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Lack of correlation between growth characteristics, agglutinability by plant lectins and the malignant phenotype.

Wheat germ agglutinin and Concanavalin A were found to agglutinate a variety of normal hamster cells as well as a number of lines of transformed or tumor cells. The normal cells included preparations from embryos and neonatal organs, and the spontaneous line, BHK-21 C13. In order to explore more fully the situation in regard to the BHK cells, three sublines were isolated and studied. A "flat revertant" selected by FUdR treatment grew to diminished saturation density and failed to produce colonies in soft agar in contrast to the parent line, which did. Yet, it was only minimally less tumorigenic and less agglutinable than the parent line. Lines derived from one and four consecutive in vivo tumor passages showed increased tumorigenicity and growth in soft agar when compared to the parent line. Although the first passage line showed increased saturation density, the saturation density of the fourth passage line was similar to or less than that of the parent cells. Agglutinability of these lines was not significantly increased. These results indicate that, in contrast to widely accepted beliefs, tumorigenicity may correlate poorly with growth characteristics and agglutination by plant lectins.

Agglutination↗

Role of ABH blood group antigens in the stimulation of a DIDS-sensitive Ca2+ influx pathway in human erythrocytes by Ulex europaeus agglutinin I and a monoclonal anti A1 antibody.

Of eleven agglutinating lectins tested, only one, Ulex europaeus agglutinin I (UEA1), stimulated Ca2+ uptake in quin2-loaded erythrocytes by about 2-fold. UEA1 is known to be an alpha-L-fucose and ABH blood group specific lectin. The 45Ca2+ influx induced by UEA1 was absent in the presence of extracellular fucose (5 and 15 mM) and depended on the ABH blood group of the donor, the stimulatory potency of the lectin decreasing in the order H greater than A2 greater than A1. Ca2+ entry blockers, such as cobalt and verapamil, did not affect the 45Ca2+ influx induced by UEA1. 4,4'-Diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS) inhibited dose-dependently with a Ki of 1-2 microM. 10 microM DIDS, 10 microM 4,4'-dinitrostilbene-2,2'-disulfonic acid (DNDS) and 20 microM dipyridamole fully blocked the 45Ca2+ influx induced by UEA1. The effect of UEA1 on 45Ca2+ influx was absent in K+ and Mg2+ media and was less pronounced in choline than in Na+ media. The 45Ca2+ influx induced by the lectin was abolished by preincubation with 12-O-tetradecanoylphorbol 13-acetate (TPA, 60 ng/ml). A monoclonal antibody raised against A1 erythrocytes (Bric 54) accelerated 45Ca2+ influx in quin2 loaded A1 erythrocytes by about 2-fold. No effect was seen in A2 and H erythrocytes. The 45Ca2+ influx elicited by Bric 54 exhibited a sensitivity towards inhibition by DIDS and TPA, as well as a dependence on the cation composition of the incubation medium similar to that observed with UEA1. The effects of UEA1 and Bric 54 were not additive. These observations suggest that the Ca2+ influx induced by UEA1 and Bric 54 is mediated by the same transport pathway. Since both the lectin and the antibody exhibit ABH blood group specificity, it appears reasonable to conclude that ABH antigens can serve as recognition sites for activation of a Ca2+ influx pathway in human erythrocytes, which is sensitive to inhibitors of the band 3 anion-exchanger.

4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid↗

Studies on the specificity of agglutinins against human erythrocytes modified by Newcastle disease virus.

The specificity of agglutinins in human sera against human erythrocytes modified by Newcastle disease virus (NDV-O agglutinins) was studied with agglutination and agglutination inhibition techniques. Virus grown in both embryonic eggs and tissue culture was used in the experiments. The findings were compatible with the existence of four specificities of agglutinins. In some sera, including those from subjects recently inoculated with vaccines prepared in eggs, the agglutinins were directed against a component of the allantoic fluid. In high-titred infectious mononucleosis (IM sera the agglutinins were directed against antigenic groupings present in the Vic strain but not the B1 strain of NDV, whereas in certain other sera they were directed against antigens present in both of them. Finally, in some sera the agglutinins probably reacted with a red cell antigen uncovered by the viral neuraminidase.

Agglutinins↗

[Reactions of hereophilic agglutinins with L-cells and different toxicity of anti-A HP to fibroblasts and L-cells in vitro].

The carbohydrate binding glycoprotein Anti-AHP binds to the plasma membrane of the (transformed) L-cell both in agglutination reactions and by the immunofluorescence technique. On the surface of (normal) mouse embryo fibroblast cells Anti-AHP receptors were not detectable. Anti-AHP is suitable as another marker for L cells. The random distribution of the binding sites at 4 degrees C changes with elevated temperatures (clustering, capping of sites). Anti-AHP inhibits the growth of L cells much more than that of fibroblasts. It does not act as a mitogen for the investigated cells.

Agglutination Tests↗

Hemolytic anemia due to a mixture of low-titer IgG lambda and IgM lambda agglutinins reacting optimally at 22 degrees C.

A 65-year-old white man had severe hemolytic anemia due to a mixture of low-titer IgG lambda and IgM lambda agglutinins showing optimum reactivity at 22 degrees C. The IgG agglutinins were detected by manual indirect antiglobulin test (IAT) using anti-IgG, and had a titer of 1 at 37 degrees C, 128 at 22 degrees C and 16 at 4 degrees C against adult type O red blood cells (RBC). The corresponding titers with cord RBC were 1 (37 degrees C), 64 (22 degrees C) and 8 (4 degrees C). Proteolytic enzyme and neuraminidase treatment of RBC did not decrease these titers. No known specificity could be assigned to these agglutinins. The isolated agglutinins (recovered by cold adsorption, warm elution) were shown by immunoelectrophoresis to be IgG lambda antibodies. They did not bind complement in vitro, consistent with the finding that the patient had negative manual direct antiglobulin test (DAT) by anti-C3d. It could be shown only by automated IAT that patient's serum also contained IgM cold agglutinins which also reacted best at 22 degrees C and appeared to be of lambda light-chain type. The patient responded to corticosteroid therapy and remains well without treatment 14 months after the hemolytic episode. The presence of IgG cold agglutinins may be predictive of a favorable response to corticosteroid therapy.

Aged↗

[The role of IgM in paratyphoid immunity].

The authors analyze the results of studies on the effect produced by the immunization of calves with paratyphoid vaccine on the production of agglutinins, changes in the blood serum immunoglobulin levels, and the specificity of IgM to Salmonella dublin and Salmonella typhimurium antigens. The highest level of agglutinins to paratyphoid antigens in the blood sera of the immunized calves was registered on days 14-21 after immunization, changes in the levels of different immunoglobulin classes being insignificant. The agglutination test and the enzyme immunoassay have revealed that antibodies to S. dublin anb S. typhimurium antigens belong to IgM.

Agglutination Tests↗

[The possibility of use of amidinated human erythrocytes as agglutinin adsorbents in hemagglutination tests].

This study is concerned with the agglutinability of amidinated human erythrocytes. Amidinated erythrocytes behave like an immuno-adsorbent. Following the adsorption of an agglutinin, amidinated erythrocytes agglutinate specifically with native red blood cells or with native eukaryotic cells, if the glycocalyx of these native cells is provided with the respective agglutinin receptor groupings.

Amidines↗

The serotypes of Bordetella pertussis isolated in Great Britain between 1941 and 1968 and a comparison with the serotypes observed in other countries over this period.

Classification, by agglutinogens, of 634 isolates of Bordetella pertussis collected from 1971 to 1968 in Great Britain demonstrated that a change from a predominantly 1,2,0,4 serotype (75% of those examined during 1941-4) to a predominantly 1,0,3,0 serotype (73% of those examined during 1966-8) occurred sometime after 1953. Furthermore, evidence from the examination of isolates collected between 1941 and 1953 suggests that the change may have been gradual. Isolates of serotype 1,2,3,4 made up 20-30% of the total of our cross-country selection for the periods 1941-4, 1946-9, 1950-3 and 1966-8, but over shorter periods in individual areas the percentage varied from negligible to as high as half of those isolated. Results from other countries show a similar drift towards a 1,0,3 sertype but more often from a 1,2,3 than from 1,2,0 serotype. The value, in epidemiological studies, of extended information obtained by monospecific typing sera to all six, rather than only two or three agglutinogens, and confirmation of the results by agglutinin production is demonstrated: for instance not all 1,0,3 isolates were identical.

Agglutination Tests↗

Isolation and EBV-transformation of a minor population of normal human tonsillar B cells bearing a cold agglutinin-associated idiotope.

Human autoantibodies which bind to red blood cells and cause agglutination in the cold, collectively comprise the cold agglutinins: a subset of these antibodies with specificity for the Ii carbohydrate antigen on the red cell surface bears cross-reacting idiotypic determinants. A monoclonal antibody has been raised which specifically inhibits the cold agglutination reaction, and has been used to probe normal lymphoid cell populations for the presence of B cells expressing the same idiotope. Although such cells comprise only 1-2% of the lymphoid cells in the tonsil, it has been possible to isolate them in good yield and purity using the antibody and magnetic beads. Released cells were infected with the Epstein-Barr (EB) virus and an idiotope-bearing line established. The IgM secreted by the line was found to agglutinate red cells in the cold indicating that cold agglutinin-producing cells were among this idiotope-positive population. The immunophenotype of the line, and the agglutinating power of the secreted IgM, have been compared with a similarly immortalized idiotope-bearing line established from neoplastic B cells of a patient with frank cold agglutination disease.

Agglutinins↗

Concanavalin A-agglutinability of membrane-skeleton-free vesicles and aged cellular remnants derived from human erythrocytes. Is the membrane skeleton required for agglutination?

Vesicles and cell remnants have been obtained by aging of erythrocytes in vitro. The vesicles lacking the membrane skeletal proteins and the remnants known to possess a rigid skeleton have been used to assess the role of membrane skeletal proteins in the process of Con A (concanavalin A)-mediated agglutination of erythrocytes. Both the vesicles and the remnants were found to bind Con A at the same density as did intact cells. The vesicles, isolated from normal as well as from the Con A-agglutinable trypsin- and Pronase-treated cells, failed to agglutinate with Con A. They were, however, well agglutinated by WGA (wheat-germ agglutinin) and RCA [Ricinus communis (castor bean) agglutinin], indicating that the vesicles are not defective in agglutination. Large, cytoskeleton-free, vesicles prepared by another procedure also gave the same results. The aged remnants from trypsin- and Pronase-treated erythrocytes showed significantly decreased agglutination with Con A, but were agglutinated as well as the fresh cells by WGA and RCA. The agglutination with Con A is thus abolished when the membrane skeleton is absent, and reduced when it is rigid, suggesting that the skeleton may play an important role in the agglutination of erythrocytes by Con A.

Concanavalin A↗

Diagnostic value of the Widal test in childhood typhoid fever.

The usefulness of the Widal test in diagnosing childhood typhoid fever in endemic areas was investigated. The test was done on 150 children with other febrile illnesses and 98 bacteriologically proved cases of typhoid fever. Of the 150 children with nontyphoidal fever, only one had an H agglutinin titer of 1:50. Using an H or O agglutinin titer of 1:50 or more as a criterion for diagnosis, a positive Widal test was found in 88% of typhoid fever cases on the first occasion on which the test was done. If the test was repeated at least 94% of the typhoid cases had a significant result. The Widal test is a useful diagnostic test in children in endemic areas, provided interpretation of the test is made against background information relating to agglutinin levels in normal children in the region.

Adolescent↗

Peanut agglutinin, a new mitogen that binds to galactosyl sites exposed after neuraminidase treatment.

Peanut agglutinin, purified by affinity chromatography, agglutinates lymphocytes from mouse, rat, guinea pig, and man only after their treatment with neuraminidase. However, it stimulates only neuraminidase-treated rat and human cells. A similar number cell surface receptors for peanut agglutinin was found on neuraminidase-treated rat and mouse lymphocytes although the latter cells were not stimulated by the lectin. Galactose specifically inhibited the agglutination and stimulation of lymphocytes by peanut agglutinin. Sequential treatment of lymphocytes with neuraminidase and beta-galactosidase markedly reduced the response of the cells to stimulation by peanut agglutinin, soybean agglutinin, and galactose oxidase. It is suggested that the same galactosyl residue may be the target for the initial step in triggering lymphocytes by the above mentioned mitogens.

Agglutination Tests↗