Antibody Production In Rabbits Research Articles

REACTION OF ESCHERICHIA COLI ENDOTOXIN WITH LITHIUM ALUMINUM HYDRIDE.

Treatment of Boivin-type endotoxin from E. coli with LiAlH4 according to the method of Noll and Braude yielded a small amount (about 10%) of an essentially pure nonphosphorylated polysaccharide which was nontoxic and nonpyrogenic. In contrast to their RE fraction this product did not stimulate the production of antibodies in rabbits, precipitate antibody nor produce tolerance to endotoxin. The bulk of the product resulting from treatment with LiAlH4 consisted of biologically unaltered endotoxin. It was isolated as a rapidly soluble, lipid-containing aluminum citrate-endotoxin complex, the infrared spectrum of which was nearly identical to that of an RE fraction with 100-fold reduced pyrogenicity. No correlation was found between fatty acid ester content, as estimated from infrared spectra, and toxic and pyrogenic properties of the endotoxin.

CHARACTERIZATION OF ENTEROCOCCUS BACTERIOPHAGES FROM THE SMALL INTESTINE OF THE RAT.

Rogers, C. G. (University of Wisconsin, Madison) and W. B. Sarles. Characterization of enterococcus bacteriophages from the small intestine of the rat. J. Bacteriol. 85:1378-1385. 1963.-Enterococcus bacteriophages were isolated by an enrichment procedure from the contents of the small intestines of Sprague-Dawley white rats. After purification by single-plaque isolation, the phages were maintained in Tryptose broth by propagation on strains of Streptococcus faecalis var. zymogenes of similar intestinal origin. The presence of two phage strains was suggested from host range studies, and confirmed by antiserum neutralization tests, whereby the bacteriophages were placed in two distinct, but serologically related, groups. When the characteristics of one phage from each group were studied, differences were observed in antigenic properties, as shown by the ability to stimulate antibody production in rabbits, and in single-step growth characteristics. At 32 C, the latent period was 21 min, and the burst size was 88 plaque-forming particles (PFP) for phage 1; and 34 min and 147 PFP for phage 2. The bacteriophages were similar in temperature of inactivation, sensitivity to pH, and in rates of adsorption to host cells (K = about 2 x 10(-9) ml/min at 32 C). In electron micrographs, the viruses were characterized by slightly elongated heads and long tails, and resembled one another closely. The dimensions of the phage particles were similar to those reported for other phage-host systems within the genus Streptococcus.

The Detection of Humoral Antibodies Directed against Salicylates in Hypersensitive States

Summary Conjugates of aspirin have been prepared which are able to stimulate antibody production in rabbits and sensitize guinea pigs to fatal anaphylactic shock. In addition, the indirect hemagglutination technique has been modified so that tanned sheep red blood cells treated with aspiryl-conjugates are able to combine with antibodies in the serum of individuals clinically sensitive to aspirin and para-aminosalicylic acid.

Allergic and Immunologic Studies of a “Purified” Fraction of Ragweed Pollen

Summary and Conclusions The allergenic and antigenic properties of pool C, a “purified” fraction of ragweed pollen, were investigated. As compared to whole ragweed pollen extract, pool C was found to be highly antigenic as measured by its ability to stimulate antibody production in rabbits, induce passive cutaneous anaphylaxis and provide indirect hemagglutination with both human and animal antisera. Pool C is not immunochemically pure as evidenced by three or four lines produced by its homologous antisera in immunoelectrophoresis. However, many of the antigens in the whole ragweed extract have been eliminated. Positive hemagglutination titers with pool C were found with the sera of 72% of untreated ragweed-sensitive individuals. This contrasted greatly with 20% positive reactions found with whole ragweed extract. On a weight basis, pool C was 100 to 1000 times more skin-reactive than whole ragweed extract, both by direct and passive transfer tests. In vitro and particularly in vivo neutralization tests indicated that some of the allergens in whole ragweed extract have been eliminated in the preparation of pool C.

Chrome typhoid vaccine

1. Experimental typhoid vaccines, treated by 1% formalin and 0·02% chrome alum (KCr(SO4)2.12H2O), as suggested by Japanese workers were prepared and tested by the usual in vitro and in vivo tests.2. Agglutination tests, antibody production in rabbits, active and passive mouse protection tests confirm the stability of the Vi antigen of the vaccine, if properly stored, and the good protection afforded to laboratory animals in both the active and passive mouse protection tests.3. It is suggested that only a full-scale field trial in a typhoid endemic area can give the answer as to the real efficacy and/or superiority of the chrome vaccine over other typhoid vaccines.

An immunologic study of two metabolically active peptides from the anterior pituitary gland.

Peptides I and II, two metabolically active peptides from porcine pituitary glands, stimulated antibody production in rabbits. The antiserum obtained had antibodies which interacted with peptides I and II from porcine pituitaries, peptides I and II from human pituitaries, and fraction H prepared by Rudman from hog pituitaries. None of the other pituitary preparations tested showed any interaction with peptides I and II. All three preparations stimulated the release of free fatty acids from rabbit adipose tissue in vitro. Antiserum inhibited this biological action to a different extent in each of the three preparations. Following an intravenous injection of peptides I and II there was a rapid rise in the plasma concentration of free fatty acids in rabbits. Injection of antiserum together with peptides I and II failed to block the rapid increase in free fatty acids. However, an injection of at least ten times the minimal amount of peptides I and II which raised the plasma free fatty acid concentration produced no increase in a rabbit with a high antibody titer to these peptides.

Immunological studies of gastrin.

Hog gastrin prepared by the Gregory method stimulates antibody production in rabbits. Rabbit anti-hog gastrin serum which was reacted with hog, dog, and human gastrins in vitro caused reduction of their gastric stimulatory effect, indicating a biological cross reaction among these species.

An immunoelectrophoretic study of the slime mold Dictyostelium discoideum

1. 1. The procedures in preparing antigens of the slime mold Dictyostelium discoideum for antibody production in rabbits and for electrophoretic separation are described. 2. 2. The technique of preparing agar plates for immunoelectrophoretic analysis of the antigens is described. 3. 3. The numbers of antigens or specific combining groups synthesized or eliminated during the transition of the slime mold from the vegetative amebas to the mature spores were determined. 4. 4. The numbers of antigens which vegetative amebas, migrating pseudoplasmodia, and mature spores share were determined by absorption of the antiserum. 5. 5. The possibility that the anterior and posterior regions of a migrating pseudoplasmodium differ in their antigenic constitution is discussed. 6. 6. It is suggested that differentiation of the slime mold from the vegetative amebas to the mature sorocarp is accompanied by the destruction of antigens and by the synthesis or change in specificity of other antigens.

Effect of variation in body temperature on antibody production in rabbits and mice.

Effect of variation in body temperature on antibody production in rabbits and mice.

Pulmonary allescheriasis.

Pulmonary allescheriasis.

SURFACE ANTIGEN DYNAMICS IN THE SLIME MOLD, DICTYOSTELIUM DISCOIDEUM

1. Methods are described whereby antigenic material of the slime mold, D. discoideum, effective in antibody production in rabbits, is prepared.2. Antisera produced in response to migrating pseudoplasmodia, mature spores and mature spore surface antigens effected maximal agglutination of amoebae. Such antisera effected only minimal agglutination of spores.3. Antisera of the above types absorbed by amoebae and spores in all combinations revealed the presence of a surface antigen(s) on the spores which is (are) not common to amoebae from early aggregates or migrating pseudoplasmodia.4. It has been suggested that a relationship exists between the adhesive properties of the amoebae and spores and their agglutinating properties in the presence of antisera.

The Cross Reactions of Antisera Prepared Against Four Human Cell Strains in Continuous Tissue Culture

Summary Four human cell strains of normal and malignant origin were used for the production of antibodies in rabbits. These antisera, when tested for cytotoxicity, showed species specificity but no tissue specificity in tissue culture test systems. Antisera prepared from any one of the 4 human strains produced cytotoxic changes in all the other strains as well as in recently explanted human cells of normal and malignant origin. Absorption of an antiserum with cells of normal or malignant origin completely removed all cytotoxicity.

Effects of corticosterone, hydrocortisone, and corticotropin on production of antibodies in rabbits.

The effects of hydrocortisone, corticosterone, and pituitary adrenocorticotropin on antibody production were studied immunochemically in rabbits. Hydrocortisone in doses as low as 2.5 mg. per day (approximately 1.0 mg. per kg. per day) markedly depressed antibody production whereas corticosterone in doses four times as large exerted no significant effect. Corticotropin in doses of 15 units daily exerted an effect similar to that of corticosterone, but its effect was more like that of hydrocortisone when given in doses twice as great. This finding is in agreement with observations that the prolonged administration of corticotropin in sufficient dosage leads to increased output of hydrocortisone by the adrenal cortex of the rabbit. Corticosterone did not antagonize the effect of hydrocortisone on antibody production, but seemed to act additively with it. Although each of the hormones tested induced some adverse effect on nitrogen balance as measured by the weights of the animals, and hydrocortisone induced the more striking decreases in weight, there was no direct correlation between the effects of these hormones on antibody production and their effects on weight.

The Use of “Incomplete” Newcastle Disease Virus as an Experimental Vaccine

NADEL and Eisenstark (1955) observed a stage in the development of Newcastle virus in chick embryos in which the allantoic fluids contained hemagglutination units but were devoid of infective power. The interpretation placed upon this observation was that Newcastle virus, in the maturation cycle, passes through an “incomplete” stage in which the virus possesses some of the properties of the mature form but lacks the property of infectivity. This phenomenon suggests the “incomplete” stage in the maturation of influenza virus reported by numerous investigators (von Magnus, 1953; and Henle, 1953).Von Magnus (1951) demonstrated that the “incomplete” PR-8 influenza virus possessed antigenic properties. Similarly Nadel (1955) showed that “incomplete” California strain 11914 Newcastle virus stimulated antibody production in rabbits, and that these antibodies gave a good hemagglutination-inhibition (HI) test (Cunningham, 1952) when checked against the mature virus form. This suggested the possibility that “incomplete” Newcastle virus might be effective as …

Studies on Vi Antigen

Summary Vi antigens derived from three V form enteric species by a uniform isolation procedure were examined in a variety of immunological tests. The antigens derived from E. coli and P. ballerup were significantly more active than the Vi antigen obtained from S. typhosa in hemagglutination, quantitative complement fixation, and quantitative precipitin tests. Antigenic activity measured in terms of stimulation of antibody production in rabbits and active immunization of mice against challenge with S. typhosa revealed that coli Vi antigen was the most potent, followed by Vi antigen from ballerup, while the Vi antigen isolated from S. typhosa was the least effective. These differences were established most clearly in the mouse protection tests. The relationship of these biological activities to specific viscosity of the antigen preparations and the possibility of this being a causal relationship was considered. Attention was drawn to certain similarities between the biological properties of the Vi antigens and the pneumococcal polysaccharides.

Disruption of influenza virus; properties of degradation products of the virus particle.

1. The hemagglutinating capacity, enzymic activity, and infectivity of several influenza viruses were destroyed by repeated freezing and thawing of dialyzed allantoic fluids containing them. 2. Influenza virus degraded by freezing and thawing, by treatment with 5 M urea, or by heating at 65 degrees C. still combined with homologous antibody and was demonstrable by blocking of the hemagglutination-inhibition and virus neutralization reactions. 3. After 50 cycles of freezing and thawing, much of the blocking antigen activity was not sedimented by centrifugation at 120,000 g for 2 hours, and electron microscopy showed complete disruption of the virus particles. So called soluble blocking antigen was obtained from four strains of influenza A, the Lee strain of influenza B, mumps, and Newcastle disease viruses. 4. Soluble blocking antigens from influenza A viruses were highly strain-specific; gave little or no reaction in complement-fixation tests; stimulated but little antibody production in rabbits and did not induce immunity in mice; caused reactivation of infective virus in neutral mixtures of homologous virus and immune serum. 5. Repeatedly frozen and thawed influenza virus preparations did not interfere with the propagation of infective virus in the allantoic sac. The blocking antigen activity they contained was precipitated by half saturated ammonium sulfate, destroyed by trypsin, chymotrypsin, or heating at 56 degrees C. for 30 minutes, but was unaffected by desoxyribonuclease or ribonuclease. 6. These findings are in accord with the view that soluble blocking antigen obtained from influenza virus particles on disruption by repeated freezing and thawing is protein in nature and represents the essential antigenic material of the intact virus.

Response to bacterial antigen and serum complement activity in rabbits treated with ACTH.

A number of investigations during recent years have shown that ACTH and cortisone can inhibit the production of antibodies in rabbits immunised by various means (Germuth & Ottinger, 1950, Bjørneboe et al., 1951, Moeschlin et al., 1951, Fagræus & Berglund, 1952, Hamburger, 1952). In a previous investigation (Teilum et al., 1950) on the influence of cortisone on plasma cells and the reticulo-endothelial system, the authors used rabbits hyper-immunised with Pfeiffer’s bacillus type b. The same antigen has been employed in this present work, the aim of which was to examine the effect of moderate doses of ACTH on the production of antibodies.

The influence of ACTH and cortisone on experimental antibody production in rabbits.

The influence of ACTH and cortisone on experimental antibody production in rabbits.

Studies with H. Pertussis

Summary A method of separation of the agglutinogen of B. parapterussis from other cellular components is described. This purified agglutinogen has been shown to be free of toxins; it is a protein-like material capable of stimulating the production of antibodies in rabbits. As shown by cross-absorptive tests, this purified fraction has retained the minor antigen common to phase I, H. pertussis. It is probably present as a prosthetic group on the molecule of the major antigen. The comparative chemical properties of the agglutinogen of B. parapertussis and phase I, H. pertussis are discussed. The possible clinical use of the material in skin testing for susceptibility to and immunization against parapertussis is suggested.

Sensitizing Capacity of Polysaccharide of Monilia tropicalis.

Polysaccharide derived from Monilia tropicalis by acetic-acid hydrolysis is capable of inducing active sensitization of guinea pigs and of eliciting the production of antibodies in rabbits. Its capacity in these respects, however, is inferior to that of the intact cell.