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The frequency of precipitins to trichloroacetic acid-extractable antigens from thermophilic actinomycetes in farmer's lung patients and asymptomatic farmers.

The hypothesis that the trichloroacetic acid (TCA)-extractable antigen fractions prepared from Micropolyspora faeni cells and other thermophilic actinomycetes are better diagnostic immunodiffusion reagents for the detection of farmer's lung disease than the crude extracellular antigen concentrates has been tested. The frequency of serum precipitins to the TCA-extractable antigens has been compared in symptomatic farmer's lung patients and in healthy farmers having positive precipitin tests to the crude antigen concentrates of the analogous thermophilic actinomycetes. Sixty-eight per cent of the symptomatic group were found to have precipitins to the M. faeni TCA-extractable antigens compared with only 22 per cent of the asymptomatic group. With regard to the other thermophilic actinomycetes tested, no significant differences in frequencies of precipitins to TCA-extractable antigens between the symptomatic and asymptomatic groups were found. However, the total number of patients available for testing with each of these antigens was small. Sufficient chemical characterization of the TCA-extractable antigens was carried out to show that they were chemically similar to the TCA-extractable antigens prepared by other workers.

Antigens, Bacterial↗

Characterization of precipitin response to Micropolyspora faeni in farmer's lung disease by quantitative immunoelectrophoresis.

Sera from 12 patients with farmer's lung disease (FLD) drawn during an acute episode and displaying precipitins to Micropolyspora faeni were analyzed by crossed immunoelectrophoresis with an intermediate gel combined with a rabbit reference precipitate system. Forty-six precipitin arcs were identified in the reference system, and FLD sera reacted with 36 of these antigens. The FLD sera displayed significantly more precipitates and higher precipitin scores when compared with sera from 16 precipitin-positive subjectw who did not have FLD. However, there was overlap between the 2 groups, and neither measurement distinguished the 2 persons who became ill after challenge from one person who did not. Three disease specific antigens were identified that may have special relevance for pathogenesis of disease. Ten exposure-relevant antigens were also identified. We propose that these antigens be important components of a standardized M. faeni antigen preparation.

Chronic Disease↗

Anti-carbohydrate precipitins and haemagglutinins in haemolymph from Tridacna maxima (Röding).

Haemolymph from the elongate clam, Tridacna maxima (Röding) readily precipitates with H-blood group substances, pneumococcus type XIV polysaccharide, human milk and salivas, and with a number of polysaccharides which contain the O-SS-D-galactopyranosyl-(1-6)-D-galactose structure. Precipitation has been demonstrated using both gel diffusion and quantitative precipitin methods. T. maxima haemolymph strongly agglutinates human erythrocytes and haemagglutination can be inhibited by the same preparations which precipitate with the clam extract. Precipitins and haemagglutinins are inhibited by N-acetyl-D-galactosamine and by D-galactose residues preferably in ss-linkage. After agar gel immunoelectrophoresis at pH 8.6, T. maxima precipitin arcs are found in the alpha-region. Precipitation and inhibition results suggest that T. maxima extract and purified precipitin, may find widespread application in the study of many biologically important carbohydrates and glycoproteins.

Agglutinins↗

Antibacterial precipitins and autoantibodies in serum of patients with cystic fibrosis.

Sera from 84 patients with cystic fibrosis, 31 patients with other respiratory diseases and 21 control persons have been investigated for the occurrence of antibacterial precipitins and autoantibodies. Preciptins were studied by means of crossed immunoelectrophoresis, antinuclear factors by an indirect immunofluorescence technique and rheumatoid factors by the latex fixation slide test. A very heterogeneous antibacterial immune response was found in patients with cystic fibrosis, notably as regards Ps. aeruginosa. None of the other patients or controls had precipitins against this bacterium. The occurrence of precipitins against St. aureus and D. pneumoniae were more frequent in cystic fibrosis patients as compared with controls, but not as compared with other respiratory disease patients. No significant differences were found as regards precipitins against H. influenzae or the occurrence of rheumatoid factors. Antinuclear factors were more frequent in cystic fibrosis patients than in the other two groups investigated. A pronounced and heterogeneous humoral immune response against Ps. aeruginosa in cystic fibrosis patients chronically infected with mucoid strains of this bacterium was found to be correlated with poor prognosis and the reason for this is discussed.

Adolescent↗

Detection by ELISA of antibodies to small RNA protein particles in systemic lupus erythematosus patients whose sera lack precipitins.

Three hundred five consecutive patients with systemic lupus erythematosus (SLE) have been evaluated for antibodies to the RNA protein particles, Sm, nRNP, Ro/SSA, and La/SSB. Precipitating antibodies were found in 224 (73.1%). With the use of ELISA methodology with affinity-purified antigens, 76 sera from precipitin-negative patients were assayed and 35 had elevated levels of one or more of the autoantibodies. Of these, 27 (36%) had elevated anti-nRNP and 17 (22%) had elevated anti-Ro/SSA. Six of the seven patients with anti-La/SSB (18%) also had anti-Ro/SSA paralleling the close association of these two antibodies noted by precipitin analysis in which 38 of 39 patients with anti-La/SSB precipitins also have anti-Ro/SSA precipitins. Thus, 258 of the 305 SLE sera (84.6%) have elevated anti-RNA protein levels. This represents a minimum estimate of the frequency of these antibodies since longitudinal study of individual patients shows that anti-RNA protein levels may vary from normal to 1000 times over background at different times. Quantitative studies over time in two patients with both anti-Sm and anti-nRNP show that anti-nRNP increased 10- to 100-fold while anti-Sm changed little. These quantitative changes may relate to clinical phenomena.

Adolescent↗

Incidence of fungal precipitins in patients treated with beclomethasone dipropionate aerosol.

The authors followed the development of serum precipitins to aspergillus and Candida albicans in 14 asthmatic patients placed on beclomethasone dipropionate inhalation at a mean duration of five months. Three sera positive for aspergillus precipitins became negative during treatment. One patient had clinical oropharyngeal candidiasis and another patient's serum became positive for candida precipitin. The authors conclude that the incidence of oropharyngeal candidiasis and candida or aspergillus precipitins are rare among patients treated with beclomethasone.

Adult↗

Clinical application of counterimmunoelectrophoresis in detection of Candida serum precipitins.

Precipitin antibodies to candida antigens were found by counterimmunoelectrophoresis (CIE) and double immunodiffusion (DID) in 100 per cent of sera from patients with significant candida infection. Such antibodies were also found by CIE in 63 per cent of individuals with candida colonization, 47 per cent of bacteremic patients, and 14 per cent of normal control subjects. No sera from these patients and normal control subjects were positive by DID. Serum precipitin titers by CIE in patients with significant candidiasis were significantly higher than in patients of the other groups. Detection of candida serum precipitins by CIE, when precipitin titers are obtained, compares favorably with DID as a diagnostic tool in significant candidiasis.

Antigens↗

[Agar-Gel Precipitin Reactions In Experimental Paragonimiasis]

In an attempt to investigate the sensitivity of immunodiagnosis in cats experimentally infected with Paragonimus westermani, agar-gel precipitin reaction were studied. Metacercariae of P. westermani were administered to cats in various doses(2~100 metacercariae per cat) and antisera were obtained at an interval of a week. 1. Precipitin bands appeared in homologous antigen-antibody in experimental paragonimiasis between 3 and 5 weeks after infection in all the cats. 2. Almost all the cases in which a large number of worms were detected, showed strong reactions as revealed by deeply stained bands. 3. Precipitin reactions did not necessarily parallel with the number of worms detected. This may be attributable to the individual difference of a cat's conditions. 4. Very weak precipitin reactions were noticed between Clonorchis antigen and Paragonimus antisera of cats, but no reactions were noticed between Paragonimus antigen and Clonorchis antisera of cats or rabbits.

Journal Article↗

High incidence of precipitins in sera of heroin addicts.

We analyzed sera of 50 consecutively hospitalized heroin-abuse patients for precipitins against several antigens. Forty-seven addicts had precipitins against extracts from moldy hay, and 34 against extracts from bagasse. Thirty-six showed precipitin bands against extracts from one or more of the following: Aspergillus, Micropolyspora faeni, and Thermoactinomyces vulgaris. These findings were significantly different from those of a control population (P less than .0001 for all antigens tested). The recent culture identification of bacteria and fungi from street heroin suggests that the addicted population may be injecting antigenic material intravenously, with subsequent antibody formation.

Adult↗

A precipitin for human serum proteins as released under stress by a marine invertebrate, the marsh snail, Littorina irrorata.

A substance released into the environment by stressed marsh snails, Littorina irrorata, precipitates with the beta globulin and albumin fractions of human serum proteins. Other diverse invertebrate species may produce a similar substance. Invertebrate precipitins have not been widely reported, and none like the one described. Some physicochemical and adaptive characteristics of the snail precipitin are presented. This precipitin can be produced in large quantities and has potential application as a diagnostic reagent.

Animals↗

Further studies on the alpha-precipitins of the Atlantic salmon (Salmo salar).

The alpha-precipitins of the Atlantic salmon have been isolated by a combination of chromatography and gel filtration. A monospecific antiserum has been raised and this has been used to measure the concentration of the antigen in 925 fish sera by CELISA. The results were analysed for the effects of sex and disease state of the animal, time of year and year of collection, and the part of the UK from which they were collected. Significant differences were found in all cases. The components of alpha-precipitin were investigated by the isoelectric focusing of pure alpha-precipitin, whole serum and serum heated for 18 h at 95 degrees C. Multiple banding occurred in the acid region of the gels; the number, relative strength and position of the bands depended upon the animal from which the serum was isolated.

Animals↗

Effect of organic solvents on lysozyme-antilysozyme precipitin reaction.

The effect of nine organic solvents and urea on hen-eggwhite lysozyme-rabbit antilysozyme precipitin reaction was studied at a ratio of the antigen to the antibody of 1:26 by weight in 70 mM sodium phosphate buffer, pH 7.0. The organic solvents used were dioxane, acetonitrile, dimethylsulfoxide, N,N-dimethylformamide, 1-propanol, propylene glycol, trifluoroethanol, ethylene glycol and glycerol. These solvents invariably caused reduction in the amount of protein precipitated during the antigen-antibody reaction. The concentration of an organic solvent, CM, required for 50% reduction in the precipitin reaction value was determined for each organic solvent. Among the nine organic solvents, dioxane was the most potent inhibitor of the precipitin reaction. The nine organic solvents did not cause irreversible inactivation of the antigen and the antibody, and at concentrations used in this study most of them would be nondenaturing. These solvents seem to destabilize the antigen-antibody complex.

Animals↗

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↗

A modification of the automated immune precipitin method for quantitation of human serum immunoglobulins.

The use of the automated immune precipitin method to measure human serum immunoglobulins has been reported by several groups. The authors encountered difficulties in evaluating the technic even when they incorporated such recent modifications as the use of polyethylene glycol to enhance the reaction. By altering the flow pattern and prediluting each specimen, they were able to increase sampling rate, decrease processing time, and decrease the frequency of line obstructions while obtaining reproducible standard curves without using polyethylene glycol. There was good correlation between the results obtained by their modification of the technic and those obtained by the Fahey radial immunodiffusion method. As with other automated immune precipitin systems, abnormally high values gave characteristic notched peaks and lipemic specimens were centrifuged in order to obtain the infranatant for quantitation. The present modification of the automated immune precipitin system provides a rapid, simple and efficient method to quantitate human serum immunoglobulins.

Automation↗

Clinical evaluation of precipitin tests for genital actinomycosis.

A precipitin test system for antibodies against Actinomyces israelii, comprising a combination of counterimmunoelectrophoretic and crossed immunoelectrophoretic gel techniques, was evaluated for its clinical usefulness in diagnosing genital actinomycosis. A total of 263 serum samples from healthy women and women with proven actinomycosis, A. israelii-associated salpingitis, other gynecological infections, or miscellaneous gynecological diseases were analyzed. Six different precipitins could be detected. Five precipitins were defined as specific for actinomycosis, whereas one was found to occur nonspecifically in women with gynecological infections. The specificity of the test system for the detection of cases of genital actinomycosis was 98%, and the sensitivity was 83%. The accuracy was 100% for negative prediction and 45% for positive prediction. Thus, the test was shown to be valuable for a noninvasive diagnosis of genital actinomycosis.

Actinomyces↗

Serum precipitin reactions in Hashimoto's thyroiditis.

Sera from certain patients with Hashimoto's disease or a related condition contain a non-precipitating antithyroglobulin which produces in double-diffusion tests with thyroglobulin a line of clearing in the agar. Two such sera are described. Other sera from examples of the disease give negative precipitin tests with thyroglobulin but combine with sera containing ;clear line', non-precipitating antithyroglobulin to give a positive precipitin test. The serum from 15 out of 26 precipitin-negative cases of Hashimoto's disease showed this latent property. Eight of the 15 were tested by the tanned red cell technique and showed substantial titres of antithyroglobulin.

Autoantibodies↗

Occurrence of specific precipitins against bovine whey proteins in serum from children with gastrointestinal disorders.

Children with cow's milk intolerance, clinically non-confirmed cow's milk intolerance, coeliac disease, non-confirmed malabsorption, other gastrointestinal disorders and normal children were investigated for the presence of precipitins against 40 individual bovine whey proteins by means of the crossed immunoelectrophoresis with intermediate gel technique. In the various groups of children the amount and specificity of bovine whey precipitins were neither connected with the various gastrointestinal disorders nor with alternating diet and challenge with milk. Bovine whey-specific IgE could not be detected in any of the patients clinically suspected for cow's milk intolerance by cutaneous test, RAST or CRIE. The immunoglobulin level of the patients investigated did not differ significantly from the normal ranges. It is concluded that investigations of serum precipitins against bovine whey proteins do not give any significant information concerning cow's milk intolerance.

Adolescent↗

Prevalence of Aspergillus and Candida precipitins in renal dialysis and transplant patients.

Serum precipitin tests to Aspergillus and Candida antigens were used to define the rate of infectivity with these fungal organisms in patients on renal dialysis and after kidney transplantation. A total of 58 studies in 53 consecutive patients, of whom 41.5% were post-transplant an average of 14.6 months, failed to detect significantly more precipitin activity than in a normal population. The specificity of this non-immunoreactivity was supported by the absence of clinical or post-mortem evidence of fungal invasion. Since serum precipitins develop despite immunosuppressive therapy, and cases of fungal invasion in transplant recipients may occur sporadically in clusters, monitoring of renal transplant patients with these relatively simple tests should detect endemic occurrences of aspergillosis and candidiasis.

Adolescent↗