Isolation and properties of a macromolecular, water-soluble, immuno-adjuvant fraction from the cell wall of Mycobacterium smegmatis.
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Trypsin- and chymotrypsin-treated delipidated cell walls of Mycobacterium smegmatis were digested overnight with lysozyme. The water-soluble products thus obtained were filtered on a column of Sephadex G-50; the first peak, excluded from the column, has immunological adjuvant activity. The material of the excluded peak is obtained after lyophilization as a snow-white, fluffy material, soluble in water and insoluble in organic solvents. It behaves as a slightly polydisperse macromolecule in an ultracentrifuge, with an approximate molecular weight of 20,000. All the constituents of this material are typical bacterial cell-wall constituents; thus, the water-soluble adjuvant is considered to be an "oligomer" of the cell wall. When added to Freund's incomplete adjuvant with an antigen (e.g., ovalbumin) and injected into hind-foot pads of guinea pigs, this water-soluble adjuvant increases the amount of precipitating antibodies and induces hypersensitivity to ovalbumin and the biosynthesis of gamma(2)-type precipitating antibodies. The water-soluble material has a stronger adjuvant activity than equal amounts of whole bacteria, cell walls, or wax D, and seems to be the first well-defined, water-soluble, adjuvant-active fraction isolated from Mycobacteria. (+info)
Candida precipitins in pregnant women: validity of the test systems used.
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Sera from 200 pregnant women, with symptoms suggestive of vaginitis and harbouring yeast in the vagina, were examined for precipitating antibodies to three antigens of C. albicans, using a gel double diffusion test. A high overall incidence of precipitin-positive sera (47.5%) was found compared with an incidence of 18% in the unselected pregnant population previously studied (Stanley, Hurley, and Carroll 1972). Using the clinicopathological criteria of Carroll, Hurley, and Stanley (1973), a final aetiological diagnosis of C. albicans mycosis was reached in 75 cases and precipitins were demonstrated in 64%. Forty-eight women harbouring C. albicans responded favourably to a single course of antifungal treatment, and probably had mycotic vaginitis. The incidence of precipitins in this group was 42%. C. albicans was isolated from a further 55 of 62 patients, in whom the incidence of precipitins was 32%.;Booking' sera were investigated from 50 of the 200 women studied. Sixty-four per cent of women had symptoms of vaginitis at booking and 32% were precipitin positive. Twenty-eight per cent had precipitins on both occasions, and a further 24% acquired candida precipitins during pregnancy. None of the seven newborn with oral or skin thrush had precipitins to C. albicans.The results indicate that the detection of precipitating antibodies to C. albicans, particularly to all three of the antigens described in this paper, would be a useful additional criterion in the diagnosis of candida vaginitis, particularly if the vaginitis were persistent, recurrent, or unresponsive to therapy. The sensitivity of the test system used was 64%, and its specificity 87%; as such, the test is valid and may be reasonably useful as a screening procedure. (+info)
Precipitins to dietary proteins in serum and upper intestinal secretions of coeliac children.
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We have used precipitin tests to detect antibodies to 10 dietary proteins in the serum (71 cases) and intestinal secretions (51 cases) of a group of children. Thirty-three of the patients had untreated coeliac disease. Our aims were to find out if, in coeliac patients, there was intestinal secretion of antibodies to wheat proteins only or if, as in coeliac serum, antibodies to many food proteins were present; and to confirm that secretion of antibodies to wheat or gluten was specific for coeliac disease.Precipitins to one or more dietary antigens were detected in the intestinal secretions of 26 out of 30 coeliacs and of 11 out of 21 children who did not have coeliac disease. Most of the positive reactions were with the antigens wheat flour, gluten, oatmeal, and egg. Though precipitins to wheat flour or gluten were present in the intestinal secretions of 22 out of 30 coeliacs this was not specific for coeliac disease for these precipitins were also present in 8 out of 21 non-coeliac children.Serum precipitins were detected in 27 out of 33 coeliacs (to the antigens wheat flour, gluten, oatmeal, rice flour, milk, bovine calf serum, sheep serum, and egg) and in 5 out of 33 non-coeliacs (mainly to milk and calf serum, but two infants aged 3 and 5 months had precipitins to several antigens). (+info)
Quantitative measurement of precipitating antibodies in streptococcal grouping antisera by the single radial immunodiffusion technique.
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A comparative study was made of the single radial immunodiffusion test and the classical quantitative precipitin test for determining the amount of precipitable antibodies present in streptococcal groups A and C antisera. The potency of 21 group A and 54 group C antisera was determined by both methods; purified group-specific carbohydrates were used as antigens. The coefficient of correlation between the results from the two methods was 0.976 for group A antisera and 0.946 for group C antisera. When the concentration of antigen, the volume of antiserum used, and the depth of the antigen-agar mixture are kept constant, the diameter of the precipitin disc is directly related to the concentration of precipitable antibodies present in the antiserum. The use of the radial immunodiffusion test for evaluating and standardizing streptococcal grouping antisera is discussed as well as the advantages and disadvantages of using a concentrated vaccine for producing these antisera. (+info)
Serological investigations of a bovine respiratory disease ("urner pneumonie") resembling farmer's lung.
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The immunological response of cattle exposed to moldy hay was examined by agar gel diffusion with standard farmer's lung hay antigens. A high incidence of precipitins against Micropolyspora faeni (60%) and moldy hay antigen (80%) was detected in exposed but apparently healthy cattle from a region with a high incidence of bovine farmer's lung. In comparison, in the plains, a low incidence area, we found only 1 animal of 164 harboring precipitins against M. faeni. We further observed that many animals from exposed populations lost their precipitins during pasturing and regained them during winter housing. Thirty-nine clinical cases of bovine farmer's lung ("Urner Pneumonie") were investigated serologically. Only 49% of these animals showed precipitins against M. faeni and 54% showed precipitins against moldy hay antigen. We discuss in this paper the probable causes of this apparent lack of immunological response. (+info)
Pathogenesis and immunological aspects of experimental histoplasmosis in cynomolgus monkeys (Macaca fascicularis).
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Studies were carried out to obtain basic information on the pathogenesis of experimental histoplasmosis in Cynomolgus monkeys (Macaca fascicularis) and to determine whether such infected primates can be used as a source of positive reference sera in serological tests for histoplasmosis. Ten monkeys were inoculated intraperitoneally with approximately 9.1 x 10(7) viable Histoplasma capsulatum yeast-form cells or cell aggregates. At periodical intervals, their sera were tested for antibodies to H. capsulatum by the complement fixation (CF), immunodiffusion, and latex agglutination tests. Selected monkeys were also sacrificed at periodical intervals for cultural and pathological evaluation of their tissues. Infection with H. capsulatum elicited high-antibody responses, and the fungus disseminated to many organs. Initially, the infected monkeys developed CF titers as high as 1:256 to both histoplasmin and H. capsulatum yeast cell antigens. Subsequent challenges boosted the CF antibody titers to levels as high as 1:1,024. All of the monkeys developed M precipitins, and some also produced H precipitins. Latex agglutination titers as high as 1:1,024 were also demonstrated. Our findings show that Cynomolgus monkeys experimentally infected with H. capsulatum by the intraperitoneal route develop a mild form of histoplasmosis and that these animals can be used as a source of reference sera in serological tests for histoplasmosis. (+info)
Chemical properties of the pili of Corynebacterium renale.
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Pili separated from the cells of Corynebacterium renale strain 46 (type II) by agitation at high speed in a homogenizer were purified by repeated cycles of ammonium sulfate precipitation, sonic treatment, and centrifugation. The preparation of purified pili formed a single antigen-antibody line in agar gel and showed an absorption maximum at 275 nm. The pili subjected to dodecyl sulfate-polyacrylamide gel electrophoresis formed a main band and corresponded to the molecular weight of 19,000. The fact that the total nitrogen of the amino acids of the pili was nearly equal to its nitrogen content, together with the absence of detectable carbohydrate, has led to the conclusion that the pili are protein. The pilial protein was composed of 20 amino acids. Preparations of pili which had been treated with 0.5 n NaOH, but not with 1 n HCl, no longer appeared filamentous and failed to form a precipitate with the antibody in agar gels. A comparison has been made of the amino acid composition and certain properties of the pili of C. renale and type I pili and F pili of gram-negative bacteria. (+info)
Specific identification of fraction I-positive Pasteurella pestis colonies on antiserum-agar plates.
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A method is described for the use of antiplague serum in Blood Agar Base plating media to detect fraction I-positive Pasteurella pestis. The antiserum was produced conveniently and in large volume in rabbits by use of Cutter plague vaccine combined with Freund's complete adjuvant. P. pestis colonies were specifically identified within 48 hr after plating by the presence of a precipitin ring surrounding each colony. The basis of the test was shown to be a precipitin reaction between fraction I antigen released from P. pestis colonies after chloroform vapor treatment and fraction I antibody present in the antiserum-agar medium. (+info)