Immunological studies on DNA-lysine photoadduct.

Calf thymus native DNA was covalently linked with lysine by irradiation with 254 nm light. Thermal melting studies indicated a decrease of 16 degrees C in the Tm of DNA-lysine photoadduct over native DNA. The decrease in Tm was dependent of irradiation dose. The photoadduct was found to be a potent immunogen in experimental animal. Thymine-lysine photoconjugate in DNA-lysine photo-monoadduct appears to be the major immunodominant portion. A strong recognition of DNA-lysine photoadduct was observed with anti-DNA autoantibodies found in the sera of patients with SLE. Thymine-lysine conjugate in DNA-lysine photo-monoadduct appears to provide an immunodominant epitope(s) for SLE autoantibody recognition. The results suggests for the possible involvement of DNA-lysine photoadduct or similar modified structure(s) as a potential trigger for anti-DNA autoantibody production.

Machine operator's lung. A hypersensitivity pneumonitis disorder associated with exposure to metalworking fluid aerosols.

Six auto parts manufacturing workers were referred for evaluation of a 6-week history of work-related dyspnea, cough, and fatigue. Two workers also reported fever and weight loss. All six worked in a machining area where a waterbased metalworking fluid was used and recirculated under high pressure, thereby creating an aerosol. Chest radiographs revealed pulmonary interstitial infiltrates in four workers. Lung function tests showed that four workers had decreased diffusing capacity. After removal from the work area, all workers recovered. The metalworking fluid was cultured for bacteria and fungi. Isolates from broth cultures were sonicated to obtain antigen extracts. Serum precipitins to one or more of the microbial isolates were identified in all six workers but not in eight of nine nonexposed control subjects. The most frequent precipitin response (six of six workers) was against antigens of Pseudomonas fluorescens, which was cultured from the metalworking fluid. In all workers, precipitins to at least one other cultured organism were detected; these included Aspergillus niger, Staphylococcus capitas, an acid-fast Rhodococcus sp, and Bacillus pumilus. This represents the first report of hypersensitivity pneumonitis associated with industrial exposure to aerosolized metalworking fluid. Observed precipitin responses to a variety of microbial contaminants in metalworking fluid strongly suggest a causative role for microbial antigens in the induction and elicitation of this manifestation of hypersensitivity pneumonitis.

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.

Localization of the tube precipitin and complement fixation antigens of Coccidioides immitis by immunoelectron microscopy with murine monoclonal antibodies.

The cellular localization of the tube precipitin (TP) and complement fixation (CF) antigens of Coccidioides immitis was examined by immunoelectron microscopy with murine immunoglobulin G1 monoclonal antibodies directed against the TP and CF antigens, respectively. Immunoelectron microscopic analyses of saprobic- and parasitic-phase cells showed that the TP antigen is present at a high concentration within the inner cell wall layer and along the plasma membrane. The antigen was also detected, at a lesser concentration, within cytoplasmic vacuoles. In contrast to the predominant localization of the TP antigen in the cell walls, the CF antigen residues primarily within the cytoplasm, where it appears to be dispersed throughout the cytoplasm rather than associated with a specific cytoplasmic organelle. A sparse amount of the CF antigen within the inner cell walls was also demonstrable. The localization of the TP and CF antigens throughout the morphogenetic phases of C. immitis has important implications in antigen production and in analyses of host response in coccidioidomycosis.

Precipitins and specific IgG antibody to Aspergillus fumigatus in a chest unit population.

BACKGROUND: The enzyme linked immunosorbent assay (ELISA) for detecting IgG antibodies to Aspergillus fumigatus is more sensitive than the measurement of Aspergillus precipitins. The relation of the results from both techniques to the clinical pattern of disease in a large unselected group of patients from a large referral centre is unknown. METHODS: The clinical relation of precipitins to Aspergillus fumigatus to clinical disease was determined retrospectively in 98 patients attending a primary referral centre. Precipitin results were compared with the specific IgG antibody to A fumigatus in 88 of the sera. Precipitins were determined by the agar gel double diffusion test and specific IgG antibody to A fumigatus by a quantitative ELISA. RESULTS: Precipitins were detected in the unconcentrated serum of 51 patients. Thirty nine of these had a mycetoma or allergic bronchopulmonary aspergillosis, 34 having specific IgG antibody to A fumigatus more than the control range. Forty seven patients had precipitins only after threefold concentration of serum or to only one of the four A fumigatus antigen extracts. Most of these had specific IgG in or near the control range. Thirty of these had A fumigatus skin test negative asthma or bronchiectasis, in which aspergillus was probably not pathogenic. There was a close relation between the level of antibody detected by the ELISA and the number of precipitin lines. CONCLUSIONS: This study reaffirmed the supportive role of aspergillus precipitins in the diagnosis of pulmonary aspergillosis. No additional benefit in the routine use of the ELISA was seen. It also showed that care should be taken in interpreting positive precipitin results from concentrated serum and that using several rather than one A fumigatus antigen extract is helpful for identifying allergic aspergillosis.

Precipitins in bird breeder's disease: how useful are they?

Precipitating antibodies to avian antigens play a controversial role in the diagnosis of bird breeder's disease (BBD). In order to establish the sensitivity, specificity and accuracy of precipitins in our laboratory, we conducted a prospective study including 128 sera received in 1988 for determination, by immunoelectrophoresis, of avian precipitins. Accurate information was obtained for 90 patients; definitive clinical diagnosis was given by the patient's own physician. We found a high sensitivity (86%), specificity (93%) and accuracy (92%) of avian precipitins in the diagnosis of BBD. Bayes' theorem was applied to determine the predictive value of the test with varying disease prevalence, and established that precipitins were particularly valuable for low or medium a priori probability. Using the receiver operating characteristic (ROC) curve we evaluated the effects of varying the positivity threshold of precipitins. The threshold used in this study appeared to offer the best compromise between sensitivity and specificity. These results suggest that the diagnostic value of avian precipitins in the work-up of BBD should be reconsidered, also because they represent a simple, cheap and non-invasive diagnostic test.

Characterization and partial purification of the cockroach antigen in relation to housedust and housedust mite (D.f.) antigens.

Allergens in crude cockroach antigens (CRa) have not been well defined. In order to characterize the CRa, the authors partially purified CRa by gel filtration (G-75) and analyzed it by polyacrylamide electrophoresis (PAGE) and allergy skin testing (AST). Hyperimmune serum was produced against CRa in rabbit (R anti-CRa) and used for immunoelectrophoresis (IEP), double immunodiffusion (DID), crossed immunoelectrophoresis (CIE), and crossed radio immunoelectrophoresis (CRIE) to further characterize the CRa. The crude CRa contained 9 and 15 precipitating arcs by IEP and CIE, respectively. Indirect CRIE revealed that CRa contained nine to 15 different allergens. There were no lines of identity between the CRa and housedust mite (D.f.) or between the CRa and commercial housedust by DID or IEP. Autologous housedust of cockroach asthmatic subjects showed lines of identity with the CRa. Fractionation of CRa on a Sephadex G-75 column chromatography showed two distinct peaks (B1 and B2) separated by a broad valley. The high molecular fraction B1 was most allergenic (100%) and contained three antigens of high molecular weight. These three antigens were identifiable by the techniques of IE, CIE, and DID. B2, the low molecular fraction (molecular weight less than 13,000 dalton) contained minor allergen(s). B2 elicited positive AST in 55% of cockroach-sensitive individuals, but it was not detected by IEP, CIE, nor by DID.

[Use of precipitins in the diagnosis of bird fancier's lung: apropos of 2 cases]. / Utilité des précipitines dans le diagnostic de la maladie des oiseleurs: à propos de deux cas.

Bird breeder's disease is a frequent diagnosis among the forms of extrinsic allergic alveolitis seen in the urban environment. We report two illustrative cases and discuss the respective roles of broncho-alveolar lavage cells and precipitating antibodies against avian antigens in the diagnostic work-up and the pathogenesis of the disease. In 1988 we evaluated over 100 sera from all over Switzerland with clinical suspicion of bird breeder's disease. In this cohort preliminary results show sensitivity of 85% and specificity of 92% for the presence of precipitins against avian antigens in the diagnosis of bird breeder's disease. The results suggest that this non-invasive test deserves revaluation as a diagnostic tool in bird breeder's disease.

The fluctuant nature of precipitating antibodies in dairy farmers.

Four hundred and forty five of 888 dairy farmers studied in an epidemiological survey four years previously were restudied to determine change in precipitins and in clinical features. Subjects answered the same questionnaire, which was filled in by the same nurse in the same winter months, and had blood withdrawn for precipitin analysis (double diffusion technique). Of the 445 subjects retested, 48 (10.8%) had been precipitin positive at the first study; of these, 34 remained positive four years later and 14 had reverted to negative. Twenty eight subjects previously negative for precipitins had become positive, so that 62 subjects in all were precipitin positive when restudied (13.9%). Of the 445 farmers, 369 were precipitin negative at both studies. The prevalence of symptoms, including cough, sputum production, wheezing, dyspnoea, and fever and chills, was similar for subjects who were consistently negative for precipitins and consistently positive and for those who changed from one to the other. Only one subject developed farmer's lung; he had precipitins in both studies. This study shows a fluctuation in the precipitin state of dairy farmers, suggesting that more farmers have precipitins at least once in their lifetime than are identified by screening at a single point in time. Relatively few develop farmer's lung or other respiratory symptoms. The presence of precipitins in a symptomless farmer appears to have no clinical importance.

Circulating immune complexes in sickle cell anaemia.

A polyethylene-glycol insoluble serum fraction was studied in patients with sickle cell anaemia during the steady state of the disease. The levels of C1q-precipitins were normal but increased amounts of proteins, IgM C3 and factor B were detected in this immune complex enriched serum fraction. These findings are a sign that circulating immune complexes can be detected even in the asymptomatic period of the disease.

Unusual precipitins in pathological human sera.

In conducting double diffusion gel precipitation tests, unexpected, strong precipitation reactions were noticed between a serum originating from a renal graft recipient and many pathological human sera. The highest frequency of positive reactions were produced by sera of patients with infectious mononucleosis (83%), chronic Epstein-Barr virus infections (72%), rheumatoid arthritis (57%), lepromatous leprosy (57%), and AIDS (44%). The precipitin in these pathological sera was identified as an antibody of IgM variety and the precipitinogen in the transplantation serum was shown to be a thermostable component with beta-globulin mobility. No explanation of the nature of reactions observed can be given at present.

A nonimmunoglobulin precipitin to tissue extracts in pathological human sera.

An alpha-globulin component was noted in pathological human sera, which produced gel precipitation reactions with extracts of human and animal liver. The highest incidence of the precipitin was found in malaria (95%), renal graft rejection (81%), and rheumatoid arthritis (57%). The precipitinogen was thermostable and ethanol soluble; of two precipitation lines formed by this component, one merged into identity reaction with a line produced by commercial lecithin of bovine origin. The possible diagnostic application of the reactions noted was considered.

Comparative analysis of precipitating antibodies in White Rock and Fayoumi hens injected with bovine serum albumin or crude mite extract with resulting effects on northern fowl mite, Ornithonyssus sylviarum (Acari: Macronyssidae) population densities.

Precipitating antibody concentration responses to crude northern fowl mite extract (CME) and bovine serum albumin (BSA) injections were compared in White Rock and Fayoumi hens with two-dimensional immunoelectrophoresis and rocket electrophoresis. The effect of CME injections on northern fowl mite population development was also determined. White Rock and Fayoumi hens developed similar antibody concentrations in response to intramuscular injections of BSA according to serum samples analyzed with two-dimensional immunoelectrophoresis. Rocket electrophoresis analyses of pooled serum samples showed significant differences between slopes of White Rock and Fayoumi pools for CME and BSA injections, suggesting differences in antibody-antigen interactions. Fayoumi hens injected with CME, 78, 50, and 14 days prior to experimental infestation with 2,000 northern fowl mites/bird supported significantly fewer mites than BSA-injected hens, although mite populations were low on both treatment groups. Injections of CME had no effect on mite population development on White Rock hens, even though CME-specific antibodies were detected. Although White Rock hens supported significantly greater mite numbers than Fayoumi hens, the difference was not attributed to anti-CME antibody activity alone.

Sequential bronchoalveolar lavage in experimental extrinsic allergic alveolitis. The influence of cigarette smoking.

We studied the alterations induced by acute experimental extrinsic allergic alveolitis (EAA) on bronchoalveolar cell population in smoking and nonsmoking guinea pigs. Sixty-two animals divided into 3 groups were studied: Group 1 (17 animals), controls; Group 2 (21 animals), extrinsic alveolitis; Group 3 (24 animals), cigarette smoking and alveolitis. Bronchoalveolar lavages (BAL) were performed on Days 1, 19, and 44 for all animals. Group 3 animals had a fourth lavage before starting cigarette smoking, that is, 28 days before the beginning of the antigen injections. The other lavages were as for the other groups. BAL results on Day 1 were similar for each group. Cigarette smoking per se did not modify BAL in Group 3. EAA induction resulted in a large increase in all BAL cells, especially neutrophils of recovered fluid, which increased from 38 x 10(3) to 1,474 x 10(3) ml-1 (p less than 0.01) in Group 2 and from 58 x 10(3) to 740 x 10(3) in Group 3 (p less than 0.01). After maintenance, BAL neutrophils.ml-1 decreased to 444 x 10(3) in Group 2 (p less than 0.01), but stayed the same in Group 3: 973 x 10(3). After EAA induction, BAL neutrophils.ml-1 were higher in Group 2 than in Group 3 (p = 0.039); however, Group 2 had less neutrophils.ml-1 than Group 3 (p = 0.035) after EAA maintenance. We conclude that EAA results in a neutrophilic alveolitis and which can be evaluated by sequential BAL, and that cigarette smoking decreases the initial neutrophilic response and retards the eventual recovery during maintenance injections.

Clinical findings in patients with SLE whose sera contain antibodies to ribosomal ribonucleoprotein.

In a clinical and serological study performed on a large series of patients with different connective tissue diseases, anti-ribosomal ribonucleoprotein (rRNP) antibodies were detected only in a small proportion of sera with systemic lupus erythematosus (SLE). SLE patients positive for anti-rRNP autoantibodies showed a significantly higher incidence of hemolytic anemia. The reasons for this surprising association are still unclear; however, this finding suggests that rRNP precipitin might be considered as a useful marker of a particular subgroup of patients with SLE.