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PubMed1977

Antibodies against Pseudomonas aeruginosa in serum from normal persons and patients colonized with mucoid or non-mucoid Pseudomonas aeruginosa: Results obtained by crossed immunoelectrophoresis.

Serum from 154 normal persons was investigated for precipitating antibodies against P. aeruginosa by crossed immunoelectrophoresis, using a polyvalent P. aeruginosa standard antigen. Ninety-four per cent of the sera contained no demonstrable precipitins, and 6% contained precipitins against only one P. aeruginosa antigen. Thirty-four per cent of sera from 122 patients colonized with P. aeruginosa in various anatomical regions contained precipitins against 2 to 41 P. aeruginosa antigens. Serum from 14 patients harbouring mucoid strains of P. aeruginsa contained precipitants against these bacteria, and the average number of precipitins was significantly higher than the average number of precipitins in serum from patients harbouring non-mucoid strains. The largest number of precipitins was found in serum from adult patients chronically colonized with mucoid P. aeruginosa in the lower respiratory tract, and from adult patients with P. aeruginosa bacteremia originating from suppurative urinary tract infections caused by mucoid strains. Serum from patients with P. aeruginosa cystitis or from patients with P. aeruginosa colonization of the middle ear or with wound infection, contained only few or no precipitins.

Adolescent↗
PubMed1976

The value of precipitating antibodies in screening for hypersensitivity pneumonitis.

To evaluate the usefulness of precipitin tests as a screening method to detect hypersensitivity pneumonitis, we compared persons with precipitins to organic antigens with precipitin-negative subjects from the same population of 1,072 office workers participating in a health survey examination. Thirty-three of the 53 persons with preciptins to Micropolyspora faeni, Thermoactinomyces vulgaris, T. candidus, pigeon serum, aspergillus, alternaria, pullularia, penicillium, cephalosporium, trichoderma, and phoma were matched according to age, sex, height, and smoking habits with precipitin-negative subjects. The subjects completed a self-administered standard questionnaire, they were interviewed and examined, chest radiograms were taken, and pulmonary functions were evaluated at rest and during mild and moderate exercise. No significant differences were found between the 2 populations in any of the pulmonary function measurements. In addition, clinical and radiologic evidence failed to distinguish between the 2 groups. No cases of hypersensitivity pneumonitis were found. In this population, precipitins had no apparent relationship to long disease. The frequency of precipitins was considerably higher than the frequency of hypersensitivity pneumonitis. We concluded that the detection of hypersensitivity pneumonitis in population surveys attempting to establish prevalence of the disease cannot be accomplished by the simple analysis of serum precipitating antibodies but requires, in addition, a more complex analysis of historical, radiologic, and immunologic data.

Antigens↗
PubMed1976

Precipitating antibodies in a midwest dairy farming population toward the antigens associated with farmer's lung disease.

A survey of the frequency of precipitins to the antigens of the thermophilic actinomycetes and Aspergillus species was conducted on serum samples from 1,045 farmers obtained at a 3-day exposition on modern farm equipment and farming practices in central Wisconsin. Each farmer filled out a questionnaire including socioeconomic information, lung disease history, exposure history, and smoking history. Precipitins were detected by the double-diffusion method. The antigen panel included eight thermophilic actinomycetes species and a mixture of Aspergillus species. Precipitins were found in the sera of 93 famers (8.9%). The distribution of positive precipitins was: Micropolyspora faeni 63 (67.7%), Thermoactinomyces vulgaris 7 (7.5%), Thermomonospora viridis 2 (2.2%), M. faeni + T. viridis 16 (17.2%), M. faeni + T. vulgaris 1 (1.1%), Aspergillus species 4 (4.3%). Of all the parameters tested for in the questionnaire, those with positive serology differed significantly from the whole population only in that a higher proportion of the positives reported exposure to silo gas and illness after uncapping silos. Comparison of the size of the farm and the number of dairy cows in the state of Wisconsin with the population samples indicated that the sample population was skewed toward those with larger farms and larger dairy herds. This study confirms that a significant proportion of the farm population in Wisconsin does have precipitins to the microorganisms associated with farmer's lung disease. Follow-up studies to establish the relationship between the positive precipitin reactions to the presence of clinical disease are now under way.

Antigens, Bacterial↗
PubMed1992

Impairment of lung mucociliary clearance in pigeon fanciers.

Lung mucociliary clearance was measured in 15 pigeon fanciers. The study group was subdivided into two: a precipitin-positive group (n = 10; mean +/- SEM age 45 +/- 5 years) with circulating blood precipitins and a precipitin-negative group (n = 5; mean +/- SEM age 40 +/- 3 years) without. Clearance was measured using an objective, noninvasive radioaerosol technique. The data for both groups were compared with those of matched control groups of healthy subjects. The mean +/- SEM area under the tracheobronchial retention curves (AUC) over the 6-h observation period was 257 +/- 27 %h for the precipitin-positive group compared with 177 +/- 16 %h for its control group (p = 0.02)--a high AUC value denoting slow clearance. That for the precipitin-negative group was 282 +/- 34 %h compared with 150 +/- 15 %h for its control group (p = 0.02). Our study illustrates in pigeon fanciers involvement of the conducting airways in that a major defense mechanism of the airways--namely, mucociliary clearance--is substantially compromised. The presence or absence of precipitins appears not to be related to the degree of mucociliary clearance impairment.

Bird Fancier's Lung↗
PubMed1981

The bovine immune response to Brucella abortus IV. Studies with a double immunodiffusion test for antibody against A2.

A double immunodiffusion test for precipitins against Brucella antigen A2 was developed and applied to a variety of samples. The A2 precipitins were produced by a heifer infected with B. abortus strain 2308, cattle vaccinated with killed B. melitensis strain H38 or live B. abortus strain 19 and by a dog infected with B. canis. Precipitins were also detected in the second International Standard for anti-Brucella abortus serum, in several anti-B. canis sera and at low levels in one anti-B. ovis serum tested. Antisera produced in calves against Yersinia enterocolitica serotype 0:9 had no anti-A2 activity despite titers greater than or equal to 1/1024 and greater than or equal to 1/80 in standard Brucella agglutination and CF tests, respectively. The test for A2 precipitins lacked specificity as weak reactions were obtained with five of 295 sera from brucellosis-free herds. This test was relatively insensitive, detecting precipitins in only 16 of 24 sera from infected cattle and 27 of 54 sera positive by complement fixation and enzyme labelled antiglobulin tests performed with whole cell and smooth lipopolysaccharide antigens, respectively. The A2 precipitins were detected in nine sera from five cattle, in two infected herds, which were negative by agglutination and complement fixation tests.

Animals↗
PubMed1995

IgM anti-A and D SnRNP proteins and IgM anti-dsDNA are closely associated in SLE sera.

U1RNP and Sm precipitins occur in systemic lupus erythematosus (SLE) and in overlap syndromes, while precipitins to Ro/SSA and La/SSB occur in SLE and its variants as well as Sjögren's syndrome. In studying IgM and IgG antibodies to these polypeptides by Western blot, we found the expected frequency of antibodies to the A protein of the U1RNP specificity and the D protein of the Sm specificity in sera with precipitins to U1RNP and Sm, but also found the frequent occurrence of both IgM and IgG anti-A and D in SLE sera with no precipitins, and in sera with anti-Ro/SSA and/or anti-La/SSB precipitins, but not in sera from patients with rheumatoid arthritis, polymyositis, scleroderma, or normals. We have studied the sera which bind the A and D polypeptides on Western blot to test their ability to bind native U1RNP (which contain the A and D proteins) in ELISA. The results indicate that most anti-Ro/SSA alone, anti-Ro/SSA, and anti-La/SSB sera as well as SLE sera with no precipitins react preferentially with the denatured A and D proteins. Further study of sera with IgM anti-A and D in Western blot reveals that these sera also frequently contain anti-double-stranded DNA antibodies. These results show that the anti-A and D responses, especially IgM anti-A and D, occur not only in patients with precipitating antibodies to SnRNPs, but in almost half of patients across the lupus spectrum. The extended prevalence of antibodies to the denatured A and D proteins in Western blot is associated with the cross-reaction of antibodies to dsDNA.

Autoantibodies↗
PubMed1994

Serum anti-Aspergillus fumigatus antibodies by immunoblot and ELISA in cystic fibrosis with allergic bronchopulmonary aspergillosis.

Allergic bronchopulmonary aspergillosis (ABPA) occurs with a prevalence of 5% to 15% in patients with cystic fibrosis (CF). Because of the frequent colonization with Aspergillus fumigatus (Af) in CF, the causative agent of ABPA, antibody reactivity to Af proteins is frequently observed, which obscures the diagnosis of ABPA. Patients with CF are also categorized according to the presence of positive skin test responses to Af and/or the presence of positive precipitins. In this study we used ELISA and immunoblot assay to detect IgE and IgG anti-Af antibodies in patients with CF and ABPA (n = 13) compared with other groups of patients with CF: those with positive skin test and positive precipitin results (n = 18), those with positive skin test and negative precipitin results (n = 14), those with negative skin test and positive precipitin results (n = 10), and those with negative skin test and negative precipitin results (n = 35). IgE and IgG anti-Af antibodies were significantly elevated in patients with ABPA as determined by both immunoblot assay (p < 0.01) and ELISA (p < 0.01). However, detection of Af antibodies by ELISA was more sensitive in discriminating patients with CF and ABPA from patients with CF who had positive skin test and positive precipitin results but lacked radiographic and clinical evidence of ABPA. In patients with CF and ABPA the immunoblot assays demonstrated a multitude of IgE, IgG, and IgA antibody responses to Af proteins, which ranged in molecular weight from 14 kd to greater than 106 kd. The level of IgE anti-Af antibody to individual proteins decreased during remissions of ABPA.(ABSTRACT TRUNCATED AT 250 WORDS)

Analysis of Variance↗
PubMedDate not supplied

Precipitating and agglutinating activity in duck anti-soluble protein immune sera.

Ducks were inoculated parenterally, up to 4 times, with bovine serum albumin (BSA), B gamma globulin (BGG), or horse gamma globulin (HGG). Their sera were tested in immunodiffusion tests for precipitins. Only BSA and BGG induced precipitins. Only 7 of 443 sera tested, obtained from 3 of 62 inoculated ducks, developed a precipitin line with homologous antigen. All 7 sera were obtained from the earliest bleeding (7 days) after inoculation. Sera were also tested for agglutinins in direct passive hemagglutination (DPHA) and direct red-cell-linked antigen tests (DRCLAT) and for nonagglutinating antibodies in indirect red-cell-linked antigen tests (IRCLAT). No duck had a passive-hemagglutination-demonstrable primary immune response. Demonstrable DPHA titers after subsequent inoculations were very low compared with responses to these antigens of other species noted by other workers. Duck immune response was greatly diversified: all inoculation regimens that induced agglutinins in some ducks left others completely unstimulated. Precipitins and agglutinins of the same ducks correlated well: those 7 sera that had precipitins also had the highest DPHA titers for the 3 donor ducks. However, hemagglutinating titer and presence of precipitins in sera of different ducks correlated poorly. Increasing the age of the ducks at the first inoculation from 6 to 10 weeks increased the number of responders and DPHA titers of their sera. Non-agglutinating antibodies were demonstrated in anti-BSA sera: in IRCLAT, the sera had titers 2-to-256-fold higher than the sera in DPHA or DRCLAT had. Duck immunoglobulins were deficient in those immunological reactions (precipitation and agglutination) that require functional bivalency.

Animals↗
PubMed1970

The extracellular antigens of Micropolyspora faeni: their significance in farmer's lung disease.

A serological analysis of the extracellular antigens of Micropolyspora faeni by immunodiffusion with a combination of sera revealed 29 individual antigens. A survey was made of the incidence of precipitins to the antigens in sera from patients with clinical farmer's lung disease (FLD) and other respiratory diseases. Precipitating antibody was found in 75% of farmer's lung cases and in 20% of other cases who had been exposed to the same environment. More precipitin reactions were seen in sera from severe forms of FLD than from milder forms. The distribution of precipitins to individual antigens was not significantly affected by severity of disease.Most of the patients with precipitins to M. faeni, but without the symptoms of FLD, were suffering from mild or moderate symptoms of other respiratory diseases with a history of chronic onset of symptoms. The distribution of precipitins to individual antigens in this group was similar to that in clinical FLD patients but the incidence was considerably lower.The significance of these results is discussed.

Actinomyces↗
PubMedDate not supplied

Identification of antibodies to nuclear acidic antigens by counterimmunoelectrophoresis.

Saline extracts of rabbit thymus were found to contain many nuclear antigens that reacted with antibodies in the sera of patients with systemic rheumatic diseases. Counterimmunoelectrophoresis (CIE) was used to detect antibodies to nuclear acidic protein (Sm), nuclear ribonucleoprotein (RNP), and antibody to nuclear antigen B, which was reported previously in Sjögren's syndrome. All these nuclear antigens behaved as anions with different mobilities in CIE and could be distinguished from one another by the locations of the precipitin lines. They could also be distinguished by the facts that the nuclear RNP precipitin lines were abolished by digestion with ribonuclease whereas others were unaffected, and that Sm precipitin lines developed later than other precipitin lines. With this technique antibody to nuclear RNP was detected in 46% of patients with systemic lupus erythematosus (SLE) compared to 26% detected by the hemagglutination technique. Similarly the increased sensitivity of the CIE technique was able to show that antibody to B antigen was present in 12% of SLE patients, whereas this antibody was not detectable in the same group of patients by immunodiffusion. This study shows that CIE is a rapid and sensitive technique for detecting precipitating antibodies to a number of nuclear acidic antigens. Methods are described to identify the immunochemical specificities of the precipitin lines by the use of standard reference sera.

Antibodies, Antinuclear↗
PubMed1983

Sporobolomyces: a possible cause of extrinsic allergic alveolitis.

A 28-year-old horseback rider presented with symptoms, chest radiograph, and pulmonary function tests suggestive of extrinsic allergic alveolitis related to exposure to a horse barn. Exposure to the barn produced symptoms, fever, and a fall in VC commencing 4 hr after exposure. Precipitins were positive against Sporobolomyces, suggesting this might be the causative agent; precipitins were negative against other fungi and horses. Lymphocyte stimulation to Sporobolomyces in vitro was positive in the patient and negative in two control subjects. Sporobolomyces was grown from straw in the barn. Cessation of exposure to this barn (but continued exposure to horses) has resulted in improvement in clinical condition. A survey for immunologic sensitivity to Sporobolomyces revealed that eight of 30 atopic subjects had positive wheal-and-flare prick skin tests to Sporobolomyces antigen, whereas none of 30 laboratory controls or 30 grain handlers had precipitins against Sporobolomyces. Sporobolomyces is a common fungus in cereal grain growing areas. Its spore size is less than 5 micron, consistent with other causative agents of this disorder. In this patient, positive precipitins and lymphocyte stimulation to Sporobolomyces and negative precipitins to other known causes of extrinsic allergic alveolitis provide circumstantial evidence that Sporobolomyces was the cause of the syndrome.

Adult↗
PubMedDate not supplied

A serologic response in human infection with Enterobacteriaceae.

Sera from patients infected with Escherichia coli, Proteus, Klebsiella, and Serratia were studied for precipitins to ultrasonic extracts of these organisms in gel-diffusion plates. Sera from 66 per cent of these patients contained precipitins when initially tested. Twenty-four per cent of sera tested in the first week after onset of infection contained precipitins, but this rose to 78 per cent by the third week. Cross-reactions of sera with Pseudomonas antigens were unusual, but were common with other enterobacterial antigen extracts. However, higher titers were usually present to homologous as compared to heterologus antigens. Sera from seven patients contained precipitins to a common enterobacterial antigen. Precipitins to E. coli, Proteus, Klebsiella, and Serratia were detected in only a small proportion of control sera.

Antibodies, Bacterial↗
PubMed1985

Factors affecting the development of precipitating antibodies in workers exposed to contaminated humidifiers.

A total of 601 sera from groups of workers exposed to heavily and moderately contaminated humidifiers respectively were examined by the double diffusion test for precipitating antibodies to humidifier extracts. Clinical information was obtained using a standardized questionnaire. Skin-prick tests to control, to three common inhalant antigens and to an extract from the humidifier were performed in 103 subjects exposed to a heavily contaminated humidifier. In this environment a strong inverse relationship between current smoking and precipitins was detected (P less than 0.001). Pack years smoked had no bearing on precipitin level. In the larger study there was a clear dose response effect of smoking on precipitins. The effect of smoking appeared to reverse within 3 years. Duration of exposure also had a major effect on the development of precipitins with a clear 'dose response' again being seen in the larger study. These relationships did not apply in the groups exposed to moderately contaminated humidifiers. Levels of airborne antigen were measured using a competitive ELISA inhibition assay and were found to be much lower in sites with moderately contaminated humidifiers than at sites with heavily contaminated humidifiers. Our results suggest that the airborne antigen level is crucial in determining the pattern and strength of precipitin response in smokers and non-smokers exposed to contaminated humidifiers.

Air Pollutants, Occupational↗
PubMed1981

Precipitating antibodies against Staphylococcus aureus in serum from patients with staphylococcal septicaemia, investigated by means of quantitative immunoelectrophoretic methods.

Crossed immunoelectrophoresis was employed in the study of precipitating antibodies against Staphylococcus aureus in sera from 46 patients with S. aureus septicaemia. The occurrence and titres of antibodies were compared to previously reported findings for normal persons. Ten infection-specific precipitins were detected. However, most of these precipitins were detected in low titres and could only be detected during convalescence. When antibodies against a single antigen were measured, titres above the normal level were found in 39.1% - 65.2% of the patients. When the total number of precipitins in each serum was compared to findings in serum from normal persons, 54.3% of the patients had a higher value. Most promising was the precipitin score, considering both the number of precipitins and their titres. This score showed an early rise and was increased above the normal level in 82.6% of the patients. The antibody response against individual S. aureus antigens varied from person to person both concerning titres and time course.

Adolescent↗
PubMed1977

Immunochemical studies on the galactomannans isolated from mycelia and culture broths of three Hormodendrum strains.

Six major antigenic galactomannans isolated from both mycelia and culture broths of three representative strains of three species of the genus Hormodendrum, H. pedrosoi IFO 6071, H. compactum IFO 6726, and H. dermatitidis IFO 6421, were investigated for their immunochemical properties by precipitin reaction and the passive cutaneous anaphylaxis test. We found that partial acid hydrolysis of the these galactomannans with 0.005 M sulfuric acid yielded free galactose and corresponding acid-resistant core moieties. In agar-gel double-diffusion reactions, no acid-resistant core moiety of galactomannan showed a corresponding precipitin line against homologous antiserum, indicating that the acid-labile galactofuranosyl residues were solely responsible for the precipitin activity of these galactomannans. All galactomannans and their core moieties gave sharp cross-precipitin lines against anti-Saccharomyces cerevisiae serum, specific for the alpha1 leads to 3- or alpha1 leads to 2-linked D-mannopyranosyl residue. Quantitative precipitin reactions of the same antigen-antibody systems showed that all acid-resistant core moieties were completely inactive in the homologous systems but were highly cross-reactive with anti-S. cerevisiae serum. Passive cutaneous anaphylaxis tests using guinea pigs sensitized with homologous antisera or with anti-S. cerevisiae serum provided results consistent with those obtained in the in vitro assay.

Animals↗
PubMed1972

Serological investigations of a bovine respiratory disease ("urner pneumonie") resembling farmer's lung.

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.

Agar↗
PubMed1972

Pseudomonas aeruginosa: immune status in patients with cystic fibrosis.

In order to have a better understanding of the clinical significance of Pseudomonas aeruginosa, circulating and secretory antibodies were measured. Of 100 patients diagnosed as having cystic fibrosis (CF) and an atypical mucoid P. aeruginosa cultured from their sputum, each possessed serum precipitins. These immunoprecipitates, however, were not detected in the sera of 40 CF patients, some of whom were chronically ill with pulmonary colonization by typically rough-smooth strains of P. aeruginosa. The sera of 46 CF patients and 27 CF patient parents not colonized by P. aeruginosa were negative for the precipitins. The sera from 15 of 45 chronically ill patients not having CF, however, but harboring P. aeruginosa, also possessed serum precipitins. The sera from 85 subjects not having CF and not clinically infected with P. aeruginosa were negative for precipitins. Serum hemagglutination titers as high as 1:4096 were measured in older CF patients having advanced pulmonary disease and who were infected with mucoid P. aeruginosa. Salivary titers ranged from 1:8 to 1:64. Increased levels of both circulating and secretory antibodies of the immunoglobulin A and G classes were demonstrated in patients with CF. Once a patient with CF becomes colonized with P. aeruginosa a process of conversion from the rough and smooth forms to the mucoid form is almost inevitable. Although the mucoid form predominates in the sputum, intermediates of the various colony types are often present. Serum precipitins were demonstrable only after the appearance of mucoid strains in the sputum of patients with CF. Although antibiotics tend to reduce the number of mucoid microorganisms, they are rarely, if ever, eradicated from these patients' lungs. Recurrent episodes of servere pulmonary infection and the evidence of increasing antibody formation to mucoid strains indicates the invasiveness of these particular strains.

Adolescent↗
PubMed1979

Extractable antigen shared by Peptostreptococcus anaerobius strains.

Extracts from several species of gram-positive cocci were prepared by a modification of the Rantz-Randall autoclave method and tested for reactions with rabbit anti-Peptostreptococcus anaerobius (ATCC 27337 and VPI 5737) sera in a capillary precipitin test. Antigen preparations from two reference strains of P. anaerobius (ATCC 27337 and VPI 5737) and six clinical isolates of P. anaerobius reacted with the P. anaerobius antisera. These extracts formed a line of identity by immunodiffusion and displayed at least one precipitin line by immunoelectrophoresis. Absorption of the antisera with either the autoclaved extract or a 10% whole-cell suspension from each of the eight P. anaerobius strains removed the precipitin line(s) observed during immunodiffusion and immunoelectrophoresis. Extracts prepared to other species of Peptococcus, Peptostreptococcus, and Streptococcus did not react with the P. anaerobius antisera in a capillary precipitin test. In addition antisera to Lancefield groups A to G did not react with the extracts from the eight P. anaerobius strains. Preliminary chemical analysis of the extracts from the eight strains showed that they contained approximately 0.2 mg of carbohydrate per ml and 3.6 mg of protein per ml. The rabbit anti-P. anaerobius sera used in this study detected a common antigen(s) shared by strains of P. anaerobius, but did not react with autoclave extracts prepared from other species of gram-positive cocci. This extractable antigen could be used in a capillary precipitin test to rapidly identify P. anaerobius strains isolated in the clinical microbiology laboratory.

Antigens, Bacterial↗