Showing papers on "Hemagglutination published in 1984"

Escherichia coli fimbriae recognizing sialyl galactosides.

Abstract: Fimbriae recognizing sialyl galactosides (S fimbriae) were purified from an Escherichia coli strain. The S fimbriae were morphologically identical to type 1 and P fimbriae of E. coli and showed a hemagglutination that was abolished when erythrocytes were treated with neuraminidase. Hemagglutination by the purified fimbriae was inhibited by orosomucoid but not by its desialylated derivative. Of the oligosaccharides tested, sialyl-(alpha 2-3)-lactose and sialyl-(alpha 2-3)-N-acetyllactosamine had the strongest inhibitory activities. It was concluded that S fimbriae have the strongest affinity for (alpha 2-3)-linked sialyl galactosides. In the enzyme-linked immunosorbent assay, the hyperimmune serum to the S fimbriae reacted strongly with the homologous antigen but not with type 1, P, or nonhemagglutinating KS71C fimbriae of E. coli. Analogously, the hyperimmune sera to the other E. coli fimbriae did not react with the purified S fimbriae. The immunoprecipitation assay showed that S fimbriae on different E. coli serotypes shared immunological cross-reactivity.

Genes of pyelonephritogenic E. coli required for digalactoside-specific agglutination of human cells.

Abstract: Most pyelonephritic Escherichia coli strains bind to digalactoside-containing glycolipids on uroepithelial cells Purified Pap pili (pili associated with pyelonephritis) show the same binding specificity A non-polar mutation early in the papA pilin gene abolishes formation of Pap pili but does not affect the degree of digalactoside-specific hemagglutination Three novel pap genes, papE , papF and papG are defined in this report The papF and papG gene products are both required for digalactoside-specific agglutination by whole bacteria cells as well as for agglutination by pilus preparations Pili prepared from a papE mutant have lost their binding ability although whole cells from this mutant retain it, implying an adhesin anchoring role for the papE gene product A mutant with lesions both in the papA and the papE genes does not mediate digalactoside-specific agglutination The implications of this finding for pilus biogenesis are discussed

Monoclonal antibodies to the S1 spike and membrane proteins of avian infectious bronchitis coronavirus strain Massachusetts M41.

Abstract: We have established four murine hybridoma cell lines which secrete specific antibody to avian infectious bronchitis virus (IBV) strain Massachusetts M41. Two monoclonal antibodies reacted with the spike protein and two reacted with the membrane protein. The specificity of the monoclonal antibodies for the external structural proteins was detected by immunoprecipitations using radiolabelled virus. The reactions of the monoclonal antibodies showed that one (S1) of the two glycopolypeptides associated with the spike protein has a strain-specific region involved in neutralization and haemagglutination, and the membrane protein has antigenic determinants which are present on the three strains of IBV tested (M41, Beaudette and D41).

The reactions of monoclonal antibodies with structural proteins of mumps virus.

Abstract: Mouse hybridomas producing antibodies against structural proteins of mumps virus were established by fusion of FO or SP 2/0 myeloma cells with spleen cells from BALB/c mice immunized with purified preparations of egg-grown mumps virus. Ascites fluids collected after i.p. inoculation of mice were characterized by different serologic tests. By immune precipitation tests with [35S]methionine-labeled mumps virus polypeptides, 17 clones were found to produce antibodies against the nucleocapsid protein (NP), 11 against the polymerase (P) protein, 10 against the membrane (M) protein, 12 against the fusion (F) protein, and 24 against the hemagglutinin-neuraminidase (HN) protein. Competitive binding enzyme-linked immunosorbent assay (ELISA) tests were performed to determine the reactivity of the monoclonal antibodies with different antigenic sites of each structural component. The monoclonal antibodies directed against the NP, P, and M proteins identified a minimum of 10, 10, and 9 separate antigenic sites, respectively. The 12 clones directed against F were directed against a minimum of eight separate antigenic determinants. These antibodies did not neutralize the infectivity of the virus either in the absence or presence of anti-gamma-globulin. Only low capacity to block hemolysis (HL) activity of the virus was detected in clones directed against three of the eight antigenic sites. Based on their serologic reactivity, the 24 clones directed against the HN protein could be divided into four groups. The first group of clones could not inhibit any biologic activity of the protein. The second group consisted of two clones that blocked HL but did not block hemagglutination (HA) or neuraminidase (NA) activity. The third group, which included five clones, blocked HA, NA, and HL activity of the virus and had high neutralizing capacity. These clones were directed against three distinct antigenic sites. Two of the clones directed against one antigenic site could block NA activity only when a large substrate, fetuin, was used, but not when a small substrate, neuraminlactose, was used in the test. The fourth group included five clones that could block NA but not HA activity of the virus. These clones could neutralize the infectivity of the virus and had high capacity to block HL activity. In blocking experiments, all these antibodies reacted with one antigenic site. The reaction of all clones was tested in ELISA with four different strains of mumps virus. Each strain had unique antigenic sites. Variations were found in four, three, and three different antigenic sites of the NP, P, and HN proteins, respectively.

Elucidation of the topography and determination of the protective epitopes on the E glycoprotein of Saint Louis encephalitis virus by passive transfer with monoclonal antibodies.

Abstract: We have identified and characterized eight antigenic epitopes on the 53,000 dalton envelope (E) glycoprotein of Saint Louis encephalitis (SLE) virus by using monoclonal antibodies. One of these epitopes (E-1c) encoded for the type-specific biologic functions of hemagglutination (HA) and neutralization (N). Injection of 50 ng of anti-E-1c antibody protected the majority of mice from peripheral challenge with 100 i.p. LD50 of SLE virus. Similar levels of protection with antibodies specific for other epitopes usually required greater than or equal to 1000-fold additional antibody. Attempts to block N or protection at the E-1c antigenic domain by using antibody to several other SLE epitopes that strongly competed for the E-1c site were unsuccessful. Enhancement of protection was observed with mixtures of the more cross-reactive antibodies. The E-1c antibody was also effective in abrogating SLE virus replication until neural invasion occurred. On the basis of these findings, the topologic arrangement and function of the eight SLE E glycoprotein epitopes on the virion spike is proposed.

Monoclonal antibodies as functional probes of the HN glycoprotein of Newcastle disease virus: antigenic separation of the hemagglutinating and neuraminidase sites.

Abstract: A panel of nine monoclonal antibodies has been used to construct a functional inhibition profile of four antibody-binding sites previously delineated on the HN glycoprotein of Newcastle disease virus. Antibodies to all four sites are capable of neutralizing infectivity and inhibiting hemolysis and neuraminidase activity with fetuin. There is a good correlation between the fractions of infectivity and neuraminidase activity which survive antibody treatment. However, the inhibition of hemolysis is in the inverse relative order of the neutralization of infectivity. Antibodies to sites 1 and 2 are capable of inhibiting hemagglutination, whereas only antibodies to site 2 can inhibit neuraminidase activity on the smaller substrate N-acetyl neuraminlactose. This antigenically separates the hemagglutinin and neuraminidase sites on the HN glycoprotein. We used these functional inhibition studies to speculate about the location of the four antigenic sites relative to the hemagglutinin and neuraminidase sites.

Hemagglutination patterns of Aeromonas spp. in relation to biotype and source.

Abstract: Aeromonas spp. show patterns of hemagglutination with human group O cells in the presence of fucose, galactose, and mannose. These patterns are related to biotype as well as to the source of isolates. There was good correlation between hemagglutination pattern and the presence of diarrhea among strains isolated in Western Australia, which was the only source with adequate data for classification of children with an without diarrhea. Most of the environmental and other nonfecal isolates produced patterns different from those in strains associated with diarrhea. These results suggest that hemagglutinins should be considered with enterotoxins as virulence factors in Aeromonas spp.

Enzyme-linked immunosorbent microassay and hemagglutination compared for detection of thyroglobulin and thyroid microsomal autoantibodies

Abstract: We have evaluated for their potential use in the routine clinical laboratory enzyme-linked immunosorbent assays (ELISA) for human thyroglobulin antibodies (hTg-Ab) and microsomal antibodies (M-Ab). Results are expressed in terms of an "ELISA Index," based on comparison with a laboratory standard. The specificity of both ELISA assays is shown by dose-dependent inhibition of the hTg-Ab and M-Ab activities of the laboratory standards by the appropriate specific antigens. Similar concentrations of ovalbumin had no significant effect on the standard activity in both assays. For consecutive samples evaluated for hTg-Ab (n = 113) and M-Ab (n = 106) by ELISA and hemagglutination, rank order analysis of the results showed a highly significant correlation between the methods (r = 0.81, p = less than 0.001 for hTg-Ab; r = 0.82, p = less than 0.001 for M-Ab). However, 8/47 (17%) of samples positive in the hTg-Ab ELISA were negative by hemagglutination, and 7/69 (12%) of samples positive in the M-Ab ELISA were negative by hemagglutination. We effectively excluded the possibility of false positivity of these specimens by ELISA by blocking specimen positivity with the specific antigens in 12 of 14 specimens investigated. We conclude that ELISA techniques for human thyroid autoantibodies are sensitive and specific, easy to initiate, objective, and capable of use in large-scale screening. They are superior to standard hemagglutination techniques by having an increased detection rate for hTg-Ab and M-Ab.

Serological cross-reactivity of porcine reference antisera to Mycoplasma hyopneumoniae, M. flocculare, M. hyorhinis and M. hyosynoviae indicated by the enzyme-linked immunosorbent assay, complement fixation and indirect hemagglutination tests.

Abstract: The enzyme-linked immunosorbent assay (ELISA) indicated significant cross-reactivity between the antigens of Mycoplasma hyopneumoniae ( HyoP ) and M. flocculare (Floc), another porcine mycoplasma of wide distribution but uncertain pathogenic significance, when porcine antisera of each specificity were tested against HyoP antigen. The titers of the anti-Floc sera ranged from threefold to 13-fold less than the titer of the anti- HyoP reference serum at different times after immunization. These values ranged from onefold less than to fourfold greater than the minimal positive titer of 80. The antisera to the other porcine mycoplasmal antigens [i.e. M. hyorhinis ( HyoR ) and M. hyosynoviae ( HyoS )] reacted less strongly to HyoP antigen but titers only slightly less than to slightly greater than the minimal positive titer were noted for some sera. Cross-reactivity was also detected by the complement fixation test, although the titers for this test were generally lower than for the ELISA, presumably reflecting lower sensitivity of the complement fixation test. Positive indirect hemagglutination titers to HyoP antigen were also observed for both anti-Floc sera obtained at one or more times during the immune response. With two exceptions (one anti- HyoR serum with a complement fixation titer of 16 and one anti- HyoR serum with an indirect hemagglutination titer of 10), none of the anti- HyoR or anti- HyoS sera had detectable indirect hemagglutination or complement fixation titers to HyoP antigen at any time after immunization. The levels of cross-reactivity detected by the complement fixation test and indirect hemagglutination and, especially, the ELISA would be of significance for the development of any practical sero-diagnostic test for mycoplasmal pneumonia of swine.

Studies of Streptococcus suis type 2 infection in pigs.

Abstract: Investigations into the immunology, pathogenesis and epidemiology of Streptococcus suis type 2 infections were carried out in experimental pigs and in naturally-occurring field outbreaks of disease. The capsular polysaccharide from Str. suis type 2 was shown to induce opsonic antibodies in pigs when injected with Freund's incomplete adjuvant, but difficulties encountered in experimental production of the disease prevented a study of their protective effects. Problems with the bactericidal tests led to an investigation of other assays for antibodies against Str. suis type 2, namely, a phagocytic test with pig neutrophils, a mixed reverse passive antiglobulin haemagglutination test and an indirect haemagglutination test. There was evidence that with modifications both the latter tests would be useful. Transmission studies in 39 conventionally-reared and 7 hysterectomy-derived, colostrum-deprived pigs yielded interesting results with regard to the distribution of the organism in relation to the disease process. Tonsil carriage in clinically-healthy pigs was demonstrated after experimental and natural infection. Detectable carrier rates varied between 0 and 59%. The organism was shown to persist in the presence of circulating opsonic antibodies and in pigs on penicillin-medicated feed. Attempts to isolate the organism from the genital tract were unsuccessful. Medicated early weaning and classical SPF techniques applied to infected herds appeared to be effective in producing pigs free from Str. suis type 2 infection.

Hepatitis-B virus infection in Dhaka, Bangladesh

Abstract: Three thousand six hundred and ten patients with acute hepatitis in two large hospitals in Dhaka city were tested for HBsAg. Besides, 780 commercial blood donors, 126 doctors and 576 apparently healthy persons were also tested. Passive haemagglutination technique was applied for this test. Patients with post-transfusion hepatitis and doctors with acute hepatitis showed the highest incidence, being 60% and 65.5% respectively. HBsAg was detected only in 15.4% of children and 27.2% of adult patients with acute hepatitis.

Oligosaccharide structures mediating agglutination of sheep erythrocytes by Staphylococcus saprophyticus.

Abstract: Agglutination of sheep erythrocytes by Staphylococcus saprophyticus was used as a model system for adherence studies. Glycolipids were isolated from sheep erythrocyte membranes, and oligosaccharides were prepared by trifluoroacetolysis. The oligosaccharides were characterized by sugar analyses, methylation analyses, gas-liquid chromatography-mass spectrometry, and nuclear magnetic resonance spectroscopy. We showed that oligosaccharides containing terminal beta-D-galactose-p-(1-4)-beta-D-2-acetamido-2-deoxyglucose-p-(1- were good inhibitors of the hemagglutination of sheep erythrocytes by S. saprophyticus.

Comparison of enzyme-linked immunosorbent and indirect hemagglutination assays for determining anthrax antibodies.

Abstract: An enzyme-linked immunosorbent assay has been established to measure anthrax antibody titers. The protective antigen component of anthrax toxin was used as the capture antigen. Two types of conjugates (protein A-horseradish peroxidase and anti-human immunoglobulin G plus immunoglobulin A plus immunoglobulin M-horseradish peroxidase) were tested. Results from enzyme-linked immunosorbent assay testing were compared with those from indirect hemagglutination titers on serum from vaccinees. The overall trend of enzyme-linked immunosorbent assay and indirect hemagglutination titers was significant. The enzyme-linked immunosorbent assay offered speed, precision, and reduced cost per test. Images

Virulence factors of Escherichia coli isolated from cows with acute mastitis.

Abstract: A total of 184 Escherichia coli isolates recovered from cows with acute mastitis were examined for recognized pathogenic mechanisms and virulence factors commonly found in pathogenic groups of E coli. A modification of the Eng procedure (for detecting complement deficiencies in serum) was used to test for resistance to different animal sera. The Sereny test (for invasiveness), infant mouse test (for heat-stable enterotoxin), and Y-1 adrenal tumor cell assay (for heat-labile enterotoxin) were used. Hemagglutination tests, using rabbit, sheep, and guinea pig RBC, were done with and without added mannose. All of the 184 isolates were serum resistant in all tested sera. None of the isolates was invasive. Only 1 isolate was positive for heat-stable enterotoxin and 2 cultures were positive for heat-labile enterotoxin. Multiple patterns of hemagglutination were observed. The majority of the isolates exhibited both mannose-sensitive and mannose-resistant hemagglutinins with guinea pig and rabbit RBC. A few strains were positive only in mannose-sensitive or mannose-resistant hemagglutination tests. A few strains were negative in all hemagglutination tests. Based on our results, E. coli from cows with acute mastitis lack the virulence factors commonly observed in other E coli groups associated with disease. Serum resistance was the only characteristic that could be related to virulence.

Antibody-mediated immune response in the bat, Pteropusgiganteus

Abstract: Among mammals, the Chiroptera (Bats) have been perhaps the least studied with respect to systematic accounts of immune responses. To resolve this omission in comparative immunology, we have begun to analyze aspects of immunity of the large frugivorous bat, Pteropus giganteus with respect to antibody secreting plaque forming cells and serum antibodies as measured by haemagglutination titres. Homologous complement was more effective than guinea pig or rabbit complement in lysing SRBC for developing plaques. Kinetic of antibody response to three different doses of SRBC has been studied. We observed a peak primary response and its decay after a single antigenic challenge which were notably delayed, and a same dose of antigen could produce 2-Mercaptoethanol resistant PFC and HA-tires.

Immunogenic properties of the small chain HA2 of the haemagglutinin of influenza viruses.

Abstract: Summary The small chain of influenza virus haemagglutinin, HA2 was isolated by a selective enzymic removal of HA1 or by preparative SDS-polyacrylamide gel electrophoresis. Anti-HA2 specific antisera and monoclonal antibodies were subtype-specific in immunodiffusion tests and radioimmunoassays. These antibodies did not inhibit haemagglutination or haemolysis, did not prevent virus release, did not neutralize infectivity, and HA2 did not induce a protective immunity. HA2-specific antigenic determinants could not be demonstrated on the surface of infected cells. Lymphocytes from pre-immunized mice could not be stimulated by HA2 to exert a cytotoxic effect.

Rapid serotyping of infectious bronchitis virus isolates with the hemagglutination-inhibition test.

Abstract: SUMMARY The hemagglutination-inhibition (HI) test was evaluated as a method of typing recent suspect infectious bronchitis virus (IBV) isolates. Hemagglutination (HA) antigen was prepared from each isolate by phospholipase C treatment of virus concentrated from allanto-amniotic fluids of infected chicken embryos. An HI test was run with the HA antigen of each isolate against a battery of 17 antisera that had been prepared against different IBV strains classified by virus neutralization (VN). The HI test identified Ark 99 and Holland isolates that were similar to strains previously classified by VN. Two isolates included in this study were not inhibited by any of the reference antisera and therefore appeared to be antigenically different. The isolates were also evaluated by VN, and the results of the VN and HI methods agreed. Therefore the results suggest that the HI test can be useful for making a rapid, presumptive identification of new IBV isolates. RESUMEN

Multiphasic interactions of Mycoplasma pulmonis with erythrocytes defined by adherence and hemagglutination.

Abstract: The mechanism(s) of interaction between Mycoplasma pulmonis and eucaryotic cells was studied by adherence to and hemagglutination of erythrocytes. Simple and complex carbohydrates and glycoproteins were unable to inhibit either adherence or hemagglutination, indicating that neither was a lectin activity. Both interactions appeared to be hydrophobic due to their requirement for salt and their sensitivity to temperature. Hemagglutination, but not adherence, was inhibited by both trypsin and glutaraldehyde treatment of the mycoplasma, suggesting that adherence and hemagglutination are qualitatively different. The erythrocyte receptor sites for the two activities were also separable since hemagglutination, but not adherence, required trypsinization of erythrocytes. The hemagglutinin was shown to be an integral mycoplasma component and not a broth contaminant. Once removed, hemagglutinating activity could not be replenished by incubation in serum or broth at 4 degrees C, but could be regenerated during protein synthesis under nonreplicative conditions. Thus, a mycoplasma membrane protein was detected which was capable of interacting with opposing membrane surfaces through hydrophobic interactions. Consequently, a multiphasic model of M. pulmonis-eucaryotic cell interactions was proposed.

Biologic and Immunologic Characterization of the Soluble Skin-Test Antigen of Varicella-Zoster Virus

Abstract: A sensitive method for assay of the potency of the skin-test antigen of varicella-zoster virus was established. Potency was determined by a reverse passive hemagglutination test, and the results correlated well with the cutaneous reaction of sensitized guinea pigs and an immune human adult to the antigen. Cutaneous reactivity was not induced by repeated inoculation of the antigen, but humoral neutralizing antibody was detected in most guinea pigs when a large amount of antigen was used. No cutaneous cross-reactivity between varicella-zoster virus and herpes simplex virus was noted; however, positive cutaneous reactions and neutralizing antibody responses to homologous virus were detected in infected guinea pigs. These results suggest that both skin tests are specific and support the clinical usefulness of the varicella-zoster skin test.

Comparison of enzyme-linked immunosorbent assay, DNA hybridization, hemagglutination, and electron microscopy for detection of canine parvovirus infections.

Abstract: Canine fecal samples were analyzed by enzyme-linked immunosorbent assays (ELISA) by using monoclonal antibodies to the canine parvovirus hemagglutinating protein. These data were compared with results obtained with DNA hybridization assays, hemagglutination assays, and electron microscopy. The highest correlation was observed between the ELISA and the hemagglutination tests, with 94.4% of samples showing agreement. Lower correlation was obtained between ELISA and DNA hybridization tests (73.3%). Correlation between ELISA and electron microscopy was 60.9%. The studies indicated that the ELISA can be used as a sensitive and specific diagnostic assay for canine parvovirus infections.

Autoantibodies in sera of Thai patients with Plasmodium falciparum infection.

Abstract: Serum from 98 Thai adults infected with Plasmodium falciparum were examined for the presence of autoantibodies. Malarial containing antibodies sera were also revealed positive for anti-nuclear antibodies with fluorescence speckled pattern, anti-smooth muscle antibodies, anti-mitochondria antibodies and rheumatoid factor. There was no detectable antibody to double stranded DNA. There was a significant relationship between high titre of malarial antibody and high frequency of speckled pattern of anti-nuclear factor or anti-nuclear antibodies. By the ELISA technique determination of serum antibodies against an extractable nuclear antigen (ENA) in patients with P. falciparum infection gave 43.8% (43 of 98) positive result. In addition, sera contained malarial antibodies gave positive antibodies to ENA in 50% (49 of 98) by tanned red cell haemagglutination. Among the positive sera with antibodies to ENA, they showed the presence of antibodies to both ribonucleoprotein RNAase sensitive (RNP) and ribonucleoprotein RNAase resistance (Sm). Also both of antibodies exhibited positive staining of speckled pattern of antinuclear factor. This observation indicated that malaria infection induces autoantibodies which were predominantly anti-nuclear antibodies.

The role of glycosylation on haemagglutination and immunological reactivity of rubella virus.

Abstract: Treatment of purified rubella virus with mixed glycosidases resulted in loss of haemagglutinating (HA) activity but the capacity to combine with haemagglutination inhibiting (HI) and other antibodies was retained Electrophoretic analysis revealed a greater effect on VPI than VPII

F41 antigen among porcine enterotoxigenic Escherichia coli strains lacking K88, K99, and 987P pili.

Abstract: Colonial morphological variations among enterotoxigenic Escherichia coli which lack K88, K99, and 987P (3P-) were shown to be correlated with expression of several surface antigens, i.e., type 1 pili, F41 pili, and somatic and capsular antigens. Such correlations were established by electron microscopy, serology, and hemagglutination of cells derived from these specific colonial types. Identification of F41 produced by the 3P- enterotoxigenic E. coli strains was made by immunodiffusion in agarose gel, immunofluorescence microscopy, subunit molecular weight determination in sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and hemagglutination of F41-piliated 3P- cells and purified F41 pili. Two of the 3P- enterotoxigenic E. coli strains were also shown to be virulent in conventional neonatal pigs and calves. Intense immunofluorescence staining by reference F41 serum of ileal villi of 3P- -infected animals indicated that F41 was expressed during the disease process.

Immunological characterisation of rubella virion polypeptides

Abstract: SUMMARY. Antibodies specific for rubella virion polypeptide, VPI, secreted by clones of hybridoma cells or produced in rabbits in response to specific antigenic stimulation, located determinants for haemagglutination inhibiting (HI) and neutralising (Nt) antibodies on this envelope component.