Removal of lipopolysaccharide from acellular Bordetella pertussis vaccine by detergent treatment.

Protective antigen was extracted from Bordetella pertussis cells with 1.0 M NaCl and precipitated with ammonium sulfate, 20-40% saturation (designated fraction 15A-1B). The protective antigen was purified further by detergent (Emulphogene BC720) treatment and adsorption to aluminum hydroxide gel (designated fraction 15A-108A). Compared with B. pertussis vaccine and fraction 15A-1B, fraction 15A-108A retained protective activity as assessed by the mouse protection test, but had reduced protein and markedly reduced endotoxin content. Fraction 15A-108A also had reduced leukocytosis-promoting, histamine sensitizing splenomegaly-inducing, and adjuvant activities. Emulphogene treatment provided a relatively simple method for removing endotoxin from a potential acellular B. pertussis vaccine.

Immunomodulation by Bordetella pertussis: antiviral effects.

Treatment of mice by intraperitoneal inoculation of pertussis vaccine or lipopolysaccharide extracted from B. pertussis will effect resistance to rabies virus, encephalomyocarditis virus, Semliki Forest virus, and Herpes simplex virus. Our previous observations indicated that treatment of C3H/HeN (+/nu) and BDF1 mice with pertussis vaccine injected i.p. five days prior to a mouse adenovirus lethal dose i.p. challenge elicited resistance to clinical disease and death. Susceptibility returned to a portion of the test population 35 days after pertussis vaccine treatment. The pertussis vaccine induced resistance developed in athymic (nude) mice also; however, the population succumbed to infection 35 days later. Titration of pertussis vaccine with respect to induction of resistance indicated the median effective dose (ED50) was approximately 25 micrograms dry weight. This report describes the antiviral activity of acellular components extracted from pertussis vaccine. Extraction of B. pertussis cells with 1.0M NaCl and ammonium sulfate fractionation (20-40% saturation) of the extract resulted in an acellular preparation that induced resistance to lethal dose mouse adenovirus infection. The resistance inducing activity was retained after treatment of the extract with detergent (GAF Emulphogene BC 720) to remove lipopolysaccharide and adsorption to alum gel. Comparison of endotoxin content of pertussis vaccine acellular fractions, polysaccharide fraction and purified lipopolysaccharide suggested that endotoxin probably plays a role in the induction of resistance. The endotoxin content of a Emulphogene-treated preparation that protected 80% of a test population was 39 ng. The lipopolysaccharide extracted from Escherichia coli, Vibrio cholerae, Salmonella typhimurium, and Salmonella minnesota did not induce a resistant state seven days after administration; however, lipopolysaccharide extracted from B. pertussis induced a resistant state.(ABSTRACT TRUNCATED AT 250 WORDS)

Detection of antibody to carcinoembryonic antigen by the hemolytic plaque assay.

A modification of the standard hemolytic plaque-forming cell assay is described. The conjugation of a soluble antigen, carcinoembryonic antigen, to the membrane of the sheep red blood cell was accomplished and used as the indicator system to detect antibody-forming cells. The specificity of the assay was determined by cross-reacting studies and competitive inhibition by antigen blocking.

Effect of prostaglandin E1-induced elevation of cyclic AMP on glucose repression in the lactic streptococci.

Cyclic adenosine 3', 5' -monophosphate (cAMP) activity was observed in Streptococcus lactis C2, Streptococcus cremoris C10, Streptococcus diacetlactis 18-16, and Streptococcus thermophilus C3. In vitro assays of cell-free extracts obtaianed from S. lactis C2 showed that the cAMP-associated enzymes adenyl cyclase and phosphodiesterase were also present. In vitro experiments showed that prostaglandin E1 (PGE) stimulation of adenyl cyclase increased cAMP concentrations approximately fivefold, and in vivo studies showed that PGE treatment of S. lactis C2 increased intracellular cAMP concentrations twofold. Futhermore, PGE-induced elevation of intracellular cAMP levels was shown to prevent the repression of ss-D-phosphogalactoside galactohydrolase synthesis by glucose.

The chicken's femoral-lymph nodules: T and B cells and the immune response.

Femoral lymph nodules (FLN), which are barely perceptible in normal birds, after a footpad injection of sheep red blood cells (SRBC) may either significantly enlarge (responder) or remain reduced in size (nonresponder). There were approximately 38% T cells and 53% B cells in the FLN of responder chickens. Significantly more plaque-forming cells (PFC) developed in the FLN than in the spleen after a footpad injection of SRBC. Total antibody, mercaptoethanol- (ME) resistant, and ME-sensitive fractions were significantly higher in birds given i.v. than in those given footpad injections. There were no differences in PFC and agglutinin titers between FLN-responders and nonresponders. The number of PFC in FLN exceeded the number of splenic PFC previously reported. The high PFC response of the FLN may reflect the large percentage of B cells in this lymphomyeloid tissue or the presence of antigen-experienced B cells in the FLN. Although FLN may influence a systemic immune response its major role appears to be restricted to a local response.

Thrombocyte migration and the release of thrombocyte inhibitory factor (ThrIF) by T and B cells in the chicken.

The elaboration of various inhibitory factors by sensitized lymphocytes has been reported for both mammals and birds. Until recently, the ability of the B cell to elaborate these had been questioned by a number of investigators. We have demonstrated the release of a lymphocyte inhibitory factor from the avian thymus and bursal lymphocyte. The avian thrombocyte is active in haemostasis as well as being a phagocytic cell. We have demonstrated that the ability of the thrombocyte to migrate, significantly exceeded that of thymic lymphocytes. Sensitized T and B cells, as well as activated supernatants from these cells, were capable of inhibiting thrombocyte migration. Treatment of sensitized B cells with anti-T-cell serum plus complement failed to eliminate this cell's ability to block thrombocyte migration. These data attribute the capability of elaborating a thrombocyte migration inhibitory factor to both the avian thymic and bursal lymphocyte, and attribute a degree of maturity to these cells not previously demonstrated.

Antigenic variation and increase in pathogenicity in Pseudomonas aeruginosa as a result of growth on glucose or N-hexadecane as sole carbon source.

When Pseudomonas aeruginosa is grown on glucose as opposed to n-hexadecane as the sole carbon source, the antigenicity, virulence, and protein composition of the outer membrane are altered. The hydrocarbon-grown cells demonstrate a 3-log increase in virulence over the glucose-grown cells (in mice). There also appears to be an additional protein present in the outer membrane of the n-hexadecane-grown cells. This protein may contribute to the observed antigenic differences between the two cell types.

A simple transporting and maintenance medium for Mycoplasma synoviae.

A simple technique for preserving and transporting Mycoplasma synoviae, which involves suspending the cells in a 20 percent glycerol medium, is described.