the mouse test but is also less than one-tenth as toxic as whole-cell vaccine as judged by leucocytosis promotion, histamine sensitisation, and endotoxicity tests. Field trials showed that component vaccine was as effective as and produced less side-effects than did conventional whole-cell vaccine. The vaccine has been used for mass immunisation in Japan since the autumn of 1981.