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PubMed1979

Microbiological and serological studies of farmers' lung in Finland.

Sera from few Finnish patients with clinical farmers' lung react in precipitin tests with extracts of the thermophilic actinomycetes that commonly cause the disease elsewhere. Hays associated with the disease in Finland showed less evidence of spontaneous heating and contained fewer actinomycete spores than British hays. Only Thermoactinomyces vulgaris was sometimes abundant. Some species of mesophilic fungi were more abundant than in Britain and one, Aspergillus umbrosus, reacted with most sera from farmers' lung patients in precipitin tests. A panel of antigens, including thermophilic actinomycetes, A. umbrosus and other species of the Aspergillus glaucus group, is recommended for screening farmers' lung sera.

Adult↗
PubMed1980

Aspergillus hypersensitivity in asthmatics in Cape Town.

An analysis of skin prick reactions in 500 consecutive patients referred for investigation and management of asthma showed that 240 patients had multiple positive reactions. Of these patients 46% had positive reactions to Aspergillus and they differed from patients with multiple positive reactions excluding Aspergillus by having onset of asthma at a younger age and being more atopic. Precipitins to Aspergillus were present in the serum of twenty-six of seventy-one consecutive patients in whom these were looked for. Thirteen of the patients with positive precipitins had chest radiograph shadows compatible with allergic bronchopulmonary aspergillosis (ABPA) and in eight of these there was sufficient evidence to make a diagnosis of ABPA. These results were obtained in a region where ABPA had never been previously recognized and they highlight the importance of routine skin prick testing with Aspergillus antigen and of awareness of ABPA in making this diagnosis.

Adult↗
PubMed1980

Extrinsic allergic bronchiolo-alveolitis in children.

Fifteen children, 6-14 years old, who were heavily exposed to pigeon droppings, presented with cough, chills and breathlessnes on mild exertion. Twelve gave immediate positive skin test reactions to pigeon droppings. Ten of the fifteen also gave a late (Arthus) reaction, and seven gave a positive delayed reaction which was maximal at 24--72 hr. Nine children had precipitins against avian droppings. In ten patients who were tested for lymphocyte transformation, five gave positive responses to both serum and pigeon droppings. Radiological findings of the chest showed parenchymatous infiltrations and diffuse interstitial reticulations, suggestive of extrinsic allergic alveolitis. In a control group of fifteen heavily-exposed children who had no symptoms, immediate and late skin reactions occurred respectively in five and two. One only had a positive precipitin test, and one of five tested had a positive lymphoblastic transformation. In a non-exposed comparable group of children, all tests were negative except for one immediate positive skin response. Children heavily exposed to the inhalation of avian antigens can develop extrinsic allergic bronchiolo-alveolitis.

Adolescent↗
PubMed1982

Antibody against a pigeon bloom-extract: a further antigen in pigeon fanciers' lung.

Pigeon bloom, a waxy powder which coats the feathers of racing pigeons, has been investigated as a potential antigen in pigeon fanciers' lung. Precipitins to a soluble bloom-extract were demonstrated in fifty-nine of ninety-seven people studied at two Scottish-pigeon shows, whilst precipitins to pigeon serum were found in thirty-seven. A radioimmunoassay was developed to allow an estimation of the intensity of antibody response to this antigen in nineteen pigeon fanciers being investigated for pigeon fanciers' lung. The mean antibody level to bloom extract was significantly higher (P less than 0.05) in those with acute symptoms compared to those with less severe symptoms. Pigeon bloom is a highly antigenic material and provides an alternative to pigeon droppings as a source for the inhalant antigens regularly encountered by those keeping pigeons.

Alveolitis, Extrinsic Allergic↗
PubMed1975

Interactions between ricinus agglutinin and human IgM and IgG.

Ricinus agglutinin, purified until homogeneous, precipitates serum glycoproteins with terminal nonreducing galactose residues including IgM and IgG. Almost 100% of IgM reacted with anti-IgM and ricinus agglutinin in quantitative precipitin tests. In similar tests, almost 100% of polyclonal IgG was precipitated by excess anti-IgG, whereas only about 10% reacted with ricinus agglutinin. In quantitative precipitin analyses and affinity chromatography experiments with insolubilized ricinus agglutinin and isolated monoclonal IgG1 and IgG3 proteins, only IgG3 proteins reacted with ricinus agglutinin.

Antibodies, Anti-Idiotypic↗
PubMed1995

Binding studies on the combining site of a GalNAc alpha 1-->-specific lectin with Thomsen-Friedenreich activity prepared from green marine algae Codium fragile subspecies tomentosoides.

The combining site of a GalNAc alpha 1-->-specific lectin (CFT) with Thomsen-Friedenreich (T, Gal beta 1-->3-GalNAc alpha 1-->Ser/Thr) activity, purified from the subspecies tomentosoides of green marine algae Codium fragile was studied by quantitative precipitin and precipitin-inhibition assays. Of 27 glycoforms tested, Tn (GalNAc alpha 1-->Ser/Thr) glycoprotein from armadillo submandibular glands, and asialo porcine submandibular glycoprotein, which contains T, Tn and GalNAc alpha 1-->3Gal(A) sequences, completely precipitated the lectin added, and less than 1 microgram glycoprotein was required to precipitate 50% 4.7 micrograms lectin nitrogen. However, CFT precipitated negligibly with Pneumococcus type-XIV polysaccharide and asialo human alpha 1-acid glycoprotein, that contain exclusively the human blood-type-II precursor sequence (II, Gal beta 1-->4GlcNAc) at the nonreducing ends. Among the sugar inhibitors tested, the human blood A-active trisaccharide [Ah, GalNAc alpha 1-->3 (LFuc alpha 1-->2)Gal] was the best inhibitor; it was about twice as active as the T disaccharide. Oligosaccharides without GalNAc alpha 1--> as part of their sequences were inactive, indicating that the acetamido group at C2 of galactose is essential for binding and that GalNAc is the main contributor in the T sequence for binding. From the data provided, it is clear that the combining site of CFT requires an alpha-anomer of GalNAc and recognizes Ah, internal GalNAc alpha 1--> of T and Tn determinants of glycans, but not the blood group I/II (Gal beta 1-->3/4GlcNAc) sequences. Consequently, CFT is a useful reagent for detecting GalNAc alpha 1-->-containing glycoconjugates.

Acetylgalactosamine↗
PubMed1977

Immune complexes in cystic fibrosis.

Eleven patients with cystic fibrosis (CF) chronically infected with mucoid P. aeruginosa and ten patients without P. aeruginosa infection were examined for occurrence of circulating immune complexes, for immune complex deposits in the dermo-epidermal junction of the skin and for precipitins against P. aeruginosa, S. aureus, H. influenzae and D. pneumoniae antigens. The serum concentrations of haptoglobin, orosomucoid, immunoglobulins, C1q, C3, C4 and total haemolytic complement, antinuclear and rheumatoid factor activities as well as white blood cell counts and erythrocyte sedimentation rates were determined also. The results indicated that 6 patients from the chronically P. aeruginosa infected group, exhibiting a spectrum of serum precipitins against P. aeruginosa antigens, also had immune complexes in the serum, while only one patient (suffering from selective IgA deficiency) in the group without P. aeruginosa infection was positive for soluble immune complexes. Granular deposits of IgM was found in the skin of 10 of the chronically P. aeruginosa infected patients and in 7 of the patients without P. aeruginosa infection. A few pactients in both groups had dermo-epidermal deposits of C1q, C3 or fibrinogen as well. Eight of the patients in the chronically infected group and five in the group without P. aeruginosa infection had organ non-specific antinuclear factors. The haptoglobin levels appeared to be elevated in the chronically infected patients (p less than 0.05). None of the other parameters showed any significant difference between the two groups.

Adolescent↗
PubMed1978

Crossed immunoelectrophoretic analysis of Neisseria meningitidis antigens and of corresponding antibodies in patients with meningococcal disease.

Sixty-three different antigens of Neisseria meningitidis groups A, B and C were demonstrated by means of crossed immunoelectrophoresis against rabbit antiserum. The group-specific polysaccharides A and C were identified in the immunoprecipitate pattern. The majority of the remaining antigens were common to all three groups of meningococci. A clearcut rise in antibodies was found in all of 27 patients with meningococcal meningitis or septicaemia. These antibodies were directed against both group-specific antigens and common antigens. No such rise was seen among the 22 patients with purulent meningitis of non-meningococcal etiology. Altogether 15 precipitins were detected in the patients with meningococcal disease. Nine of these antibodies could be identified by comparison with the rabbit antiserum. The antibody rise was semiquantified by means of a scoring system. The rise in the precipitin score of antibodies against common antigens in patients infected with group B meningococci was significantly higher than in patients infected with group A meningococci.

Antibodies, Bacterial↗
PubMed1980

Quantitative immunoelectrophoretic analysis of Salmonella typhi antigens and of corresponding antibodies in human sera.

By quantitative immunoelectrophoretic methods, 86 different antigens were found in sonicated preparations of a Salmonella typhi, using corresponding rabbit antiserum. Lipopolysaccharide was identified, and 5 other antigens characterized. Antibodies against a total of 19 S. typhi antigens were found in human sera. The antibody response in patients with typhoid fever was significantly more pronounced compared to that in normal persons. When the antibody response was expressed as a precipitin score, the predictive value of both a positive (precipitin score greater than or equal to 8) test and a negative test was 100%, when sera from patients suffering from typhoid fever were compared with sera from normal persons.

Animals↗
PubMed1966

Chloramphenicol-specific antibody.

Antibody to the antibiotic chloramphenicol was obtained by immunizing rabbits with a chloramphenicol derivative coupled to bovine gamma globulin. Production of antibody was demonstrated by the precipitin and complement fixation reactions with "reduced chloramphenicol" coupled to rabbit serum albumin as the test antigen. Specificity of the antibody was confirmed in that crystalline chloramphenicol completely inhibited complement fixation and precipitin reactions. "reduced chloramphenicol" coupled to human serum albumin provides an antigen for the detection of antibody to chloramphenicol, if it occurs in human serum in dyscrasias. With quantitative complement fixation, as little as 10(-5) microg (4 x 10(-14) mole) of chloramphenicol was detectable by the inhibition assay.

Animals↗
PubMed1969

Time, cost, and efficacy study of identifying group A streptococci with commercially available reagents.

During the 12-month period primary throat, wound, and skin cultures, tentatively identified as B streptococci, were submitted by 10 different clinical laboratories for evaluation. A total of 692 beta-hemolytic streptococci were isolated from cultures submitted and examined in parallel by the fluorescent-antibody, precipitin, and bacitracin techniques. An evaluation of the specificity and sensitivity in conjunction with basic and personnel costs was determined for each method. The standard Lancefield precipitin method was established as the standard by which the bacitracin and flourescent antibody techniques were compared. With some variation depending on the commerical source of the disc, approximately 7% of the strains examined produced false reactions with the bacitracin disc. False-negative reactions were rarely noted by the group A fluorescent antibody technique (0.5%), but an appreciable number of other Lancefield groups (B, C, and G) were nonreactive with homologous conjugates.

Bacitracin↗
PubMed1970

Antigenic variation in group A streptococci: types 11 and 9.

A strain of group A Streptococcus which was virulent but M-nontypable was isolated from patients in a hospital nursery during an epidemic. This strain, Boston 11, reacted in T-agglutination tests with antisera for types 9 and 11, an unusual combination. A comparison of this strain with Lancefield's M-11 strain (NCDC SS-721) and Alabama 11 (Provisional 61) revealed three serologically related but distinct strains. Antiserum produced with the Boston 11 strain exhibited similar reactivity with all three "11" strains as well as with M-9 (SS-501) as demonstrated in precipitin tests. Immunodiffusion studies indicated that the Boston 11 antigen was partially identical with the M-11 and M-9 strains and shared at least one antigen with the Alabama 11 strain. The Boston 11 antiserum could be made specific for precipitin tests, but bactericidal activity for the Alabama 11, M-11, and Boston 11 strains was essentially negative.

Agglutination Tests↗
PubMed1972

Comparison of a newly developed latex agglutination test and an immunodiffusion test in the diagnosis of systemic candidiasis.

A newly developed latex agglutination (LA) test and a modified immunodiffusion (ID) test were evaluated. The antigen used was a homogenate of Candida albicans. A total of 167 antisera were employed in the evaluation. They included 36 sera from clinically well persons; 78 from patients with various clinical forms of candidiasis; 52 from patients with proven cases of aspergillosis, blastomycosis, coccidioidomycosis, cryptococcosis, histoplasmosis, nocardiosis, paracoccidioidomycosis, sporotrichosis, and tuberculosis; and one serum from a patient with toruloposis. Use of the LA test in conjunction with the ID test permitted the detection of more than 90% of 43 proven candidiasis cases. Of all the heterologous cases and normal human sera tested, LA reactions were noted with 3 of 10 cryptococcosis case specimens, 1 of 9 tuberculosis case specimens, and with the torulopsemia case serum. In contrast, the only heterologous serum reactive in the ID test was that from the patient with torulopsemia. Torulopsis glabrata and C. albicans antisera gave identical reactions in LA and ID tests with T. glabrata or C. albicans antigens. ID tests with selected antigens, however, permitted differentiation of rabbit and human T. glabrata antibody from that of C. albicans antibody. Six different precipitins were recognized with the C. albicans antigens. The occurrence of multiple precipitin lines and high LA titers was suggestive of severe candidiasis. The LA test, in contrast to the ID test, appeared to have prognostic value. Together, the LA and ID tests provided a simple, rapid, and accurate means of detecting and monitoring infections by species of Candida.

Agglutination Tests↗
PubMed1973

Specific immunodiffusion test for blastomycosis.

A specific immunodiffusion test for blastomycosis has been developed. The test permitted the detection of approximately 80% of 113 proven cases of blastomycosis. Two diagnostically important precipitins designated A and B were frequently recognized in patients with blastomycosis. Routine use of reference sera containing the A and B precipitins in immunodiffusion tests would permit the specific diagnosis of blastomycosis without the need for parallel tests with coccidioidin and histoplasmin.

Antigens, Fungal↗
PubMed1975

Immunochemistry of purified polysaccharide type antigens of group B streptococcal types Ia, Ib, and Ic.

The HCl-extracted purified polysaccharide type antigens of group B Streptococcus types Ia and Ib were composed of galactose and N-acetylglucosamine in a molar ratio of 3:1 for the Ia antigen and 2:1 for the Ib antigen. Immunochemical data were the same for the Ia antigens of type Ia, purified in this study, and type Ic, purified earlier. Glucosamine inhibited the Ib quantitative precipitin reactions more effectively than did N-acetylglucosamine, whereas the reverse was true of the Ia reaction. Ouchterlony studies were consistent with these observations and also revealed two type-specific precipitin bands with the HCl-extracted Ia antigens. All saline-extracted type antigens, however, formed single Ouchterlony bands that were only partially identical to the corresponding HCl antigens. Purification of the saline antigens was accomplished by treatment with cold trichloroacetic acid and by fractional ethanol precipitation. Immunoelectrophoresis experiments showed that the saline antigens were more negatively charged than the HCl antigens. The presence of sialic acid in the saline antigens probably accounted for their net negative charge and the fact that they were partially degraded by mild acid hydrolysis. The same serological specificities were observed with saline- and with HCl-extracted antigens.

Absorption↗
PubMed1976

Purification and immunochemical characterization of type e polysaccharide antigen of Streptococcus mutans.

The type-specific antigen of Streptococcus mutans strain MT703, serotype e, has been chromatographically purified and characterized. Two chromatographic fractions were obtained from saline extracts which reacted with both anti-MT703 whole-cell serum and Lancefield group E serum. The major fraction (eI) was identified as a polysaccharide composed of 37% glucose, 56% rhamnose, 5% protein, and 0.3% phosphorus, whereas the minor fraction (eII) contained 66% protein in addition to 10% glucose and 17% rhamnose. The immunological specificity of these antigens was found to be the same by immunodiffusion in agar gel. Another fraction with a negative charge (eIII) reacted with polyglycerophosphate antisera from Streptococcus mutans and Streptococcus pyogenes. For comparison, the MT703 antigen in a hot trichloroacetic acid extract (eA) and the group E antigen from a saline extract of cells of strain K129 (EI) were similarly purified by anionic ion-exchange chromatography. Although the ratio of glucose and rhamnose in eA was 1:0.9 and in eI and eII approximately 1:1.5, reactions of identity were obtained in gel diffusion against specific anti-e serum. This difference in ratio is probably a result of the extraction procedures. Both the type e and group E antisera were reactive with both eI and EI antigens. The adsorption of group E antiserum with MT703 cells removed all E antibody, whereas type e-specific antibody remained after adsorption with K129 cells. These results suggest that eI antigen possesses both e and E specificities, whereas EI possesses E only. These findings were supported by the quantitative precipitin test and immunodiffusion and/or immunoelectrophoretic patterns in agar gel. Methyl-beta-D-glucopyranoside markedly inhibited the precipitin reaction in both type e and group E sera. However, a significantly stronger inhibition by cellobiose of type e serum than of group E serum indicates that a beta-linked glucose-glucose dimer is the predominant antigenic determinant of the e specificity. The presence of both e and E specificities on a single polysaccharide molecule was demonstrated by the use of purified e antigen released from a specific e-anti-e complex. This antigen reacted with a group E-specific serum as well as a type e-specific serum. An examination of five S. mutans type e strains showed the presence of group E specificity also, whereas the I, II, and IV serotypes of group E streptococci only possessed the group E specificity.

Antigens, Bacterial↗
PubMed1983

Crossed immunoelectrophoretic analysis of Legionella pneumophila serogroup 1 antigens.

By crossed immunoelectrophoresis, 85 different antigens were demonstrated in sonicated preparations of Legionella pneumophila serogroup 1 (Lp1). The precipitin patterns of 82 anodic-migrating antigens were numbered and were designated the Lp1 reference system. Eleven antigens were stable to boiling, and seven of these were shown to be surface antigens. One heat-stable surface antigen (antigen no. 61) was highly reactive with limulus amoebocyte lysates and formed a precipitin resembling lipopolysaccharide. Serum from an isolation confirmed case of Lp1 infection and serogroup-specific rabbit antiserum reacted specifically with antigen no. 61, which was designated the serogroup-specific antigen. Normal human and rabbit sera commonly had antibodies to antigen no. 66 of the Lp1 reference system. This antigen is antigenically related to the "common antigen" of Pseudomonas aeruginosa.

Animals↗
PubMed1972

Pathogenesis and immunological aspects of experimental histoplasmosis in cynomolgus monkeys (Macaca fascicularis).

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.

Animals↗