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.

Effects of glucose and fructose loading on glycogenesis in chicks infected with avian tuberculosis.

The chick was used as a rapid metabolic model to determine the fate of ingested fructose vs. glucose in noninfected chicks and in those subjected to the stress of avian tuberculosis. The chicks were crop-loaded with either a 72% fructose or glucose solution 21 and 28 days post TB inoculation and killed 2 and 4 hr after loading.

Role of bovine serum albumin in the nutrition of Mycobacterium tuberculosis.

Bovine serum albumin promotes the growth of small inocula of Mycobacterium tuberculosis in media containing unesterified fatty acids. Albumin binds fatty acids present in concentrations toxic for the organisms. In the present study, additional roles of albumin were investigated.

Characterization of the immunoprotective antigen of ribosomal preparations from Haemophilus influenzae.

This investigation was designed to characterize the immunoprotective antigen of ribosomal preparations from Haemophilus influenzae. The ribosomes that elicited 80 to 90% protection contained 25% protein and 75% ribonucleic acid but did not contain any detectable hexoses. The immunodiffusion and hemagglutination inhibition tests also failed to demonstrate that the capsular material (polyribose phosphate) was in ribosomal preparations.

Adoptive transfer of immunity from mice immunized with ribosomes or live yeast cells of Histoplasma capsulatum.

This investigation was designed to compare the role of lymphoid cells and immune serum in protective immunity induced by immunization with ribosomes or live yeast cells of Histoplasma capsulatum. Spleen cells, peritoneal cells, and serum from C3H mice immunized with Histoplasma ribosomes or live cells were transferred intravenously to separate groups of syngeneic recipients. All recipients along with a set of immunized and control mice were challenged intravenously with 4 x 10(6) yeast cells of H.

Immunoprotective activity of ribosomes from Haemophilus influenzae.

Immunization with ribosomal preparations from Haemophilus influenzae type b elicited protective immunity in mice. Ribosomes from disrupted cells where isolated by differential centrifugation using sodium dodecyl sulfate. The washed ribosomes contained 25% protein and 75% ribonucleic acid and sedimented as a single peak on sucrose density gradient analysis with a sedimentation coefficient of 67S, using Escherichia coli ribosomes as a 70S marker.

Ribonucleic acid synthesis in normal and immune macrophages after antigenic stimulus.

Macrophage ribonucleic acid (RNA) synthesis is an important metabolic process intimately related to the function of these cells. Mouse peritoneal macrophage RNA was extracted with phenol in the presence of bentonite and electrophoresed on composite agarose-polyacrylamide gels. The pulse-chase technique was used to follow the precursor relationships in macrophage ribosomal RNA (rRNA) maturation.

Comparative chemotherapeutic activity of amphotericin B and amphotericine B methy ester.

The comparative efficacy of amphotericin B and amphotericin B methyl ester (AME) against experimental histoplasmosis, blastomycosis, cryptococcosis, and candidosis in mice was assessed by determining the effect of daily intraperitoneal therapy on 21-day survival and persistence of organisms in internal organs. AME, like amphotericin B, was effective against each of the experimental infections, but the efficacy was lower than the parent compound. For Histoplasma and Blastomyces infections the mean effective dose (ED(50)) of amphotericin B was 0.

Comparative in vitro antifungal activity of amphotericin B and amphotericin B methyl ester.

The in vitro antifungal activity of amphotericin B methyl ester (AME), a water-soluble derivative of amphotericin B, was compared to that of the parent compound against a variety of pathogenic and potentially pathogenic fungi. AME has a significant antifungal activity, but the activity of AME was slightly lower than that of amphotericin B. Among the yeast-like organisms, only the yeast cells of Sporothrix schenckii were more resistant than others to both antibiotics, with a minimal fungicidal concentration of 5 to 10 mug/ml.

Biological activity of heated diphtheria toxin.

Diphtheria toxin splits into two fragments when heated at 100 C for 10 min in a phosphate buffer. The separated fragments have molecular weights of 24,000 and 39,000, respectively. These molecular weights are similar to those of the A and B fragments found in diphtheria toxin preparations after thiol reduction.

Quantitation of the bioenergetics of a tuberculosis infection in chicks.

Interaction of an avian tuberculosis infection with a known metabolizable energy yield of dietary corn oil in chicks was used to quantitate total host energy expenditure necessitated by the infectious process. Three trials in which two doses of inoculum were used resulted in mild and severe involvements. Trial 1 (mild) indicated that 6% and trials 2 and 3 (severe) that 96 and 93% of the energy supplied by known quantities of corn oil were utilized by the tuberculosis process.

Site in cell-free protein synthesis sensitive to diphtheria toxin.

The effects of diphtheria toxin on cell-free protein synthesis in a bacterial system, and preparations obtained from animals that were sensitive and resistant to toxin were examined. In the presence of nicotinamide adenine dinucleotide (NAD), toxin inhibited the incorporation of amino acids by endogenous and synthetic polynucleotides in both rat liver and guinea pig liver cell-free systems that were exposed to 6 Lf units per ml of toxin. A cell-free system derived from Streptococcus faecalis was resistant to high concentrations of toxin.

The response of cultured mammalian cells to diphtheria toxin. I. Amino acid transport, accumulation, and incorporation in normal and intoxicated sensitive cells.

The response to diphtheria toxin of two sensitive cell lines, KB and HeLa, was investigated. Inhibition of the incorporation of radioactively labeled amino acids into protein was the earliest detectable effect of diphtheria toxin. It was observed that, during the period of intoxication, the cell membrane was morphologically intact and retained its semi-permeable character, although it was rendered fragile and more easily disrupted by mechanical manipulations than the normal cell.

DEVELOPMENT OF RESISTANCE TO POLYENE ANTIBIOTICS IN CANDIDA ALBICANS.

Hebeka, Elias K. (Rutgers, The State University, New Brunswick, N.J.