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PubMed1995

Hypersensitivity pneumonitis associated with home ultrasonic humidifiers.

We describe five patients with hypersensitivity pneumonitis (HP) that was related to using home ultrasonic humidifiers. All patients had micronodular infiltrates on their chest radiograph, and their lung biopsy specimens revealed alveolitis with or without epithelioid cell granulomas. Challenge tests were performed on two patients with the humidifier water and three patients using the humidifier. All patients tested exhibited a positive response. Tests for precipitating antibodies against an extract of the humidifier water gave strongly positive reactions in all patients tested. Precipitins to Cephalosporium acremonium and Candida albicans were also present in all cases, whereas precipitins to thermophilic actinomycetes were not detected. Although cultures of the water grew a variety of fungal and bacterial organisms, thermophilic actinomycetes could not be detected. These findings suggest that thermophilic organisms may not be the causative antigens of HP associated with ultrasonic humidifiers. All five patients had an increase in the bronchoalveolar lavage (BAL) lymphocytes that were predominantly CD4+ lymphocytes. The T helper cell count (CD4) to suppressor T cell count (CD8) ratio was significantly higher than that observed in summer-type HP, and lower than that observed in bird fancier's lung, indicating that the phenotypes of the BAL lymphocytes may vary with the type of HP.

Adolescent↗
PubMed2005

Sensitization to Aspergillus antigens and occurrence of allergic bronchopulmonary aspergillosis in patients with asthma.

BACKGROUND: Allergic bronchopulmonary aspergillosis (ABPA), which is predominantly a disease of asthmatic subjects, is caused by hypersensitivity to Aspergillus antigens. Screening for Aspergillus sensitization in asthmatic subjects could identify those who are at risk for ABPA. Few studies have shown that fungal sensitization could be an important risk factor for asthma severity. We sought to determine the frequency of sensitization to Aspergillus antigens in asthmatic subjects and its effect on disease severity. We also determined the occurrence of ABPA in these subjects. DESIGN: Prospective study of consecutive patients with asthma. SETTING: Tertiary university referral hospital, outpatient department. PATIENTS AND METHODS: One hundred five asthmatic subjects and 26 volunteers underwent skin testing with aeroallergens, including Aspergillus, serum precipitins against Aspergillus antigens, and specific IgG against Aspergillus fumigatus, total serum IgE levels, and routine blood and radiologic investigations. ABPA was diagnosed when all eight major criteria were fulfilled. RESULTS: Thirty patients (28.5%) had a positive skin reactivity to Aspergillus antigens. Eleven patients (10.4%) had positive specific reactions to IgG, and 8 patients (7.6%) demonstrated positive reactions to serum precipitins. Eight of these 30 patients (26.6%) received diagnoses of ABPA, which was 7.6% of the total. None of the control subjects were sensitized to Aspergillus antigens. The patients were classified into the following four groups: negative skin test results; positive reactions to aeroallergens other than Aspergillus; positive reactions to aeroallergens including Aspergillus antigens; and patients with ABPA. Based on clinical and serologic parameters, patients with Aspergillus-sensitive asthma and ABPA had a significantly more severe form of the disease. CONCLUSIONS: Sensitization to the mold Aspergillus increases the severity of asthma. ABPA should be excluded in all patients with Aspergillus-sensitive asthma.

Adolescent↗
PubMed1984

Establishment of a human pancreatic cancer cell line and detection of pancreatic cancer associated antigen.

We succeeded in an establishment of a human pancreatic cancer cell line (PK-1) from liver metastasis of pancreatic cancer. Primary pancreatic cancer cells grew as islands surrounded by fibroblastic cells. However, these fibroblastic cells were gradually omitted by the polygonal shaped cancer cells. This cell line contained neither zymogen granules nor trypsin indicating that this pancreatic cancer originated from pancreatic duct cells. Modal chromosome numbers of this cell line were 42 and 72 and the doubling time was 48 hr. This cell line was transplantable in athymic nude mice to form progressive tumors which had histology similar to that of the original cancer (papillotubular adenocarcinoma). Neither AFP nor ferritin but CEA was detected on the surface and in the cytoplasm of this cell line in indirect immunofluorescence. Rabbit antiserum against this pancreatic cancer cell line detected pancreatic cancer associated antigen besides CEA in the culture supernatant. This antiserum reacted with sera from patients with pancreatic cancer to form a distinct precipitin line in agarose gel which fused with the precipitin line formed between the culture supernatant of this cell line and the antiserum.

Animals↗
PubMed1984

Serological tests in the diagnosis of pulmonary aspergillosis.

Sera from 44 patients with clinically suspected pulmonary aspergillosis (mainly of an allergic type) were examined for antibodies to Aspergillus fumigatus using an enzyme-linked immunosorbent assay (ELISA) and counter-immunoelectrophoresis (CIE). Of the sera, 15 were considered to contain Aspergillus antibodies using ELISA; 11 of these also contained precipitins by CIE. In no instance was a CIE-positive serum negative by ELISA. The serum of 1 patient with autopsy-verified invasive pulmonary aspergillosis was negative in both tests. Protein-enriched antigens derived by ammonium sulphate precipitation of crude hyphal homogenates seemed of most use in ELISAs. In addition, 4 of 8 sera obtained from patients with possibly invasive candidosis also revealed significant levels of Aspergillus antibody by ELISA (but not CIE). All of the 8 sera contained readily detectable Candida precipitins by CIE. Problems of potential cross-reactivity obviously need careful consideration with ELISAs. Our results suggest that ELISA procedures under appropriately controlled conditions are more sensitive than CIE for detecting Aspergillus antibodies. However it seems that some patients with invasive aspergillosis will be antibody-negative even with sensitive tests such as ELISA.

Antibodies, Fungal↗
PubMed1985

Rat Clq: isolation and purification from normal serum and development of a sensitive hemolytic assay.

Rat Clq was isolated and purified from normal rat serum by a two-step procedure: affinity chromatography on a human IgG-Sepharose 4B column and Bio-Gel A 5m column chromatography. From 1.8 l of normal serum, the procedure yielded 4.5 mg of homogeneous and hemolytically active rat Clq as shown by three bands of peptide in SDS polyacrylamide gel electrophoresis under reducing conditions and by a single precipitin line between rat Clq and rabbit anti-rat Clq antibody. Rabbit anti-rat Clq antibody prepared by immunizing a rabbit with 225 micrograms of rat Clq was specific as shown by a single precipitin line between the antibody and normal rat serum and rat Clq with complete identity. The amino acid composition of rat Clq was very similar to that of human Clq. The purification procedure also yielded Clq-depleted rat serum which was used with the homogeneous Clq to establish a sensitive hemolytic assay: 4.5-13.5 ng of rat Clq can be reproducibly quantitated. The concentration of Clq in Brown Norway rats was estimated to be 41.5 +/- 3.0 micrograms/ml serum.

Amino Acids↗
PubMed1986

Protein and antigenic composition of adult and microfilarial stages of Setaria cervi.

The protein and antigenic pattern of adult (female/male) and microfilarial stages of Setaria cervi, a bovine filarial parasite, was studied using certain immunochemical techniques. SDS-polyacrylamide gel electrophoretic analysis showed the presence of 35-40 protein bands in adults and 25-29 protein bands in microfilariae in molecular weight range of 10,000-200,000. Immunoelectrophoresis revealed the presence of 9-10 precipitin lines in adult and only 4 precipitin lines in microfilarial antigenic preparations. Crossed immunoelectrophoresis resolved these antigenic preparations further, and revealed the presence of 22-24 antigens in adults and 12-14 in microfilariae. These results demonstrate complex nature of somatic extracts of adult stage as compared to microfilariae and also reveal some qualitative and quantitative differences between these stages.

Electrophoresis, Polyacrylamide Gel↗
PubMed1983

Alteration of seminal proteins during freeze-drying of bovine semen.

Bovine semen in TEST-yolk extender was frozen, freeze-dried to 50, 25, 12, 6, and less than 2% residual moisture, and stored at -196 degrees C. The freeze-dried semen was rehydrated, sampled for protein analysis, and used to inseminate cattle. Agar gel electrophoresis revealed no moisture was reduced to less than 6%. At that point, the percent of cationic proteins and neural proteins decreased with a concurrent increase in anionic migrating proteins. Immunodiffusion data with antisera against spermatozoa and seminal plasma revealed no difference in formation of precipitin lines if residual moisture was at least 6%. However, semen freeze-dried to 2% residual moisture was modified antigenically as certain precipitin lines were lost, new lines appeared, and concentration of other seminal antigens decreased. The absence of fertility with semen freeze-dried to 2% residual moisture is hypothesized to 2% residual moisture is hypothesized to be from alteration of the tertiary structure of certain essential seminal proteins.

Animals↗
PubMed1986

[Fractionation of antigen for ELISA of bovine fascioliasis]

In order to obtain the most specific and sensitive antigen from crude antigens of Fasciola hepatica for the immunodiagnosis of bovine fascioliasis by the enzyme linked immunosorbent assay(ELISA), phosphate buffered saline extract of F. hepatica was prepared. The crude extract was fractionated into 7 antigens using Sephadex G-100 column chromatography. Seven fractionated antigens were applied to ELISA, precipitation test and intradermal test, respectively. Results obtained are as follows: The specificity (95 per cent confidence interval in negative sera of bovine fascioliasis; Mean+2 x SD of absorbance ) of the first (MW>150,000) and the second antigens (MW 120,000) were 93.7 per cent, but those of others including crude antigen showed 100 per cen.t. The sensitivity (positive sera of bovine fascioliasis having higher values with compared to the criterion) of the first, the sixth (MW 16,000) and the seventh antigen (MW<5,000) were 91.6 per cent, 87.5 per cent and 0 per cent, respectively, but those of others showed all 100 per cent. The absorbance by ELISA using the fifth antigen (MW 26,000) was 8.43-folds higher in the positive sera than that in the negative sera. This could be used as one of the most specific antigens for the immunodiagnosis of bovine fascioliasis. In Ouchterlony test, precipitin lines were not found in the sera naturally infected with F. hepatica, but some were found in the sera of rabbits immunized with the crude antigens. The numbers of precipitin lines in the sera of rabbits were different in the different fractionated antigens. They were 6 in the crude, 2 in the second and the third antigens, 1 in the forth, the fifth and the sixth antigens and absent in the seventh antigen, respectively. The wheal size for the bovine infected with F. hepatica was 2.46+/-0.15 cm in the intradermal test antigen (saline extract of F. hepatica) supplied by the Veterinary Research Institute, Rural Development Administration, Korea. The wheal size of the first, the second and the third antigens were larger than that of intradermal test antigen, whereas those of the forth, the fifth, the sixth and the seventh antigens showed smaller than that of the intradermal test antigen. The results suggest that the fifth antigen may be specific antigen for the immunodiagnosis of bovine fascioliasis.

Journal Article↗
PubMed1982

An immunological study of the diverse forms of vitellogenin identified in quail plasma.

Quail vitellogenins (which exist in three different native forms designated Vg alpha, Vg beta and Vg gamma) can be separated into two fractions by DEAE-cellulose chromatography: peak III (containing Vg alpha, Vg beta, and Vg gamma) and peak IV (containing Vg alpha and Vg beta). In the present study, antibodies were prepared against peak III and peak IV vitellogenin fractions, which were then compared with each other and with vitellogenin-containing plasma by immunodiffusion, rocket immunoelectrophoresis, and two-dimensional immunoelectrophoresis. Peak III and peak IV vitellogenin fractions behaved similarly on immunodiffusion and gave an apparently homogeneous precipitin band. However, rocket immunoelectrophoresis revealed at least three or four antigenic determinants in peak III or peak IV vitellogenin fractions. Two-dimensional immunoelectrophoresis showed that peak III vitellogenin fractions contained an antigenic determinant not present in peak IV vitellogenin. In addition, some constituents within any vitellogenin-containing sample had antigenic determinants in common, as revealed by the continuity of certain precipitin peaks, indicating a possible structural relatedness between Vg alpha, beta, and/or gamma. Plasma from untreated male quail had an antigenic determinant in common with all vitellogenin-containing plasma samples.

Animals↗
PubMed1991

Type III hypersensitivity reaction in mushroom growers.

Some respiratory symptoms in mushroom growers such as mushroom worker's lung develop by inhalation of certain agents arising from the environment of mushroom cultivation. Recently we observed mushroom workers who had respiratory symptoms which might be type III hypersensitivity reaction to the antigen of Pleurotus floridae. We gave questionnaires to all the mushroom growers at one of the biggest cultivation areas of mushrooms, Pleurotus floridae in Pocheon, Kyunggi Province. Those with respiratory symptoms were subjects for the study. CBC, chest X-ray, pulmonary function test, skin test with Pleurotus floridae extract, and precipitin antibody test to Pleurotus floridae were performed in the study subjects. Out of a total 308 mushroom workers, 23 workers (14 males, 9 females) had respiratory symptoms. Their mean age was 45 years, and their mean duration of engagement was 3.4 years. Their main symptoms were cough (100%), sputum (82.6%), dyspnea (43.5%), and fever with chills (13.0%). Two cases showed increased interstitial lung markings on chest X-ray films. Sixteen cases (73.9%) showed precipitin antibodies against P. floridae extract by counterimmunoelectrophoresis. Antibodies against Micropolyspora faeni and Thermoactinomyces vulgaris were not detected in any subject.

Adult↗
PubMed1977

Immunodiffusion, counterimmunoelectrophoresis and immunofluorescence in diagnosis of Sudan mucosal leishmaniasis.

Sera from 16 patients suffering from oral, nasal, oro-nasal or laryngeal types of Sudan mucosal leishmaniasis were tested against leishmanial antigen by three immunodiagnostic methods. Reactive soluble antigen was prepared from sonicated promastigotes. When tested against undiluted sera, in all 16 cases it gave easily visible precipitin lines in agar gel in both immunodiffusion and counterimmunoelectrophoresis tests. In immunofluorescence tests, with promastigote slide antigen and using the indirect antibody technique, the sera were positive at serum dilutions ranging from 1:200 to 1:6,400. There was no correlation between fluorescent titer levels and the strength of precipitin reactions in the immunodiffusionand counterimmunoelectrophoresis tests. Sixteen normal sera gave negative results in all three tests.

Adult↗
PubMed1980

A seroepidemiological study of amebiasis in Surinam by the enzyme-linked immunosorbent assay (ELISA).

Sera from nine populations in Surinam, South America, were screened for antibodies against Entamoeba histolytica by the enzyme-linked immunosorbent assay (ELISA) technique and a precipitin test (gel diffusion or counterimmunoelectrophoresis). In two small surveys in 1974 and 1978, a high rate of clinically significant ELISA titers was found in a rather isolated group of Amerindians living near the Brazilian border, predominately in the youngest age groups. Amerindians living near the coast showed a low level of seropositivity. Low levels were also found in Bush Negro villages in the interior of Surinam and in Hindustani school children from the savannah area. A high frequency of ELISA titers greater than or equal to 320, indicating a high rate of invasive amebiasis, was detected in persons in a mental institution in Paramaribo. It is concluded that the ELISA, which is much more sensitive than the precipitin tests, is a useful screening test for assessing the prevalence of invasive amebiasis on an epidemiological scale.

Amebiasis↗
PubMed1980

Comparison of counterelectrophoresis the enzyme-linked immunosorbent assay, and Kato fecal examination for the diagnosis of fascioliasis in infected mice and rabbits.

The enzyme-linked immunosorbent assay (ELISA), counterelectrophoresis (CEP), and Kato thick-smear stool examinations were used to follow Fasciola hepatica infections in mice and rabbits given or not given chemotherapy. In both mice and rabbits antibody levels were evident by ELISA within 2 weeks of infection, rising to high levels 2 weeks later. Precipitins monitored by CEP were also evident by 2 weeks of infection in mice, but developed more slowly (4-6 weeks) in rabbits. Eggs of F. hepatica were detected at 6-8 weeks of infection in mice and 8-10 weeks in rabbits. In mice with fascioliasis both ELISA and CEP antibody levels tended to drop by 8 weeks of infection, but in rabbits they remained strongly positive for 34 weeks. Mice treated with rafoxanide at 5 weeks of infection rapidly (3-4 weeks) became Kato-, CEP-, and ELISA-negative. Rabbits treated at 26 weeks of infection still had high antibody levels by ELISA 8 weeks later but were Kato-negative and had low precipitin (CEP) levels. ELISA was most sensitive in detecting early infection but CEP was the best indicator of chemotherapeutic success.

Animals↗
PubMed1979

Humidifier-associated extrinsic allergic alveolitis.

Three cases of allergic alveolitis due to indoor humdification systems are described. Thermoactinomyces vulgaris precipitins were detected in the serum of a 37-year-old female patient who had typical febrile attacks during exposure to cool-mist from a home humidifier. When the cause was detected and eliminated, the symptoms and signs disappeared and the woman's gas transfer factor improved from 56% to normal within six months. In a printing office a 60-year-old woman had had febrile attacks with cough for more than a year. The patient herself associated the respiratory disease with a cool-mist humidifier sometimes used at work. The water reservoir was heavily contaminated with amoebas (Amoeba proteus), which might have been the causative organisms in this case. Aspergillus fumigatus precipitins were found in the serum of a 53-year-old female printer with the clinical picture of occupational allergic alveolitis. The same organism was detected in the ambient air of the printing office.

Adult↗
PubMed1979

Susceptibility of mink to certain viral animal diseases foreign to the United States.

Mink (Mustela vison) were inoculated with viruses: African horse sickness (AHS), African swine fever (ASF), bovine herpes virus II (BHV2), foot-and-mouth disease (FMD), goat pox (GP), hog cholera (HC), peste des petits ruminants (PPR), rinderpest (RP), swine vesicular disease (SVD), vesicular exanthema of swine (VES) and vesicular stomatitis (VS). Their susceptibility was measured by development of clinical signs, virus isolation and detection of precipitin and/or virus neutralizing antibodies. SVD virus produced a lesion in one mink. Virus was isolated from mink inoculated with SVD, FMD and BHV2. Neutralizing and/or precipitin antibodies were detected in mink inoculated with ASF, FMD, GP, RP, SVD and VS viruses. Mink were not susceptible to AHS, HC, PPR and VES viruses.

African Horse Sickness↗
PubMed1983

Prevalence of serologic types of Pasteurella multocida from 57 species of birds and mammals in the United States.

The serologic types of 265 isolates of Pasteurella multocida collected from 50 species of wild birds and seven species of wild mammals over a 22-yr period were determined with the gel diffusion precipitin test. Antigens prepared from these isolates reacted with reference P. multocida antisera representing serotypes 1, 2, 3, 4, 5, 6, 7, 8, 12, and 15. Antigens from some isolates reacted slightly with antisera from more than one serotype. Overall, gel precipitin reactions involving serotype 1 (65%) and 3 (20%) were the most prevalent.

Animals↗
PubMed1992

Insects feeding on desert bighorn sheep, domestic rabbits, and Japanese quail in the Santa Rosa mountains of southern California.

Desert bighorn sheep (Ovis canadensis cremnobates), a domestic rabbit (Oryctolagus cuniculus), and Japanese quail (Coturnix japonica) were used as bait animals to collect blood-feeding flies in an area of active blue-tongue and epizootic hemorrhagic disease virus transmission. Precipitin tests were used to confirm the blood source where feasible. Eight species of Culicoides, members of the Leptoconops kerteszi group, Simulium spp., Anopheles franciscanus, and Stomoxys calcitrans were collected from the bighorn sheep. Feeding on the bighorn sheep by Culicoides brookmani (n = 25), C. variipennis (n = 6), C. cacticola (n = 1), and Simulium spp. (n = 3) was confirmed by precipitin testing. Primary species attacking the rabbit were C. brookmani, C. variipennis, and the L. kerteszi group. The quail were attacked primarily by members of the C. copiosus group and the L. kerteszi group.

Animals↗
PubMed1978

Immunoglobulin M synthesized by human lymphoblastoid cells: interaction with Staphylococcus aureus and protein A.

Immunoglobulin M synthesized by a human lymphoblastoid cell line, LA173, was found to bind specifically to the protein A-bearing Cowan I strain of Staphylococcus aureus. The (3H)-leucine-labeled, secreted IgM from these LA173 cells also formed precipitin complexes with purified protein A. Soluble complexes formed at high protein A/IgM ratios retained the ability to bind to the bacterial surface. Precipitin complexes also were observed in double diffusion Ouchterlony gels with a line of identity formed between the IgM, protein A, and anti-IgM in adjacent wells. Intracellular IgM species from detergent-lysed LA173 cells were bound to S. aureus. Labeled 19S pentamers, 8S monomers, and HL subunits were eluted from the bacteria and identified by velocity sedimentation and SDS agarose-acrylamide gel electrophoresis. In addition, several intermediates migrating between 8S and 19S were detected and shown to contain authentic H and L chains. Binding of the labeled IgM 19S pentamers to staphylococci was not inhibited by prior treatment of the bacteria with an excess of unlabeled human IgG. However, S. aureus saturated with unlabeled IgG did not bind either labeled IgM monomers or labeled IgG. The interaction of this human IgM with S. aureus exhibited a high degree of specificity with quantitative recovery of secreted 19S IgM. Intracellular IgM species were bound selectively by the bacteria with little if any contamination by other cytoplasmic proteins.

Cell Line↗