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At least 19 recordsLinked to original sources
PubMed1976

Titration of Newcastle disease virus and its neutralizing antibodies in microplates by a modified hemadsorption and hemadsorption inhibition method.

Using the microtiter system, titration of Newcastle disease virus infectivity and neutralizing antibodies was carried out in chicken embryo fibroblasts grown in "U" or flat-bottomed plates. Infectivity was detected by a combined hemadsorption-hemagglutination method. Inhibition of that reaction indicated the presence of neutralizing antibodies. A 24-h microneutralization test was developed and compared to the plaque neutralization and microhemagglutination inhibition test. Reproducibility of the microneutralization test was statistically analyzed.

Animals↗
PubMed1975

Detection of cell-bound immunoglobulins by a radioisotopic micro-mixed hemadsorption reaction with technetium-99m-labeled erythrocytes.

The mixed hemadsorption (MHA) reaction detects antibodies reactive with cell surface antigens by means of antiglobulin-coated indicator erythrocytes. We have developed a radioisotopic modification which employs sheep erythrocytes (SRBC) that have been prelabeled with technetium-99m (99mTc), a high specific acitivity metastable gamma-emitter of short half life. The 99mTc MHA reaction was performed on human and murine cells cultured in Micro-test II plated with six replicate wells per serum dilution. Antibody activity in species-specific xenoantisera and mono- and polyspecific alloantisera was detected in high titer. The sensitivity of 99mTc micro-mixed hemadsorption was 2 times that of the visual assessment of mixed hemadsorption, 100 to 200 times that of the 125-I-mixed antiglobulin reaction and 500 to 1000 times more sensitive than indirect immunofluorescence. The assay system was applied successfully to confirm the species of origin of a panel of previously karyotyped human and mouse cell lines. Our results indicate that the 99mTc micro-mixed hemadsorption method is a rapid, sensitive, quantitative test for the detection of cell surface antigens and membrane reactive antibodies.

Animals↗
PubMed1975

Detection of antibody and complement complexed in vivo on membranes of human cancer cells by mixed hemadsorption techniques.

The mixed hemadsorption (MHA) techniques demonstrated antibody and complement fixed in vivo to the surface of human cancer cells. Tumors from 12 cancer patients and normal tissues from 5 cancer patients and 8 patients with cerebrovascular or cardiac diseases were collected from biopsy and autopsy for in vitro testing. Antiserum to human whole immunoglobulins and antiserum to human C3 were used in the MHA techniques. Positive MHA patterns were demonstrated on the surface of cancer cells by both methods. Positive reactions ranged from 12 to 32% in mixed hemadsorption for anitbody detection and from 10 to 34% in mixed hemadsorption for complement component 3 detection. Normal tissues obtained from cancer patients or from patients who died of causes other than cancer rarely exhibited distinct MHA reactivity. Collectively, the data suggest that most human cancers are antigenic in the autologous host and that tumor-associated antigens of cancer cells react in vivo with their humoral antibody to fix complement.

Antibodies, Neoplasm↗
PubMed1975

Temperature-dependent rosette formation by mouse lymphoma cells as a result of viral hemadsorption.

Cells from several mouse lymphomas formed rosettes with nonsensitized foreign erythrocytes through C-type virus particles clustered on the cell surface in serum-free medium held at 4 degrees C. This type of rosetting was found most typically in a lymphoma induced by Rauscher leukemia virus in tissue culture (RD-12), but it also occurred in 23 of 61 spontaneous thymic lymphomas in AKR mice. Chemically or X-ray-induced leukemias and spontaneous reticulum cell sarcomas did not form rosettes. The nature of the rosette formation may be interpreted as viral hemadsorption, with a possible relationship to hemagglutination by murine leukemia viruses. The receptor on virus particles was trypsin sensitive and showed high affinity to serum inhibitors (RIF). Serum rosette-inhibiting activity was assessed by a quantitative rosette inhibition test; rosette inhibition proved widely distributed among species. Physicochemical properties of serum RIF and their function both in vivo and in vitro were described. Rosette formation with similar temperature requirements, previously reported in a mouse lymphoma carrying membrane-bound heterophile cold hemagglutinin, was readily distinguished from viral hemadsorption by its insensitivity to mouse serum RIF.

Animals↗
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↗
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↗
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↗
PubMed1977

Electron microscopic study of hemadsorption on vaccinia virus infected cells.

Hemadsorption (HAD) induced in HEp-2 cells infected with vaccinia virus was observed. In ultrathin sections, binding of 36 red blood cells (RBCs) was examined in detail and 3 types of HAD were observed: (1) direct and close binding of RBCs to infected HEp-2 cells (cyto-HAD) was observed in cross sections of 27 RBCs (2) binding of RBCs through microvilli of infected cells was found in 11 RBCs, and (3) five RBCs were distorted to form tentacle-like projections by which they were bound to the HEp-2 cell surface. Scanning electron microscopy revealed that more than 30% of the RBCs were bound to microvilli of vaccinia virus-infected HEp-2 cells, and that the number of microvilli twined round each RBC was over ten, RBCs were attached to certain microvilli through swollen sucker-like tips which were not observable in non-infected HEp-2 cells. RBCs sometimes revealed a polygonal shape at regions of binding to microvilli. Virion-mediated RBC HEp-2 cell binding could not be observed.

Binding Sites, Antibody↗
PubMed1978

Tissue typing of cells in culture. II. Histocompatibility typing of diploid fibroblastic cultures by the mixed hemadsorption (MH) test.

Skin fibroblasts derived from persons who had been HLA-typed by conventional lymphocytotoxocity tests were subjected to HLA-typing by the mixed hemadsorption (MH) test. The agreement between the two methods was good, provided sera for the MH test were carefully selected. Minor discrepancies found in this study can probably be corrected by further serum selection. There was no difficulty in typing late passages of cultured diploid cells (22 doublings) and in fact it was easier to type a culture at the 33rd doubling than at the 11th. Thus no signs of disappearance of HLA-antigens were seen in these tests.

Cells, Cultured↗
PubMed1976

Use of the hemadsorption phenomenon for determining virus and neutralizing antibody titers of rabies.

Chicken embryo cells infected with the HEP Flury strain of rabies virus adapted to tissue culture produced a hemadsorption (HAD) phenomenon by using goose erthyrocytes. The optimal conditions for HAD included the incubation of cell cultures at 37C for 3 days after virus inoculation, the use of a 0.4% suspension of goose erythrocytes in phosphate buffer adjusted at pH 6.2, and adsorption of erythrocytes at 4C. This phenomenon was inhibited with anti-rabies serum. Virus titer obtained with the HAD technique was almost the same as with the fluorescent antibody technique or the intracerebral inoculation of suckling mice. Results of the neutralization test by using the HAD technique could be easily determined 3 days after inoculation of chicken embryo cells with the mixture of 100 mean tissue culture infective doses of virus and diluted serum. The neutralizing antibody titers coincided with those obtained in mice.

Animals↗
PubMed1978

The advantage and adequacy of the mixed hemadsorption test for rabies antibody determination.

The goal of rabies vaccination is an early development of protecting antibodies. The mixed hemadsorption (MH) test is shown to be an adequate test to evaluate the immunization response. It measures antibodies combining with antigens on rabies-infected cells equivalent to the glycoprotein antigen at the exterior of the budding virus (1). Further, the MH test used measures antibodies solely of IgG class, thus avoiding positive results being obtained by IgM antibodies which can be produced during a prolonged period because of antigen persistence due to daily immunization doses (2). Antibodies of IgM class may play only a minor role in the early defence against rabies as probably only antibodies of IgG class can reach a locally introduced virus and, so far, are the only immunoglobulins shown to induce immune lysis in rabies-infected cells (3). The MH test is described. It is a sensitive, reliable and technically convenient method as it has a capacity for testing many sera in the same run. Results obtained are compared with neutralization values obtained from different types of rabies vaccines (4).

Antibody Formation↗
PubMed1978

Concanavalin A-mediated hemadsorption by normal and malignant human mammary epithelial cells.

The concanavalin A (Con A) reactivity of malignant and normal human mammary epithelial cells in culture was determined with a hemadsorption assay. Human erythrocytes were treated with various concentrations of Con A, and these indicator Red Blood Cells were incubated with the test cells in situ in culture dishes. The Con A concentration at which approximately 50% of the test cells adsorbed erythrocytes ([Con a] 1/2 max) was determined. Five malignant epithelial cell lines and the primary cultures derived from 3 pleural effusions and 20 solid tumors were tested. Primary cultures of normal epithelial cells were established from human milk samples obtained from 3 separate donors. The average [Con A] 1/2 max value for the 5 cell lines and the pleural effusion cultures was 6 and 5 microgram/ml, respectively. The average [Con A] 1/2 max value for the 20 solid breast tumors was 20 microgram/ml. In contrast to the malignant cells, normal mammary epithelial cells did not adsorb erythrocytes coated with as much as 100 microgram Con A per ml. These results show that Con A reactivity distinguishes normal from malignant human mammary epithelial cells.

Breast Neoplasms↗
PubMed1976

Cell surface antigens of human malignant melanoma: mixed hemadsorption assays for humoral immunity to cultured autologous melanoma cells.

We studied how frequently patients with malignant melanoma have specific antibody to cell surface antigens of cultured autologous melanoma cells as demonstrated by mixed hemadsorption assays. Of 35 patients studied over periods ranging from 1 to 36 months with Stage II, III, and IV disease, two showed consistent and high titered reactivity against autologous melanoma cells, two showed less consistent and intermediate reactivity, seven showed sporatic, low titered reactivity, and the remainder were consistently negative. A detailed analysis was carried out with the sera of one patient with sufficiently high titer against autologous melanoma cells. By direct tests and by absorption analysis with a variety of melanoma and nonmelanoma cell lines which included autologous fibroblasts, the antigen could not be demonstrated on any cell type other than the autologous melanoma.

Antibodies, Neoplasm↗
PubMed1975

Inhibition of the growth of hemadsorption 2 virus by three acyl derivatives of amino acids.

Three acyl derivatives of amino acids, dicarbobenzoxy-l-lysine sodium, carbobenzoxy-l-aspartic acid-beta-benzyl ester potassium, and N-3-phenylpropionyl-S-benzyl-l-cysteine potassium inhibit the growth of parainfluenza 1 (hemadsorption 2) virus. The growth of simian virus 40, vaccinia, poliomyelitis type 1, Semliki Forest, Eastern equine encephalitis, and Western equine encephalitis viruses was not affected by these compounds. Four other acyl derivatives of amino acids did not inhibit the growth of any of the viruses tested.

Amino Acids↗