Enhancement of immunogenic activity of ribosomal preparations from Haemophilus influenzae by various adjuvants.

Ribosomes from Haemophilus influenzae type b have been reported to have immunoprotective activity in animals that can be enhanced by adjuvants. In this report we evaluated the adjuvant activity of several compounds in conjunction with ribosomes from the b and c serotypes of H. influenzae.

Induction of active immunity with membrane fractions from Haemophilus influenzae type b.

Using Escherichia coli strain E-1 as a model, we developed procedures for the preparation of outer- and inner-membrane-enriched fractions as structural units. These procedures could be used to prepare relatively pure inner and outer membrane fractions as determined by succinate dehydrogenase activity, ketodeoxyoctonate levels, and polyacrylamide gradient gel electrophoresis. The use of these procedures to fractionate membrane components from Haemophilus influenzae type b strains H-2 and H-E led to good separation of outer- and inner-membrane-enriched fractions as determined by succinate dehydrogenase and ketodeoxyoctonate levels but incomplete separation as determined by polyacrylamide gradient gel electrophoresis.

Opsonization and phagocytosis of Haemophilus influenzae type B organisms by mouse polymorphonuclear leucocytes and antiribosomal serum.

Sera from rabbits immunized with ribosomes passively protect mice challenged with Haemophilus influenzae type b. The protective antibody interacted with organisms in the blood and possibly at the sites of dissemination, but not at the site of inoculation. Macrophages did not phagocytize oposonized bacteria in our system.

Kinetics of Haemophilus influenzae type B infection in normal and ribosome-immunized mice using intraperitoneal and intracerebral routes of inoculation.

The kinetics of infection was studied in normal and ribosome-immunized mice challenged with Haemophilus influenzae Type b organisms. Ribosomal preparations extracted by the differential-centrifugation and sodium-dodecyl-sulphate treatment or ammonium-sulphate-precipitation procedures were highly immunoprotective when mice were challenged by the i.p.

Serological and biological activities of anti-Haemophilus influenzae ribosomal serum.

The antibody content in serum from rabbits immunized with ribosomes from Haemophilus influenzae type b was determined by passive hemagglutination, enzyme-linked immunosorbent assay, and complement fixation. Attempts to use passive hemagglutination to assay anti-ribosomal antibodies were unsuccessful. In the enzyme-linked immunosorbent assay tests, rabbit antiserum was allowed to react with ribosomes that adhered to microtiter plates.