Agglutinin production in normal and splenectomized rabbits.
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The red alga, Agardhiella tenera was found to contain a glycoprotein which agglutinates mouse leukemia cells, L5178Y but not L1210. It also agglutinates guinea pig and rabbit erythrocytes, and has weak activity against human A, B and O, mouse, horse and sheep erythrocytes and hamster and mouse lymphocytes. The agglutination was not inhibited by simple sugars. The major active component was purified and determined to be a beta-structure protein containing 2.7% glucose as sugar moiety. The molecular weight was estimated to be 12,000 by gel filtration and 13,000 by polyacrylamide gel electrophoresis, in the presence of sodium dodecylsulfate. Its isoelectric point was 6.1, and it contained high amounts of glycine, serine and threonine, but no half cystine or histidine. It had no subunit structure, and the C- and N-terminal amino acids were threonine and arginine, respectively.
Cell surface alterations occurred during murine erythroleukemia cell (clone 745) differentiation that were detected by both agglutination and lectin binding. Agglutination of erythroleukemia cells was produced by wheat germ agglutinin; whereas, concanavalin A, Ricin, soybean agglutinin and fucose-binding protein were either ineffective or much less efficacious. Treatment of leukemia cells with the inducer of erythroid differentiation dimethylsulfoxide (DMSO) caused a progressive accumulation of hemoglobin-containing cells in culture and a decrease in the rate of agglutination by wheat germ agglutinin, which began at 24 h after exposure to the polar solvent, reached a nadir at 48 h, and remained essentially constant thereafter. The binding of radioactive wheat germ agglutinin by untreated control erythroleukemia cells increased with time in culture, reaching a maximum value at 48 h, and decreased progressively thereafter. Although an increase in 3H-labeled wheat germ agglutinin binding also occurred in DMSO-treated cells, the level bound was significantly lower than that observed in control cells at 24-96 h. The treatment of erythroleukemia cells with various concentrations of DMSO resulted in a decrease in the number of wheat germ agglutinin receptor sites. Other inducers of differentiation (i.e., dimethylformamide, bis(acetyl)diaminopentane) also inhibited the rate of wheat germ agglutinin-induced agglutination of erythroleukemia cells while, in contrast, the inducer tetramethylurea did not. These studies indicate that membrane changes occur during differentiation and suggest that there may be more than one mechanism involved in the initiation of maturation which ultimately leads to the common pathway of erythroid development.
A separation procedure has been developed for mouse splenic T and B lymphocytes which is based on their differential agglutination by wheat germ agglutinin (WGA). In the presence of 50-100 micrograms/ml of WGA, multicellular aggregates are formed which are enriched in B cells. These aggregates can be separated from monodisperse T cells by gravity sedimentation and subsequently dissociated into single cells by treatment with N-acetylglucosamine (NAG). Immunocytochemical analyses and mitogenic assays indicate approximately 10-15% cross contamination of the resultant B and T cell fractions. The separation procedure is not only convenient and rapid but also allows the simultaneous recovery of viable T and B cells from the same spleen preparation.
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The study of 26 B. bronchiseptica strains with typical morphological and biochemical properties resulted in the detection of 8 strains having the main specific agglutinogens of 3 Bordetella species (serovars) in different combinations. The presence of the agglutinogens was confirmed in the agglutination test and the agglutinin adsorption test with the use of monospecific antisera to the main agglutinogens. The comparison of natural B. bronchiseptica serovars and artificial convertants (resulting from the conversion of B. parapertussis by B. pertussis phages) revealed their identical biochemical activity, their capacity for causing necrosis when injected intradermally into rabbits and for the formation of two types of colonies, differing in size and serological activity. In contrast to B. parapertussis convertants, B. bronchiseptica serovars had no lysogenic properties and were sensitive to B. pertussis and B. bronchiseptica phages.
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Serum from each of five persons with pulmonary mycobacteriosis due to Mycobacterium intracellulare contained agglutinins specific for the serotype of the causative organism. The findings suggest that the direct agglutination test may prove useful in diagnostic mycobacteriology.
Normal sera and sera from burned patients were examined for Candida agglutinin titers, precipitin titers, and the ability to disperse germ tubes of Candida albicans in an attempt to determine whether germ tube dispersion is correlated with Candida infection as animal models have indicated. Other investigators have reported that immunoglobulin G antibody to Candida interferes with a serum clumping factor resulting in germ tube dispersion. Germ tube dispersion in sera from burned patients with varying degrees of Candida infection is significantly greater than that found in uninfected controls. In addition, the germ tube dispersion test indicated the presence of Candida infection in several patients who had clinical evidence of infection but no detectable agglutinins or precipitins.
T and B lymphocytes of the Syrian golden hamster were separated from spleen cell preparations on the basis of their differential agglutinability with the lectin soybean agglutinin. Only B lymphocytes were agglutinated by this lectin, and they could be separated from the unagglutinated T lymphocytes by sedimentation through 50% heat inactivated calf serum at unit gravity. The B lymphocyte aggregates could be dissociated into single cells that were viable and functional after treatment with 0.5 M galactose. The isolated cell fractions were characterized by their blastogenic response to various T cell and B cell specific mitogens and by the presence or absence of cell surface IgG.
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A series of modified (field) tests were compared to a crossmatch between mare and foal for their reliability in predicting neonatal isoerythrolysis (NI) in eight foals born to experimentally alloimmunized mares. In the field tests, mare's serum, plasma and colostrum were combined with foal erythrocytes washed by a modified procedure to determine which combination was the best predictor of impending NI. A consistent grading system for agglutination and hemolysis was employed. The field tests using mare's plasma demonstrated less agglutination and hemolysis than tests where serum was employed. Immediate assessment of agglutination failed to demonstrate agglutinin activity when compared to tests where incubation was included. Rouleaux formation posed a problem in interpretation of minor agglutination, however the grading of hemolysis was simpler, quicker, and more accurate. The field test that was most reliable when compared to the crossmatch and presuckle anti-foal erythrocyte titers in demonstrating agglutinins was the combination of mare's serum and foal's erythrocytes. The tests for hemolysin detection in serum and colostrum which incorporated rabbit sera as a complement source were also reliable.
In this paper we report the effect of cis-dichlorodiammine platinum(II) (cis-platin) on the agglutination of normal and transformed lymphocytes with lectins conconavalin A (Con A) and wheat germ agglutinin (WGA) which is a sensitive test to study the expression of cell surface components. Normal splenocytes showed small degree of cell agglutination with Con A and WGA. This cell agglutination of normal splenocytes with these lectins increases significantly after cis-platin treatment at 37 degrees C. The Dalton's lymphoma cells which show a high degree of cell agglutination with Con A and WGA when treated with cis-platin at 37 degrees C results in decrease in the degree of their cell agglutination with these lectins. Agglutination inhibition studies with alpha-methyl-D-glucopyranoside for Con A and N-acetyl-D-glucosamine for WGA shows that this increase and decrease of cell agglutination after platinum treatment of cells is a specific reaction and not non-specific. Incubation of cis-platin treated normal splenocytes with excess of neuraminic acid results in decrease of cell agglutination with both Con A and WGA. cis-Platin treatment of the cells results in the release of sialic acid from the surface of the cells. A correlation between the release of sialic acid from the cell surface and cell agglutination is also discussed.