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Lipopolysaccharide-binding lectin from the horseshoe crab, Limulus polyphemus, with specificity for 2-keto-3-deoxyoctonate (KDO).

A lipopolysaccharide (LPS)-binding lectin was recovered from the serum of Limulus polyphemus by ion-exchange chromatography. Electrophoretic analysis of this lectin preparation revealed three poorly migrating bands. When whole serum was incubated with glycolipid obtained from the Rc mutant of Salmonella minnesota prior to electrophoresis, bands corresponding to those seen in the partially purified lectin were missing, suggesting that the recovered material was composed of isolectins. Qualitative precipitin tests revealed no reactivity of this purified lectin with lipid A fractions or with LPS devoid of 2-keto-3-deoxyoctonate (KDO). The agglutination of chicken erythrocytes by this lectin was inhibited by both N-acetyl-neuraminic acid and KDO. Erythrocytes complexed with glycolipid from the Re mutant of S. minnesota were strongly agglutinated by this lectin. We conclude that this LPS-binding lectin is specific for the KDO portion of the molecule and that it is identical to the previously described sialic acid-binding lectin from L. polyphemus. This lectin may play a role in the host defense mechanisms of Limulus.

Animals↗

Variables of pulmonary allergy and inflammation in grain elevator workers.

The purpose of this study was to examine possible mechanisms of the obstructive and restrictive ventilatory defects of grain elevator workers. We found that grain handlers whose skin tests were positive to grain or fungal antigens had no excess of respiratory symptoms or pulmonary function abnormalities. Also, those having a possible work-related respiratory problem showed no response to inhalation challenge tests with grain dust extract. In addition, grain handlers exhibited neither an increase in positive serum precipitin tests with fungal antigens nor abnormal serum levels of C3 and C4, C-reactive protein, rheumatoid factor or antinuclear factor. However, their serum a1-antitrypsin level was elevated. The latter finding may be indicative of a pulmonary inflammatory process, but we have found no clear indication of a type 1 or 3 allergic state in grain elevator workers.

Adult↗

[Bronchial asthma and methemoglobinemia caused by milk allergy].

Bronchial asthma and methemoglobinemia due to the allergy on milk was described in 34-year old man. Very marked cyanosis with 15.3% methemoglobin, E--7.2, 272% hemoglobin, 52% hematocrit, severe form of bronchial asthma with PaO2--7.63 kPa, 89% saturation, PaCO2--4.46 kPa were found. Sternal and bone biopsy showed myeloproliferative disease. Allergy testing showed positive early and late reaction on milk. Serum IgE was 1670 mu/ml, RAST and precipitin test were positive on milk. Bronchoprovocation test on milk showed positive asthmatic reaction. After elimination of milk products from food all findings were soon normalised and the patient has not had bronchial asthma and cyanosis for two years now.

Adult↗

Antibody responses in patients with farmer's lung disease to antigens from Thermoactinomyces vulgaris.

A serological analysis of mycelial antigens of Thermoactinomyces vulgaris in immunodiffusion with human sera revealed five individual antigens. Three antigens were proteins, sensitive to pronase and soluble in phenol. Two were cationic polysaccharides, sensitive to sodium periodate, and containing glucosamine and muramic acid. Latex coated with mycelial antigens was compared with precipitin tests in detecting antibodies to T. vulgaris; the number of positive results detected by each test differed slightly, and a combination of the two tests detected the highest number. Counterimmunoelectrophoresis (CIE) was shown to be a very sensitive method for detecting precipitins, but not for their measurement. A prospective evaluation of immunodiffusion, latex agglutination and CIE as potential serodiagnostic techniques for farmer's lung disease is suggested.

Actinomycetales↗

[Evaluation of the reliability of the diagnostic methods in farmer's lung].

Following a study of the diagnosis of farmer's lung (PDF) carried out in 1763 active farm workers in Doubs, 294 (16.7%) were suspected of PDF and they were invited to a secondary consultation in hospital for a further work up which consisted of a clinical examination for crepitations (RC) on pulmonary auscultation (AP) and for evidence of an interstitial lung disorder (SI) on pulmonary radiograph (RP) and for evidence of a restrictive syndrome (SR) on pulmonary function tests (EFR). 238 patients (80%) accepted the additional assessment and were divided into two groups on the basis of the history and serology (precipitin tests), group 1: probable PDF (n = 65): symptoms suggestive of PDF in the presence of serum precipitins; group 2: possible PDF (n = 173): non specific symptoms with or without serum precipitins. RC were present in 21.9% of patients in group 1 and 6.4% of patients in group 2 (p less than 0.001). There was an SI in 9.2% of patients in group 1 and 4% of patients in group 2 (non significant). For the elevated radiology scores for SI there was a significant difference: 7.7% for SI in group 1 and 1.73% in group 2 (p less than 0.05). There was 38.1% of abnormal EFR in group 1 and 14.7% in group 2 (p less than 0.001) but the difference of the frequency of SR in the 2 groups was not statistically significant (9.5% in group 1 and 4.7% in group 2). In this study, an additional assessment (AP, RP, EFR) carried out at a difficult moment during winter does not appear to confirm great benefit.(ABSTRACT TRUNCATED AT 250 WORDS)

Auscultation↗

The use of precipitin analysis in agar for the study of human streptococcal infections. II. Ouchterlony and Oakley technics.

It has been shown by agar precipitin tests (Ouchterlony and Oakley) that human sera may contain from 0 to 5 antibodies against antigens present in a partially purified streptolysin O preparation, and from 0 to 7 antibodies against antigens in a crude ammonium sulfate concentrate of the streptococcal culture supernate used. These antigens were prepared from a Group A hemolytic streptococcus (strain C203S). Strong evidence was presented suggesting that some of the bands seen with streptolysin O concentrate represented antibody reponses to streptococcal antigens heretofore undescribed. Tests were also carried out with other streptococcal antigens, including streptokinase-desoxyribonuclease mixture from Group C streptococci (varidase-Lederle), crystalline proteinase, proteinase precursor, C carbohydrate, and sonic vibrated streptococcal cell extracts (group A, C203S). Fewer bands were seen with these preparations, and with some they were quite uncommon. The observations indicated that the predominating antibody responses in human streptococcal infections were to extracellular products of the micro-organisms, and only very slightly and infrequently to intracellular antigens. The human sera studied included sera from patients with active or convalescent rheumatic fever, and non-rheumatic subjects suffering from a variety of illnesses. As was expected, the rheumatic subjects showed antibody responses to many more of the antigens present in these preparations than did the nonrheumatic group. Pooled normal human gamma globulin was found to contain many of the antibodies found in potent human sera. This finding confirmed the antigen-antibody nature of the bands seen with individual sera. The epidemiological significance of these findings with gamma, globulin was briefly discussed. It was found that rabbit, guinea pig, and human antibody precipitin bands join quite readily in the Ouchterlony tests. This finding adds another tool for the identification of the precipitin bands found with human sera. Evidence was obtained which indicated differing immunological specificities of two samples of streptococcal desoxyribonuclease, one from Group A, the other from a Group C streptococcus. The value of these technics as representing a new approach to the study of human infectious disease was discussed.

Agar↗

Serology and semen culture for the diagnosis of Brucella ovis infection in chronically infected rams.

The serological response to Brucella ovis and the shedding of the organism in semen was followed for a period of 13-14 months in 42 naturally infected rams. Most rams remained chronically infected and excreted the organism in their semen throughout the investigation. B. ovis was isolated from 87.9% of the semen samples from the infected rams. The most common sites from which B. ovis could be isolated at necropsy were the epididymides and accessory sexual glands. In one ram the organism was isolated from lung, spleen, kidney and iliac lymph nodes. Three rams ceased to shed B. ovis in their semen during the course of the investigation. Seventy-five (11%) of 686 sera from infected rams were negative in the complement fixation test (CFT) although 76% and 77% of CFT-negative sera were positive in the gel diffusion precipitin test (GDT) and enzyme labelled immunosorbent assay (ELISA) respectively. The high incidence of CFT-negative infected rams was due to the selection for the investigation of many rams with histories of negative or vacillating CFT titres. Sera from five rams which never shed B. ovis in their semen reacted erratically in the three serological tests. The five rams were from heavily infected flocks and were kept in contact with infected rams throughout the investigation.

Journal Article↗

Comparisons of the serologic response of chickens to Pasteurella multocida and its lipopolysaccharide.

Chickens were inoculated with serotype 3 Pasteurella multocida cells or purified lipopolysaccharide (LPS), and their serologic responses to LPS and heat-stable antigens of 16 serotypes were compared. Chickens inoculated with cells or LPS had antibodies against LPS as determined by indirect hemagglutination tests; titers were highest 2-4 weeks after the initial inoculation. Sera from chickens inoculated with cells reacted with unheated and heated cell antigen in a tube-agglutination test. Sera from chickens inoculated with LPS reacted only with heated cell antigen in the tube-agglutination test. Nonspecific reactions with heat-stable antigens of other serotypes occurred in the gel-diffusion-precipitin test with sera from chickens inoculated with cells but not with sera from chickens inoculated with LPS. Antisera prepared against LPS could be used for serotyping field isolates of P. multocida.

Agglutination Tests↗

Allergic bronchopulmonary aspergillosis and sensitization to Aspergillus fumigatus in chronic bronchiectasis in adults.

Fifty adult subjects for whom a diagnosis of idiopathic bronchiectasis (excluding those secondary to tuberculosis or hypogammaglobulinaemia) had been confirmed previously were investigated by: questionnaire; blood eosinophil count; sputum culture for Aspergillus fumigatus and eosinophil count; chest radiography; skin-prick tests with several aeroallergens and four preparations of A. fumigatus, including a reference extract; measurement of specific IgE antibodies; precipitin testing and self-crossed immunoelectrophoresis with A. fumigatus. Five subjects were possible cases of allergic bronchopulmonary aspergillosis in whom the condition had been previously misdiagnosed or in whom sensitization to A. fumigatus had occurred after the onset of bronchiectasis. These five subjects had positive immediate skin reactions to A. fumigatus and a history of recurrent pneumonias. Four had a previous history of asthma and the others showed increased bronchial responsiveness to inhaled methacholine. At the time of the survey, A. fumigatus grew in the sputum of one out of five subjects. These subjects had increased levels of specific IgE. Two had precipitins by double diffusion and three subjects were positive on self-crossed immunoelectrophoresis. It is concluded that allergic bronchopulmonary aspergillosis or evidence of sensitization to A. fumigatus can be identified in a significant proportion of adult subjects with so-called idiopathic bronchiectasis.

Adult↗

Comparison of the API 20S Streptococcus identification system with an immunorheophoresis procedure and two commercial latex agglutination tests for identifying beta-hemolytic streptococci.

The API 20S Streptococcus identification system and a new immunorheophoresis procedure were evaluated as means for determining the Lancefield serogroup of beta-hemolytic streptococci recovered from human clinical specimens. The serogroup of 96 strains was determined by these methods and by two commercially available latex agglutination tests. Streptex and SeroSTAT. The results of all four procedures were compared with the results of a classical precipitin test. The API 20S system correctly categorized 92.7% of the isolates; 94.8% were correctly identified by the immunorheophoresis procedure. The latex agglutination procedures were of comparable accuracy, yielding correct identifications with approximately 92% of the strains tested.

Bacteriological Techniques↗

Group and strain-specific neutralization sites of infectious bursal disease virus defined with monoclonal antibodies.

Two somatic cell hybridizations were performed utilizing splenocytes from mice immunized with one or more strains of infectious bursal disease virus (IBDV). Supernatants from hybridoma cell lines were initially screened by the enzyme-linked immunosorbent assay (ELISA) against multiple strains of IBDV. Cell lines that secreted antibodies with ELISA reactivity patterns of interest were cloned, and their monoclonal antibodies (MCAs) were subsequently tested in cross-virus-neutralization tests. Two of the nine MCAs selected exhibited strong neutralizing activity and precipitated IBDV antigens in agar gel precipitin tests as well. MCA B69 significantly neutralized only the cloned D78 strain of IBDV, whereas MCA R63 neutralized all IBDV strains (representing both serotype I and II viruses) against which it was tested. Results of competitive ELISAs that used the R63 and B69 MCAs showed that the two neutralization sites on the D78 strain were not overlapping.

Animals↗

Serum precipitins against microbes in mouldy hay with respect to age, sex, atopy, and smoking of farmers.

This study was based on a sample of 3,065 farmers from a larger survey population of 12,056 Finnish farmers. Data were collected in a postal survey conducted by the Social Insurance Institution of Finland. Serum samples for determination of precipitating antibodies were taken at local health centres. Precipitins were determined by the method of microplate immune diffusion. The antigen panel consisted of mycelial antigens of Micropolyspora faeni, Thermoactinomyces vulgaris, Aspergillus fumigatus, and Aspergillus umbrosus. Precipitins to any one of the four microbes were detected in 8.6% of the sera. The most common causes of positive precipitin tests were Aspergillus umbrosus and Thermoactinomyces vulgaris, which agrees with previous findings reported from Finland. In general, the precipitins were more prevalent among women, which corresponds to local cultural traditions, and in older farmers. The prevalence of precipitins did not differ between non-atopic and atopic (defined as past or present co-existence of atopic dermatitis including infantile eczema and/or hay fever or other allergic rhinitis) subjects. In contrast, the prevalence of precipitins was about 1.5-2 times larger among non-smokers than among smokers, which confirms the findings in previous reports. In future studies on occurrence of precipitins, the data should be controlled with respect to age, sex, and smoking.

Adult↗

Application of a micro enzyme-linked immunosorbent assay (ELISA) to detection of anti-amebic antibody in various forms of amebic infection.

Techniques of a micro enzyme-linked immunosorbent assay (ELISA) used for the serodiagnosis of schistosomiasis were applied to amebic infection. Test sera were divided primary on the basis of serologic diagnosis and stool examination as follows; (I) gel diffusion precipitin test (GDP) positive and stool examination positive: 9 specimens, (II) GDP positive and stool examination negative: 29 specimens, (III) GDP negative and stool examination positive: 32 specimens. Virtually all of the individuals belonging to (III) were asymptomatic, while more than 75% of (I) and (II) were symptomatic. The upper limit of 99% critical range was calculated from the data of 70 serum specimens from healthy adult Japanese and was employed as the cut-off value. All of the specimens of (I) and (II) were judged positive by ELISA, generally with a much higher absorbance than the cut-off value; whereas, approximately 80% of (III) were judged positive. The average absorbance of (III) was lower than that of (I) and (II). These findings suggest that the ELISA is well in accord with GDP qualitatively as far as GDP-positive individuals are concerned, and that even asymptomatic cyst carriers with negative serology by GDP may often be producing anti-amebic antibodies, although the titers are low.

Amebiasis↗

Pigeon-breeder's lung. Extensive workup clarifies the clinical picture.

Pigeon-breeder's lung should be included in the differential diagnosis when patients present with interstitial lung disease or intermittent cough and dyspnea. No single test confirms the diagnosis. Rather, careful attention must be given to results of history taking, physical examination, pulmonary function and precipitin testing, chest radiography, and, if indicated, lung biopsy. Symptoms usually resolve when exposure to suspected antigens is avoided.

Adult↗