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Isolation of hemagglutinin and neuraminidase subunits of hemagglutinating virus of Japan.

When purified hemagglutinating virus of Japan (HVJ) was treated with trypsin, two major surface antigens were released from the virus. The "hemagglutinin" subunits obtained by this method were reactive with homologous hemagglutination-inhibition antibody and could be detected by an antibody-blocking test. They adsorbed to but did not agglutinate red cells and thus appeared to be "monovalent." The neuraminidase subunits were obtained in fully active form and did not adsorb to red cells. This finding suggests that these two activities of HVJ are associated with different subunits of the virus particle. The hemagglutinin and neuraminidase subunits could be partially separated by zonal rate centrifugation or gel filtration on Sephadex G-200. The molecular weights estimated for these subunits were approximately 124,000 and 114,000, respectively. After treatment with trypsin, virus-associated hemagglutinin and neuraminidase activities were both reduced significantly. The electron micrographs of such trypsinized virus particles showed complete or partial loss of surface projections. These results suggested that the subunits obtained by this method seemed to be those projections liberated from the virus by the action of trypsin.

Animals↗

The amino-terminal half of rotavirus SA114fM VP4 protein contains a hemagglutination domain and primes for neutralizing antibodies to the virus.

We have previously reported the synthesis in Escherichia coli of polypeptide MS2-VP8', which contains the amino-terminal half of the SA114fM VP4 protein fused to MS2 bacteriophage polymerase sequences (C. F. Arias, M. Lizano, and S. López, J. Gen. Virol. 68:633-642, 1987). In this work we have synthesized the carboxy-terminal half of the VP4 protein also fused to the MS2 polymerase. This protein, designated MS2-VP5', was recognized by sera to the complete virion and was able to induce antibodies to the virus when administered to mice; however, these antibodies had no neutralizing activity. The two chimeric polypeptides were tested for their ability to agglutinate erythrocytes and to prime the immune system of mice. Bacterial lysates enriched for the MS2-VP8' hybrid polypeptide, but not those enriched for the MS2-VP5' protein or those containing proteins from the host E. coli strain, had hemagglutinating activity. This hemagglutination was inhibited by sera to SA114fM rotavirus. In addition, a single dose of the MS2-VP8' polypeptide was able to prime the immune system of mice for an augmented neutralizing antibody response when the animals were subsequently immunized with purified SA114fM virus.

Amino Acid Sequence↗

The homocytotropic and hemagglutinating antibody responses to Brugia pahangi infection in the multimammate rat (Mastomys natalensis).

Homocytotropic and hemagglutinating antibody responses were followed in multimammate rate (Mastomys natalensis) infected with Brugia pahangi. Homocytotropic antibodies were detected by both active and passive cutaneous anaphylaxis using an antigen prepared from Dirofilaria immitis. The homocytotropic antibody response was first evident at 3 weeks after infection and increased progressively until after patency. It then waned gradually and was absent at 33 weeks after infection (or 20 weeks after patency). During this time microfilaremia continued to increase steadily. Homocytotropic antibody induced by the infection was heat (56 degrees C) labile, sensitive to 2-mercaptoethanol reduction, and was detected in the skin of recipients following a 72-hour latent period. Hemagglutinating antibodies to the infection were detected by the D. immitis antigen. These were present by 9 days after infection and persisted throughout the course of the infection. Only 2-mercaptoethanol labile antibody was detected during the prepatent period of infection.

Agglutinins↗

[Functional activity of hemagglutinins from Hantaan viruses strains and use of hemagglutinating antigens for serodiagnosis hemorrhagic fever with kidney syndrome].

Hemagglutinins of Hantaan virus strains isolated in Southern Far East of Russia from patients with hemorrhagic fever with the renal syndrome and from rodents, natural vectors of the virus, were studied in the cross kinetic hemagglutination inhibition test with immune sera to the studied strains. The strains differed by the functional activity index. Use of hemagglutinins of functionally active Hantaan strains in serologic studies notably improves the diagnostic efficacy of the hemagglutination test in Hantavirus infection.

Animals↗

Usefulness of repeated direct intratumoral gene transfer using hemagglutinating virus of Japan-liposome method for cytosine deaminase suicide gene therapy.

To investigate the feasibility of repeated gene transfection in suicide gene therapy against human solid tumors by a combination of 5- fluorocytosine (5-FC) and its converting enzyme, cytosine deaminase (CD), we repeatedly transfected the yeast CD gene into the human pancreatic cancer cell line BXPC3 using the hemagglutinating virus of Japan-liposome in a new gene transfer method. The in vivo growth of the s.c. transplanted BXPC3 tumor in nude mice given CD-gene transfection was significantly suppressed by i.p. injection of 5-FC when compared with tumors treated with the control vector. Furthermore, the tumor transfected with the CD gene during a 7-day interval was suppressed much more than that of a single transfection. These results suggest that repeated transfection of the suicide gene together with the combination of 5-FC and the yeast CD gene using the hemagglutinating virus of Japan-liposome gene transfer method may be useful for the treatment of human solid tumors, including pancreatic cancer.

Animals↗

[Evaluation of an indirect hemagglutination test for the serodiagnosis of amebiasis].

167 sera have been tested to appreciate the value of an indirect hemagglutination test (Amibiase HAI FUMOUZE) comparatively to an agglutination test of sensibilized particles of latex (Bichro latex Amibe Fumouze BLA) Amibiase HAI test comes out as sensitive and specific for the detection of antibodies in patients suffering from visceral amoebiasis. But some antibodies are also detected in patients with an antecedent of amoebiasis, as it is usually the case with some other techniques. A high positivity of the indirect hemagglutination test, and the concordance between the test HAI and the BLA one are in favour of a visceral amoebiasis. While lower rates or discrepancy between the two tests may evoke an hidden infestation in patients coming out or originated from endemic zones.

Amebiasis↗

Determination of hemagglutination activity recovered from oil-emulsion Newcastle disease vaccines as a prediction of efficacy.

Hemagglutination (HA) activity was recovered from the aqueous phases of commercially and experimentally prepared oil-emulsion (OE) Newcastle disease virus (NDV) vaccines using two methods: aqueous partition and freeze-thaw. Quantitation of the HA activity retrieved by the aqueous partition technique was directly related to the degree of protection conferred to chickens against a strain of velogenic viscerotropic (VV) NDV in nine of the ten vaccines analyzed. One of the ten vaccines yielded high levels of retrieved HA activity but induced low hemagglutination-inhibition (HI) antibody levels and low levels of protection. Therefore, the retrieval and quantitation of HA activity from OE NDV vaccines using the partition technique provides a useful adjunct to vaccine testing methods that doesn't require vaccination and challenge trials. The freeze-thaw method did not yield measurable HA titers for all vaccines with high efficacy, so its use should be restricted to only those vaccines for which it has been demonstrated to be suitable.

Animals↗

Rotavirus. I. Hemagglutination and complement fixation inhibitor antibodies in individuals of Mexico city.

Hemagglutination inhibiting antibodies were studied in 1,278 sera of individuals of different ages in Mexico City representative of four different socioeconomic levels. It was found that 1,261 (98.7 per cent) had antibodies against the Nebraska strain (calf rotavirus). An unusual finding was that more than 95 per cent of children from one to four years of age, had antibodies. For this reason five year old children were studied utilizing two viral strains as antigens in hemagglutination inhibition tests. It was found that in 918 sera, 821 (89.4 per cent) had antibodies against SA-11 samples (monkey rotavirus); percentage increased from 87.2 per cent in neonates to 97.5 percent in children from four to five years of age. A similar phenomenon was observed when Nebraska strain hemagglutinin was utilized. Also complement fixation antibodies against SA-11 strain in 583 sera were studied finding that 260 (44.6 per cent) had antibodies. Utilization of this technique showed that seropositivity percentage decreased from 47.6 per cent in the neonate group to 14.5 per cent in the one ot three month old children increasing progressively until reaching 65.1 per cent in children three to four years of age. These results show that rotavirus infection takes place at early ages, as has been confirmed by other authors that have used electron microscopy techniques, RNA electrophoresis, etc., regardless of the presence of transplacental antibodies.

Adolescent↗

[Experience in using the indirect hemagglutination test to detect Shigella Sonnei in market milk].

The results obtained by analyzing 231 samples of market milk during the period of 1977-1978 are presented. The microbial index, E. coli titer, the presence of pathogenic and opportunistic microflora, as well as the presence of Sh. sonnei antigen were determined. In 39.3% of the milk samples the microbial index was found to be 4 x 10(6), and E. coli titer was 0.001 and lower. No pathogenic microflora was detected in bacteriological assays, but the indirect hemagglutination test with the use of diagnostic antibody reagent revealed the presence of Sh. sonnei antigen in 12 samples. The examination of 28 members of the families which had supplied this milk to the market revealed 3 cases of dysentery infection. The authors propose to widely use the indirect hemagglutination test with diagnostic antibody reagent as an express method for the detection of Sh. sonnei in milk.

Agricultural Workers' Diseases↗

Characterization of in vivo gene transfer into the arterial wall mediated by the Sendai virus (hemagglutinating virus of Japan) liposomes: an effective tool for the in vivo study of arterial diseases.

Recently, much attention has been paid to the arterial gene transfer technique using several vector systems. In the present study, we experimentally examined the transfection efficiency of the exogenous gene, the duration of gene expression, and the cytotoxic effect of the hemagglutinating virus of Japan (HVJ) liposome on the intact arterial wall to evaluate its effectiveness for the study of arterial diseases. To evaluate the transfection efficiency and duration of gene expression, pSV beta-galactosidase was transferred into the carotid arterial wall of rabbits. The cytotoxic effect of HVJ liposomes on the vascular cell components was also evaluated by a neutral red assay in vitro and scanning electron microscopy in vivo. HVJ liposomes could achieve highly efficient gene transfection into the medial smooth muscle cells of intact arteries at 150 and 760 mmHg of pressure (mean = 85.3% and 93.5% of total smooth muscle cells, respectively) without any inflammatory reaction. The introduced exogenous gene was expressed for at least 14 days. In addition, the cytotoxicity for the arterial smooth muscle cells and endothelial cells induced by HVJ liposome vehicles at routinely used concentrations (5,000 to 10,000 hemagglutinating activity units/ml) was minimal both in vitro and in vivo. As an example, we introduced human vascular endothelial growth factor cDNA, which was driven with cytomegalovirus enhancer and beta-actin promoter, into the rabbit carotid arteries, and it induced not only angiomatoid proliferation of endothelial cells forming irregular vascular channels but also intimal hyperplasia. Based on these findings, we conclude that HVJ liposome-mediated arterial gene transfer is a highly efficient, noninvasive, and effective gene delivery method for the study of vascular disorders.

Animals↗

Hemagglutination with transmissible gastroenteritis virus.

Transmissible gastroenteritis virus grown in primary swine kidney cell cultures agglutinated erythrocytes from chicken, guinea pig and cattle but not erythrocytes from mouse and goose. The optimal incubation temperature was at 4 degrees C. The hemagglutination (HA) reaction was inhibited by specific antiserum. Some factors involved in the HA and HA-inhibition (HI) were investigated and standard HA and HI tests were established. HI antibody titers of individual pig sera showed a significant positive correlation with their neutralizing antibody titers.

Animals↗

Hemagglutination with avian infectious laryngotracheitis virus.

Avian infectious laryngotracheitis virus grown in primary chicken kidney cell cultures was tested for hemagglutination (HA) with erythrocytes of a variety of species at 4 degrees C, 22 degrees C, and 37 degrees C. HA was observed at all temperatures with mouse erythrocytes but not with cattle, sheep, goat, swine, rabbit, guinea pig, chicken, and goose erythrocytes. A strain variation between mice in the agglutinability of their erythrocytes necessitated selection of mice to obtain erythrocytes. The HA reaction was inhibited by specific antiserum. Some factors involved in the HA and HA-inhibition (HI) were investigated and standard HA and HI tests were established. HI antibody titers of individual chicken sera had a significant positive correlation with their neutralizing antibody titers.

Animals↗

Hemagglutination by herpes simplex virus type 1.

The McIntyre and HSZP strains as well as clinical isolate of herpes simplex virus type 1 were found to agglutinate C57Bl/10su and CBA mouse red blood cells. The hemagglutinating activity was inhibited by antisera that neutralized the infectivity of the virus.

Animals↗

Hemagglutination with rhabdoviruses related to bovine ephemeral fever virus.

Three different bovine ephemeral fever group viruses were tested for hemagglutination (HA). One of them, Tortilla Flat virus (CSIRO 368), agglutinated erythrocytes from geese, pigeons, horses, hamsters, mice and guinea-pigs when the concentrated infectious culture fluid was used as a hemagglutinin. The HA was dependent on the pH of phosphate buffered saline used as the erythrocyte diluent when using borate buffered saline (pH 9.0) with 0.4% bovine serum albumin as the antigen diluent. The optimal pH of the phosphate buffer was from 5.2 to 5.8. The HA, however, was not dependent on salt concentration. The incubation temperature did not affect the HA titer significantly. This HA reaction was inhibited by serotype specific antiserum.

Animals↗

An improved hemagglutination test for study of canine parvovirus.

Optimal conditions for hemagglutination (HA) by canine parvovirus (CPV) strains were investigated using several buffers. Porcine erythrocytes often agglutinated spontaneously in phosphate-buffered salt solution, isotonic saline solution or barbitone-complement-fixation buffer. Results were reproducible when borate-buffered saline (BBS) was used as the diluent for antigen, and "virus adjusting diluent" (VAD), containing 0.15 M NaCl and 0.3 M phosphate was used as the diluent for erythrocytes. Highest HA titers were obtained at pH 6.0 using BBS and VAD. Specific HA with CPV was observed not only at 4 degrees C but at 37 degrees C, and erythrocytes from horse, shrew mouse, hamster, cat, sheep and dog, as well as pig and African green monkey were agglutinated by CPV using the improved method.

Animals↗

Humoral aspects of the immune response to homografts. II. Relationship between the hemagglutinating and cytotoxic activities of certain isoimmune sera.

Isoimmune sera, prepared in BALB/c mice by immunization with living or lyophilized tissue from C57Bl/6 mice, were found to possess both hemagglutinating and cytotoxic activity for donor cells. Comparative titrations indicated a good correspondence between these two activities. Absorbtion with living or dead donor tissue caused removal or proportionate reduction of both activities. The results are consistent with the existence in isoimmune sera of a single class of isoantibodies, capable of agglutinating donor cells or of sensitizing them for complement lysis. Living and lyophilized tissue differ quantitatively, but not qualitatively, in their capacity to induce formation of these antibodies.

Allografts↗

Quantitative assay of diphtherial toxin and of immunologically cross-reacting proteins by reversed passive hemagglutination.

A reversed passive hemagglutination (RPHA) assay for diptherial toxin has been developed. Antitoxic antibodies were isolated from commercially available equine diptherial antitoxin by immunoabsorption using highly purified diphtherial toxin covalently linked to Sepharose 4B. Formalinized, tanned sheep erythrocytes sensitized with the purified antitoxic antibodies are specifically agglutinated by diphtherial toxin but are not agglutinated by extracellular antigens of Corynebacterium diptheriae that are unrelated to toxin. The RPHA assay described can detect less than 20 pg of diphtherial toxin and is comparable in sensitivity to intracutaneous tests for toxin. The RPHA assay was shown to be at least 1,000 times more sensitive than quantitative immunological assays for diptherial toxin performed by single radial immunodiffusion or by one-dimensional double diffusion in agar gels. Fragment A prepared from purified diphtherial toxin and nontoxic mutant proteins that cross-react immunologically with toxin can be assayed directly by RPHA, but the sensitivity of the assay for these proteins is less than for native diphtherial toxin. Inhibition of RPHA was also shown to be a sensitive quantitative method for measuring diptherial antitoxin in vitro.

Bacterial Proteins↗