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Correlation between ANA determinations on tissue substrate, WiL-2 cell substrate, and precipitin antibody by double diffusion.

Four hundred fifty-five sera from patients with various connective tissue diseases were screened for the presence of fluorescent antinuclear antibodies (ANA) on two different substrates-mouse kidney sections and WiL-2 cell smears-and precipitin antibodies by using WiL-2 cell extracts as an antigenic source. The antigen for the precipitin was prepared as for extractable nuclear antigen tests. Of 258 sera from systemic lupus erythematosus cases, 73% were positive for ANA on mouse kidney sections, 98% on Wil-2 cell smears, and 47% by precipitin tests. However, 33 sera which were ANA negative on mouse kidney sections were positive on WiL-2 cell smears and also positive for precipitin antibodies. WiL-2 cells used as ANA substrates increased the frequency of ANA in connective tissue diseases, and there was a strong correlation between the WiL-2 cell ANA and the presence of precipitin antibodies. Twenty-four of these 33 sera had anti-SSA/Ro antibody. Some sera containing anti-SSB/La or anti-nRNP antibodies were ANA negative on mouse kidney sections, but all were positive on WiL-2 cells.

Antibodies↗

Pseudomonas aeruginosa infection in cystic fibrosis. Diagnostic and prognostic significance of Pseudomonas aeruginosa precipitins determined by means of crossed immunoelectrophoresis.

A total of 133 patients with cystic fibrosis have been followed for up to 5 years with monthly examinations including bacteriological examinations of sputum. Sera from the patients were examined by means of crossed immunoelectrophoresis for the occurence and number of precipitating antibody specificites against Pseudomonas aeruginosa. Poor prognosis in cystic fibrosis was associated with chronic colonization (9 months - more than 5 years) of the respiratory tract with mucoid Pseudomonas aeruginosa, and with an onset of the chronic colonization before puberty. Among the patients with chronic Pseudomonas aeruginosa colonization, poor prognosis was associated with high numbers of precipitins against antigens from these bacteria (up to 61). The number of Pseudomonas aeruginosa precipitins increased on an average with five per year in chronically colonized patients. Rapidly increasing number of precipitins was associated with poor prognosis. Patients with any degree of impairment of the ventilatory function and any changes on the chest radiographs could contract chronic Pseudomonas aeruginosa colonization. Poor ventilatory function and severe changes on the chest radiographs was associated with high numbers of Pseudomonas aeruginosa precipitins and with poor prognosis. Although many O groups of Pseudomonas aeruginosa were found in the chronically colonized group of patients, 53% of the patients harboured strains belonging to O group 3 or 3/9, and the highest numbers of precipitins were found in serum from these patients.

Antibodies, Bacterial↗

The candida precipitin test in an immunosuppressed population.

Ninety-one immunosuppressed, high-risk patients suspected of having invasive candidiasis were tested during their septic episode for the presence of candida precipitin (CP) antibody using the counterimmunoelectrophoresis (CIE) method. In 12 patients, systemic candidiasis was confirmed but only 6 of these (50%) had candida precipitins. In 14 "questionable" cases who died without a definitive diagnosis, 8 (57%) had precipitin antibody. Of the 65 patients who did not have serious candida infection, 20 (31%) had candida precipitins. In this population we found the presence of candida precipitins to be neither sensitive (50%) nor specific (69%) for the diagnosis of invasive candidiasis.

Antibodies, Fungal↗

Conversion of C5 precipitin line in the serum treated with activating substances of complement system.

The hemolytic activity of C5 in the serum treated with zymosan, immune precipitate, or C1s was measured, and the C5 precipitin line on immunoelectrophoresis and the protein concentration of C5 in these serum specimens were also analyzed. A marked decrease in the hemolytic activity of C5 and a complete conversion of C5 precipitin line from beta- to alpha-globulin region were observed in teh serum treated with more than 1 mg/ml of zymosan. The elongation of C5 precipitin line from beta- to alpha-globulin region and the decrease in C5 hemolytic activity were observed in the serum treated with the immune precipitate. But neither change in C5 precipitin line, nor a decrease in hemolytic activity of C5 was observed in C1s treated serum. C5 protein concentrations in these serum preparations were essentially the same as those of control. From these results, it was concluded that the immunoelectrophoretic change of C5 precipitin line might express the grade of the decrease in C5 hemolytic activity in the serum treated with the activating substances of the complement system.

Antigen-Antibody Complex↗

Milk precipitins, circulating immune complexes, and IgA deficiency.

Twenty-two patients with selective IgA deficiency were studied for the presence of serum precipitins to bovine milk, bovine and fetal calf serum, and circulating immune complexes. Fifty-nine percent had circulating immune complexes, 50% had milk precipitins, 23% had precipitins to bovine serum, and 13% had precipitins to fetal calf serum. All patients with precipitating antibodies against milk or against bovine or fetal calf serum had circulating immune complexes and the precipitin titers correlated with the amount of circulating immune complexes. After one IgA-deficient patient had drunk 100 ml of milk, studies of sequential serum samples showed the presence of casein in the circulation at 60 min and the appearance of increasing amounts of immune complexes for 120 min. These findings are interpreted to indicated that in human beings the IgA system may provide a major barrier to absorption of immunogens from the gastrointestinal tract.

Adult↗

Precipitin system detected in sera from patients with non-A, non-B hepatitis.

A precipitin system detected by immunodiffusion was identified using sera from patients with non-A, non-B hepatitis. This system resembled those described by other authors as serological tests for non-A, non-B antigens and antibodies. However, testing of sera from a large number of patients with acute and chronic liver diseases revealed that this precipitin system was not specific for non-A, non-B hepatitis. Further characterization of the precipitin system demonstrated that it did not represent a true antigen-antibody reaction. Precipitin lines detected by immunodiffusion during the course of acute and chronic non-A, non-B hepatitis may no represent reactions specific for this disease.

Antigen-Antibody Reactions↗

Specific anti-amoebic immunoglobulins and the cellulose acetate precipitin test in Entamoeba histolytica infection.

Using the indirect fluorescent antibody test and sera from 10 proved cases of invasive amoebiasis, the effects of absorption of IgG on observed titres of amoeba-specific IgM and IgA were investigated. In addition, results of cellulose acetate precipitin tests were compared with anti-amoebic antibody levels. Antiamoebic IgM was found at titres of 1:14 to 1:112 after IgG absorption in four cases. Anti-amoebic IgA was detected in sera from four cases, but the maximum titre was 1:28. There was no relationship between the presence of amoeba-specific IgM or IgA and the result of the precipitin test, but a raised anti-amoebic IgG level was consistently found where the precipitin test was positive. However, sera with raised amoeba-specific IgG levels did not invariably give a positive precipitin reaction.

Amebiasis↗

A precipitin for human serum proteins as released into the environment by stressed bait shrimp, Penaeus duorarum.

1. A precipitin for human serum proteins is released into the environment by stressed bait shrimp, Penaeus duorarum. 2. Two-dimensional crossed immunoelectrophoresis revealed that the precipitin reacts (a) primarily with proteins belonging to the major group, alpha-1 globulin; and (b) with more proteins than a standard mammalian antiserum. 3. This study extends the variety of species known to liberate precipitins and suggests this response may be widespread among invertebrates. 4. The precipitins are easily collected in saline solution and, by virtue of their unique specificities, are potentially useful in diagnostic testing.

Animals↗

Salt-induced inhibition of the precipitin reaction of concanavalin A with polysaccharides and glycoprotein.

The time course of the precipitin reactions of concanavalin A with glycogen, dextran and ovalbumin was investigated by a light-scattering method near 30 degrees C in 10 mM-Tris/HCl buffer, pH 7.4, containing neutral salts, i.e. NaCl, KCl, NaBr, KI and NaClO4. With 0.8 microM-lectin and 0.36 mg of glycogen/ml, the half-life, t 1/2, of the precipitin reaction was independent of salt concentration between 0.1 M and 1.5 M, and was the same (175s) in the presence of NaCl, KCl, NaBr and KI but was significantly (27%) higher in NaClO4. In contrast, the five salts caused significant to marked enhancement in t 1/2 for the reactions of concanavalin A with dextran and ovalbumin. Likewise, whereas the turbidity produced in 1 h as a result of lectin-glycogen precipitation remained unchanged, those measured for the binding of dextran and ovalbumin were decreased in the presence of three salts. The increase in t 1/2 and decrease in turbidity were found to be higher with NaClO4, followed by KI; NaBr produced moderate and NaCl (or KCl) small but generally significant inhibition of the precipitin reactions with dextran and ovalbumin. The results showed that the lectin-ligand precipitin reactions involve salt-sensitive polar interactions that are less pronounced with compactly folded ligands such as glycogen.

Bromides↗

CLq precipitin in the sera of patients with allergic vasculitis (Gougerot-Ruiter Syndrome).

The sera from two well-documented cases of allergic vasculitis were examined for the presence of C1q precipitins. Both sera contained material capable of precipitating C1q in agarose gel. The material from one of the sera was partially purified using ammonium sulfate precipitation. Sephadex G-200 filtration, and DEAE-cellulose chromatography. Attempts to identify the nature of C1q precipitin were unsuccessful, but it resembled the high-molecular-weight precipitins of sera from patients with systemic lupus erythematosus. In a lesion, deposition of fibrinogen, immunoglobulins, and complement were noted mainly in the vessel walls. No correlation between immunoglobulin and complement deposition in the skin and the presence of C1q precipitins in the blood could be established.

Aged↗

Prevalence of precipitins in groups at risk of developing hypersensitivity pneumonitis.

A study was made of the prevalence of serum precipitins to Micropolyspora faeni, Thermoactinomyces vulgaris, Aspergillus fumigatus, and pigeon serum in population groups suspected to be at high risk for the development of hypersensitivity pneumonitis. Pigeon breeders' sera contained precipitins mainly to pigeon serum (38%) and A. fumigatus (18%). Occupants of homes in which forced air heating systems were investigated for the presence of microorganisms reacted mostly with M. faeni (13%) and A. fumigatus (8%). Individuals from environments where several cases of hypersensitivity pneumonitis were discovered reacted largely with M. faeni (28%) and T. vulgaris (21%). Sera supplied by physicians from patients with respiratory symptoms reacted primarily to A. fumigatus (24%) and to a lesser extent to M. faeni (16%) and T. vulgaris (9%). The results indicate that the prevalence of precipitins to the tested antigens is not uniform and may be influenced by the environment. Furthermore, the prevalence of precipitins in groups at risk is greater than previously reported for the normal population.

Alveolitis, Extrinsic Allergic↗

Precipitin to mite detected in asthmatic patients.

Mite antigen from dermatophagoides farinae was labelled with 125I, and used for detection of precipitin to mite antigen in sera from asthmatic patients allergic to mite and house dust. By double gel diffusion using this labelled antigen, it was found that 16 out of 37 sera formed precipitin bands which were detected by radioautography. Only one serum formed a precipitin band when non-labelled mite antigen was used in double gel diffusion. The main immunogenic component in mite extract migrated to the beta region in immunoelectrophoresis. The intensity of the precipitin line did not correlate with the relative concentration of IgE antibodies as determined by RAST. IgG antibodies to mite were detected by radioimmunoelectrophoresis in three out of 37 sera.

Allergens↗

Characterization of the precipitin bands detected in the immunodiffusion test for paracoccidioidomycosis.

In order to characterize the precipitin bands detected in the immunodiffusion test for paracoccidioidomycosis, a study was undertaken in 54 patients with the disease. On the basis of the pattern of known control sera, the three commonly observed lines of precipitate were designated as 1, 2, and 3 according to their location in the immunodiffusion plate. At time of diagnosis, 28 of the patients exhibited all three bands, 16 gave two bands, and 10 showed only one precipitin line. Over 50 of the sera with three bands had high complement fixation titers (above 1:512), whereas those with one band exhibited lower titers. A similar picture was obtained with the quantitative agar-gel techniques, where titers of 1:64 and above were more commonly observed in sera with three precipitin lines. Follow-up studies carried out in 18 patients revealed that band 3 disappeared first, followed by band 2, and, finally, by band 1. At the end of 2 to 3 years, 85.7% of the patients had lost band 3, 75% band 2, and only 27.7% band 1. Cross-reactions with histoplasmin were found in eight patients who gave the M precipitin line with this antigen. It was found that the latter band and our paracoccidioidin band 3 fused, producing lines of identity. Bands 1 and 2 were specific. The implications of these findings are discussed.

Blastomycosis↗

Isolation of a coccidioidin component that reacts with immunoglobulin M precipitin antibody.

Detection of immunoglobulin M (IgM) precipitin antibody to coccidioidin, the autolysate of mycelial-phase cells of Coccidioides immitis, is an important serologic aid in establishing a diagnosis of primary coccidioidomycosis. In the present study, the component of coccidioidin that reacts with IgM precipitin antibody was isolated by a combination of immunoaffinity and anion-exchange chromatography. Antigenic analysis of the purified antigen in two-dimensional immunoelectrophoresis against goat anti-coccidioidin revealed a precipitinogen characterized by a complete cathodal leg and a partial anodal leg. The reactivity of this incomplete precipitating antigen with anti-C. immitis IgM was established by serologic assays and by the adsorption of reference IgM precipitin antibody on solid-phase immunosorbents containing the purified precipitinogen. The isolation of the coccidioidin component that reacts with IgM precipitin antibody and the production of monospecific antibody will provide the necessary reagents for the development of a sensitive immunoassay for detecting this serodiagnostic response.

Antigens, Fungal↗

Classification of Neisseria meningitidis group B into distinct serotypes. II. Extraction of type-specific antigens for serotyping by precipitin techniques.

Over 10 distinct serotypes of group B Neisseria meningitidis have been found to date by using a sensitive microbactericidal assay developed by the authors. The serotype antigens have now been extracted by hot acid or saline extraction procedures. It was found that these extracted serotype antigens may be used in a simple capillary precipitin method. This method uses unadsorbed, undiluted rabbit antisera. In the capillary precipitin method a 3+ to 4+ reaction was considered significant. The microbactericidal assay and capillary precipitin methods for serotyping group B meningococci show excellent agreement. Group B meningococci were also serotyped by an agar gel double diffusion technique. The latter technique conserves reagents and has the further advantage that it does not show the minor cross-reactions observed in the capillary precipitin method. Thus two simple, reproducible methods for serological typing of group B meningococci have been developed. These methods developed for the serological typing of group B meningococci will aid in epidemiological studies of meningococcal disease. They may also be of value for selecting suitable strains for vaccine production.

Adsorption↗

Detection of Aspergillus fumigatus serum precipitins by counterimmunoelectrophoresis.

Detection of serum precipitins to Aspergillus fumigatus by counterimmunoelectrophoresis is compared with the immunodiffusion technique. Eight of nine (89%) sera from patients with proven A. fumigatus infection were positive by both methods. No serum from subjects with other systemic mycoses, bacterial infections, or healthy controls had detectable precipitins. The highest serum precipitin titers were found in sera of patients with the mycetomal and invasive forms of the disease. Detection of A. fumigatus serum precipitins by counterimmunoelectrophoresis compares favourably with immunodiffusion and has the advantage of significantly reducing the time required for results.

Aspergillosis↗

Counterimmunoelectrophoretic detection of a high incidence of precipitin reactions in normal human sera against staphylococcal teichoic acids and protein A.

The use of counterimmunoelectrophoresis (CIE) for detection of serum antibodies to staphylococcal teichoic acids was evaluated against teichoic acids prepared by sonic treatment or lysostaphin extraction of Staphylococcus aureus (Lafferty strain). Of 54 patient sera from suspected cases of staphylococcal endocarditis, osteomyelitis, or septicemia, 33 (61.1%) were positive by CIE analysis; however, 128 of 291 sera (44.0%) from normal adult donors were also positive. Selected CIE-positive sera from patient and control groups were titered by Ouchterlony gel diffusion. In the control group of normal sera, 65% were also positive by gel diffusion, but only 15% had titers of >/=1:2. Of the patient sera, 44.4% had gel diffusion titers of >/=1:2. In addition to the specific teichoic acid band, a second precipitation band could be demonstrated with both patient or normal sera by CIE or gel diffusion. This second precipitin band was shown to involve interactions of test sera with staphylococcal protein A present in the teichoic acid extracts. The protein A precipitins were detected at high concentrations of the antigen extracts, whereas the anti-teichoic acid precipitins were optimally detected at lower antigen concentrations. The formation of protein A precipitin bands did not correlate with the presence of anti-teichoic acid antibodies, as most sera tested were positive for protein A regardless of anti-teichoic acid activity. This study suggests that a high incidence of normal people have levels of antibodies to teichoic acids which are detectable by the highly sensitive, but nonspecific, technique of CIE.

Adult↗

Predictive value of bronchoalveolar lavage cells and serum precipitins in asymptomatic dairy farmers.

In 1981 to 1982, we reported that many (19/43) asymptomatic dairy farmers, especially those with positive serum precipitins, had a lymphocytic alveolitis. We reevaluated, six and seven years later, 33 of the initial 43 farmers to verify their outcome. The restudied group included 31 men and 2 women between 24 and 67 years of age. In both studies, 24 were nonsmokers and 9 were exsmokers. Each farmer answered a questionnaire, had a physical examination, blood withdrawal for precipitin analysis, a chest roentgenogram, and pulmonary function tests (forced expiratory flows and diffusion capacity). At restudy, one subject had developed symptoms suggestive of sub-acute farmer's lung and now had inspiratory crackles; six had chronic morning cough and sputum production. Serum precipitins were positive in 10 subjects, whereas 16 had been positive at the initial study. Precipitins reverted from positive to negative in eight subjects and from negative to positive in six. Chest roentgenograms were normal in 23 subjects, while discrete interstitial abnormalities were noted in nine. One subject had significant pulmonary infiltrates. Results of current pulmonary function tests were (% predicted, mean +/- SEM) DL(CO) 114.8 +/- 3.0, FEV1 108.1 +/- 2.4, FVC 105.4 +/- 1.1. No correlations were found between these data and the lymphocytes or mast cells found in the bronchoalveolar lavage (BAL) at the initial study of 1981 and 1982. We conclude that bronchoalveolar lavage cell counts of asymptomatic farmers have no long-term clinical significance.

Adult↗