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PubMed1969

Rabbit antibodies to streptococcal carbohydrates. Influence of primary and secondary immunization and of possible genetic factors on the antibody response.

In a search for possible genetic factors which may influence the immune response to the streptococcal carbohydrates, over 100 rabbits have been immunized with streptococcal vaccines, and representative examples of high and low response pairs mated. The concentration of precipitins to the group-specific carbohydrates has been measured in the antisera following primary intravenous immunization with heat-killed streptococcal vaccines, Group A, Group A-variant, and Group C. For the majority of rabbits, the concentration of precipitins varied between 1 and 10 mg/ml of antiserum; while in the minority, it was between 11 and 32 mg/ml. The offspring of rabbits with high antibody levels had a significantly higher concentration of antibody than was seen in the offspring of rabbits of low response parents. Such data suggest that the magnitude of the immune response to these carbohydrate antigens is under some form of genetic control. Not uncommonly in rabbits with hyper-gamma-globulinemia following primary immunization, the group-specific precipitins are the predominant component of the gamma-globulin. An unusual feature of such components is that they are electrophoretically monodisperse, and possess individual antigenic specificity. In this respect they resemble the myeloma proteins. When a response of this sort is not seen after primary immunization, it may occur after secondary immunization. Therefore, prior exposure to the same or closely related antigen may also have an influence on the occurrence of high concentrations of such uniform antibodies.

Animals↗
PubMedDate not supplied

Serological reactions to Nocardia antigens.

Sera from 23 patients with nocardiosis were tested for the presence of precipitating antibodies against an extract of Nocardia asteroides. Sera from nine of 20 patients infected with Nocardia asteroides and one of three sera from patients with Nocardia brasiliensis produced one to three precipitin lines with the antigen. Sera from eight of 55 patients with tuberculosis and 11 of 13 patients with leprosy also contained Nocardia precipitins. None of 33 sera from patients with cryptococcosis, blastomycosis, actinomycosis, histoplasmosis, coccidioidomycosis, mucormycosis or candidiasis contained Norcardia antibodies. None of the sera from ten patients infected with Staphylococcus aureus, ten patients infected with Pseudomonas, or 16 sera from uninfected patients contained Nocardia precipitins.

Actinomycosis↗
PubMed1991

Serum antibodies to Pseudomonas aeruginosa outer-membrane proteins and iron-regulated membrane proteins at different stages of chronic cystic fibrosis lung infection.

Serum samples collected over periods up to 15 years from nine patients with cystic fibrosis (CF) were investigated by immunoblotting and crossed immuno-electrophoresis (CIE) for antibodies to Pseudomonas aeruginosa outer-membrane proteins (OMPs). The earliest antibody response to OMPs was directed against proteins G, H1 and I. Detection by immunoblotting sometimes preceded the CIE response; the appearance of antibodies to the other major OMPs was co-incident with an increase in CIE precipitins. Isolation of the mucoid form of P. aeruginosa was associated with a rapid increase in both precipitin numbers and antibodies detected by immunoblotting. Antibodies to iron-regulated OMPs could be detected in all the serum samples that showed eight or more CIE precipitins but their presence became pronounced only in the advanced stages of disease. The clinical strain used in this study and other isolates from CF patients showed several atypical OMPs, perhaps as a consequence of antibiotic therapy or related to the serum sensitivity of mucoid P. aeruginosa. Their expression in vivo was confirmed by detecting antibodies to them in patients' serum.

Antibodies, Bacterial↗
PubMed1977

Antigens in pigeon breeder's disease: the use of pigeon dropping antigens in detecting antibody activity.

Sera from patients with pigeon breeder's disease were analysed for precipitating antibodies by immunodiffusion and immunoelectrophoresis using whole pigeon dropping extract (PDE) and a purified fraction of PDE (PDE1) as antigens. For comparison, sera from asymptomatic pigeon breeders and normal individuals were also tested for precipitating antibodies. Whereas whole PDE formed precipitin lines with normal serum as well as with serum from symptomatic and asymptomatic pigeon breeders, PDE1 formed precipitin lines only with serum from individuals exposed to pigeons, This suggests that whole PDE forms non-specific (non-antigen-antibody reactions) as well as specific precipitin lines, while PDE1 appears to form only specific lines. These data indicate that whole PDE has limited usefulness in studies of pigeon breeder's disease. It is also possible that PDE1 will be useful in studies of this disease.

Alveolitis, Extrinsic Allergic↗
PubMed1982

Studies of Candida serology in denture stomatitis patients.

The aim of the study was to determine the humoral antibody response to Candida by means of hemagglutinin and precipitin tests and to correlate these results to yeast colonization in the oral sites and feces and to the relapse tendency after treatment; and furthermore, to evaluate whether Candida serology can serve as a tool to predict a denture stomatitis risk group. The serologic tests were performed by means of passive hemagglutination (HA) and immunoelectroosmophoresis (counterimmunoelectrophoresis, CIE) tests in 51 denture stomatitis patients and compared with 25 individuals in the reference group. There were significantly more patients with increased hemagglutinin titers (pos. greater than or equal to 1:160) to Candida albicans polysaccharide (mannan) antigen and/or precipitin to crude C. albicans cytoplasmic antigen in serum (P less than 0.01) than in the reference group. In the denture stomatitis group there was no correlation between the presence of hemagglutinating and precipitating Candida antibody in serum and the parameters yeast colonization of the oral sites/feces, yeast score on the denture base and palatal erythema score. After local oral antimycotic treatment there was no significant change in the serum Candida hemagglutinin titers and serum precipitins. Serologic tests might be useful as a guidance in a selected group of patients for continued prosthetic treatment and as a prognostic instrument.

Adult↗
PubMed1982

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

By means of crossed immunoelectrophoresis, precipitating antibodies against 55 Staphylococcus aureus antigens were investigated in serum from 11 patients with acute osteomyelitis and 47 patients with chronic osteomyelitis. Patients with acute osteomyelitis had an increase antibody response, expressed as a precipitin score, compared to normal persons, while patients with chronic osteomyelitis had an almost normal response. The precipitin score in patients with acute osteomyelitis was significantly higher than in patients with chronic osteomyelitis (p less than 0.01). However, patients with chronic osteomyelitis increased their precipitin score during an active phase of the infection.

Acute Disease↗
PubMed1970

Positive fluorescent treponemal antibody reactions in diabetes.

In a survey of 129 diabetic patients and 142 normal individuals, a significantly higher percentage of positive reactions in the fluorescent treponemal antibody-200 (FTA-200) test was found among diabetic patients than in the normal population. Absorption of all FTA-200-reactive sera with an extract of Reiter's treponeme eliminated most of the positive reactions in sera from diabetic patients, and three of the five positive reactions detected in sera from apparently normal subjects. On immunoelectrophoresis, precipitin bands developed most frequently between the Reiter sorbent and sera from diabetic patients positive in the FTA-200 test. Serum components responsible for FTA reactivity and precipitin reactions against the sorbent were resistant to treatment with mercaptoethanol, suggesting antibody of the IgG class. Cross-reacting antibodies produced in response to normal treponemal flora, and perhaps acquiring enhanced reactivity by means of nonspecific interacting substances in sera peculiar to the altered physiological state of diabetes, are suggested as possible causes of positive reactions of unabsorbed sera. No correlation could be made between age of the diabetic patient, treatment or duration of the disease, and FTA or precipitin reactivity of the patient's serum.

Cross Reactions↗
PubMed1971

Nonspecific precipitation of serum proteins by sodium lauryl sulfate in agar diffusion and immunoelectrophoresis.

The anionic detergent sodium lauryl sulfate (SLS), in a final concentration of 0.1% and greater, reacted with whole serum in agar diffusion and immunoelectrophoresis to form artifactual precipitin lines. These lines occurred when either Ionagar or agarose was used as the supporting gel and were not affected by the presence of urea and 2-mercaptoethanol. Analytic chemical tests confirmed that the precipitating agent is SLS, and staining techniques showed that the detergent precipitates both protein and lipoprotein components of whole serum. Multiple artifactual precipitin lines occurred with a wide variety of animal sera, and a single line formed with human 7S immunoglobulin. Hence, in agar diffusion studies in which SLS is present in the test system, these artifactual lines may be easily misinterpreted as true antigen-antibody precipitin reactions.

Agar↗
PubMed1981

Fractionation of Schistosoma japonicum soluble egg antigen glycoproteins by hydrophobic interaction chromatography.

We are currently studying the soluble egg antigens of Schistosoma japonicum in an attempt to determine which antigens are potent immunogens. Previously, we demonstrated by Ouchterlony immunodiffusion and inhibition of the circumoval precipitin test that the glycoprotein fraction of soluble egg antigens contains the antigens which are most immunogenic in natural infections. The soluble egg antigen glycoproteins have now been further fractionated via hydrophobic interaction chromatography on phenyl Sepharose. We found that there were at least two antigens involved in the circumoval precipitin reaction. Both the hydrophilic antigen which we call japonicum antigen glycoprotein II (JAG II) and a mixture of hydrophobic antigens (JAG III and the JAG IV complex) were capable of causing a 50% inhibition of the COP reaction around S. japonicum eggs. JAG II was not a major serological antigen of S. japonicum since it gave only a weak precipitin line upon Ouchterlony immunodiffusion analysis with pooled sera from Filipino patients with chronic S. japonicum infections. Hydrophobic interaction chromatography yielded preparations which were sufficiently pure for use in radioimmunoassays. By radioimmunoassay, the best antigens among the glycoproteins were moderately hydrophobic JAG III and the JAG IV complex. They had large amounts of antibody directed toward them in patients with schistosomiasis japonica and exhibited little reactivity with S. mansoni. The hydrophilic glycoproteins JAG I and II were poor immunogens and extensively cross-reacted with S. mansoni. This cross-reactivity means that diagnostic tests with crude soluble egg antigens would run the risk of potential false-negative results in patients with other trematode infections.

Animals↗
PubMed1972

Characterization and comparison of mycobacterial antigens by two-dimensional immunoelectrophoresis.

Two-dimensional immunoelectrophoresis (2D-IEP), in which a complex of antigens is subjected to electrophoresis first through an agarose matrix in one direction and secondly through an antiserum-agarose matrix at right angles to the first direction, was evaluated as a tool for analysis of mycobacterial antigens. Cell extracts from four species of mycobacteria, Mycobacterium tuberculosis (four strains), M. bovis strain BCG, M. scrofulaceum, and M. phlei, were assayed by 2D-IEP with four anti-mycobacterial antisera. Besides displaying the precipitin curves in a more easily interpreted format than did conventional immunoelectrophoresis (IEP), 2D-IEP offered greater sensitivity in terms of numbers of precipitin curves when like reactions were compared with IEP patterns. As many as 60 immunoprecipitates were observed on 2D-IEP slides compared to 18 on comparable IEP plates. Technical reproducibility of patterns from run to run was excellent. Other parameters, such as the influence of using different batches of antigen on the pattern, are discussed. Each of the cell extract antigens gave a unique pattern of precipitin peaks which could be easily differentiated from the patterns given by the other mycobacterial cell extracts when reacted with any of the antisera in 2D-IEP. Since both the species and strains of mycobacteria could be easily and reproducibly differentiated solely on the basis of two-dimensional immunoelectrophoretic patterns obtained with any of the antisera employed in this study, it may be possible, by using IEP, to differentiate and identify all species and strains of mycobacteria with one standard, highly sensitive antiserum, rather than a battery of antisera.

Animals↗
PubMed1973

Immunochemical properties of anti-cord factor antibody.

Sera of man and animals infected with tubercle bacilli were tested for precipitating antibody to trehalose-6, 6'-dimycolate (cord factor) of Mycobacterium tuberculosis. None of the sera of mice and rabbits infected with M. tuberculosis H37Rv reacted with cord factor, whereas the sera of animals vaccinated with cord factor-methylated bovine serum albumin complex demonstrated a precipitin reaction with cord factor. Antibody against cord factor was not detected in the sera of cases of active, convalescent, and pneumonectomized human tuberculosis, indicating that the anti-cord factor antibody is not associated with tuberculous infection in man and animals. Precipitin activity against cord factor was localized in 19S macroglobulin (immunoglobulin M) fraction of the serum of rabbits vaccinated with cord factor-methylated bovine serum albumin complex. Analytical gel filtration and immunodiffusion studies indicated that the anti-cord factor precipitin dissociated from the cord factor-antibody complex is an immunoglobulin M antibody.

Adult↗
PubMed1970

Composition and properties of a group A streptococcal teichoic acid.

Teichoic acid-like material extracted by cold trichloroacetic acid from lyophilized whole cells of streptococci from groups A,D,E,O, and T was shown to give a positive precipitin reaction with group antisera. Similar material from cells of groups B,C,F,G,H,K,L,M,N,P,Q,R, and S did not give a positive reaction with group antisera. The group A material also reacted with anti-E serum; however, the opposite did not occur. A similar result was also obtained on the group T material and anti-O serum. The group A teichoic acid was purified by Sephadex column chromatography, and was shown to be free of cell wall peptidoglycan and polysaccharide, and ribitol teichoic acid. It was composed of glycerol, phosphate, alanine, and glucosamine. Alkaline hydrolysis showed the presence of ester-linked alanine and glucosaminylglycerol. Phosphorus was released from ester linkage by alkaline phosphatase. N-acetylglucosamine produced a 72% inhibition of the precipitin test at a level of 10 mumoles, and d-alanine methyl ester was significantly stronger than the l-alanine ester. A single precipitin band was seen with group A serum. The data indicate that teichoic acid of group A streptococci is a polymer composed of glycerol phosphate and containing N-acetylglucosamine and alanine. Antisera to these streptococci contain antibodies specific for the alanine and the glucosamine linkages. The use of serum containing antibodies to alanine-polyglycerophosphate shows that the occurrence of this type of teichoic acid is widespread among the streptococci.

Acetone↗
PubMed1969

Immunochemistry of the cell walls of Listeria monocytogenes.

The antigenic specificity of Listeria monocytogenes types I, II, III, IVa, and IVb was studied by immunochemical techniques. Immunologically active carbohydrates of the various types were extracted from cell walls and were chemically analyzed. Types I and II contained predominantly glucosamine and rhamnose; type III, galactose, rhamnose, and glucosamine; and types IVa and IVb, glucose and galactose. Quantitative precipitin inhibition tests with purified monosaccharides indicated that the major antigenic determinant of types I and II is rhamnose. Precipitin reactions could not be detected with type III carbohydrate and homologous or heterologous antisera. The major determinants of types IVa and IVb were found to be galactose and glucose, respectively. As much as 87% inhibition of the quantitative precipitin test for types I and II was obtained with rhamnose, 72% for type IVa with galactose, and 72% for type IVb with glucose. The immunochemical basis for the antigenic specificity of L. monocytogenes types I, II, IVa, and IVb was further confirmed by using agar gel diffusion. Cross-reactions among the various type-specific carbohydrates and heterologous antisera were also studied. Type II carbohydrate was found to contain galactose and react with type IVa antisera. This reaction could be blocked by galactose. Type I carbohydrate did not contain galactose nor did it react with antiserum prepared from type IVa cells. Therefore, the somatic antigens of type I and type II L. monocytogenes, previously thought to be identical, appeared to differ. The dominant immuno-specific group in the cross-reaction between type IVb carbohydrate and type IVa antisera was found to be galactose. Type IVa absorbed antisera did not produce a significant cross-reaction with type IVb carbohydrate. The results obtained from this investigation indicate a lesser degree of antigenic relationship between type IVa and type IVb L. monocytogenes than was previously believed to exist.

Alanine↗
PubMed1980

Improved diagnostic accuracy using monoclonal antibody group A streptococcal carbohydrate.

Monoclonal mouse antibody to streptococcal group A carbohydrate was evaluated in diagnostic microbiology laboratory. A total of 262 isolates of beta-hemolytic streptococci were classified with commercial reagents and the monoclonal antibody to group A carbohydrate by using immunofluorescence and Lancefield precipitin tests. Both nonoclonal and commercial antibodies were identifically reactive by the precipitin test but significantly different by immunofluroescence. Monoclonal antibody gave the same reactivity as seen in the precipitin test, but commercial antibodies gave both false-positive and false-negative results. These results suggest that monoclonal reagents may be superior to conventional antisera.

Animals↗
PubMed1976

Serological diagnosis of infection of the urinary tract by yeasts.

Precipitins to mannan and cytoplasmic antigens of Candida albicans and Torulopsis glabrata were determined in 25 patients with colonization of the bladder urine by yeasts and in 25 control patients with bacteriuria. Precipitins were present in 64% of the patients with funguria and in 56% of the patients with bacteriuria. There was no correlation between the reactions obtained and the clinical significance of the yeast colonization. It is suggested that a single precipitin test is unhelpful in the assessment of the significance of urinary colonization by yeasts in hospital patients.

Antibodies, Fungal↗
PubMed1980

Distribution of pathogenic yeasts and humoral antibodies to candida among hospital inpatients.

The frequencies of the carriage of yeast pathogens and of serum precipitins to a variety of candida antigens among 254 patients generally tended to increase with the length of the patient's stay in hospital. This trend was observed even though none of the patients investigated showed signs or symptoms of superficial or systemic candidosis. The extent of the general trend varied considerably between subgroups of patients within the general categories of 'surgical' and 'nonsurgical' inpatients. Increases in both frequencies and quantities of yeasts in the mouth were most evident postoperatively among patients who underwent open-heart surgery and among nonsurgical patients who received antibiotics or steroids in hospital. The frequency of precipitins to Candida albicans cytoplasmic antigens in the absence of candidosis rose overall from 11% of 217 sera obtained within 24 hours of admission to 35% of 85 sera obtained five to 11 days after admission or operation. These 'false positive' antibodies were thought to arise after transient yeast overgrowth in the gut at the time of an acute illness or immediately after surgery. The study adds further data to documented examples of 'false positive' candida antibodies and indicates the need for care in the diagnostic interpretation of candida precipitin test results among groups of patients at risk of yeast overgrowth during their hospital stay.

Antibodies, Fungal↗
PubMed1975

Chest symptoms in farming communities with special reference to farmer's lung.

Surveys were carried out on random samples of the farming population in Devon and Wales in order to estimate the prevalence of respiratory symptoms and of positive precipitin reactions to thermophilic fungi. Bronchitis, as defined, was common among the Welsh hill farmers, and the proportion of positive serological tests was higher in both the areas surveyed in Wales as compared with Devon. All three surveys confirmed a previous finding that the proportion of positive precipitin tests was higher among non-smokers than smokers. Although the numbers were small there was some indication that measurement of peak expiratory flow showed different relationships with age in non-smokers according to the presence or absence of positive precipitin tests. The difficulty of determining prevalence rates for farmer's lung is discussed, but the results suggest a rate not dissimilar to those found in two areas of Scotland which were more than 20 times higher than any figure previously reported in Britain.

Adolescent↗
PubMed1976

Serological investigations in 15 cases of bird fanciers disease.

Serological studies in 15 patients with bird fanciers disease are reported. Quantitative analysis of serum proteins showed elevation of IgG in almost every case. High IgE was observed in three patients with isolated asthmatic reactions. In four patients alpha1-antitrypsin was transiently diminished. Two of these had heterozygous deficiency of this protein (genotype MS and MZ). Precipitins against avian sera and avian droppings were found in all patients, but also in some exposed and nonexposed controls. Cross-reactions to avian antigens from different species were observed quite regularly in patients but not in controls. One case is presented suggesting that these cross-reactions may be of clinical importance. Precipitins against other inhaled antigens such as Aspergillus fumigatus, cereals or Micropolyspora faeni were observed significantly more frequently in patients than in controls. These precipitins showed no cross-reactions with those against avian antigens.

Adult↗