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PubMed1979

Human prostatic acid phosphatases: purification of a minor enzyme and comparisons of the enzymes.

The minor enzyme of human prostatic acid phosphatases (pI 5.5) with high specific activity (orthophosphoric monoester phosphohydrolase, acid optimum, EC 3.1.3.2) has been purified for the first time as a pure enzyme protein. The enzyme was a single protein when examined by polyacrylamide gel electrophoresis and isotachophoresis. The specific activity was 1080 micromole per (min X mg) for hydrolysis of 5.5 mmole per liter of p-nitrophenylphosphate at pH 4.8 and 37 C. The purification coefficient was 540 and the recovery of enzyme activity was 2 per cent. The molecular weight of the enzyme subunit when measured by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate was 54,000. The Km of the purified enzyme was 3 X 10(-4) mole per liter for p-nitrophenylphosphate. An antiserum to this enzyme was prepared. The enzyme was cross-reactive with the main enzyme (pI 4.9) of human prostatic acid phosphatases in immunoelectrophoresis. No precipitin arc with the acid phosphatase in the serum of a prostatic carcinoma patient could be shown. Antiserum to the main enzyme caused a precipitin line with the same serum sample.

Acid Phosphatase↗
PubMed1973

Host-feeding patterns and behaviour of 4 Culex species in an endemic area of Japanese encephalitis.

Studies were made on the host-feeding patterns and behaviour of 4 Culex species in China (Province of Taiwan) to further assess their importance as vectors of Japanese encephalitis. A relatively unbiased, non-attractant technique (vacuum sweep-net) was used for collecting mosquitos resting outdoors on rural premises; blood meals from engorged specimens were identified by precipitin tests. Additional information was obtained by analysing data from related studies. Precipitin-test results indicated that C. annulus, C. tritaeniorhynchus, and C. fuscocephalus feed principally on mammals, the first species mainly on swine and the latter two largely on bovine hosts. Mosquito forage ratios suggested that the apparent preference of C. annulus for swine over bovine hosts might be a function of host density. C. annulus was found to be zoophilic, exophilic, and exophagic; however, in view of its abundance during the JE epidemic season, contact with human hosts may reach a significant level, even indoors. The other 3 species of Culex were considered to be of lesser importance as JE vectors during the 1971 epidemic season because of their host-feeding patterns and/or population densities.

Animals↗
PubMed1979

Protective immune response in mice immunized with antigens from Trypanosoma gambiense-infected mouse blood.

Immunogenicity and property of antigens obtained from Trypanosoma gambiense-infected mouse blood (IMP) were examined. A strong vaccine effect against intravenous challenges with 3 x10(3) parasites given on study day 3, 5, or 14 (day 0 = immunization) was observed in mice immunized with a combination of IMP (2 mg protein/mouse) and Freund's complete adjuvant (FCA). But when the challenge was given on day 21 or 30, per cent survival in mice dropped to the 20- and 40-per cent level, respectively. Among fractioned components of IMP, IMP-1, IMP-2, and imp-3, by gel filtration with Sephadex G-200, all of the mice immunized with IMP-1 antigen alone or together with FCA and challenged on day 5 were able to conquer intraperitoneal challenges with 1 x10(2) parasites. Mice immunized with IMP-2 or IMP-3 died within 6 days after challenge. Moreover, protection efficacy shown by IMP-1p (144,000 xg sediment of IMP-1) antigen in mice was similar to that by IMP and IMP-1 antigens. IMP-3 yielded a single precipitin line against mouse anti-IMP serum by Ouchterlony double diffusion method but this response was eliminated when the antiserum was absorbed by IMP-1p. No precipitin line was identified between mouse anti-IMP serum and IMP-1 or IMP-2. From electron microscopic observations, elements of IMP-1 and IMP-1p are possibly corresponded to the fragments of filopodia of the parasites.

Agglutination Tests↗
PubMed1975

Immunochemical analysis of a Smith-like antigen isolated from two human strains of Staphylococcus aureus.

A surface antigen consisting of aminoglucuronic acid and N-acetyl-L-alanine was isolated from the culture filtrates of two human strains of Staphylococcus aureus. Double diffusion analysis in agar suggested that the antigen is immunologically similar to the alanyl-aminoglucuronic acid capsule of the Smith strain of S. aureus. Quantitative precipitin inhibition studies indicated that N-acetyl-L-alanine is the immunodominant determinant of the acidic antigen. In addition, conjugates consisting of N-acetyl-L-alanine coupled to bovine serum albumin gave a significant precipitin reaction with anti-staphylococcal serum which is rich in alanyl-aminoglucuronic acid polymer antibodies. Antibodies with N-acetyl-L-alanine specificity were isolated from N-acetyl-L-alanine-Sepharose immunoabsorbent columns. Double diffusion analysis in agar indicated that the eluted antibodies were serologically reactive and belonged to the IgG class of immunoglobulins.

Absorption↗
PubMed1975

The heterogeneity of anti-DNA antibodies in systemic lupus erythematosus and other diseases.

The antigenic specificities of antinucleic acid antibodies occurring in systemic lupus erythematosus (SLE), chronic active liver disease, and progressive systemic sclerosis (PSS) have been studied by means of haptenic nucleosides and nucleotides coupled to human serum albumin. SLE sera were also tested with dinucleotides. SLE and chronic active liver disease sera showed marked heterogeneity, producing precipitin lines with nucleosides or nucleotides, or both. The reaction might occur with a nucleoside and not with the corresponding nucleotide, or vice versa. The SLE sera reacted to dinucleotides with marked specificity, being able to recognize base sequences or to react with a dinucleotide despite the absence of a reaction with the individual bases. All sera from patients with PSS showed precipitins with RNA, uridine and UMP. PSS sera which reacted with a nucleoside also reacted with the corresponding nucleotide. Antibodies to DNA were found in a smaller proportion of PSS sera than in sera from SLE or chronic active liver disease. Their presence was confirmed by reactivity with thymidine and TMP.

Antibodies, Anti-Idiotypic↗
PubMed1975

[Studies on the production of anti-human-lymphocyte serum (ALS) in bulls].

The applicability of bulls as productive animals was considered for the preparation of anti-humans ALS. The course of immunologic response was studied by lymphoagglutination, lymphocytotoxicity, rosette inhibition, hemagglutination tests and by precipitin formation in two experimental groups immunized by different amounts of lymphocytes from peripheral blood of normal donors. The animals were found to respond well already after the second application of very small amounts of antigen (on day 0-4 times 10(7), on day 21-2 times 10(8) lymphocytes). They showed lymphoagglutination titre 1 : 512-2000, lymphocytotoxic titre being higher than 1 : 4000 and the rosette inhibition test gave a minimum titre of 1 : 65000. On the other hand, further application of a high amount of antigen (2 times 10(9), or 4 times 10(9) lymphocytes) did not lead to further increase in the titre; on the contrary - hyperimmunization resulted in a lower titre in the case of the rosette inhibition test, which is known to correlate best with the in vivo immunosuppressive activity. The hemagglutinin titre was also acceptable under the above conditions and the formation of undersirable precipitins against human serum proteins was negligible. Good response reached by a simple and economical immunization scheme speaks for the suitability of bulls for the production of ALS.

Animals↗
PubMed1975

Immunochemical studies of lipids. IV. Chemical modification of Forssman globoside and immunological activities.

N-deacylated and N-deacetylated Forssman globoside obtained from Forssman globoside by partial alkaline hydrolysis had no Forssman activity. After re-N-acetylation of the N-deacylated and N-deacetylated Forssman globoside, the N-acetylsphingosyl Forssman oligosaccharide thus obtained recoverd its activity. Moreover, after ozonolysis and reduction of the N-acetylsphingosyl Forssman oligosaccharide, the 2-N-acetoamido-1,3,4-trihydroxybutanoyl Forssman oligosaccharide thus obtained still had a haptenic activity, but couldn't show a precipitin reaction with Forssman antibody. On the other hand, the Forssman globoside and the N-acetylsphingosyl Forssman oligosaccharide gave rise to precipitin line on agar gel double diffusion, it was thus concluded that the aqueous solution of these substances formed spherical micelles to behave like a fairly macromolecular multivalent antigen.

Acetylation↗
PubMed1978

Occupational hypersensitivity lung disease.

A case of occupational hypersensitivity pneumonitis caused by an air conditioning unit is presented. Aspergillus fumigatus and aureobasidium pullulans were cultured from this unit and antibodies to these organisms were found the patient's blood. The case shows that this disease may present in a very subtle form and may be diagnosed by a culture from the patient's environment and an easily performed blood precipitin test. The importance of searching for this type of lung disease and the need for obtaining precipitin tests are stressed.

Air Conditioning↗
PubMed1977

Studies on the feeding patterns of the sheep headfly Hydrotaea irritans (Diptera, Muscidae) in Great Britain.

Serum-precipitin tests carried out on the stomach contents of laboratory maintained and field-caught Hydrotaea irritans showed that a positive result could be obtained for at least seven to 10 days after feeding. Eye, nasal and salivary secretions, even if undiluted, did not give a precipitin response in these tests. Field results from over 3000 flies thus indicate that 20 to 55% of the active fly population had taken a blood meal within the previous 10 days: 75 to 100% of these had fed on cattle blood and 15% or less on sheep blood. Significantly lower feeding levels were obtained from flies collected within plantations, indicating a protracted resting phase in woodland. From "fly feeding areas" high blood-feeding levels were shown from early July, as ovarian development begins. Early feeding rates were similar in both sexes.

Animals↗
PubMed1980

Human granulocyte-specific nuclear antigen(s). II. Detection of antigens in human proliferative syndromes.

Antisera raised to dehistonized chromatin from isolated normal human granulocytes revealed the presence of chromatin-associated antigens specific for the human neutrophils that appear during late stages of myeloid cellular differentiation. Immunological specificity was demonstrated by C fixation, immunodiffusion, and immunocytochemical reactions. Chromatin prepared from both normal granulocytes and specimens of myeloid leukemia showed immunologic reactivity. Although the normal antigens were detected in a specimen of CML, the position of immunodiffusion precipitin lines was different from that obtained with normal granulocyte chromatin. In addition, chromatin prepared from the myeloid leukemic cell line HL-60 expressed only one of the three precipitin bands normally found in immunodiffusion. The immunocytochemical staining reaction was confined to the nucleus of mature neutrophils in normal peripheral blood smears. Greater than 90% of cells in peripheral blood specimens of CML showed positive immunocytochemical nuclear staining. In other types of leukaemia, the normal mature granulocyte reacted with antiserum, but the nonmyeloid leukemic cells in these specimens did not. The specificity of immunologic reactions described here suggests the usefulness of nuclear antigens as cell markers.

Animals↗
PubMed1980

Purified Candida albicans proteinase in the serological diagnosis of systemic candidosis.

An inducible, exocellular proteinase enzyme, purified from culture filtrates of Candida albicans, was tested as an antigen for the detection of serum anti-Candida precipitins. The purified proteinase was a more specific antigen for the serological diagnosis of systemic candidosis than traditional cytoplasmic extracts of the fungus. Precipitin titers to purified proteinase exceeded 1:4 in 75% of serum samples from 12 patients with confirmed systemic Candida infection, in 23% of serum samples from 22 patients positive for Candida anticytoplasm antibodies but without corroborative diagnostic evidence of systemic candidosis, and in none of 28 serum samples from patients without either candidosis or anticytoplasm antibodies.

Antigens, Fungal↗
PubMed1982

Demonstration of two distinct antigenic determinants on hepatitis B e antigen by monoclonal antibodies.

Mice were immunized against hepatitis B e antigen (HBeAg) isolated from sera of asymptomatic carriers of hepatitis B virus. Their spleen cells were fused with mouse myeloma (NS-1) cells, and 5 clones of hybridoma cells secreting antibody against HBeAg (anti-HBe) were isolated. For the production of anti-HBe in large scale, cells were cultivated both in vitro and in the peritoneal cavity of ascitic mice. Although monoclonal antibodies produced by these clones showed a strong reactivity of anti-HBe in hemagglutination tests, individual monoclonal anti-HBe did not reveal any precipitin line in immunodiffusion. When 2 of the 5 monoclonal antibodies were mixed together, however, some combinations showed a precipitin line against HBeAg, whereas others did not. Utilizing solid-phase radioimmunoassay involving a number of combinations of monoclonal antibodies used for solid-phase and radiolabeling, the 5 antibodies were classified into 2 groups. Three of the anti-HBe antibodies were found to be directed to 1 determinant of HBeAg (determinant a); the remaining 2 to the other determinant (determinant b). Determinants a and b were detected on HBeAg in the serum, as well as on the polypeptide of 19,000 daltons (P19) derived from the nucleocapsid of hepatitis B virus. Monoclonal anti-HBe antibodies with different specificities may provide useful tools in delineating the antigenic structure of HBeAg and also in evaluating immune responses of the host directed to its subdeterminants.

Animals↗
PubMed1983

Antigens/allergens of Aspergillus fumigatus. Identification of antigenic components reacting with both IgG and IgE antibodies of patients with allergic bronchopulmonary aspergillosis.

Evidence that both IgE and IgG antibodies present in the sera of allergic bronchopulmonary aspergillosis patients (ABPA) combine with several of the antigenic components of Aspergillus fumigatus has been obtained using a self-crossed radioimmunoelectrophoresis test. In this test, patient's serum was used to develop its own crossed immunoprecipitation pattern to a reference antigen before proceeding to autoradiography with 125I-anti-human IgE in the usual way. Comparison of autoradiograph with immunoprecipitate showed a varied pattern of reactivity, i.e. precipitin peaks with and without associated radioactivity, as well as some peaks appearing only in the autoradiograph. Control sera, including sera from A. fumigatus skin test positive patients and a skin test negative aspergilloma patient with multiple precipitins showed no such combined activity. The specificity of the test was further demonstrated using an ABPA serum pool, both by absorbing the IgE from the serum before incorporation in the gel and also by heat inactivation (56 degrees C, 4 h) of the serum to destroy the heat labile determinants on IgE molecules, thus preventing the uptake of the anti-IgE specific for these determinants.

Allergens↗
PubMed1980

Investigations on live vaccines against infectious bursal disease of chicks.

Four live virus vaccines against Infectious Bursal Disease (IBD) were studied with regard to their safety, immune response and applicability. None of the vaccines caused clinical symptoms or had an adverse impact on bodyweight. Differences between these vaccines were observed in their effect on the Bursa/Bodyweight Ratio and the severity of the microscopical lesions of the bursa Fabricii. The immunosuppressive effect of IBD vaccination at one day of age on the response to Newcastle disease vaccine applied was rather low. Three of the four vaccines induced antibodies associated with protection against challenge. Vaccination of SPF rearing chickens by drinking water at an age of 15 weeks produced an antibody response (Agar Gel Precipitin Test) whereas at 15 weeks produced an antibody response (Agar Gel Precipitin Test) whereas at an age of 23, 32 and 60 weeks it did not. Chickens of all age groups responded serologically to an intramuscular vaccination. A correlation was found between the immunological response and the effect of the vaccines on the bursa Fabricii.

Animals↗
PubMed1983

Non-A, non-B hepatitis: positive and negative considerations on 138 cases.

In 138 patients with NANB H no correlations were observed regarding types of exposure (PTH: 19.5%, DA: 18.1%, Sporadic cases: 62.4%), age groups and periods of incubation. Precipitin lines were detected assaying by ID and/or CEP onset and recovery sera with heterologous reference sera positive for Ab or Ag, respectively, or with autologous sera; "Ag"' was present during acute phase of illness, "Ab" appeared with decrease of aminotransferases, and in most cases seroconversion was observed in accord with clinical course. In few patients double precipitin lines were evidenced (heterogeneity of Ag?), while one patient reacted with all reference sera and many others with two/three sera (two NANB viruses or two Ag/Ab systems?). Tendency towards chronicity emerged in four cases, and in these Ag/Ab systems had a different behaviour.

Antibodies, Viral↗
PubMed1984

Urinary cholesterol: its association with a macromolecular protein-lipid complex.

The cholesterol-containing complexes in the urine of normal subjects and patients with diseases accompanied by hyperexcretion of urinary cholesterol were characterized. In normal subjects, the major portion of the recovered urinary cholesterol was eluted in the void volume fractions after gel chromatography on Bio-Gel A-5m; this suggested an association with a macromolecular complex above 5 X 10(6) daltons. A comparable elution pattern was seen in most of the urines of the patients with benign or malignant diseases of the kidneys or the urogenital tract. However, in single patients with hyperexcretion of urinary cholesterol, considerable amounts of cholesterol were detected in the included volume of the column. This was caused by additional excretion of high density lipoproteins or both high and low density lipoproteins in the urine which could be identified in these fractions by agarose electrophoresis and immunodiffusion. These results indicate that the macromolecular complex represents the majority of the recovered urinary cholesterol in normal subjects and in disease states with known hyperexcretion. Macroscopically, the isolated cholesterol-containing complex in the void volume fractions was turbid, and electron microscopy showed lipoprotein-like particles with diameters ranging from 300 to 700 A. The chemical analysis revealed median values of protein (46.0%), triglycerides (16.3%), cholesterol (8.2%), and phospholipids (29.5%) in normal subjects and comparable results in the patients with benign or malignant diseases of the kidney and the urogenital tract. Ethanolamine glycerophospholipids, phosphatidylcholine, sphingomyelin, and phosphatidylserine were the main phospholipid components. After ultracentrifugation in a CsCl gradient, the cholesterol-containing complex was found between densities 1.1 and 1.3 g/ml. By SDS polyacrylamide electrophoresis, up to 17 protein subunits in the molecular weight range of 14,000 to 87,500 were separated. Immunodiffusion studies showed in about 40% precipitin lines against anti-human albumin, but no reactions against anti-human apoHDL and anti-human apoLDL. However, immunodiffusion of the macromolecular complex against anti-liver-specific and anti-kidney-specific lipoproteins revealed single precipitin lines. In conclusion, the isolated cholesterol-containing urinary complex showed many characteristics of membrane-associated protein-lipid particles of the human kidney and even the liver. These proteolipids are the major source of urinary cholesterol in normal and disease states.(ABSTRACT TRUNCATED AT 400 WORDS)

Adolescent↗
PubMed1984

[Immunological specificity and cytoplasmic location of delta 6-desaturase in microsomal membrane].

The enzymatic properties of the three types of microsomal acyl-CoA desaturases, delta 6-, delta 9- and delta 5-desaturase, were immunologically compared using a monospecific antibody raised against the purified linoleoyl-CoA desaturase (delta 6-desaturase). By the double immunodiffusion technique, the anti-delta 6-desaturase antibody showed a single precipitin line to the purified delta 6-desaturase and microsomes treated with Triton X-100, but no line was observed with the partially purified delta 9-desaturase. The antibody even inhibited definitely delta 6-desaturase activity in microsomes, but neither stearoyl-CoA (delta 9-) nor eicosatrienoic acid (delta 5-) desaturations were inhibited. By these immunological investigations it was confirmed that terminal delta 6-desaturase is different enzyme from desaturases delta 9- and delta 5. The intramembrane localization of delta 6-desaturase in rat liver microsomes was examined by various methods, such as digestion by proteases, effect of detergents and inhibition by the antibodies against purified terminal desaturase. Exposure of the desaturase on the surface of microsomal vesicles was suggested by the fact that the enzyme activity in the intact microsomes was susceptible to tryptic digestion and considerably inhibited by anti-desaturase antibodies. When microsomes were previously treated with trypsin, the enzyme became more susceptible to the antibodies. Furthermore, it was demonstrated that the protein fragments cleaved from microsomal membranes by tryptic digestion formed a single precipitin line with the antibodies by the double immunodiffusion test. These findings suggest the presence of delta 6-desaturase on the cytoplasmic surface in the endoplasmic reticulum, since tryptic digestion liberates only the protein components situated on the surface area of membranes. In addition, desaturase activity in the intact microsomes was not stimulated by addition of the detergent, indicating the further outside location of the active site of the enzyme in microsomal vesicles. The previous exposure of microsomes to a low concentration (0.05%) of sodium deoxycholate, which destroys the permeability barrier for macromolecules whichout membrane disassembly, did not increase the susceptibility to tryptic digestion and the antibodies. These results show that delta 6-desaturase is not present in a latent state in the membrane.

Animals↗
PubMedDate not supplied

Comparisons of serologic responses of white Leghorn and New Hampshire red chickens to purified lipopolysaccharides of Pasteurella multocida.

White leghorn and New Hampshire red chickens were inoculated with purified lipopolysaccharides of 14 serotypes of Pasteurella multocida to determine their ability to produce serotype-specific antisera for somatic antigen typing. Specific antisera were made by both breeds of chicken to lipopolysaccharides of serotypes 1, 3, 4, 6, 8, and 16. No specific antisera were made against lipopolysaccharides of serotypes 2, 5, 7, 12, and 14. Lipopolysaccharides of serotypes 10 and 11 failed to stimulate antibody production. White leghorns were more responsive than New Hampshire red chickens. White leghorn antisera had higher titers to lipopolysaccharides in passive hemagglutination tests and produced more intense precipitin reactions with heat-stable antigens in the gel-diffusion-precipitin test.

Animals↗