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PubMedDate not supplied

Hemagglutination inhibition studies for the evaluation of blood group antigens in ethanol soluble substances (ESS) obtained from human, baboon and vervet monkey red blood cells.

Soluble blood group substances, isolated from the red blood cells of humans, baboons, and vervet monkeys by ethanol extraction, possessed serologically active specificities for the following antigens: A, B, H, Lea, LebL, P, P19 Pk and I. Human red blood cells lacking any of these specificities by the direct hemagglutination test also lacked the related antigens in their soluble extract. The only exception was in "Bombay" Oh cells, from which soluble H substance could be readily isolated. Soluble substances obtained from baboon and vervet monkey red blood cells, which lack the human variety of A, B, and H antigens on their red blood cells, inhibited both human and lectin anti-H reagents. The detection of "hidden" H activity in Oh cells will pose some important questions regarding membrane characteristics and the role of immune surveilance.

ABO Blood-Group System↗
PubMedDate not supplied

The effects of red blood cell suspending media on hemagglutination and the antiglobulin test.

Evaluation of the effects of four suspending media for test cells in antibody detection and identification is reported. The media under investigation are normal saline, low ionic strength salt solution, 22% bovine albumin and polymerized albumin. Hemagglutination at 37 C was enhanced in the test systems employing polymerized albumin. With shortened incubation times, the antiglobulin test was enhanced in tests against red blood cells suspended in the low ionic strength salt solution. When incubation times were extended to 30 minutes, all suspending media were apparently equally effective.

Antibodies↗
PubMed1999

Rapid phenotyping of HPA-1a using either diabody-based hemagglutination or recombinant IgG1-based assays.

BACKGROUND: The HPA-1 system is carried on the beta3 integrin. HPA-1a (Zw(a), Pl(A1)) is immunogenic in an HPA-1b homozygote (HPA-1b1b). In pregnancy, 1 of 365 women forms anti-HPA-1a, which causes severe thrombocytopenia in 1 in 1100 neonates. Identification of women at risk of forming anti-HPA-1a and the screening of donors to obtain HPA-1a-negative platelets for therapy need reliable, low-cost, automated assays. STUDY DESIGN AND METHODS: A diabody with dual specificity for HPA-1a x D and an IgG1 anti-HPA-1a have been constructed by the use of the genes encoding the first anti-HPA-1a fragment. With these reagents, two complementary HPA-1a phenotyping assays have been developed. RESULTS: This diabody was used in a simple hemagglutination technique to perform HPA-1a phenotyping on soluble glycoprotein IIb/IIIa from EDTA plasma samples. Over 1000 unselected donors have been correctly HPA-1a-phenotyped by use of the diabody. The human recombinant IgG1 anti-HPA-1a was produced in a rat myeloma cell line and was fluorescein labeled for use in a whole-blood flow cytometric HPA-1a phenotyping assay. This IgG1 anti-HPA-1a shows a clear differential between HPA-1a-positive and HPA-1a-negative platelets at nM antibody concentrations. CONCLUSIONS: The two recombinant reagents described are highly suitable for screening and confirmatory HPA-1a phenotyping. They permit rapid determination of the HPA-1a phenotype and are amenable to automation.

Antigens, CD↗
PubMed2000

Inhibition of Helicobacter pylori hemagglutination by polysaccharide fractions from roots of Panax ginseng.

Polysaccharides from the roots of Panax ginseng were extracted by hot water and fractionated by using ethanol precipitation and ion exchange chromatography. Fractions FC (crude extract), F1 (fraction precipitated by ethanol), F1N (fraction unbound to DEAE-Sepharose CL-6B), and F1A (bound fraction) were obtained. Their carbohydrate analyses showed that acidic fraction F1A contains higher amounts of galactose, arabinose and uronic acids, in comparison to FC and F1. Fraction F1N mainly consists of glucose. The inhibition of Helicobacter pylori-induced hemagglutination revealed different inhibitory activities of these fractions. In particular, acidic fraction F1A showed a remarkable inhibitory activity (minimum inhibition concentration was 0.25 mg/ml) among the polysacharide fractions. However, digestion of the fraction F1A with pectinase resulted in a lower molecular weight oligosaccharide fraction F1AP which was non-inhibitory at the concentration of 4 mg/ml. Comparison of inhibitory activities and carbohydrate compositions of isolated fractions indicates that the activity correlated with the contents of galactose, arabinose, and uronic acids. These data suggest that acidic polysaccharides may be responsible for the inhibitory activity.

Helicobacter pylori↗
PubMed1997

Bispecific rabbit Fab'-bovine serum albumin conjugate used in hemagglutination immunoassay for beta-microseminoprotein.

A polyclonal bispecific (bifunctional) antibody was prepared to develop a hemagglutination immunoassay for beta-microseminoprotein (beta-MSP), a predominant seminal protein. Three types of F(ab')2 fragments of rabbit IgG, affinity-purified anti-human red blood cell (RBC) F(ab')2 nonaffinity-purified anti-beta-MSP F(ab')2 and nonspecific (nonimmunized) F(ab')2, were mixed to obtain a F(ab')2 mixture containing 10% anti-RBC molecules and 10% anti-beta-MSP molecules. Fab' was obtained from the F(ab')2 mixture, and then reacted with maleimide-activated bovine serum albumin (BSA) at a molar ratio of 10:1. As estimated by the decrease in the maleimide content, approximately 7 Fab' molecules were introduced per one BSA molecule. The bispecific (anti-beta-MSP and anti-RBC) Fab'-BSA conjugate thus prepared was incubated successively with a human RBC suspension and with samples. In the presence of beta-MSP, RBCs become agglutinated, providing a test simple for forensic semen identification.

ABO Blood-Group System↗
PubMed1992

Effect of biological and synthetic polymers on BK virus infectivity and hemagglutination.

The effect of several biological and synthetic polymers, chosen on the basis of different physical and chemical properties, was investigated on BK virus infectivity and hemagglutination. It was observed that polyanions like mucin, dextran sulfate and heparin depressed the viral binding, whereas polycations had no significant activity, with the exception of poly-L-lysine, which enhanced it. The effect of the active polymers was studied in different experimental conditions and the results obtained suggested that polyanions may act directly on the virus particle, whereas the target of polycations could be at the level of cell membranes. However, the effect shown by the active compounds did not appear to be simply related to the electric charge since neutral compounds, such as tamarind gum and locust bean gum, showed a marked inhibitory effect on BK virus binding to the cells.

BK Virus↗
PubMed1993

Reverse passive hemagglutination tests for rapid diagnosis of snake envenomation.

Reverse passive hemagglutination (RPHA) tests for the detection of six major poisonous snake venoms of Thailand were studied. Three different species of red blood cells i.e., sheep (SRBC), human (HRBC) and chicken (CRBC) were sensitized with protein A-affinity purified rabbit antivenom IgG using chromic chloride as a coupling reagent. The properties of these sensitized erythrocytes with regard to sensitivity, specificity, stability to venom enzymes and storage etc., were studied and compared. The sensitivities of the RPHA tests in venom detection were 2 to 635 ng/ml. Cross-reactions were observed with heterologous venoms at concentrations at least 62 times higher than those observed with homologous venoms. After treatment with glutaraldehyde, the coupled red blood cells showed reduced sensitivity but were stable at 4 degrees C from 1 to 12 months depending upon the antibody and the species of erythrocytes. The entire test required 60 to 120 min. The RPHA using fresh SRBC correctly identified various venoms in 48 of 59 (81.3%) serum samples and 16 of 26 (61.5%) wound swabs. Venom mis-identifications were made in 2 sera (3.4%). In a comparison of 24 paired serum and wound swab samples, more positive identifications were made with serum than with swab samples but the difference was not statistically significant (p > 0.05).

Animals↗
PubMed1959

Studies on the immunochemistry of human low density lipoproteins utilizing an hemagglutination technique.

Hemagglutination is a specific and sensitive technique for investigating the purity of lipoproteins and the immunologic relationships between low density lipoprotein fractions. The S(f) 10-400 and S(f) 3-9 lipoprotein fractions, isolated from human serum by dextran sulfate-density gradient centrifugation procedure and repurified by centrifugation appeared to contain only lipoprotein antigens since these fractions did not stimulate the production of antibodies against other serum proteins. Cross-absorption experiments with lipoproteins carried on "tanned" cells demonstrated that the S(f) 3-9 lipoprotein fraction contains all the antigenic components of the S(f) 10-400 lipoprotein fraction together with additional antigenic components not found in the S(f) 10-400 lipoprotein fraction. Thus S(f) 3-9 and S(f) 10-400 lipoprotein fractions are immunologically similar but not identical. Low density lipoproteins contain no antigens in common with the high density lipoproteins. An S(f) 3-9 antiserum can be used to detect both S(f) 3-9 and S(f) 10-400 antigens. The S(f) 3-9 lipoprotein fraction used as an antigen will detect antibodies against both S(f) 3-9 and S(f) 10-400 lipoprotein fractions. The S(f) 3-9 and S(f) 10-400 antisera did not contain immune antibodies against erythrocytes of the different blood groups or against sheep, guinea pig, dog, calf, pig, horse, and chicken erythrocytes. Normal subjects and subjects with recent myocardial infarctions had no circulating autoantibodies against the S(f) 3-9 and S(f) 10-400 lipoprotein fractions.

Animals↗
PubMed1959

Studies of mouse polyoma virus infection. IV. Evidence for mucoprotein erythrocyte receptors in polyoma virus hemagglutination.

Treatment of guinea pig erythrocytes with types A and B influenza viruses rendered them inagglutinable by polyoma virus; also, the inhibitory effect of ovomucin on polyoma virus hemagglutination was destroyed by pretreatment of the ovomucin with various myxoviruses. These results indicate that polyoma virus and myxovirus erythrocyte receptor sites are identical. However, no destruction by polyoma virus of its own or of myxovirus receptors or inhibitors was detected. No serologic relationship was detected between polyoma virus and members of the myxovirus group; differences in size and stability further indicate their distinctness. No evidence was found of biologic or serologic relationship of polyoma virus with encephalomyocarditis virus or mouse encephalomyelitis virus.

Animals↗
PubMed1977

Hemagglutination by purified type I Escherichia coli pili.

Many enterobacteria can cause agglutination of erythrocytes, but previous investigations have not proven which components of the bacteria are responsible. We used a strain of Escherichia coli K12 which causes mannose-sensitive hemagglutination (HA) of guinea pig cells. Common pili were purified from these bacteria by shearing them from the bacteria followed by selective precipitation in acid and ammonium sulfate. Isopycnic centrifugation in cesium chloride removed the remaining outer membrane protein contaminants. These pili are pure by electron microscopy and gel electrophoresis. By amino acid analysis, they have a mol wt of 17,099 and consist of 45% nonpolar residues. These purified pili agglutinate guinea pig erythrocytes, a reaction that is inhibited by anti-pili antibodies and by saccharides related in structure to D-mannose. Proteolytic treatment of erythrocytes does not diminish HA but rather increases the pili-induced HA of human cells. Neuraminidase enhances HA and mannosidase slightly diminishes it. It is concluded that purified pili alone cause HA of erythrocytes by binding to mannose-like molecules on the erythrocyte surface. Thus HA by bacterial pili serves as a useful model system for the mechanism of bacterial pili attachment ot cell membranes.

Antibodies, Bacterial↗
PubMed1990

Monoclonal antibodies to bluetongue virus define two neutralizing epitopes and a hemagglutinating epitope.

Monoclonal antibodies were used to characterize neutralizing epitopes on VP2 of bluetongue virus serotype 10 (BTV-10). Six neutralizing monoclonal antibodies that immune precipitated VP2 demonstrated two distinct patterns of reactivity in the competitive enzyme-linked immune absorbent assay (ELISA). These results suggest that there are at least two distinct domains of neutralization on VP2 of BTV. Monoclonal antibodies defining the two domains were serotype-restricted in plaque neutralization, immune precipitation or ELISA. One of the two neutralizing domains also demonstrated significant hemagglutinating activity. Both cattle and sheep infected with BTV-10 produce antibodies to the two neutralizing epitopes.

Animals↗
PubMed1977

Hemagglutination as an alternate technic for the detection of melanoma-specific cytoplasmic antigens.

The extensive use of the single technic of immunofluorescence has resulted in the detection of an array of melanoma-specific antigens. Those antigens residing in the cytoplasm of the melanoma cell have been shown to cross-react across various melanoma cell lines in tissue culture, as well as across all fresh tumor cells isolated directly from patients. The ubiquitous nature of this antigenic activity, as defined by immunofluorescence, suggests that the presence of this activity may be important diagnostically. Isolation of one or more antigens from the cytoplasm, however, requires the development of an assay for antigenic activity in cell fractions. Data presented here indicate that passive hemagglutination provides as effective a means of visualizing antigenic activity as does immunofluorescence. Evidence showing that melanoma-specific antigenic activity is present as a soluble component in the cytosol and as a membrane-associated component of the endoplasmic reticulum is also presented.

Animals↗
PubMed1987

A new solid phase passive hemagglutination test for IgM antibody to hepatitis B core antigen.

There is a need for a simple, sensitive, specific, and inexpensive test for immunoglobulin M antibody to hepatitis B core antigen (anti-HBc IgM). A solid phase passive hemagglutination test (SP-PHA) was developed for this purpose and compared with the enzyme-linked immunosorbent assay (ELISA) test. Hepatitis B core antigen (HBcAg) used in PHA and SP-PHA was synthesized in Escherichia coli. Human IgM was captured to a microtiter plate coated with anti-human IgM, and the presence of anti-HBc IgM was demonstrated by the adherence of HBcAg-sensitized erythrocytes to the bottom of a U-shaped microtiter plate. ELISA and SP-PHA were made at 1:100 and 1:1,000 serum dilution, respectively. Both were positive in 100% of 36 cases of acute hepatitis B, 68.18% of 22 cases of chronic hepatitis B, and 20% of 75 healthy carriers of hepatitis B surface antigen (HBsAg) but none in 65 anti-HBc-positive blood donors that had negative results for HBsAg. Results of both tests were identical but were false positive because rheumatoid factor was found only in ELISA. End-point titration by SP-PHA and PHA was also found useful for the differentiation of acute hepatitis B from chronic hepatitis B and HBsAg carriers.

Enzyme-Linked Immunosorbent Assay↗
PubMed1977

Measurement of hemagglutination-inhibiting antibody to influenza virus in the 1976 influenza vaccine program: methods and test reproducibility.

Titers of hemagglutination-inhibiting (HAI) antibody were determined for all sera obtained from participants in the 1976 Influenza Vaccine Test Program. At least eight control sera were included in each test during the vaccine trial period for the purpose of monitoring HAI test reproducibility. Estimates of day-to-day reproducibility were defined as the percentages of duplicate aliquots of the same sera, tested on two separate days, having HAI antibody titers that did not differ by more than one twofold dilution. These reproducibility estimates ranged from 89% to 97% with influenza A/New Jersey/76 and A/Mayo Clinic/74 antigens. In contrast, within-day reproducibility estimates obtained from all sets of control sera ranged from 96% to 98%. Estimates of day-to-day test reproducibility obtained with selected sera taken after vaccination that were titrated on two differen days ranged from 90% to 98%. Geometric mean titers of these sera tested weeks or months apart differed on some occasions during the test period.

Antibodies, Viral↗
PubMed1977

Comparison of the hemagglutination-inhibiting and neutralizing antibody responses of volunteers given 400 chick cell-agglutinating units of influenza A/New Jersey/76 split-virus vaccine.

Thirty-two volunteers 25 years of age or older who were employees of the Laboratory of Infectious Diseases (National Institute of Allergy and Infectious Diseases, Bethesda, Md.) were given 400 chick cell-agglutinating units of influenza A/New Jersey/76 virus vaccine (Wyeth Laboratories, Philadelphia, Pa.) intramuscularly. Sera were assayed for hemagglutination-inhibiting (HAI) and neutralizing antibody before and after inoculation. All 32 subjects had a fourfold rise in titer of HAI antibody, and 25 of 32 had a similar rise in titer of neutralizing antibody. The average log2 rise was 4.5 for HAI antibody and 4.1 for neutralizing antibody. Therefore, this vaccine is believed to be capable of stimulating HAI and neutralizing antibodies to an equal extent.

Adult↗
PubMed1978

Neutralizing and hemagglutination-inhibiting antibodies to rubella virus as indicators of protective immunity in vaccinees and naturally immune individuals.

In general, hemagglutination-inhibiting (HAI) and neutralizing (NT) antibodies to rubella virus tend to parallel one another, and the presence of either antibody has been interpreted as reflecting protective immunity. It has recently been shown that persons who have HAI antibody but lack NT antibody may be subject to reinfection. In the present study these two antibodies were compared with regard to their appearance and persistence in vaccines and in those who have experienced natural infection. NT antibody appeared more slowly following immunization with HPV77DE5 or Cendehill vaccines than after natural infection or immunization with RA 27/3 vaccine. As with natural infection, initial NT antibody responses to RA 27/3 vaccine were of higher titer and persisted at higher levels for three years than was the case with either of the other vaccines. NT testing procedures were found to differ in sensitivity, depending on several factors including the cells in which the virus was grown.

Adult↗
PubMed1979

Evaluation of indirect hemagglutination and its inhibition in the differentiation between antibodies to herpes simplex virus types 1 and 2 for seroepidemiologic studies: use of a II/I index threshold of 85 and an assay of type-specific antibodies.

Two parameters, a II/I index threshold value of 85 and an assay of type-specific antibodies to herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), were assessed, for seroepidemiologic purposes, for their efficacy in the establishment of past infections with HSV-1 and HSV-2 as indicated by the results of tests for indirect hemagglutination and its inhibition. Neither criterion alone established past infection with either HSV-1 or HSV-2. However, by combination of the two parameters, antibodies to HSV-1 and HSV-2 could be differentiated for seroepidemiologic studies.

Adolescent↗