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Solid-phase hemadsorption assay for IgM antibody to Treponema pallidum in serum and umbilical cord blood.

A solid-phase hemadsorption assay (SPHA) for Treponema pallidum was evaluated using serum and umbilical cord blood. A total of 133 sera reactive using immunofluorescent antibody assay were tested by SPHA for IgM antibody against T. pallidum. They were categorized into clinical stages of syphilis, including eight sera with primary, 32 sera with secondary, 68 sera with latent, 21 sera with unclassified, and four sera with congenital syphilis. Sera were reactive for 75%, 90.6%, and 33.8% using SPHA in the primary, secondary, and latent phases, respectively. The method gave 57.1% reactivity for the unclassified phase of syphilitic sera and 100% reactive for the congenital syphilitic sera. All 80 immunofluorescent nonreactive sera were nonreactive by SPHA. Of 19 reactive cord blood samples tested, seven samples were reactive by SPHA. However, all 20 nonreactive umbilical cord blood samples were nonreactive by SPHA. The SPHA assay for the determination of IgM antibody to T. pallidum in serum appears to be sensitive for primary, secondary, and congenital syphilis. The SPHA assay in the cord blood is sensitive and specific.

Antibodies, Bacterial↗

Identification of a 32-kilodalton protein of Mycoplasma pneumoniae associated with hemadsorption.

Monoclonal antibodies directed against a 32-kilodalton (kDa) protein of Mycoplasma pneumoniae have been used to characterize a hemadsorption-negative (HA-) mutant class whose protein profile was previously indistinguishable from the wild-type, hemadsorbing (HA+) strain. Electron microscopy and colloidal gold labeling techniques were applied for ultrastructural analysis of the 32-kDa protein. Results indicate that this protein clusters in the tip structure of M. pneumoniae (HA+) wild-type organisms. Additionally, the protein is precipitated by infected hamster sera.

Animals↗

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↗

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↗

Solid-hase hemadsorption: a method for rapid detection of Treponema pallidum-specific IgM.

A solid-phase hemadsorption (SPHA) technique for the rapid detection of Treponema pallidum-specific IgM antibodies is described. The demonstration of these immunoglobulins permits differentiation between a recent and a long-lasting infection and facilitates the decision as to whether or not a patient should receive therapy for syphilis. Errors that occur in the fluorescent treponemal antibody-absorbed test with IgM conjugate (IgM-FTA-Abs test) because of high titers of T. pallidum-specific IgG (competitive inhibition), rheumatoid factors, or 7S IgM do not occur with the SPHA technique. This test is easy to perform; it requires only microtiter plates coated with antisera to human IgM in addition to the standard T. pallidum hemagglutination reagents. As compared with the Veneral Disease Research Laboratory (vdrl) test, SPHA test has the advantage of being specific for T. pallidum; furthermore, IgM antibodies can be demonstrated earlier after infection and disappear sooner after adequate treatment of syphilis.

Antibody Specificity↗

Cell typing in lymphoproliferative disorders. Use of a sensitive hemadsorption technique for sections of normal and abnormal lymphoid tissue.

We have adapted a hemadsorption technique originally devised to detect Fc receptors on human cancer cells for categorizing different lymphoid cells in frozen sections of normal and abnormal human biopsy specimens. The procedure is sufficiently sensitive to demonstrate T-lymphocytes and receptors on the cells in diffuse non-Hodgkin's lymphomas in sections which has been difficult, if not impossible, using other techniques. It demonstrates the expected distribution of B- and T- lymphocytes and mononuclear phagocytes in normal lymphoid organs. When used to type the cells of non-Hodgkin's lymphomas, it furnishes results comparable to more tedious methods requiring cell separation: 63 of 74 non-Hodgkin's lymphomas were classified as neoplasms of B-cells. In 16 cases of Hodgkin's disease of the nodular sclerosis and mixed cellularity types, all major classes of lymphoid cells were present. The proliferating cells were typed as B-cells in four cases of immunoblastic lymphadenopathy.

Adult↗

[The SPHA technic (solid phase hemadsorption) in syphilis serology. A year's experience in routine laboratory procedure].

The first results in routine syphilis serology with the new solid-phase hemadsorption assay (SPHA) are presented. Sera (63019) were screened by the VDRL test and by the automated hemagglutination assay with Treponema pallidum antigen (AMHA-TP). Samples reactive to one or both of these tests were further examined by the FTA-ABS test, the IgM-FTA-ABS test, the 19S IgM-FTA-ABS test (IgM-FTA-ABS test performed with the 19S fraction of the serum after gelfiltration) and the IgM-SPHA test. IgM-SPHA reactive sera derived from patients who were in the early stage or in the phase of latency (of undefinable duration), further from patients with active neurosyphilis and from 26 patients after reinfection. Erroneous results in the IgM-FTA-ABS test caused by a competitive inhibition of IgM by IgG can be eliminated by the IgM-SPHA method. The agreement between 19S IgM-FTA-ABS test and IgM-SPHA test is above 96.3%. As compared to the fluorescence test, the IgM-SPHA test is simpler to perform and can be easily automated.

Adult↗

[Observation on application of solid phase indirect hemadsorption assay for detection IgM antibodies to tuberculosis in non-serum specimens].

We used solid phase indirect hemadsorption assay (SPIHA) to detect specific IgM antibodies in specimens from cerebrospinal fluid, pleural fluid and ascitic fluid in patients with tuberculosis and compared with the enzyme linked immunosorbent assay (ELISA) and indirect hemagglutination assay (IHA). The result showed that detection of tuberculous specific IgM antibodies in cerebrospinal fluid by SPIHA is important for diagnosing the tuberculous meningitis.

Antibodies, Bacterial↗

African swine fever virus EP153R open reading frame encodes a glycoprotein involved in the hemadsorption of infected cells.

The open reading frame EP153R, located within the EcoRI E' fragment of the African swine fever (ASF) virus genome, is predicted to encode a membrane protein of 153 amino acids that presents significant homology to the N-terminal region of several CD44 molecules. EP153R contains multiple putative sites for N-glycosylation, phosphorylation, and myristoylation, a central transmembrane region, a C-type animal lectin-like domain, and a cell attachment sequence. Transcription of EP153R takes place at both early and late times during the virus infection. The disruption of the gene, achieved by insertion of the marker gene LacZ within EP153R, did not change either the in vitro virus growth rate or the virus-sensitive/resistant condition of up to 17 established cell lines, but abrogated the hemadsorption phenomenon induced in ASF virus-infected cells. As the sequence and expression of the ASF virus protein pEP402R, a CD2 homolog responsible for the adhesion of erythrocytes to susceptible cells, was unaffected in cultures infected with the EP153R deletion mutant, we conclude that the gene EP153R is needed to induce and/or maintain the interaction between the viral CD2 homolog and its corresponding cell receptor.

African Swine Fever Virus↗

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↗

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↗

Colony opacity, hemadsorption, hemolysis, and mitogenicity are not associated with virulence of Mycoplasma pulmonis.

Colony opacity, hemadsorption and hemolysis of erythrocytes, and the ability of whole mycoplasmal cells to induce a blastogenic response when incubated with C3H/HeN or C57BL/6 mouse lymphocytes were examined for 18 strains of Mycoplasma pulmonis to determine if any of these characteristics could be associated with virulence in vivo. Although there were differences among strains in each of these characteristics, none of these parameters were associated with virulence.

Animals↗

A spontaneous hemadsorption-negative mutant of Mycoplasma pneumoniae exhibits a truncated adhesin-related 30-kilodalton protein and lacks the cytadherence-accessory protein HMW1.

A spontaneous, hemadsorption-negative mutant of Mycoplasma pneumoniae lacks the cytoskeleton-forming HMW1 protein and exhibits a truncated adhesin-related 30-kDa protein. Genetic analyses revealed deletion of one nucleotide in the hmw1 gene and loss of eight repeated sequences comprising 144 nucleotides in the gene for the adhesin-related 30-kDa protein.

Adhesins, Bacterial↗

Hemadsorption of mumps virus examined by light and electron microscopy.

The hemadsorption (HAD) reaction of chick embryo cells infected with mumps virus was studied by means of light and electron microscopy, with special reference to the plasma membrane of the infected cell. The concomitant observation of membrane-free aggregates of viral nucleocapsid in the cytoplasm and attached red blood cells on the surface of the same cell indicated that only infected cells hemadsorbed and that hemagglutinin is confined within the infected cell. The attachment of red blood cells to morphologically intact cell membrane prior to its differentiation into viral envelope suggested that the HAD phenomenon, dependent on the presence of hemagglutinin, was independent of the viral maturation process. The gap of low electron density normally separating the morphologically intact membrane of the tissue culture cell and that of the red blood cell at the binding site was replaced by newly formed surface projections in HAD involving a segment of differentiated plasma membrane.

Adsorption↗

An adhesion-hemadsorption inhibition test for the detection of serum antibody to Mycoplasma gallisepticum.

A simple adhesion-hemadsorption inhibition (AHAI) test was developed for the detection of antibodies to Mycoplasma gallisepticum in the chicken sera. The AHAI antibody was detected simultaneously with HI antibody from sera of chickens intratracheally inoculated with viable cells of M. gallisepticum. A good correlation between HI and AHAI antibody titers was obtained with 382 (84.7%) of 451 sera from chickens reared on farms spontaneously contaminated with M. gallisepticum, whereas the remainder, 69 sera, was positive for HI but negative for AHAI test. It was not apparent whether the latters exhibited a non-specific reaction or the discrepancy was due to the lower sensitivity of AHAI reaction. The AHAI test does not require a great amount of antigen, special reagents or instruments, or pre-absorption treatment of test sera, and, therefore, it may serve as a simple serological test for detecting antibodies to M. gallisepticum.

Animals↗

The Point Mutations in Hemagglutinin Gene of Measles Virus Are Responsible for Alteration in Hemadsorption.

Three current strains of measles virus (MV) Fu, IMA and SMD, isolated using B95a cell line, were adapted to Vero cells. After adaption, these strains obtained the ability to agglutinate African green monkey red blood cells (AGM-RBC). We compared hemagglutinin gene sequences and fusion gene sequences of these two kinds of viruses. Fu strain has an Asn (HAD negative) to Tyr (HAD positive) substitution at position 481 in the hemagglutinin glycoprotein. IMA strain has an Asp(HAD negative) to Asn(HAD positive) substitution at position 14 and a Ser(HAD negative) to Gly (HAD positive) substitution at position 546 in the protein. F gene was confirmed identical between viruses. Our data show that two mutations, Asn to Tyr at position 481 and Ser to Gly at position 546 in hemagglutinin glycoprotein are responsible for alteration in hemadsorption.

Journal Article↗