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PubMed1984

Effect of anti-thrombospondin antibodies on the hemagglutination activities of the endogenous platelet lectin and thrombospondin.

The proposal that thrombospondin is the endogenous platelet lectin was evaluated using antisera and monoclonal antibodies to thrombospondin. The platelet-bound hemagglutinin activity of human platelets stimulated with A23187 was inhibited by rabbit anti-thrombospondin sera and by a monoclonal anti-thrombospondin IgG. A second monoclonal IgG did not inhibit platelet-bound agglutinin activity. Preparations of purified platelet thrombospondin differed in their hemagglutination activities. The hemagglutination activity of an active preparation of thrombospondin was inhibited by the monoclonal antibody that inhibited platelet-bound lectin activity. The hemagglutination activity of an almost inactive preparation of thrombospondin was enhanced by the anti-thrombospondin monoclonal antibody that did not block platelet-bound lectin activity. The results demonstrate that expression of the platelet-bound form of the endogenous lectin is thrombospondin-dependent and suggest that thrombospondin must become part of a larger complex, either by binding to the platelet surface or by becoming aggregated in solution, before hemagglutination activity can be expressed.

Antibodies, Monoclonal↗

PubMed1984

Hemagglutination by human rotavirus strains.

Human rotavirus isolates, KUN , MO, and Wa strains were found to agglutinate erythrocytes of the day-old chicken and adult goose, optimally at pH 6.6. Only those fractions containing double-shelled rotavirus particles isolated by isopycnic centrifugation in cesium chloride had hemagglutinating activity. Trypsin treatment decreased the hemagglutination titer of human rotavirus strains and conversely increased their infectivity. Hemagglutination inhibition tests with antisera against type-specific antigens demonstrable by neutralization showed no type specificity.

Animals↗

PubMed1985

Hemagglutinating activity associated with bovine herpesvirus type 1.

Using C57BL/HPB mouse erythrocytes, hemagglutination has been observed with the Los Angeles and Colorado-1 strains of bovine herpesvirus type 1 and with 12 other Canadian field isolates as well. The specificity of the hemagglutination observed with the viral strains has been confirmed by a hemagglutination-inhibition assay.

Animals↗

PubMed1988

Hemagglutination by Staphylococcus saprophyticus and other coagulase-negative staphylococci.

Hemagglutination tests were performed to specify surface lectins (hemagglutinins) of four coagulase-negative staphylococcal species: S. saprophyticus (31 strains), S. epidermidis (5 strains), S. haemolyticus (3 strains), and S. warneri (3 strains). All strains of S. saprophyticus agglutinated sheep red blood cells (RBC) and the hemagglutination was inhibited by N-acetylglucosamine (GlcNAc) plus either N-acetylgalactosamine (GalNAc, 15 strains) or N-acetylneuraminic acid (NANA, 16 strains). Those strains showing inhibition by GalNAc also agglutinated horse RBC while those inhibited by NANA agglutinated rabbit RBC. The former type was more common among urinary tract isolates (10/15) and the second one among respiratory isolates (9/14). The eleven strains of other staphylococci agglutinated rabbit (and not sheep or horse) RBC; this hemagglutination was never inhibited by GlcNAc but instead by NANA alone or together with another sugar (7 strains) or by other sugars (4 strains, 3 different patterns of inhibition).

Animals↗

PubMed1961

The specificity of the passive hemagglutination methods used in serology of tuberculosis.

The Middlebrook-Dubos hemagglutination test with normal red cells and the Boyden hemagglutination test with red cells tanned with tannic acid were shown to be strictly specific, for tubercle polysaccharide and for the protein antigens. The former test detects polysaccharide antibodies and the latter protein antibodies. Convincing evidence is given that polysaccharide does not adhere to the surface of tanned red cells nor protein to the normal red cell surface. Hemosensitizing ability and in vitro antigenicity were found to be two distinct properties of tubercle polysaccharide and protein antigens. The phosphatide hemagglutination test with normal red cells sensitized with tubercle phosphatide is also specific; it detects only phosphatide antibodies. The three different kinds of antibodies, antipolysaccharide, antiprotein, and antiphosphatide, were shown to be completely distinct from each other in tuberculous serum. Each of them can be completely removed from serum by absorption with red cells or kaolin particles coated with its corresponding antigen. The reliability of the cross-inhibition test is discussed.

Antibodies↗

PubMed1963

PRESERVATION OF ANTIGEN-COATED SHEEP ERYTHROCYTES BY FREEZING FOR USE IN INDIRECT HEMAGGLUTINATION PROCEDURE.

Hubert, Earl G. (Veterans Administration Center, Los Angeles, Calif.), George M. Kalmanson, and Lucien B. Guze. Preservation of antigen-coated sheep erythrocytes by freezing for use in indirect hemagglutination procedure. J. Bacteriol. 86:569-572. 1963.-Preservation of sheep erythrocytes, coated with various protein and lipopolysaccharide antigens, was accomplished by rapid freezing and storage at -70 C in a dextrose-lactose solution. The stored cells were tested at intervals in an indirect hemagglutination procedure. Sensitivity of these cells was maintained, and antibody titers were consistently reproducible throughout the period of testing. These observations suggest that sheep erythrocytes may be coated with a variety of antigens and may be employed satisfactorily in an indirect hemagglutination test, even after storage at -70 C for a period up to 6 months.

Animals↗

PubMed2004

Hemagglutination of preoperative blood donation in patients with rheumatoid arthritis.

We gave preoperative blood transfusions to 37 patients with rheumatoid arthritis (RA) and 35 patients with osteoarthritis (OA), including some whose baseline hemoglobin level was less than 10 g/dl. Transfusion packs can preserve whole blood containing citrate phosphate dextrose (CPD) for 3 weeks. The baseline hemoglobin level of RA cases was 10.4 g/dl (range 8.4-13.1 g/dl), and that of OA cases was 11.9 g/dl (range 10.4-15.0 g/dl). By collecting 200-400 g every week before the operation, the total was 800-1200 g. Erythropoietin was given to patients intramuscularly when their hemoglobin was less than 13 g/dl after blood had been collected. Hemagglutination, with diameters of more than 1 cm, made filter occlusions in 11 RA cases (30%) and one OA case (3%) (P << 0.0031) after retransfusion. There were no differences between hemagglutination patients (agglutination group) and nonhemagglutination patients (nonagglutination group) regarding baseline C-reactive protein (CRP), white blood cells, platelets, or fibrinogen. We could not predict the formation of macrohemagglutination in the packs collected during the clinical course. In RA cases, allogenic transfusions were performed for four cases (36%) in the agglutination group and for one case (12%) in the nonagglutination group. Preoperative transfusion for the RA patients showed hemagglutination in some cases, and highlighted the need for modifications to reduce these hemagglutinations.

Journal Article↗

PubMed1997

Efficient gene transfer method into the whole heart through the coronary artery with hemagglutinating virus of Japan liposome.

OBJECTIVE: To confirm gene transfer techniques especially into the whole heart, we tried out a gene transfer method involving liposome with the viral envelope hemagglutinating virus of Japan liposome as an alternative to existing techniques such as cationic lipofection or other viral vectors. METHOD: For this study, hemagglutinating virus of Japan liposome (H group) or cationic liposome (L group) was used to compare the efficacy of gene transfection of oligonucleotide labeled with fluorescein isothiocyanate and cDNA of beta-galactosidase and human manganese-superoxide dismutase. Fluorescein-labeled oligonucleotide, cDNA of beta-galactosidase, or manganese-superoxide dismutase was complexed with liposomes, DNA-binding nuclear protein, and the viral protein coat of hemagglutinating virus of Japan. After donor rat hearts arrested by cardioplegia had been harvested, the coronary artery during cardioplegic arrest was infused via an aortic cannula with the liposome-gene complex. Next, the hearts were transplanted into the abdomen of recipient rats of the same strain, and all recipients were put to death after 3 days of transfection. RESULTS: Fluorescein isothiocyanate was detected in the nuclei of more than 70% of the myocytes (75% +/- 14%, n = 5) in the H group compared with fewer than 10% in the L group (7% +/- 5%, n = 5). The intensity of fluorescein isothiocyanate was significantly higher in the H group (979 +/- 112 FI) than in the L group (116 +/- 68 FI). beta-Galactosidase was expressed in the cytosol of more than 50% of the myocytes in the H group (61% +/- 7%, n = 5) compared with none in the L group (0%, n = 5). After 3 days of gene transfection, and when exposed to ischemia (30 minutes, 37 degrees C) and reperfusion (30 minutes, 37 degrees C) with Langendorff apparatus, the hearts transfected with manganese-superoxide dismutase (S group, n = 5) showed a significantly higher percentage of recovery of left ventricular end-diastolic pressure (S vs C, 86% +/- 3% vs 54% +/- 12%) and coronary flow (98% +/- 2% vs 66% +/- 12%) than did the control hearts (C group, n = 5). Western blotting analysis showed an apparent increased expression of manganese-superoxide dismutase in the hearts transfected with manganese-superoxide dismutase compared with the control hearts. These results clearly demonstrated that the donor hearts were transfected with fluorescein-labeled oligonucleotide and the beta-galactosidase gene as a result of coronary infusion of the hemagglutinating virus of Japan liposome during cardioplegic arrest at the time of harvest. Furthermore, the hearts transfected with manganese-superoxide dismutase showed significant improvement in tolerance against ischemia reperfusion injury. CONCLUSION: We believe that this method represents a novel in vivo gene transfer technique for the heart and thus may provide a new tool for research and therapy of heart transplantation.

Animals↗

PubMed1985

Characterization of Lotus tetragonolobus fucolectin components for differences in hemagglutinating and macrophage activating activities.

The fucose binding proteins (FBP) extracted from Lotus tetragonolobus seeds were isolated by affinity chromatography and compared with affinity purified commercial preparations for physical, antigenic, and biological properties. All preparations contained three protein components as determined by polyacrylamide gel electrophoresis, each with a subunit molecular weight of approximately 27 Kd. FBP preparations were also found to be antigenically identical by immunodiffusion analysis and possessed similar biological activities for hemagglutination of group 0 erythrocytes and macrophage activation in the migration inhibition assay. A reversible temperature dependent hemagglutination characteristic was found; FBP agglutinated erythrocytes at 4 degrees and 22 degrees C but not at 37 degrees C, which was reversed by decreasing the incubation temperature from 37 degrees C to 22 degrees C. Temperature dependent binding of FBP for macrophages was also demonstrated. Adsorption of crude FBP by group 0 erythrocytes preferentially removed hemagglutinin without loss of macrophage activating properties. Similarly adsorption of FBP with macrophages preferentially removed macrophage activating component. Separation of the lectin components by DEAE cellulose chromatography yielded two major fractions: a potent hemagglutinin with weak macrophage activating properties and a potent macrophage activator with weak hemagglutinating activity. Separation of the crude lectin by ultrafiltration indicated that the macrophage activating component exists in a highly aggregated form which may determine its macrophage activating properties. Our results indicate that L. tetragonolobus consists of two distinct classes of components which correspond to tetrameric glycoproteins of 118-120 Kd with potent temperature dependent hemagglutinating activity and a highly aggregated dimeric component of 58 Kd with macrophage activating properties.

ABO Blood-Group System↗

PubMed1984

Genetic transformation in Streptococcus sanguis. Simultaneous variation of surface-spreading, competence, hemagglutination and polar fimbriation in selected strains.

Strains of Streptococcus sanguis expressing spreading zones around the colonies and competence in genetic transformation, were subcultivated in the laboratory, and in three strains a variation to non-spreading colony morphology on human blood agar plates was observed. These variants were isolated and compared with the original strains by genetic transformation, hemagglutination and electron microscopy. In genetic transformation with streptomycin resistance as genetic marker, the non-spreading variants showed a decrease to 1/50 or less of the transformation frequencies compared to the wild types. Hemagglutination of guinea-pig erythrocytes is a common characteristic of S.sanguis strains, but the non-spreading variants did not hemagglutinate. In electron microscopy studies the wild-type strains of S.sanguis, but not the non-spreading variants, presented long, polar fimbriae. One of the non-spreading variants had short, peritrichous fimbriae. The co-variation in surface-spreading, competence in genetic transformation, hemagglutination and polar fimbriation is discussed.

Fimbriae, Bacterial↗

PubMed1987

Hemagglutinating activity of lipopolysaccharides from subgingival plaque bacteria.

Lipopolysaccharide (LPS) preparations from the Bacteroides species B. endodontalis, B. intermedius, B. denticola, and B. melaninogenicus and from Fusobacterium nucleatum, Haemophilus actinomycetemcomitans, Capnocytophaga gingivalis, and Eikenella corrodens directly agglutinated erythrocytes of some kinds of animals. LPSs from the Bacteroides species B. gingivalis, B. asaccharolyticus, B. corporis, and B. loescheii and from Capnocytophaga ochracea did not possess any hemagglutinating activity. Pretreatment of LPSs with lipase and phospholipase C completely eliminated the hemagglutinating activity. The hemagglutinating activity was also affected by human serum, saliva, colistin, and polymyxin B but was not affected by sugars, amino acids, EDTA, or proteolytic enzymes. The role of this hemagglutinating activity in colonization by these microorganisms of the periodontal region is discussed.

Dental Plaque↗

PubMed1987

In vivo adherence and colonization of Vibrio cholerae strains that differ in hemagglutinating activity and motility.

A scanning electron microscopic study was carried out to compare the in vivo pathogenicity of two strains of Vibrio cholerae in an adult rabbit ligated-gut test model. V. cholerae C5 (serotype Ogawa, biotype El Tor), a motile strain possessing hemagglutinating activity in vitro, and C21 (serotype Ogawa, classical biotype), a nonmotile strain possessing no hemagglutinating activity, were tested. Tissue samples from small intestinal loops were examined 3, 6, 9, and 12 h postinoculation. Contradictory to most published data, neither hemagglutinating activity nor motility appeared to be essential prerequisites for the pathogenesis of cholera in the experimental animal model used: nonmotile hemagglutinin-negative strain C21 adhered to and colonized the small intestine at least to the same extent as did motile hemagglutinin-positive strain C5. Maximum colonization was seen at 9 h postinoculation for both strains. C5 and C21 vibrios caused comparable damage to the villi of the small intestine. The villous epithelium showed only mild changes during the first 9 h postinoculation. However, after 12 h the epithelium was seriously damaged concomitant with a decrease in the number of vibrios. Many villi showed partial or total denudation, owing to repelled epithelium, leaving a bare basal lamina with only some to moderate numbers of vibrios attached. Since similar changes were induced by pure cholera enterotoxin, these changes were likely the result of excessive fluid accumulation. From this study it is concluded that, at least in the animal model used, factors other than hemagglutinating activity and motility may also play a role in the association of V. cholerae with the small intestinal surface.

Animals↗

PubMed1998

Hemagglutinating properties of apolipophorin III from the hemolymph of Galleria mellonella larvae.

In search for factors that cause encapsulation of foreign bodies in insect hemolymph we discovered that larval hemolymph of Galleria mellonella caused aggregation of mammalian erythrocytes. The hemagglutinating agent was identified as an 18-kDa protein that did not react with lectins. The sequence of 81 amino acids in three protein fragments and the properties of the protein revealed that it was Galleria homologue of apolipophorin III (apoLp-III). ApoLp-III was found in high amounts in the hemolymph of Galleria larvae, pupae, and adults, as well as in the molting fluid. The hemagglutinating action of the whole hemolymph or the purified apoLp-III was independent of the presence of sugars in the medium. This indicated that it was not mediated by carbohydrates on the erythrocyte surface. The hemagglutination was inhibited at low pH (3.0), in the absence of calcium ions, and in the presence of certain bacterial lipopolysaccharides or their essential component, the 2-keto-3-deoxyoctonate-3-deoxyoctulosonic acid (KDO). It is suggested that interaction of apoLp-III with lipopolysaccharides in bacterial cell walls may play a role in insect immune reactions.

Amino Acid Sequence↗

PubMed1995

Hemagglutination by Staphylococcus aureus strains responsible for human bacteremia or bovine mastitis.

Although hemagglutination by Staphylococcus aureus has been associated with the pathogenesis of bovine mastitis, this trait has not been characterized with regard to human disease. In this study, the prevalence of hemagglutination in 100 strains of S. aureus responsible for bovine mastitis or human bacteremia, was characterized. Under optimum conditions hemagglutination was noted in 23% of the bovine strains, but only 13% of human strains, leading us to conclude that this trait is not a significant virulence determinant in human systemic infection. Additional studies indicate the hemagglutinin of S. aureus strains responsible for human bacteremia is proteinaceous in character.

Animals↗

PubMed1983

DNA structure, and hemagglutination properties of bovine adenovirus type 2 strains which bypass species specificity.

Bovine adenovirus type 2 (Ad bos 2) strains were examined which had been isolated during natural outbreaks among calves, and lambs in Hungary [Belák, S., Pálfi, V.: Arch. ges. Virusforsch. 46,366-369 (1974)]. Differences were detected in hemagglutination properties, and in the restriction site maps of the DNA, which seem to be sufficient to group isolates of Ad bos 2 into two subtypes (subspecies). Some of the strains, recovered from cattle including prototype strain No. 19 are suggested to be separated as subtype A. These viruses hemagglutinate bovine red blood cells, and the physical map of the DNA is similar to, or identical with that of the prototype strain. Virus strains tentatively grouped into subtype B are pathogenic for both cattle, and sheep under natural conditions. Members of subtype B hemagglutinate only rat erythrocytes, and characteristic differences may be detected with BamHI, EcoRI, KpnI, and SalI restriction endonucleases in comparison to the physical maps of the DNA of prototype virus. The genome size of all isolates tested was measured to be of Mr 19.5 to 20.0 X 10(6), similar to Ad ovi 1, 4, and Ad bos 4, and 6. All isolates of subtype B characteristics were shown to encapsidate heterogeneous genome populations which could be distinguished from those of subtype A by the presence of specific restriction endonuclease cleavage fragments with molar ratios of less than 1.0.

Adenoviridae↗

PubMed1987

Bovine herpesvirus type 1 gp87 mediates both attachment of virions to susceptible cells and hemagglutination.

The 87,000-dalton glycoprotein (gp87) of bovine herpesvirus type 1 (BHV-1) was found to selectively attach to susceptible cells. The attachment of gp87 to the cells was markedly decreased by the prior adsorption of intact virions. Anti-gp87 (site Ia) monoclonal antibody, which inhibited BHV-1 adsorption to the cells and neutralized the virus without complement [Okazaki et al., Virology 250: 260-264], was effective in inhibiting the adsorption of gp87. Only the same antibody was able to inhibit the hemagglutination activity of BHV-1. Other monoclonal antibodies to the glycoproteins of BHV-1, including antibodies directed to sites Ib and Ic on gp87, were ineffective in inhibiting either virus adsorption or hemagglutination. The results of this study indicate that site Ia of gp87 molecule is the critical site of virus attachment for initiation of infection as well as the hemagglutination of BHV-1.

Animals↗

PubMed1987

Hemagglutination inhibition and virus neutralizing response of rabbits inoculated with bovine herpesvirus 1 subunit vaccine.

The immunogenicity of bovine herpesvirus type 1 (BHV-1) hemagglutinin has been investigated. Both live and nonionic detergent solubilized vaccines were prepared and 5000 hemagglutinating units (HAU) were injected subcutaneously into rabbits. Both types of vaccine induced a good antibody response but live virus was four times more efficient in inducing hemagglutination inhibiting and neutralizing antibodies than either Triton X-100- or octylglucoside-solubilized subunit vaccine. Blotting analysis revealed that five proteins, of 105,000, 90,000, 74,000, 64,000 and 54,000 mol. wt, were recognized by the serum of vaccinated animals. Triton X-100-solubilized vaccine did not induce antibodies against the 105,000 and 64,000 mol. wt proteins, indicating the important role of VP 90,000 and VP 74,000 in hemagglutination and neutralization. The order in which antibodies to the different viral proteins were induced was VP 90,000, (VP 105,000, VP 64,000, VP 54,000) and VP 74,000. Our data indicate that VP 90,000 is the hemagglutinin. Using convalescent serum from intranasally infected animals, we could identify nine structural proteins for BHV-1; VP 105,000, VP 90,000, VP 74,000, VP 64,000, VP 54,000, VP 50,000, VP 47,000, VP 40,000 and VP 31,000.

Animals↗

PubMed1950

Lack of identity in neutralizing and hemagglutination-inhibiting antibodies against influenza viruses.

There is an exponential linear relationship between the quantity of influenza virus neutralized and the quantity of immune serum employed in in ovo neutralization. The slope of the neutralization line is extremely steep. The concentration of neutralizing antibody can be measured with considerable precision in ovo if the constant virus-varying serum technique is utilized. The amounts of hemagglutination-inhibiting and neutralizing antibodies which are absorbed by a given quantity of influenza virus (PR8) were found to be predictable and the degree of reactivity of these two antibodies was shown to be directly related to the extent of immunization. It was demonstrated that there are marked discrepancies in correlation between antibody titers obtained by in vitro hemagglutination-inhibition and in vivo neutralization techniques and that neutralizing antibody is preferentially absorbed by a given quantity of virus. Inasmuch as the results were found not to be attributable to peculiarities of the techniques employed, it appears that the antibodies measured by hemagglutination-inhibition in vitro and by neutralization in vivo are not identical.

Antibodies↗