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PubMed1978

Tissue typing of cells in culture. III. HLA antigens of established human cell lines. Attempts at typing by the mixed hemadsorption technique.

Thirty established cell lines from solid human tumors were typed by the mixed hemadsorption technique with respect to the HLA-A and HLA-B and partly also the HLA-C series. All cultures exhibited unique antigenic patterns. The discriminatory potential was further increased very much taking into consideration one as yet undefined antigen series tentatively named Ek-1 to Ek-11. Sixteen cell lines not designated as HeLa cells but on various non-immunological indications suspected to be HeLa cell contaminants, all gave, with our present test, the same antigenic composition as HeLa cells or clones thereof. The mixed hemadsorption reaction is a highly sensitive and convenient method for immunological typing of cells in monolayer cultures. Some important implications of this are discussed.

Antigens, Neoplasm↗
PubMed1981

Characterization of hemadsorption-negative mutants of Mycoplasma pneumoniae.

Previously isolated mutants of Mycoplasma pneumoniae incapable of hemadsorption were characterized with respect to specific protein content, tracheal ring attachment capability, and virulence for both in vitro and in vivo model systems. Two-dimensional gel electrophoresis revealed both quantitative and qualitative differences between the protein complements of two different mutant strains and that of the virulent parent strain. Studies of mycoplasma attachment to hamster tracheal rings in vitro demonstrated that only one of these mutant strains still possessed the ability to attach to the respiratory epithelium via neuraminidase-sensitive receptors. Measurement of [3H]orotic acid uptake in mycoplasma-infected tracheal rings indicated that infection with the hemadsorption-negative mutants resulted in only slight reductions of ribonucleic acid synthesis, similar to levels observed for tracheal rings infected with an avirulent strain of M. pneumoniae. The virulence potential of the two mutant strains was further investigated by utilizing the hamster model system. Both mutant strains were rapidly cleared from the lungs of infected animals and produced little or no microscopic pneumonia.

Animals↗
PubMed1981

Hemadsorption immunosorbent technique for determination of rubella immunoglobulin M antibody.

A highly specific and sensitive hemadsorption immunosorbent technique for measuring rubella immunoglobulin M (IgM) antibody is described. IgM from human sera was absorbed into anti-human IgM-coated wells in plates and rubella-specific IgM was detected by adding rubella virus hemagglutinin and a small quantity of sheep erythrocytes. Centrifugation of the plates facilitated reading of the test. Specific IgM-positive sera showed hemadsorption, whereas negative sera showed hemagglutination. Rheumatoid factor and rubella-specific IgG antibody did not interfere with the results. The test was clearly more sensitive than the solid-phase immunosorbent technique for detection of rubella IgM antibody by hemagglutination inhibition and at least as sensitive as the hemagglutination inhibition test on IgM fractions from a sucrose density gradient and the indirect immunofluorescence test for IgM antibody with absorbed serum. All of 40 sera from 17 rubella patients taken 4 to 49 days after the onset of rash were positive in the new test, with antibody titers ranging from 2,560 to 81,920 between 4 and 28 days. The test is reliable, practical, and suitable for general diagnostic use.

Antibodies, Viral↗
PubMed1982

Use of a hemadsorption technique to evaluate the stability of the hemagglutination reaction of Escherichia coli cultures possessing human colonization factor antigens.

Two strains of Escherichia coli, producing different colonization factor antigens (CFA), were monitored for the population density of CFA-producing bacteria after repeated subculture. The production of CFA was estimated by flooding agar plates containing isolated colonies with suspensions of human or bovine erythrocytes. The erythrocytes were suspended in a low-ionic-strength buffer and were fixed to CFA-positive colonies with a 1.0% tannic acid solution. Strain H-10407, possessing CFA/I fimbriae, showed a rapid loss of the hemagglutinin when subcultured, whereas strain CL-9699, producing CFA/II, was very stable. By using the hemadsorption assay, we could rapidly and easily distinguish CFA- positive colonies from the CFA-negative variants. A survey of additional E. coli strains demonstrated the utility and specificity of the hemadsorption technique used.

Animals↗
PubMed1983

Immune adherence hemadsorption: specific method for rapid identification of adenovirus antigens.

An immune adherence hemadsorption test for the rapid group-specific identification of adenoviruses is described. The test was performed directly on the cell culture isolate in less than 1 h, by the sequential addition of the antiserum of any adenovirus serotype, complement, and then C3b receptor-rich human erythrocytes. Microscopic examination revealed the presence of erythrocytes adhering firmly to the infected cell sheet. The immune adherence hemadsorption test proved a specific and rapid diagnostic procedure for the detection of adenovirus group antigen in tissue cultures and was more simple than conventional tests.

Adenoviridae↗
PubMed1986

Hemadsorption immunosorbent technique for the detection of dengue immunoglobulin M antibody.

We developed a highly specific, sensitive, and economical hemadsorption immunosorbent technique for the detection of dengue-specific immunoglobulin M (IgM) antibody. The technique is based on the reaction of human sera with anti-human IgM immobilized onto a solid phase followed by the detection of dengue-specific IgM by the addition of a known quantity of dengue virus hemagglutinin and goose erythrocytes. Dengue-specific IgM-positive sera showed hemadsorption. IgM antibody specific for dengue virus was detected in 22 of 39 (56%) convalescent-phase sera from primary dengue infections and 8 of 10 (80%) convalescent-phase sera from secondary dengue infections. Additionally, 32 of 76 single sera from patients were positive for dengue IgM; these sera were previously uninterpretable by the hemagglutination inhibition test, as only a single serum specimen was available. No false-positive results were obtained with sera that were negative by the hemagglutination inhibition test for dengue virus. Crude dengue virus hemagglutinin preparations could be used without purification. Dengue-specific IgG did not interfere with the results, nor was there any cross-reactivity between dengue hemagglutinins and IgM specific for other viruses. Some cross-reactivity of the dengue-specific IgM was observed with Japanese encephalitis virus hemagglutinins, but this did not present any problems in the interpretation of results. This test is specific, inexpensive, highly reproducible, and simple to perform.

Antibodies, Viral↗
PubMed1993

Solid-phase antibody capture hemadsorption assay for detection of hepatitis A virus immunoglobulin M antibodies.

A solid-phase antibody capture hemadsorption (SPACH) assay was developed to detect hepatitis A virus (HAV)-specific immunoglobulin M (IgM) antibodies in sera from humans recently infected with hepatitis. The assay is performed with microtiter plates coated with anti-human IgM antibodies to capture IgM antibodies from the test sera. HAV-specific IgM antibody is detected by the addition of HAV hemagglutinating antigen and goose erythrocytes. Hemadsorption of erythrocytes to antigen-antibody complexes attached to the solid phase indicate the presence of IgM antibodies. The SPACH assay was compared to a commercial radioimmunoassay and was found to be equally or more sensitive and specific for the detection of HAV IgM antibodies. The SPACH assay is an alternative, rapid assay that doesn't require hazardous substrates or radioactivity for the detection of HAV-specific antibodies.

Evaluation Studies as Topic↗
PubMed1977

New tests for characterization of mumps virus antibodies: hemolysis inhibition, single radial immunodiffusion with immobilized virions, and mixed hemadsorption.

Hemolysis inhibition (HLI), single radial immunodiffusion (SRID) with immobilized virions, and mixed hemadsorption tests were used for measuring antibodies against mumps virus. Rabbit hyperimmune sera against mumps and early and late human convalescent sera were analyzed. All three tests identified antibodies against both hemagglutinin and the second major envelope component, hemolysin (fusion factor). The sensitivity of the HLI test corresponded to that of the hemagglutination inhibition (HI) test, but in some sera HLI antibodies occurred in greater quantity than HI antibodies. The SRID test readily identified rises in antibody titers in connection with acute infection. Due to its simplicity and lack of sensitivity to nonspecific inhibitors, it is recommended for use in this context. The mixed hemadsorption test showed a high sensitivity for specific identification of mumps antibodies. It therefore may be suitable for use in screening for immunity to mumps.

Antibodies, Viral↗
PubMed1978

Electron microscopic study of measles virus infection: cell fusion and hemadsorption.

Virus-induced cell fusion has been studied after infection of Vero cells with measles virus. Scanning and transmission electron microscopy were combined with immunoperoxidase labeling of measles antigens to correlate viral production and distribution of virus-induced erythrocyte binding sites with progress of fusion. Release of infectious virus started before syncytia were detected and decreased while the number and size of syncytia were increasing. Most virions were seen budding from mononucleated cells or from the periphery of syncytia where cells were being recruited. Moving inward, the surfaces of syncytia where cells were being recruited. Moving inward, the surfaces of syncytia were covered with numerous ridges containing viral antigen, but few viral buds were seen, suggesting that syncytia might be sites of defective viral formation. Hemadsorption occurred predominantly within the confines of syncytia. Erythrocytes were scattered sparsely over immature syncytia but were densely packed in the center of mature syncytia. Active binding sites for erythrocytes were located on cell villi and ridges covered with measles antigens. Hemadsorption was completely inhibited in measles virus-infected cultures pretreated with virus-specific immunoglobulin G for 1 h at 4 degrees C. However, when these cultures were shifted to 37 degrees C, hemadsorbing sites were recovered at the periphery of enlarging syncytia. Virus-induced sites for erythrocyte adsorption were found to move centripetally on syncytium membranes as fusion progressed.

Antigens, Viral↗
PubMedDate not supplied

[Detection of oncornavirus D-induced antigen in cells by a mixed hemadsorption method].

Further study of a new oncornavirus D-induced antigen was done. Its immunological specificity and localization in cells was determined by a sensitive method of mixed hemadsorption. A continuous HEp-2 cell culture was used as the source of the new as well as virus antigen. Adsorbed specific sera to the new antigen and to oncornavirus D obtained by the previously developed method were used for the antigens detection. This method demonstrated localization of both virus antigens and oncornavirus D-induced antigens not included into the virion structure on the surface of oncornavirus D-infected cells. Mixed hemadsorption test as well as complement-fixation test gave the results identical in their specificity indicating the presence in HEp-2 cells of a new non-virion antigen.

Animals↗
PubMed1983

The detection of defective interfering Rubella virions by a modified hemadsorption technique.

Rubella virus (RV) has been titrated in murine fibroblasts in monolayer cultures using a modified hemadsorption assay. Infected monolayers were incubated in the presence of sheep erythrocytes for 24 h: after the tissue culture slides were washed, the hemadsorption foci were clearly visible on microscopic examination. When serial doubling dilutions of virus suspensions were assayed in this way the decline in the number of hemadsorbing units was proportional to the fractional power of the concn. These data suggest that this simple titration of Rubella virus may be useful in the detection of defective interfering (DI) virions in Rubella populations. The detection of DI Rubella virions by more time-consuming and expensive techniques including isopycnic centrifugation, ribonucleic acid extraction, 125I-labelling, polyacrylamide slab gel electrophoresis, and autoradiography have been described. The clinical importance of defective virions and the need for an inexpensive, rapid means of detecting them are discussed.

Animals↗
PubMed1991

Hemadsorption by colonies of Ureaplasma urealyticum.

Hemadsorption by colonies of Ureaplasma urealyticum and Mycoplasma pneumoniae differed quantitatively and qualitatively. Using standard methodology, few strains of U. urealyticum hemadsorbed; with a modified method, most strains hemadsorbed, indicating a second type of association. Scanning electron microscopy of tannin-osmium-stained preparations showed guinea pig erythrocytes embedded in ureaplasma colonies and craters left when erythrocytes were dislodged.

Animals↗
PubMed1982

Detection of parainfluenza IgM antibody by hemadsorption immunosorbent technique.

A hemadsorption immunosorbent technique (HIT) was developed for the detection of immunoglobulin M (IgM) to parainfluenza virus types 1, 2, and 3. Twenty-six (90%) of twenty-nine patients under 6 yr of age from whom parainfluenza virus was isolated showed parainfluenza IgM antibody in one or both of their paired sera, with titres ranging from 320 to 81,920. In about one third of the cases IgM antibody was demonstrated in the initial sera taken 1 to 3 days after the onset of illness. Heterotypic IgM antibody responses were observed in about 20% of the patients. The HIT test was more sensitive than the hemagglutination inhibition (HI) and complement fixation tests in detecting a seroresponse in the 29 virus-positive children. The results of studies in older patients with HI titre rises to parainfluenza virus suggested that reinfection probably induced IgM antibody in a proportion of cases. The HIT test proved to be specific for the IgM class of antibody and avoided false-positive results due to rheumatoid factor. It permits an early presumptive diagnosis in a proportion of patients with parainfluenza infection.

Adolescent↗
PubMed1977

An attempt to detect cell surface antigens in cultured human brain tumors by mixed hemadsorption test. First report.

The mixed hemadsorption test using rabbit anti-human glioma serum was applied to investigate cell surface antigens of nervous tissue tumors, non-nervous tissue tumors, epithelial and mesenchymal cells. Gliomas, neurinomas, and fetal brain cells exhibited a strongly positive reaction. Meningiomas, a metastatic brain tumor originated from the lung and HeLa cells exhibited a moderately positive reaction. No positive reaction was detected in human skin or dural fibroblasts, or in kidney cells even at a high concentration of rabbit anti-human glioma serum.

Antigens, Neoplasm↗
PubMed1978

An electron microscopic study of the surface structures and hemadsorption on chick embryo cells infected with rabies virus.

Characteristic alterations at the surface of chick embryo cells infected with the HF-TC strain of rabies virus and the binding sites of hemadsorption were studied employing both scanning and transmission electron microscopes. The initial alteration of the cell surface structure revealed by scanning electron microscopy was an appearance of elongated and reticulated microvilli on the 2nd day after virus inoculation. On the 3rd day, numerous bullet-shaped virions could be seen budding as single, tetrapod-like structures and as radial projections both from the perikarya and microvilli. Thereafter, elongation of microvilli, formation of numerous blebs in various sizes, disappearance of filopodia, and rounding up of infected cells were observed as characteristic cytopathic effects by rabies virus infection. The attachments of goose erythrocytes to the infected cells occured in two forms. The one was adsorption of erythrocytes to the cell surface involving microvilli and filopodia in the absence of detectable virus, and the other was adsorptio n of erythrocytes to the virus particles budding from cell surface. The former could be seen from the early stage of infection through the end of observation period, while the latter was observed only on and after the 3rd day after virus inoculation. These findings were also confirmed with transmission electron microscopy.

Cell Membrane↗
PubMed1977

Micro mixed hemadsorption assay--a sensitive method for detecting antibodies against tumor-associated antigens.

Antibodies against methylcholanthrene (MCA)-induced sarcomas in Fischer (F344) rats were detected by a modified micro mixed hemadsorption (MHA)-assay. The assays detected anti-tumor antibodies as titers up to 1 : 320 in sera from hyperimmunized rats and at titers up to 1 : 160 in sera from rats bearing a 5-8 cm3 progressively growing tumor. MHA-titers decreased when sera were absorbed with sarcoma cells prior to MHA-assays. IgG antibodies in sera from tumor bearing rats showed titers of 1:20 and 1:5. These anti-tumor sera formed rosettes on the corresponding sarcoma cells as well as other sarcomes induced by MCA in F344 and Lewis strain rats tested. The assay was modified for a micro technique using a microtest plate (No. 3034, Falcon, CA). This modification yielded as assay requiring only 10 microliter of test sera. The test is quantitative and highly sensitive and results are reproducible. Several critical factors which influence test results in this assay were examined.

Animals↗
PubMed1980

Rubella immunity testing by mixed hemadsorption.

A mixed hemadsorption (MH) test for naturally acquired rubella immunity and immune responses after rubella vaccination has been compared with the hemagglutination-inhibition (HI) test, the immunodiffusion (ID) test, the neutralization test (NT), the hemolysis-in-gel (HIG) test and the indirect immunofluorescence (IF) test. The MH test was approximately equivalent to the HI test and the NT as to sensitivity for the early response to rubella vaccination, and all three tests are thought to measure protective antibodies, i.e. antibodies supposedly directed against envelope antigens, to about the same extent. The MH test for rubella antibodies is especially suited for laboratories in which this type of test is routinely used for other purposes, e.g. rabies, respiratory syncytial virus, herpes simplex, varicella and certain auto-antibodies. Where the method is already standard it may be preferable to the NT which, although of similar clinical value requires tedious standardization. Both tests may be necessary in cases where the HI test is equivocal or hampered by non-removable on-specific inhibitors.

Antibody Formation↗