Skin Reactions In Rabbits Research Articles

The delayed-type hypersensitivity reaction is dependent on IL-8. Inhibition of a tuberculin skin reaction by an anti-IL-8 monoclonal antibody.

Cell-mediated immune reactions are essential to our immune response toward foreign organisms such as microorganisms, or in the response toward foreign tissue Ags, as seen in the rejection of allogeneic transplanted organs. Similar reactions form the basis for the development and the progression of delayed-type hypersensitivity (DTH) reactions. We found that the alpha-chemokine IL-8 plays an important pathophysiologic role for the development of a DTH reaction because infusion of a neutralizing anti-IL-8 mAb (WS-4) was able to suppress the development of a tuberculin skin reaction in rabbits, as judged by histologic, biochemical, and clinical examinations. Thus, the number of neutrophil granulocytes and lymphocytes at the site of tuberculin injection was decreased considerably, and the clinical signs of inflammation were suppressed almost completely at 24 h after intracutaneous injection of tuberculin, as judged by the size of the infiltrates. In contrast, we did not see any effect on the visible erythema of the skin. We found that the tissue content of myeloperoxidase (MPO), reflecting the number of infiltrating neutrophils, was lowered significantly. Furthermore, immunohistochemical analysis confirmed that IL-8 immunoreactivity is actually enhanced in the skin of positive tuberculin reactions. The results indicate that IL-8 plays an important role for the early accumulation of leukocytes in the skin and for the clinical signs of a DTH reaction.

Effect of Rumex nepalensis extracts on histamine, acetylcholine, carbachol, bradykinin, and PGs evoked skin reactions in rabbits

The antihistaminic, anticholinergic, antibradykinin, and/or antiprostaglandin activity of aqueous and the alcoholic leaf extracts of the plant Rumex nepalensis were tested on the prepared site on the back of rabbits, using a control and a test site. Aqueous extract was found to reduce the mean size of the wheal produced by histamine, acetylcholine, carbachol, and bradykinin. The alcoholic extract reduced the size of the wheal produced by histamine, acetylcholine and carbachol. The results indicate that Rumex nepalensis may have antihistaminic, anticholinergic, and/or antibradykinin activity.

Infection, immunity and the development of local skin reactions in rabbits infected with cyclically-transmitted stocks of Trypanosoma congolense.

The development of local skin reactions following bites by tsetse flies (Glossina morsitans) infected with Trypanosoma congolense was observed in normal rabbits, in rabbits with concurrent infections with T. congolense and in rabbits challenged after immunization by infection and treatment. Local reactions developed in normal rabbits within seven days of fly bite and were resolved by approximately 19 days after infection. In rabbits with concurrent infections no typical local reactions developed when they were bitten by infected tsetse flies. By allowing a single infected fly to bite a rabbit sequentially it was established that suppression of reaction development to homologous secondary challenge occurred within three to eight days of primary infection. Suppression of reaction development also occurred in infected rabbits challenged with an antigenically heterologous trypanosome stock within 14 to 18 days of primary infection, suggesting that reaction suppression is not associated with the specific immune response. It was also found that no neutralizing antibodies had developed against the parasites used for secondary infection by 28 days post-challenge. Rabbits were immunized by allowing an infected tsetse fly to feed on a rabbit and seven days later, when local reactions had appeared, treating it with homidium chloride. All rabbits challenged within 75 days of immunization by a tsetse fly infected with a homologous trypanosome stock resisted re-infection and some animals were immune up to 300 days after treatment. In most of the immunized animals specific neutralizing antibody activity against homologous parasites was low, suggesting that local expression of immunity in the skin might be more important in resistance to cyclically transmitted infections than serum antibodies. No local reactions developed on immune rabbits challenged with a homologous trypanosome stock but immunized rabbits challenged with heterologous trypanosome stocks developed typical local reactions.

Development of local skin reactions in rabbits infected with metacyclic forms of Trypanosoma congolense cultured in vitro.

Development of local skin reactions in rabbits infected with metacyclic forms of Trypanosoma congolense cultured in vitro.

Endotoxic activity of cell-free rumen fluid from cattle fed hay or grain.

The cell-free rumen fluid from cattle fed hay or grain exhibited the following biological characteristics which strongly suggest the presence of endotoxin or a toxic principle similar to endotoxin of gram-negative bacteria: proved lethal to mice when injected with actinomycin D; proved extremely lethal to chick embryo; induced biphasic pyogenic response in rabbits; enhanced susceptibility to bacterial infection in mice; evoked positive epinephrine skin reaction in rabbits and phenol-water or aqueous ether proved lethal to mice and chick embryos. A quantitative difference in concentrations of endotoxin was observed on LD50 in mice and chick embryos and response to the epinephrine skin test in rabbits. Cell-free rumen fluid of grain-fed cattle contained at least twice as much endotoxin as that of hay-fed cattle. Endotoxin in cell-free rumen fluid and in higher concentration in cattle fed grain than in those fed hay support the hypothesis that rumen bacterial endotoxins may participate in the pathogenesis of diseases associated with high grain feeding such as lactic acidosis and the sudden-death syndrome.

Immunochemical and Biologic Studies of the Lipopolysaccharide of Bacteroides Melaninogenicus Subspecies Asaccharolyticus

Abstract Lipopolysaccharide (LPS) was isolated from the outer membrane complex of Bacteroides melaninogenicus by gel chromatography by using sodium deoxycholate (NaD), an endotoxin-disaggregating detergent in the running buffer. Reaggregation occurred after removal of residual NaD. LPS was also prepared by the phenol/water method for comparison. The LPS was composed of loosely bound lipid (62%) and carbohydrate (32%), with less than 5% protein. Glucose, galactose, and glucosamine were the major sugars as detected by gas-liquid chromatography (GLC). Heptose and 2-keto-3-deoxyoctonate (KDO) were found to be absent when using colorimetric tests. Fatty acid analysis by GLC disclosed an unusual pattern; β-OH myristic acid was notably absent. Biologic activity was assessed with the dermal Shwartzman test, the Limulus lysate assay, and the chick embryo lethality test. LPS preparations gave no positive skin reactions in rabbits in doses up to 1 mg, compared to Salmonella typhi 0:901 endotoxin which was positive at 12.5 µg. Limulus lysate gelation and chick embryo lethality occurred at doses 30-fold or more different from Salmonella endotoxin controls. Hemagglutination inhibition (HAI) studies demonstrated the presence of capsular polysaccharide as a contaminant found in the phenol/water extracted LPS. This polysaccharide could be removed partially by further purifying the LPS with ultracentrifugation. The LPS of B. melaninogenicus lacks two of the core sugars characteristic of aerobic Gram-negative endotoxins, has a unique fatty acid composition, and displays low biologic potency. These findings may explain the rarity of septic shock in patients infected with this organism. The presence of capsular polysaccharide contaminating the phenol/water-extracted LPS may explain partially the serologic heterogeneity which has been described previously in studies of the LPS of this organism.

Endotoxic activities of lipopolysaccharides of microorganisms isolated from an infected root canal in Macaca cynomolgus

Lipopolysaccharides (LPS) prepared from a strain of Bacteroides oralis, a strain of Fusobacterium necrophorum, and a strain of F. nucleatum, all isolated from an infected root canal in monkey (Macaca cynomolgus), were examined for endotoxic activities using primary skin reactions in rabbits and induction of leukocyte chemotaxis in rats. LPS of B. oralis showed considerably lower ability to cause skin inflammation than LPS of the fusobacteria. However, the leukotactic effect of the LPS preparations as determined by the wound chamber method in rats was approximately of the same proportion. In both tests the reactions were compared with those of commercial LPS of Salmonella typhi. This study shows that endotoxic LPS can be isolated from oral Gram-negative bacteria, which have infected the root canal. Therefore LPS may play a role in the development and maintenance of chronic inflammation of the periapical tissues.

Chemical and Immunological Properties of Protein and Polysaccharide Fractions Extracted from Aspergillus fumigatus

Protein and polysaccharide fractions were extracted from the cells of Aspergillus fumigatus and chemical and immunological properties of these fractions were investigated. Protein fraction which was designated AAP fraction was purified by the column chromatography on DEAE-cellulose. In the purified protein fraction (AAP-A fraction), about 3.5 per cent of carbohydrate were contained. By the intracutaneous injection of 10 μg of AAP fraction in the rabbits or guinea pigs which were immunized with A. fumigatus, Arthus and delayed type skin reactions were elicited. In the patients with pulmonary aspergillosis, delayed type and wheal-erythema skin reactions were elicited by the intracutaneous injection of 1 to 10 μg of AAP fraction. The activity of delayed type skin reaction of AAP fraction was lost completely by the treatment of AAP fraction with proteolytic enzyme. The AAP fraction showed high specificity by the skin reaction in the rabbits which were immunized with aspergillus and other fungal species. Polysaccharide fraction was further fractionated into two fractions, glucan and galactomannan, and they were purified by the column chromatagraphy on Dowex 50 and DEAE-cellulose and gel filtration on Sephadex. Galactomannan which was designated APS-66 was active by the skin and serological reactions in the immunized rabbits, guinea pigs and patients with pulmonary aspergillosis. The APS-66 (galactomannan) was available for the antigen of precipitation, complement fixation and passive hemagglutination reactions with rabbits antisera. By the intracutaneous injection of 10 μg of APS-66 in immunized rabbits or guinea pigs, Arthus type skin reaction was elicited. In case of the patients with pulmonary aspergillosis, wheal-erythema skin reaction was elicited by the intracutaneous injection of 1 to 10 μg of APS-66. Glucan was inactive immunologically in immunized rabbits. Above results show that protein (AAP) and polysaccharide (APS-66) fractions purified from A. fumigatus were useful for the antigens of the skin and serological reactions in the patients with pulmonary aspergillosis and immunized rabbits or guinea pigs.

Skin reactions in rabbits sensitized with typical and atypical mycobacteria and tested with homologous and heterologous protoplasm.

Skin reactions in rabbits sensitized with typical and atypical mycobacteria and tested with homologous and heterologous protoplasm.

PROLONGED SURVIVAL OF KIDNEY HOMOTRANSPLANTS IN DOG TREATED BY EPSILON-ACETAMIDE CAPROIC ACID.

IN previous experiments the inhibiting influence of epsilon-acetamide caproic acid (EAACA) on Sanarelli-Schwartzmann haemorrhagic skin reaction in rabbits, as well as on rejection of skin transplants in rats, was demonstrated1–3.

Studies on Synthetic Polypeptide Antigens

Summary The majority of the skin reactions in rabbits immunized with a variety of synthetic polypeptide antigens which elicited the production of circulating antibody were the classical active Arthus reactions (Arthus type I). However, the polypeptide preparation Glu56Lys43Tyr1-1 elicited a skin reaction which followed the course of the delayed, or tuberculin, reaction despite the fact that it elicited the production of circulating antibody (Arthus type II). On the basis of the skin reactions elicited by the various synthetic polypeptides, the hypothesis was advanced that the character and intensity of the skin reactions are determined in part by the amino acid composition of the antigen.

Chemical studies on cellular components of Bordetella pertussis. I. Purification and properties of agglutinogen.

Onoue, Kaoru, (Kyushu University, Fukuoka, Japan), Masayasu Kitagawa, and Yuichi Yamamura. Chemical studies on cellular components of Bordetella pertussis. I. Purification and properties of agglutinogen. J. Bacteriol. 82:648-656. 1961.-A method is described for the preparation of the agglutinogen of Hemophilus pertussis in phase I by methanol precipitation and ion-exchange chromatography. High homogeneity of this agglutinogen was evidenced by chromatography, but a trace of impurity was detected by ultracentrifugal analysis and by an agar gel diffusion test.The agglutinogen produced an allergic skin reaction in rabbits which had been immunized with whole phase I organisms. Chemical studies indicate that the agglutinogen should be a simple protein in nature. The approximate molecular weight was determined to be 10,000 by Archibald's method.

Purification of factor I and recognition of a third factor of the anthrax toxin.

SUMMARY Factor I of the anthrax toxin was isolated and showed one major component in the ultracentrifuge and on paper electrophoresis; it contained less 5.5% of extraneous antigens detectable by serological precipitation in gels. The final preparation contained all the usual amino acids (N = 10.1%) and some carbohydrate (6%, calculated as glucose) and phosphorus (0.7%). The most striking aspects of its analysis were a high ash (10-13%) and a light absorption at 260 mμ. The high ash was not due to one element but to a highly variable metal content (mainly Ca, Mg, Ni, Cu) indicating a powerful and indiscriminate chelating action of factor I. This chelating action might have been due to the chemical entity which absorbed light at 260 mμ and which was not RNA or DNA. The final preparation of factor I was not toxic when injected alone but when mixed with purified factor II it evoked oedema in the skin of a rabbit and killed mice. However, the concentrationof this mixture which killed mice formed a much larger skin reaction in rabbits than a comparable dose (based on mouse LD 50) of either curde toxin or a mixture of crude factors I and II. An investigationof this fact led toteh demonstraion and partial purification of a third factor (III) of the anthrax toxin which: (1) was different serologically from factors I and II; (2) was present in anthrax toxin produced in vivo; (3) was non-toxic when injected alone; (4) was lethal for mice when mixed with factor II but not with factor I; (5) increased the lethality of mixtures of factors I and II for mice and decreased their capacity to produce oedema in the skin of rabbits. A mixture of factors I, II and III showed synergic action in toxicity tests in mice; the mixture killed guinea pigs which showed signs of ligaemic secondary shock (as did guinea pigs killed by anthrax infection).

Further study of skin reactions in rabbits with experimental allergic encephalomyelitis: reactions to tests with purified white matter fractions; use of a quantitative technique for evaluating sensitivity production of sensitization and disease by intradermal injection of antigen without adjuvants.

Further study of skin reactions in rabbits with experimental allergic encephalomyelitis: reactions to tests with purified white matter fractions; use of a quantitative technique for evaluating sensitivity production of sensitization and disease by intradermal injection of antigen without adjuvants.

水産食品の腐敗中毒に関する研究-VI

As shown in our previous paper, V, neither trimethylamine (TMA) nor trimethylamine oxide (TMAO) was described to have any recognizable synergism on the action of histamine or “saurine” to contract intestine or uterus of guinea pigs. This paper also deals with the effect of those substances when added to histamine or “saurine” tested by shock symptoms in guinea pigs, skin reaction in rabbits and lethal dose for mice (LD50). The addition of TMA or TMAO resulted in no appreciable change in the action of histamine of “saurine” Per se in each experiment. Therefore, the presence of TMA or TMAO, if any, coald not be regarded to stand as a cooperative factor to either histamine or “saurine” in a foodstuff responsible for allergylike food poisoning.

A Skin Reaction in Rabbits Produced by Intradermal Inoculation of Suspensions of Killed Pasteurella tularensis

A Skin Reaction in Rabbits Produced by Intradermal Inoculation of Suspensions of Killed Pasteurella tularensis

Hemorrhagic Skin Reactions in Rabbits with Cutaneous Sensitization to Streptococci and Pneumococci (Shwartzman Phenomenon).

SummaryWhen an intracutaneous injection of bacterial filtrates or heat-killed organisms into sensitized rabbits was followed by intravenous injections of sufficient amounts of killed or living organisms, a hemorrhagic reaction frequently developed at the site of the intracutaneous injection.The most intense hemorrhagic reactions were observed in animals with the highest degree of cutaneous sensitization. We did not observe hemorrhagic phenomena in non-sensitized animals treated in a similar manner.

Diagnosis of Echinococcal (Hydatid) Disease in Man by Intradermal Reaction to Rabbit Cysticercus Antigen

SummaryAn extract of cysticerci of the common rabbit cestode, Taenia pisiformis, has been used as an antigen for skin testing in cases of echinococcal infection. An antigenic relationship between Taenia pisiformis and Taenia echinococcus is suggested by the precipitin-reaction, the alexin fixation, and the skin reaction in rabbits harboring Taenia pisiformis.Four patients known to have hydatid disease gave immediate positive skin reactions with the rabbit antigen. Twenty normal subjects used as controls exhibited at most but slight reactions, and were considered negative.

The Use of Trypan Blue to Demonstrate the Immediate Skin Reaction in Rabbits and Guinea Pigs

Summary and Conclusions By the intravenous administration of trypan blue, the immediate skin reaction to a homologous antigen may be demonstrated in serum treated rabbits and guinea pigs in the usual testing for direct, passive and reversed passive sensitization in both animals. The same type of reaction may be shown to an injection of sera of sensitized animals, of sera showing so-called primary toxicity for the guinea pig, and to very high dilutions of histamine. It is believed that the infiltration of the dye into these reaction sites is, at present, to be looked upon as no more than a sort of indicator of the presence of an edematous change which is an expression of a toxic action upon the capillary endothelium.

The skin response of rabbits to non-hemolytic streptococci.

A study of the skin reactions in rabbits which were induced by intra-dermal injection of non-hemolytic streptococci demonstrated an unusual phenomenon. Following the injection of these microorganisms a local reaction took place which reached its maximum within 24 to 36 hours. The lesion then diminished until the eighth day, when an exacerbation occurred which at times approached the original lesion in intensity. This persisted for about 2 days and disappeared slowly. The late lesion is referred to as the “secondary reaction.” This reaction was first encountered while studying a green streptococcus isolated from a rheumatic subcutaneous fibroid nodule; with it definite secondary reactions were induced in 50 per cent of normal adult rabbits and doubtful reactions in an additional 13 per cent. On further extending the strains of green streptococci studied, the reaction was found to be incited by 5 out of 10 strains from nodules, blood or heart of rheumatic fever patients; by 1 out of 4 strains from the blood of subacute bacterial endocarditis patients; by 1 strain from the urine of a nephritic patient and by 1 anomalous strain isolated from a rheumatic nodule. It never followed the injection of green streptococci from normal throats, nor of hemolytic streptococci, staphylococci, micrococcus catarrhalis, B. coli, and B. influenza. The material used for intra-cutaneous injection consisted of the centrifuged sediment from 5 cc. of an 18-hour growth of the bacteria in blood broth. It was subsequently found that the reaction could be obtained with heat killed organisms. Lesions were excised daily and studied bacteriologically and histologically. In the early stages an abundant growth of streptococci was obtained, but the number of viable organisms progressively diminished until the 7th day, when none was present, although many could still be seen in the smears.