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PubMed1990

Immunogenicity of glutaraldehyde inactivated pertussis vaccine.

The immunogenicity of different types of glutaraldehyde inactivated pertussis vaccine (GIPV) preparations made by inactivation of Bordetella pertussis organisms with different concentrations of glutaraldehyde for variable periods at room temperature and conventional heat inactivated pertussis vaccine (HIPV) was evaluated with regard to production of agglutinins and neutralizing antibodies against pertussis toxin (PT). The different types of GIPV preparations had variable intracerebral mouse potency which depended upon the conditions of inactivation with glutaraldehyde. The agglutinin production in mice against various types of GIPV preparations and HIPV were very similar irrespective of the potency of the preparations. The agglutinins were produced against all the three major agglutinogens 1, 2 and 3 as the preparations were made from B. pertussis strain 10536 (serotype 1,2,3). The titres of agglutinins produced against adjuvanted preparations were slightly higher than those against non-adjuvanted preparations. Neutralizing antibodies against PT were produced for eight out of nine types of GIPV preparations while these antibodies were not produced for conventional HIPV.

Agglutination Tests↗

PubMed1979

Serological response of patients with non-gonococcal urethritis to causative organism of contagious equine metritis 1977.

The presence of allugtinins to the causative organism of contagious equine metritis (C.E.M.) in human serum has been confirmed. Agglutinins were found in the serum of 84 (37.6%) of 223 patients with non-gonococcal urethritis (N.G.U.), and in 12.5% of these patients there was a four-fold or greater rise in titre during the course of their illness. There was no evidence that these agglutinins were the result of infection by chlamydiae or ureaplasmas. Certain patients with these agglutinins seemed to respond better to therapy with antibiotics to which the C.E.M. bacterium is susceptible in vitro than did patients in whom these agglutinins were not found. The findings suggest that the C.E.M. bacterium or a microorganism related to it may be aetiologically involved in a proportion of patients with N.G.U. A search for such an organism in these patients is in progress.

Agglutination Tests↗

PubMed1979

Cross-protection against Salmonella enteritidis infection in mice.

Mice were immunized subcutaneously with live vaccines and live vaccines with complete adjuvant incorporating Salmonella enteritidis Se 795, Salmonella typhi-murium M206, Salmonella gallinarum 9R or Salmonella pullorum Sp223. They were challenged along with unvaccinated controls with 100 LD50 of virulent S. enteritidis 5694 SMR subcutaneously on the 21st day post-vaccination. The humoral immune response was studied by assessing the sequential level of agglutinins, complete and incomplete somatic antibodies, opsonophagocytic antibodies, cytophilic antibodies and bactericidal antibodies before and after challenge. The level of these antibodies and the protection afforded by the particular vaccine is correlated and the possible involvement of a humoral mechanism is discussed.

Adjuvants, Immunologic↗

PubMed1982

Study of agglutinins to Brucella abortus, B canis and Actinobacillus equuli in horses.

Horses at a veterinary teaching hospital and a slaughterhouse were surveyed for antibodies to Brucella abortus, B canis and Actinobacillus equuli. Four of the 141 hospitalised horses and none of the 73 slaughtered horses had titres of 1:100 or greater to B abortus. Six horses of both populations reacted to the card test. One was culture positive. A card test using B canis antigen was positive in 38 per cent of the sera from hospitalised horses and all of the slaughtered horses. Twenty (27.4 per cent) of the latter group had high tires in a tube agglutination test. High titres could not be reduced by 2-mercaptoethanol serum treatment. The titres appeared to be associated with advanced age but not to sex. Adsorption of sera with B canis did not affect titres to A equuli but the reverse was true.

Abattoirs↗

PubMed1992

An agglutinin with unique specificity for N-glycolyl sialic acid residues of thyroglobulin in the hemolymph of a marine crab Scylla serrata (Forskal).

A novel agglutinin with specificity for sialic acid sequence of sugars in thyroglobulin is identified in the hemolymph of Scylla serrata. The physico-chemical characteristics of its binding affinity, such as pH and temperature optima, and cationic requirements are defined. N-glycolyl neuraminic acid (NeuGc) (at 0.6 mM), in contrast to N-acetyl neuraminic acid (NeuAc) (at greater than 5.0 mM), is the potent inhibitor of hemagglutination. Bovine and porcine thyroglobulins containing NeuGc, inhibited the agglutination. NeuGc-acid glycoprotein fraction (bovine) but not NeuAc-acid glycoprotein fraction (human) inhibited the hemagglutination. The inability of other NeuGc-glycoproteins (bovine submaxillary mucin) to inhibit the agglutination suggests that the agglutinin may also recognize glycosidic linkage associated with NeuGc. The potential of the agglutinin in identifying NeuGc containing human tumor associated antigens is discussed.

Adsorption↗

PubMedDate not supplied

An EDTA-associated anti-B agglutinin: the role of ionized calcium.

BACKGROUND: It is believed that EDTA-dependent panagglutination is associated with free carboxylic acids that support reactions of rare autoagglutinins. CASE REPORT: An ABO typing discrepancy occurred in an 88-year-old patient. The specificity of his autoagglutinin was demonstrated by panel cell study and absorption tests using normal donors' red cells or immunoadsorbents coated with A, B, or O substances. Inhibition assays were performed to determine whether the autoagglutinin was inhibited by ionized calcium or carboxylic acids. The autoagglutinin had anti-B specificity when tested in the presence of EDTA. It was neutralized by group B secretor saliva and adsorbed by crystalline silica coated with simple B substances with or without EDTA, although it was absorbed by group B red cells only in the presence of EDTA. The agglutinating activity was stronger at 25 degrees C (titer 64) than at 37 degrees C (titer 16) and was destroyed by treatment of the serum with dithiothreitol, which suggests that the autoagglutinin is IgM. This activity also appeared in the patient's serum after dialysis and in an eluate obtained after adsorption with simple B substances, and it was inhibited by the addition of CaCl2 at 0.5 mM or higher concentrations. This suggests that the agglutination is not dependent on EDTA but, rather, on the concentration of ionized calcium. The autoagglutinin failed to react with group B red cells treated with glutaraldehyde for 10 minutes. CONCLUSION: An anti-B autoagglutinin was shown to have caused an ABO typing discrepancy in the presence of EDTA. These results suggest that autoagglutination requires an environment with low levels of ionized calcium, but not the presence of carboxyl groups.

ABO Blood-Group System↗

PubMed1973

The relationship of concanavalin A binding to lectin-initiated cell agglutination.

We have investigated the relationship of concanavalin. A binding to the cell surface of normal and transformed cells and the subsequent agglutination of the transformed cells. At room temperature almost no differences could be detected in agglutinin binding between transformed and untransformed cells. At 0 degrees C, however, where endocytosis was negligible, the transformed cells bound three times more agglutinin. However, transformed cells and trypsin-treated normal cells do not agglutinate at 0 degrees C although the amounts of agglutinin bound at 0 degrees C are sufficient to permit agglutination when such cells are shifted up to room temperature. Both transformed and trypsin-treated normal cells show a marked increase in agglutination at 15 degrees C as compared to agglutination at 0 degrees C. From this, as well as the observation that mild glutaraldehyde fixation of the cell surface inhibited agglutination but not agglutinin binding, it was concluded that concanavalin A-mediated cell agglutination requires free movement of the agglutinin receptor sites within the plane of the cell surface.

Animals↗

PubMed1983

The biochemical and ultrastructural effects of tunicamycin and D-glucosamine in L1210 leukemic cells.

Tunicamycin was found to specifically inhibit the incorporation of a number of sugars into L1210 leukemia cell glycoproteins. This inhibition of glycoprotein biosynthesis led to a cessation of cell growth which was reversible in a dose-dependent and time-dependent manner. After removal of the antibiotic from L1210 cell cultures resumption of sugar incorporation preceded that of thymidine incorporation and the recovery of cell growth. The treatment of cells with tunicamycin resulted in a significant increase in the intracellular pool of UDP-N-acetylglucosamine which occurred concurrently with alterations in cell ultrastructure including distentions of the endoplasmic reticulum and nuclear membranes. Similar ultrastructural changes and increases in the intracellular pools of UDP-sugars were observed in L1210 cells exposed to 5 mM D-glucosamine, which suggested that the antiproliferative effects of tunicamycin may be related to the accumulation in the endoplasmic reticulum of one or more nucleotide sugar precursors of asparagine-linked glycoprotein biosynthesis. However, the biological effects of tunicamycin could be distinguished from those caused by D-glucosamine. Exposure of L1210 cells to tunicamycin resulted in specific alterations in the biochemical composition of the plasma membrane and in the inhibition of cellular agglutination by wheat germ agglutinin which were not apparent following exposure to equitoxic concentrations of the aminosugar. These studies, together with those which demonstrated that recovery of the cellular capacity to synthesize glycoproteins was obligatory for the recovery of cellular proliferation in tunicamycin-treated cells, suggested that inhibition of the synthesis of glycoproteins was the major factor limiting L1210 leukemic cell proliferation.

Agglutination↗

PubMedDate not supplied

Separation of sub-populations of sperm with higher fertility potential from normal and pathological semen by peanut agglutinin.

Incubation of human sperm from semen with counts below 20 x 10(6)/ml with peanut agglutinin (PNA) resulted in agglutination of about 70% of sperm. An inverse correlation was found between non-agglutinating (PNA) sperm and sperm density (r = 0.48, p less than 0.01) and a direct correlation with acrosome-damaged sperm (r = 0.83, p less than 0.001). Binding of 125I-PNA to sperm revealed high, and possibly also low affinity binding sites on sperm from both normozoospermic and oligozoospermic origins. Quantitative transmission electron microscopy analysis revealed that agglutinated, PNA+, sperm had lower frequency of acrosomeless spermatozoa than PNA(-)-sperm (44% versus 77%) and higher fertility score (+0.18 +/- 0.1 and -1.5 +/- 0.87 respectively, p less than 0.01). Removal of PNA- sperm from oligozoospermic semen may increase the fertility score of the remaining sperm.

Cell Separation↗

PubMed1987

Metacyclogenesis is a major determinant of Leishmania promastigote virulence and attenuation.

The in vivo virulence patterns of promastigote populations defined on the basis of agglutination by the lectin peanut agglutinin (PNA) were studied for various cloned lines of Leishmania major. Promastigotes derived from logarithmic-phase cultures, which were routinely 100% agglutinated at 100 micrograms of PNA per ml, were relatively avirulent for BALB/c mice. The relative virulence of stationary-phase promastigotes appeared to be attributable to the proportion of nonagglutinable (PNA-) promastigotes contained within these populations. Purification of PNA- organisms from stationary cultures provided for each clone the most virulent inoculum, supporting the view that this change in lectin binding accurately reflects the development of infective metacyclic stage promastigotes. By studying this marker, we found that there was considerable variation in the degree to which different strains and clones underwent metacyclogenesis during growth. Examination of a reportedly avirulent L. major clone revealed that metacyclogenesis was unusually delayed and inefficient for this clone, but that those PNA- promastigotes which could be recovered from late-stationary-phase cultures were virulent for BALB/c mice. The loss of virulence associated with frequent subculture could also be attributed to a drastic diminution in metacyclogenesis potential over time. A clone which yielded over 90% PNA- promastigotes during growth within passage 1 generated fewer than 10% PNA- promastigotes during growth by passage 94. Subcloning of late-passage attenuated promastigotes yielded a clone for which no PNA- promastigotes could be generated during growth, and an infective population could not be derived from this clone. Thus, metacyclogenesis does not appear to be stable for even cloned lines of Leishmania promastigotes, and virulence comparisons between different strains and clones can be meaningfully made only if the metacyclic populations contained within the respective inocula are determined.

Agglutination↗

PubMed1979

Lectin-binding proteins in central-nervous-system myelin. Binding of glycoproteins in purified myelin to immobilized lectins.

The capacities of immature and mature rat brain myelin, bovine myelin and human myelin to be agglutinated by soya-bean agglutinin, Ricinus communis agglutinin, wheatgerm agglutinin, and Lotus tetragonolobus agglutinin were examined. The first two lectins, which are specific for galactose and N-acetylgalactosamine, strongly agglutinated immature and mature rat myelin, weakly agglutinated bovine myelin, but did not affect human myelin. The other myelin and lectin combinations resulted in very weak or no agglutination. [(3)H]Fucose-labelled glycoproteins of purified adult rat brain myelin were solubilized with sodium dodecyl sulphate and allowed to bind to concanavalin A-Sepharose and each of the other lectins mentioned above, which had been immobilized on agarose. About 60% of the radioactive fucose was in glycoproteins that bound to concanavalin A-Sepharose and these glycoproteins could be eluted with solutions containing methyl alpha-d-mannoside and sodium dodecyl sulphate. Periodate/Schiff staining or radioactive counting of analytical gels showed that most of the major myelin-associated glycoprotein (apparent mol.wt. approx. 100000) bound to the concanavalin A, whereas the glycoproteins that did not bind were mostly of lower molecular weight. Preparative polyacrylamide-gel electrophoresis of the glycoprotein fraction that was eluted with methyl alpha-d-mannoside yielded a relatively pure preparation of the myelin-associated glycoprotein. Similar results were obtained with each of the other lectins, i.e. the myelin-associated glycoprotein was in the fraction that bound to the immobilized lectin. Double-labelling experiments utilizing [(3)H]fucose-labelled glycoproteins from adult myelin and [(14)C]fucose-labelled glycoproteins from 14-day-old rat brain myelin did not reveal any difference in the binding of the mature and immature glycoproteins to any of the immobilized lectins. The results in this and the preceding paper [McIntyre, Quarles & Brady (1979) Biochem. J.183, 205-212] suggest that the myelin-associated glycoprotein is one of the principal receptors for concanavalin A and other lectins in myelin, and that this property can be utilized for the purification of this glycoprotein.

Agglutination↗

PubMed1986

Role of galactosyltransferase activity in phage sensitivity and nodulation competitiveness of Rhizobium meliloti.

A stock culture of Rhizobium meliloti 102F51 contains colonies of two distinct phenotypes (Handelsman et al., J. Bacteriol. 157:703-707, 1984); one colony type is agglutinated by high dilutions of the alfalfa agglutinin, is sensitive to phage F20, and is resistant to phage 16B, and the other is agglutinated only by low dilutions of the alfalfa agglutinin, is resistant to phage F20, and is sensitive to phage 16B. Cells of the latter phenotype have an inner-membrane-bound galactosyltransferase activity that transfers galactose from UDP-galactose to a water-insoluble anionic polymer. This enzymatic activity is absent in cells of the first phenotype. All of the phage 16B-resistant mutants selected from a sensitive strain were agglutinated by high dilutions of the alfalfa agglutinin, were sensitive to phage F20, and lacked galactosyltransferase activity. The galactose-containing polymer prepared in vitro was immunologically cross-reactive with the cell surface.

Agglutination↗

PubMed1974

Pertussis agglutinins in vaccinated children: better response with adjuvant.

Children were immunized with a single batch of pertussis vaccine, either adsorbed on aluminium hydroxide or plain. With a primary course of three injections, adsorbed vaccine produced higher titres of pertussis agglutinins in the serum than did plain vaccine. There was no obvious difference in response between those who received the three doses at intervals of 1-2 months, starting at 3-4 months of age, and those in whom the third dose was delayed until about 6 months after the second, but the number of children in each group was small.

Adjuvants, Pharmaceutic↗

PubMed1984

Induction in rats of serum agglutinins to Eubacterium, Peptostreptococcus and Coprococcus species by the faecal flora from patients with Crohn's disease and healthy subjects.

Sera from patients with Crohn's disease more often than those from other subjects contain agglutinins against anaerobic coccoid rods, identified as Peptostreptococcus productus, Eubacterium contortum (two strains) and Coprococcus comes. The presence of antigens of the four coccoid rods in faeces of patients with Crohn's disease and healthy subjects was investigated by inoculation of rats with faecal suspensions. Antigens of the coccoid rods were detected in faeces from both patients with Crohn's disease and healthy subjects.

Agglutination Tests↗

PubMed1977

Evaluation of candida precipitin and agglutinin tests for the diagnosis of systemic candidiasis in burn patients.

Candida agglutinin and precipitin tests were performed on a group of burned patients without evidence of systemic candidiasis to determine whether tests could be used to distinguish between colonization and systemic disease, as has recently been reported. There were considerable numbers of false-positive results using both tests. These false-positive results occurred regardless of the candida colonization status of the patient. We conclude that these tests are of little diagnostic value in burn patients.

Agglutination Tests↗

PubMed1982

Abnormal blood group galactosyltransferase in blood type A1B-subjects whose sera contain anti-B agglutinin.

A blood type A1B female, whose plasma agglutinates B red blood cells from other subjects but not her own, was found. Her sister with blood type A1B (sister-1) exhibited the same peculiarity. The agglutination titer of red cells from these two subjects is lower than that of normal B. Her father is blood type B, and his plasma did not agglutinate B red cells from other subjects and his own. Her mother and another sister (sister-2) were usual blood type A1. Blood group galactosyltransferase (B-enzyme) activity of plasma from the propositus, sister-1, and their father was very low. Km for 2'-fucosyllactose of B-enzyme of these subjects was 1.3 mM, which was more than two times higher than that of the normal value. Km for UDP-Gal was similar, but the pH-activity profile differed for the two enzymes. Red cell membranes from her father contained about 70% ungalactosylated H-sites, whereas, virtually all H-sites are galactosylated in the usual B red cell membranes. The blood group ABH components are known to be heterogeneous. Because of the abnormal B-enzyme with low activity and low substrate affinity, some H components might not be galactosylated, particularly in A1B cases (i.e., the propositus and her sister-1), due to competition by A1 enzyme. The lack of certain B components is likely to be the cause of the existence of the anti-B agglutinin in their sera.

ABO Blood-Group System↗

PubMed1988

Monoclonal antibodies directed against the sexual binding site of Chlamydomonas eugametos gametes.

Monoclonal antibodies were raised against the mt- sexual agglutinin of Chlamydomonas eugametos gametes. Those that blocked the agglutination site were selected. They were divided into two classes dependent upon whether they gave a weak (class A) or clear positive (class B) reaction with mt- flagellar membranes in an ELISA and an indirect immunofluorescence test using glutaraldehyde-fixed mt- gametes. Class A antibodies were shown to be specific for the agglutinin in an extract of mt- gametes, based on results from immunoblotting, immunoprecipitation, affinity chromatography, and the absence of a reaction with nonagglutinable cells. Surprisingly, class A mAbs also recognized two mt+ glycoproteins, one of which is the mt+ agglutinin. Class B antibodies were shown to bind to several glycoproteins in both mt- and mt+ gametes, including the mt- agglutinin. Fab fragments from class A mAbs blocked the sexual agglutination process, but those from class B did not, even though the parent antibody did. We conclude that the class A epitope lies in or close to the agglutination site of the mt- agglutinin, whereas the class B epitope lies elsewhere on the molecule. We also conclude that the mt- agglutinin is the only component on the mt- flagellar surface directly involved in agglutination. Class A mAbs were found to elicit several reactions displayed by the mt+ agglutinin. They bound to the mt- agglutinin on gamete flagella and induced most of the reactions typical of sexual agglutination, with the exception of flagellar tip activation. None of these reactions was induced by Fab fragments. High concentrations of class A mAbs completely repressed the sexual competence of live mt- gametes, but low concentrations stimulated cell fusion.

Agglutination↗

PubMed1976

The stability of the serotypes of Bordetella pertussis with particular reference to serotype 1,2,3,4.

Strains of Bordetella pertussis in which all the organisms contain agglutinogens 1 and 3 or 1,2 and 4 are easy to identify as serotypes 1,0,3,0 and 1,2,0,4 respectively; and similarly, stable strains of serotype 1,0,3,4 are occasionally found. During repeated subcultures, passage in vivo, and lyophilization and preservation for many years, these serotypes do not change. Mixing 1,0,3,0 and 1,2,0,4 serotypes and culturing them together in vivo and in vitro produces cultures from which organisms of the same two serotypes can be isolated. In contrast, strains which type as 1,2,3,4 are often a heterogeneous group. We have attempted to classify these as "stable", "variable" and "mixed" cultures. Some strains comprise organisms all of which contain the four agglutinogens and are as easy to type as the strains described above. These we have called "stable" 1,2,3,4 strains. Other 1,2,3,4 strains are made up of colonies possessing all four agglutinogens, as shown by agglutinin production, but in amounts varying from day to day so that direct typing is inconsistent. These we have called "variable" 1,2,3,4 strains. The last category, "mixed", is made up of organisms most of which give rise to stable 1,2,3,4 cultures; a few of the component organisms, however, have one or two of the four agglutinogens missing. The importance of the "variable" cultures is emphasized for work on apparent change of serotype, e.g. during infection.

Agglutination Tests↗