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PubMed1975

The quantitation of antibody in farmer's lung syndrome using a radioimmunoassay. Results of a clinical survey and comparison of three serological methods.

Three serological tests for antibody to Micropolyspora faeni, the precipitin test (Ouchterlony double diffusion test in two dimensions) the fluorescent antibody test (double-layer technique using patient's serum in middle layer) and a quantitative radioimmunoassay for IgG antibody were applied to cases of farmer's lung syndrome and compared as to their value in diagnosis and management of these cases. Of twenty-one cases with clinical symptoms only ten (48%) had positive precipitin tests, while all (100%) showed a positive fluorescent antibody test. Sixteen (76-4%) were found by the radioimmunoassay to have antibody in excess of the normal controls. The remaining five cases with symptoms had a range of antibody levels which overlapped with those of symptom-free individuals (with sera negative to fluorescent antibody tests). The precipitin test was positive above a level of 75 mug/ml of IgG antibody as determined by the radioimmunoassay, whilst the cases judged by the fluorescent antibody test to be positive were all above a level of 35 mug/ml of IgG antibody. Application of the radioimmunoassay to a group of farmers in the West of Scotland successfully detected high antibody levels in all with clinical evidence of farmer's lung syndrome; farmers who were symptom-free also had measurable antibody. The advantages of a quantitative test for antibody levels in the management of cases of this disease are discussed.

Antibodies, Bacterial↗
PubMed1994

[A case of chronic necrotizing pulmonary aspergillosis effectively treated with miconazole inhalation].

A 63-year-old male was hospitalized due to persistent cough on August 19, 1991. He was suspected to have pulmonary tuberculosis based on the findings of chest roentgenograms. He was therefore given anti-tuberculous drugs and antibiotics as of the first hospital day. Approximately two months later, he developed high fever, and the infiltrative shadows on his chest roentgenograms worsened. Therefore, he was readmitted to our hospital on October 31, 1991. Despite the use of several antibiotics, his high fever persisted and the infiltrative shadows on chest roentgenography continued to worsen after admission. Although no pathogenic microorganisms were isolated, in spite of frequent sputum examinations, we strongly suspected pulmonary aspergillosis because serum precipitin antibody against aspergillus antigens was positive. Intravenous administration of miconazole was thus initiated and lead to reduction in the fever. The fever recurred but, after the start of miconazole inhalation, gradually diminished. Serum precipitin antibody subsequently became negative. We diagnosed this case as chronic necrotizing pulmonary aspergillosis based on the clinical course. It is often difficult to confirm the presence of aspergillus hyphae. Therefore, this disease should be suspected on the basis of serum precipitin antibody. In such cases, a therapeutic diagnosis should be attempted.

Administration, Inhalation↗
PubMed1976

Allergic bronchopulmonary aspergillosis: a study of 46 cases with special reference to laboratory aspects.

A study of 46 cases of allergic bronchopulmonary aspergillosis has been presented with special reference to laboratory aspects. Episodic airways obstruction and dual skin reaction to aspergillin were present in all the cases. Furthermore, there were pulmonary infiltrations in 91.1%, blood eosinophilia in 80.4%, precipitins in 91.3%, and sputum cultures repeatedly positive for A. fumigatus in 82.6% of the patients. It was observed that the multiplicity of precipitin bands corresponded to the higher recovery of A. fumigatus in culture. A. flavus and A. niger were recovered from 58.7 and 80.4% cases of allergic bronchopulmonary aspergillosis respectively but dual skin reactions to these species occurred only in 47.6 and 26.2% and precipitins in 21.7 and 4.3% of cases, respectively. The findings are compatible with the poor allergenic and antigenic properties attributed to these two species in comparison with A. fumigatus. It is suggested that allergic bronchopulmonary aspergillosis may not be too uncommon in India and it deserves greater attention in the differential diagnosis of chest diseases.

Adolescent↗
PubMed1996

[Diagnosis of pulmonary aspergillosis in cases with cavity as sequela of tuberculosis].

Non-invasive pulmonary aspergillosis is frequently seen in cases with cavity as sequela of tuberculosis. Diagnosis of non-invasive pulmonary aspergillosis is not easy except for cases whose chest X-ray findings show a typical fungus ball. How to diagnose pulmonary aspergillosis was studied in 21 patients who showed changes in the radiological findings of cavity as sequela of tuberculosis. We made sputum culture of Aspergillus, measurement of precipitin antibody titer to Aspergillus fumigatus and measurement of PASTOREX ASPERGILLUS in serum and sputum on the 21 patients (group A). PASTOREX ASPERGILLUS in sputum was evaluated in three grades: 1+ approximately 3+. We diagnosed 14 patients as pulmonary aspergillosis (PA group) among the 21 patients taking into account the chest X-ray findings and the results of the above-stated investigations. The types of chest X-ray findings of these 14 patients were fungus ball type (FB) in 2 patients, productive aspergilloma on the inner wall of a cavity (PAIC) in 8, mixed type with FB and PAIC in 2 and non-specific change in 2. However, there were 3 patients with the chest X-ray findings suggestive of PAIC in 7 patients (non-PA group) who were not diagnosed as pulmonary aspergillosis. Sputum culture were positive in 11 patients of PA group (79%) and negative in all patients of non-PA group. Precipitin antibody were positive in 8 patients of PA group (67%) and negative in all patients of non-PA group. PASTOREX ASPERGILLUS in serum were negative in all 21 patients. PASTOREX ASPERGILLUS in sputum were 3+ (+3) in all 14 patients of PA group and in 2 patients of non-PA group and 2+ (+2) in 2 patients of non-PA group. 12 patients of PA group were treated by antifungal agents and 11 patients responded well to the treatment. We also made sputum culture of Aspergillus and measurement of PASTOREX ASPERGILLUS in sputum on 14 patients (group B) who had respiratory diseases with stable cavities and 17 patients (group C) who had respiratory diseases without a cavity. In group B, sputum culture were positive in 1 patient and PASTOREX ASPERGILLUS in sputum were 3+ (+3) in 2 patients, 2+ (+2) in 1 and 1+ (+) in 2. In group C, sputum culture were negative in all patients and PASTOREX ASPERGILLUS in sputum were 3+ (+3) in 1 patient, 2+ (+2) in 3 and 1+ (+) in 2. Measurement of PASTOREX in sputum was a useful tool to diagnose non-invasive aspergillosis in addition to sputum culture and measurement of precipitin antibody titer. The sensitivity of PASTOREX in sputum was high but its specificity was low, however, its specificity could be raised by applying semi-quantitative analysis of PASTOREX in sputum.

Adult↗
PubMed1998

Fasciola hepatica: partial characterization of circulating antigens.

Antigens present in sera from Fasciola hepatica-infected goats were partially characterized by means of polyacrylamide gel electrophoresis of immunoprecipitates obtained by counterimmunoelectrophoresis. Agar containing the 2 immunoprecipitin lines, agar containing no precipitin lines, a crude extract of F. hepatica, and normal rabbit IgG were assayed. The electrophoretic pattern of 1 precipitin line showed, apart from the light and heavy chains of IgG and other faint bands, 2 prominent polypeptides of about 70 and 85 kDa. The other precipitin line showed a similar pattern, but with the 70-kDa band absent.

Animals↗
PubMed1964

IMMUNOLOGIC RELATION OF STREPTOCOCCAL AND TISSUE ANTIGENS. III. PRESENCE IN HUMAN SERA OF STREPTOCOCCAL ANTIBODY CROSS-REACTIVE WITH HEART TISSUE. ASSOCIATION WITH STREPTOCOCCAL INFECTION, RHEUMATIC FEVER, AND GLOMERULONEPHRITIS.

Sera from patients with recent streptococcal infection or non-suppurative sequelae exhibit with variable frequency a precipitin reaction in agar gel with a partially purified streptococcal antigen which has been shown previously to be immunologically related to human heart tissue. This precipitin could be absorbed from sera with normal human heart tissue homogenates but not with homogenates of other organs. Demonstration of this cross-reaction by heart absorption was found dependent both upon the serologic properties of individual sera and the nature or state of purification of the streptococcal product employed as test antigen. Evidence was obtained of a close association of heart-related and non-heart-related antigenic determinants in partially purified preparations of the streptococcal antigen by both gel diffusion and immunoelectrophoresis. On immunoelectrophoretic analysis, cross-reactive antigen exhibited a more rapid mobility toward the anode than M protein. It was destroyed by digestion with trypsin, pepsin, and chymotrypsin. Based on specific absorption tests with a Type 5 and Type 19 strain, the antigen was localized to cell walls and to a lesser extent to cell membranes of these strains. Precipitating activity related to cross-reactive antibody was localized to the immunoglobulin zone in immunoelectrophoresis. Reactive sera showed diminution or loss of serological activity following heat inactivation at 56 degrees C or after prolonged storage at 4 degrees C. Sera containing cross-reactive precipitating antibody exhibited an immunofluorescent reaction with sarcolemma of cardiac myofibers, which was inhibited by streptococcal cross-reactive antigen. By this inhibition test, the immunofluorescent reaction related to cross-reactive antibody could be distinguished from that due to other heart-reactive factors. Antibody to streptococcal cross-reactive antigen defined by precipitation-absorption tests was observed in 24 per cent of patients with recent history of uncomplicated streptococcal infection and in the majority of patients with acute rheumatic fever, rheumatic heart disease, or acute glomerulonephritis. It was observed rarely in patients with non-streptococcal related disease. These data provide evidence that induction of cross-reactive autoantibody to heart in certain individuals is associated with streptococcal infection.

Aged↗
PubMed1965

FIBRINOGEN PRECIPITATION BY STREPTOCOCCAL M PROTEIN. I. IDENTITY OF THE REACTANTS, AND STOICHIOMETRY OF THE REACTION.

Evidence confirming the identity of fibrinogen-precipitating factor and streptococcal M protein is provided by the demonstration of bactericidal, mouse protective, and long chain-producing antibodies in the sera of rabbits immunized with washed M-fibrinogen precipitates. Two precipitin lines were observed in immunoelectrophoresis of human plasma vs. rabbit anti-M-fibrinogen antiserum; no precipitin reactions were observed between human serum and rabbit anti-M-fibrinogen antiserum. The reaction between M protein and fibrinogen was stoichiometric; a constant equimolar ratio of reactants was observed in precipitates formed with a wide range of M protein concentrations.

Animals↗
PubMed1968

Specific identification of fraction I-positive Pasteurella pestis colonies on antiserum-agar plates.

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.

Agar↗
PubMed1972

Quantitative measurement of precipitating antibodies in streptococcal grouping antisera by the single radial immunodiffusion technique.

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.

Agar↗
PubMed1965

NEW METHOD OF PREPARING IMMUNIZING ANTIGENS FOR THE PRODUCTION OF ANTI-M SERA AGAINST CERTAIN SEROTYPES OF GROUP A STREPTOCOCCI.

Ashworth, Helen (Communicable Disease Center, Atlanta, Ga.), and William K. Harrell. New method of preparing immunizing antigens for the production of anti-M sera against certain serotypes of group A streptococci. J. Bacteriol. 89:141-145. 1965.-Antisera against types 4, 29, 41, and 46 streptococci have been prepared by use of a vaccine killed by adjusting the pH to 11.7 and heating for 1 hr at 56 C. Heating at the high pH destroyed the antigenic material responsible for the major reciprocal crosses present in cells of types 4, 29, and 46 without affecting the M protein. Antisera prepared with the "alkaline-treated" vaccine did not contain the major reciprocal crosses. Antibodies against the M protein were shown to be present by the bactericidal test, failure to react in the precipitin test with acid extracts of homologous trypsinized cells, and retention of the precipitin reaction after absorption with homologous trypsinized cells. The antisera did not react with the "B" antigen of the provisional type B 3264. Type 11 antisera prepared with alkaline-treated vaccine did not contain antibodies against the M protein. The usefulness of this type vaccine, particularly for the preparation of type 4 antiserum, is discussed, as well as its limitations.

Animals↗
PubMed1984

Farmer's lung serologies. An early manifestation of acute histoplasmosis.

Three cases of acute histoplasmosis demonstrated, in addition to the usual serologic findings, strongly positive precipitins against farmer's lung antigens. Twelve additional serums from patients with acute histoplasmosis were subsequently studied. Forty percent of these patients demonstrated positive precipitins against organic dust antigens.

Adolescent↗
PubMed1974

Quantitative measurement of precipitating antibodies and standardization of Streptococcus group B antisera by the reverse radial immunodiffusion technique.

A comparative study was made of the reverse radial immunodiffusion (RRID) technique and the quantitative precipitin test for determining the amount of precipitable antibodies present in streptococcus group B antisera. The coefficient of correlation between the 2 tests was 0.963 when a purified carbohydrate antigen was used. The results of this study show that: (1) a group B antiserum should contain at least 3 mg of precipitable antibodies per ml to ensure that it will react with acid extracts of group B diagnostic cultures; and (2) the RRID technique is an accurate and simple procedure for determining the amount of precipitable antibody present in an antiserum, and it could be used for the standardization of these diagnostic reagents. A method for preparing the group B carbohydrate antigen used in the RRID and quantitative precipitin tests is described.

Animals↗
PubMed1977

Longitudinal studies of immunological parameters in Farmer's lung.

A group of patients with serum precipitins against M. faeni culture filtrate antigens and with clinically proven farmer's lung disease was examined. A group of precipitin-negative farmers with comparable antigen exposure and with unrelated forms of pulmonary disease served as control. Immunoglobulins G and A were elevated at first consultation in the majority of the acute cases of farmer's lung. Immunological parameters normalized during corticosteroid medication and antigen avoidance. Autologous complement (C) consumption by M. Faeni antigens proved valuable for diagnostic purposes. Despite normalizing immunological factors and subsiding symptomatology, C-consuming antibody levels remained constant.

Actinomycetaceae↗
PubMed1980

Antibody responses to mycobacterial antigens in patients immunized with BCG for the treatment of neoplastic disease.

Precipitin and agglutination antibody response to antigens of Mycobacterium tuberculosis was measured in 43 patients with neoplastic disease who were treated with weekly BCG or intratumor BCG vaccination. Resuspended freeze-dried Tice vaccine was shown to contain soluble mycobacterial antigens. Fourteen patients receiving weekly BCG vaccination developed intense precipitin and agglutination antibody response over the course of several months. Twenty-nine patients receiving intratumor and oral vaccination developed less intense antibody response. Twenty-one of 28 purified protein derivative (PPD)-negative patients converted their PPD. No correlation between antibody, PPD response and antitumor effect could be identified. The importance of identifying the soluble antigen in BCG and of defining the humoral response to purified antigens is discussed.

Agglutination Tests↗
PubMed1980

Heterogeneity of precipitating antibodies in polymyositis and dermatomyositis. Characterization of the Jo-1 antibody system.

Precipitating antibodies occurring in 26 "pure" polymyositis (PM) patients and 22 "pure" dermatomyositis (DM) patients were studied by double immunodiffusion using calf thymus nuclear extract and whole calf thymus extract. Seventeen of 26 (65%) PM sera and 13 of 22 (59%) DM sera showed positive reactions, and 18, 10, and 2 sera exhibited 1, 2, and 3 precipitin lines, respectively. Common specificities were established by immunodiffusion for the 11 PM and 6 DM sera with strong precipitin lines. These were classed into three groups of shared specificity and six reactions specific for individual sera. These 9 calf thymus antigens were shown to be different from the known soluble tissue antigens Sm, nuclear RNP, Ro, and La. A 600-fold purification of the most frequently appearing system designated Jo-1 was achieved and its clinical specificity for patients with PM syndromes was established.

Antibodies↗
PubMed1976

Detection of tumor-specific antigens in human mucinous cystadenocarcinoma of the ovary by immunodiffusion.

Rabbit antiserum to a tissue extract of human mucinous cystadenocarcinoma of the ovary reacted with tissue extracts of normal ovary and various ovarian malignancies, and ascitic or cystic fluids of ovarian origin by Ouchterlony double gel diffusion and precipitin inhibition techniques. The tumor-associated antigen(s) of mucinous cystadenocarcinoma, which were demonstrated by Ouchterlony double diffusion, were not present in tissue extract of pooled normal ovaries and cystic fluid of benigh tubo-ovarian cyst. An organ-associated tumor antigen as well as the type-specific tumor antigen may exist in mucinous cystadenocarcinoma of the ovary. The mucinous cystadenocarcinoma was not very immunologically different but was distinguishable from serous cystadenocarcinoma and other types of ovarian cancer by double gel diffusion. Precipitin-inhibition reactions demonstated that the adsorbed antiserum to human ovarian mucinous cystadenocarcinoma mixed with tissue extracts of dysgerminoma and serous cystadenocarcinoma, and ascitic fluid of papillary embryonal adenocarcinoma of the ovary could not eliminate the specific precipin line developed with tissue extract of mucinous cystadenocarcinoma.

Antigens, Neoplasm↗
PubMed1995

Interaction of mistletoe toxic lectin-I with sialoglycoproteins.

The binding properties of mistletoe toxic lectin-I (ML-I) with sialo-N- and O-glycans were investigated by quantitative precipitin and precipitin inhibition assays. Human alpha 1-acid glycoprotein reacted strongly with ML-I, precipitating over 82% of the lectin nitrogen tested, while the precipitability of its asialo product decreased by 30%. Native fetuin precipitated 50% of the ML-I added, and its reactivity was reduced by 20% after desialylation. On the contrary, the poor reactivity of rat sublingual sialoglycoprotein with ML-I increased substantially after removal of sialic acid and completely precipitated the lectin added. The glycoprotein-lectin interactions were inhibited by NeuAc alpha 2-->3/alpha 2-->6Gal beta 1-->4Glc and/or Gal beta 1-->4Glc (NAc) residues. From the above results, it is concluded that ML-I is specific for sialic acid. However, sialic acid of some O-glycans also acts as masking molecule as the precipitability of rat sublingual and bovine submandibular glycoproteins with ML-I increased after desialylation.

Animals↗
PubMed1987

Immunochemical and partial chemical characterization of fractions of membrane-bound smooth lipopolysaccharide-protein complex from Brucella abortus.

Smooth lipopolysaccharide (sLPS) of Brucella abortus was prepared and fractionated by a modification of the procedures of Moreno et al. (J. Bac. 138:361-369, 1979). Washed B. abortus cells were disrupted by 21 freeze-quick thaw cycles with ultrasonication to separate the non-membrane-bound material. Ultrasonicated bacteria were used for preparation of membrane-bound sLPS (approximately f5, the main crude sLPS fraction described by Moreno et al.). Phenol extraction was repeated 3 times and then washed with H2O 10 times to remove most of the chromogen, polysaccharides and nucleic acids, eliminating the need for enzyme treatment as described previously. The membrane-bound sLPS was fractionated into 3 to 5 groups according to the extent of dialysis and centrifugation, these fractions required only 80 ng for positive ELISA, about 0.2 ng for positive Limulus lysate tests, and reacted well with precipitating antibodies in the serum from a strain 2308 infected cow. They had marked differences in precipitin curves and chemical composition. The protein content varied from 16% to 42% as determined by dye binding test and 17 to 60% by Lowry phenol method using bovine serum albumin as the standard, which implies that the proteins associated with LPS may also play important roles in the complex for the immunochemical interactions and the heterogeneity of B. abortus lipopolysaccharide protein complex. As compared with previous reports, a higher yield of sLPS, ranging from 3.6% to 7.7% of dried bacteria, was obtained. Group f5A, which had a standard bell shaped curve in the precipitin assay, is one of the major fractions in all three strains (1119.3, 19 and 2308). The amount of other subfractions obtained varied with batches or strains of B. abortus. These results provide a new profile of the immunochemical reactivities and the heterogeneity on B. abortus smooth membrane-bound endotoxins.

Antigen-Antibody Complex↗