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PubMed1976

Concanavalin A-induced agglutination of human leukemic and lymphoma cells.

With a newly developed turbidometric method, concanavalin A was shown to agglutinate normal lymphocytes, lymphoma cells, and leukemic cells from chronic lymphocytic leukemia and from acute myelocytic and lymphocytic leukemia. However, there was a marked difference in the kinetics of this agglutination process. Leukemic blast cells and cells from a patient with convoluted lymphoma agglutinated poorly in this system. Conversely, the degree of agglutination for chronic lymphocytic leukemia cells was greater than that for the blast cells and also slightly greater than that for normal lymphocytes. Cultured cells from a Burkitt's lymphoma (Raji) and from a patient with poorly differentiated lymphoma agglutinated very rapidly with concanavalin A. Prior incubation of all cell types with neuraminidase markedly enhanced the agglutination process similar to that of trypsinization. Thus, these studies illustrate the usefulness of this method in quantitating the kinetics of agglutination of various human neoplastic cell types by concanavalin A.

Agglutination↗
PubMed1998

Serotyping of Flavobacterium meningosepticum by co-agglutination method.

The purpose of this investigation was to evaluate the usefulness of a co-agglutination procedure for the typing of Flavobacterium meningosepticum. The sensitivity and specificity of the co-agglutination test was compared to the slide agglutination test using reference strains of the bacterial species. Antisera were characterized by both technics to determine their titer and working dilution. The specificity of the sera was assessed by performing tests which include strains of other species and serotypes. A collection of 47 strains of F. meningosepticum isolated from clinical specimens were typed by both co-agglutination and slide agglutination methods. Co-agglutination proved to be markedly more specific than the slide procedure although both methods were similar in sensitivity. It was concluded that co-agglutination proved to be an excellent method for the serotyping of F. meningosepticum.

Agglutination Tests↗
PubMed1999

Evaluation of direct agglutination test (DAT) as an immunodiagnostic tool for diagnosis of visceral leishmaniasis in Nepal.

Before field application of the direct agglutination test (DAT) for leishmaniasis, it was assessed as a diagnostic tool. Fifteen confirmed visceral leishmaniasis cases (bone marrow aspiration positive), 120 tuberculosis, 58 leprosy, 15 malaria, 26 intestinal parasitic infection cases, 24 endemic healthy controls from adjacent to the study area, and 18 controls from Kathmandu (who had never visited the VL endemic areas) were tested for anti-leishmanial antibody agglutination titers. Two of the tuberculosis cases were positive for anti-leishmanial agglutinating antibodies at 1:800. All the visceral leishmaniasis confirmed cases were reactive to anti-leishmanial antibody at > or = 1:3,200. Other specimens were negative for serology. The sensitivity of the direct agglutination test was 100% and the specificity was 99.2%. The direct agglutination test had positive and negative predictive values of 100% and 99.2% respectively. The direct agglutination test has been found to be simple, rapid, reliable, economic, safe and adaptable to micro-techniques using microtiter plates. It is specific and sensitive. The direct agglutination test is simple enough for it to be performed in a field laboratory.

Agglutination Tests↗
PubMed1992

An evaluation of the staphylococcal co-agglutination test for the detection of group A rotavirus in human faeces.

The group A rotavirus staphylococcal co-agglutination test was evaluated and its sensitivity and specificity compared with an in-house enzyme-linked immunosorbent assay (ELISA) and a commercial latex agglutination test (Rotalex). In addition, the storage stability of the staphylococcal reagents was ascertained. Examination of 136 clarified suspensions of diarrhoeal faeces by the staphylococcal co-agglutination test revealed a high proportion of false positives (26%) and uninterpretable results (34%) due to non-specific agglutination. Non-specific agglutination could be removed effectively by prior absorption of the clarified faecal specimens with unsensitized staphylococci. The staphylococcal co-agglutination test was less sensitive and specific than the in-house enzyme-linked immunosorbent assay but was comparable to the Rotalex slide latex agglutination test. The staphylococcal reagents have a shelf life of at least 29 weeks.

Agglutination Tests↗
PubMed1992

[Further studies of the acid plate agglutination reaction in the diagnosis of brucellosis in humans].

Using the methods of acid platelet agglutination, agglutination, agglutination with 2-mercaptoethanol, Coombs' reaction, and complement binding reaction, 152 serum specimens taken from patients with diagnosed brucellosis, were subjected to studies. The following per cent of seropositive results was shown: acid platelet agglutination--11.2%, agglutination--40.1%, agglutination with 2-mercaptoethanol--34.4%, Coombs' reaction--42.8%, complement binding reaction--7.9%. The acid platelet agglutination reaction cannot be used for screening for brucellosis in humans.

Abattoirs↗
PubMedDate not supplied

[Lipid particles agglutinated in hyperlipidemias with opalescent sera (author's transl)].

During the last five years, we observed under the microscope all the hyperlipidemic sera received in our laboratory, to search a lipid particle agglutination. We have retained 24 sera with obvious agglutination (9 types V, 9 types IV and 6 types IIb), the lipidemia of which was compared, type to type, to 56 control sera (16 types V, 16 types IV and 24 types IIb), in which no agglutination has been found. We observed significant differences in the means of cholesterol (CT) triglycerides (TG), vitamin A (VA) and CT/TG, VA/TG and VA/CT ratios, comparing agglutinated and control sera. These differences suggest that the agglutination phenomenon is in relation with the nature of the lipid particles present in the hyperlipidemic sera, which could be "chylomicron remnants" in type V hyperlipidemias, and other forms of "remnants" in type IV and IIb hyperlipidemias. These "remnants" could result from a blockade of lipolysis. This blockade of lipolysis in the agglutinated sera might be related to what is known about the authentificated autoimmune hyperlipidemia (AIH). Thus we can suppose that hyperlipidemias with obvious serum lipid particle agglutination are autoimmune hyperlipidemias.

Agglutination Tests↗
PubMed1990

[Sperm agglutination antibodies studied in infertile couples at the Women's Clinic in Plzen 1980-1988].

The authors assessed sperm-agglutination antibodies in infertile couples in 1980-1988, using the Kibrick test: in sera from 699 female patients, a positive result was found in 142 (20%) women, in sera from 352 men, in 6 (1.7%) sperm-agglutination antibodies were detected. Using the microagglutination test, they examined sera of 3501 sterile women; in 273 (6.8%) they found sperm agglutination up to 1:16, above titre 1:32 in 12 (0.4%) of the patients. Using the same test, they examined sera of 2803 men and found sperm-agglutination up to 1:16 in 21 (0.7%) and above 1:32 in 6 (0.3%). The most frequent type of sperm agglutination is aggregation of the heads; aggregation by tails or mixed agglutination are less frequent. In 3252 sterile women where no sperm-agglutination antibodies were found Kremer's test was negative in 1145 patients. The authors analyzed in 196 sterile women the ovulation mucus by the microagglutination test and found a high anti-spermatozoidal activity in 24 (12%) women. Concurrently an indirect MAR test was made. Antibodies against sperm cells were found in 24 (12.2%) of specimens in sIgA, in 8 (4.1%) in IgG, in 2 (2%) in IgM and in one sample (0.5%) in IgE. The indirect MAR-test in sera of 325 sterile women revealed a predominance of anti-spermatozoidal antibodies in IgG, in men in IgA. In seminal plasma the authors detected antibodies against sperm cells in sIgA. In seminal plasma the authors detected antibodies against sperm cells in sIgA and IgE.

Antibodies↗
PubMed1990

Inhibition of von Willebrand factor-induced platelet agglutination by ADP does not result from reduced binding of total von Willebrand factor or its larger multimers.

Earlier experiments showed that platelet agglutination induced by von Willebrand factor (vWf) plus ristocetin was greatly diminished if adenosine diphosphate (ADP) was added first in the presence of ethylenediaminetetraacetic acid (to prevent aggregation). Platelets treated with ADP and then fixed also agglutinated less than control fixed platelets. The studies reported here demonstrate that ADP did not decrease ristocetin-induced binding of vWf whether binding was measured on suspended platelets with iodine 125-labeled vWf or on suspended or agglutinated platelets with the use of any of three 125I-labeled monoclonal antibodies that bind to vWf but that do not interfere with ristocetin-induced agglutination. Equal amounts of vWf were eluted from ristocetin/vWf-treated platelets when they were resuspended without ristocetin, whether or not the platelets had been exposed to ADP, and the vWf recovered in either case was composed only of large multimers. No evidence for an agglutination site other than glycoprotein Ib could be demonstrated by measuring agglutination of a mixture of platelets fixed after inhibition with antibody against glycoprotein Ib and platelets fixed after inhibition with ADP. We conclude that inhibition of agglutination by ADP must involve the way in which vWf is bound, because it does not result from a decreased amount or from a difference in multimer size of bound vWf.

Adenosine Diphosphate↗
PubMed1988

[Agglutination of intravenous lipid emulsions by acute phase proteins of inflammation].

Agglutination of intravenous fat emulsions (IVFE) by sera of acutely ill children was studied in vitro in 23 patients. C-Reactive protein (CRP) has been showed to agglutinate with IVFE; after discarding CRP, serum of acutely ill patients still agglutinate with IVFE. The agglutination score in generalized sepsis is significantly highest than in localized sepsis. We studied agglutination during three periods: first days of treatment by antibiotics (period 1), between the fourth day and the end of treatment (period 2), and after the end of treatment (period 3). In period 2, when orosomucoid concentrations; nevertheless, the observation of an agglutination in periods 1 and 3 where the mean orosomucoid level is within normal range, strongly suggests that one or more others acute phase proteins are also involved in occurrence of agglutination.

Agglutination↗
PubMed1977

[Agglutination reaction of T. cruzi, T. cruzi like Strains, T. rangeli and T. conorhini with Soja hispida lectin and Aaptos papillata protectin (author's transl)].

Protectin from the sponge Aaptos papillata (Keller) was used in the characterization of five strains of T. cruzi (Venezuela, Guatemala, Y. Brasilien, Peru, Wien) and six T. cruzi like strains (Triatoma, Maryland, ITMAP 943, FH4, FH5, LN). Based upon their membrane receptors, these T. cruzi and T. cruzi like isolates could be differentiated from rangeli (Venezuela Strain) and T. conorhini (Hawai Strain) by agglutination reaction to the proctectin. Furthermore, after pronase treatment T.rangeli could also be distinguished from T. conorhini by agglutination test with A. papillata protectin and also Soja hispida lectin. It is not possible to differentiate the T. cruzi complex with S. hispida lectin, because it did not agglutinate T. cruzi (Vienna Strain) and T. cruzi like (Maryland Strain). However, after treating this human pathogenic strain with pronase the pseudocrypt antigen of the first order is made available to the S. hispida lecting thereby producing agglutination. The T. cruzi like strain however did not agglutinate with this treatment. On the other hand, while T. rangeli did not agglutinate even after pronase treatment, T. conorhini showed the agglutination reaction. This observed reaction is explained by the availability of the pseudocrypt antigens of the first order after pronase treatment.

Agglutination Tests↗
PubMed1986

[A new agglutination reaction for the diagnosis of the developmental stage of acquired toxoplasmosis].

Agglutination of acetone treated toxoplasma (AC) is different from that one of formalin fixed parasites (HS). Sera from patients with a recently acquired ("acute") infection agglutinate both HS and AC parasites suspensions as well; contrary to sera from patients with past infection ("chronic stage") in which high titers of HS agglutination are often present, while the titres of AC agglutination are lower even negative. This is markedly observed in patients with local lesions (relapsing chorioretinitis, patients with AIDS and brain abscesses). The reason might be that different membrane toxoplasma antigens may induce the synthesis of agglutinating IgG. For example, antigens 35 KD and 27 KD described by E. Handman et al. The antibody specific for 27 KD is apparently present mainly during acute infection, contrary to the antibody specific for 35 KD which might be responsible of the high HS agglutination titre in sera from patients with chronic infection. Even if these hypotheses were not confirmed in the future, comparison of the titre in the HS and AC agglutination test might actually be helpful for practical diagnosis of the stage of toxoplasma infection.

Acute Disease↗
PubMed1979

Isolation of human spermatozoa membrane antigens binding sperm-immobilizing and sperm-agglutinating antibodies.

Peptides of human spermatozoa were dissolved with Hyamine 2389 and Triton X-100 and separated by chromatography on Biogel P4 columns into seven fractions. The antigenic activities of the separated sperm-membrane fractions were tested according to their capacity to inhibit sperm agglutination and sperm immobilization (immune inhibition test) in human sera of sterile patients. Sperm-agglutinating and sperm-immobilizing activity was tested by the microtray agglutination and microtray immobilization test. A titer reduction was achieved only in sperm-immobilizing sera. Four sperm antigenic fractions revealed in the majority of the repeatedly tested sperm-immobilizing sera an inhibition of the antigen-antibody reaction. No reaction was observed after exhaustive absorption of the tested seven antigen fractions with sperm-agglutinating sera. Therefore the conclusion can be drawn that sperm-agglutinating and sperm-immobilizing antibodies react with different sperm antigens. Normal human sera without sperm antibodies served as control. As no sperm agglutination or sperm immobilization was obtained after absorption of these control sera our antigen fractions do not produce sperm agglutination or sperm immobilization.

Antigens, Surface↗
PubMed1978

[Lipid particles agglutinated in hyperlipidemias with opalescent sera (author's transl)].

During the five last years, we observed under the microscope all the hyperlipidemic sera received in our laboratory, to search a lipid particle agglutination. We have retained 24 sera with obvious agglutination (9 types V, 9 types IV and 6 types IIb), the lipidemia of which was compared, type to type, to 56 control sera (16 types V, 16 types IV and 24 types IIb), in which no agglutination has been found. We observed significant differences in the means of cholesterol (CT) triglycerids (TG), vitamin A (VA) and CT/TG, VA/TG and VA/CT ratios, comparing agglutinated and control sera. These differences suggest that the agglutination phenomenon is in relation with the nature of the lipid particles present in the hyperlipidemic sera, which could be "chylomicron remnants" in type V hyperlipidemias, and other forms of "remnants" in type IV and IIb hyperlipidemias. These "remnants" could result from a blockade of lipolysis. This blockade of lipolysis in the agglutinated sera might be related to what is known about the authentificated autoimmune hyperlipidemia (AIH). Thus we can suppose that hyperlipidemias with obvious serum lipid particle agglutination are autoimmune hyperlipidemias.

Agglutination↗
PubMed1987

An evaluation of agglutination and coagglutination techniques for serotyping of Haemophilus pleuropneumoniae isolates.

A comparative evaluation of rapid slide agglutination, tube agglutination, 2-mercaptoethanol tube agglutination, and coagglutination tests was made for serotyping isolates of Haemophilus pleuropneumoniae. The results indicated that a majority of the isolates could be serotyped by any of these tests. But, it was not uncommon to find isolates which were inagglutinable or poorly agglutinable in homologous sera. Heat treatment of whole-cell suspensions of such isolates was essential to unmask the serotype-specific antigenic determinants; however, in the process of heat treatment, cross-reactive common antigens of minor nature were also exposed. The antibodies involved in such cross-reactions were mainly of immunoglobulin M type, because the cross-reactivities were completely abolished in coagglutination and 2-mercaptoethanol agglutination tests. Thus, both these tests were satisfactory for serotyping inagglutinable mucoid strains. For serotyping strains which were either polyagglutinating or autoagglutinating, agglutination tests could not be used, but the coagglutination test proved to be satisfactory. The coagglutination test was serotype-specific, sensitive, simple, rapid, reproducible, and easier to read and interpret than rapid slide or tube agglutination tests. This test could be used to serotype mucoid, smooth, or rough isolates.

Agglutination Tests↗
PubMed1997

[Evaluation of the usefulness of the agglutination test with Mangifera indica extract for the identification of pathogenic Yersinia enterocolitica strains].

The study was performed on 137 Y. enterocolitica strains belonging to various serological groups, including 75 03 group strains isolated form human clinical material. The agglutination test on slides was carried out on this strains using Mangifera indica extract of own production. Agglutinating preparation obtained from the seeds of M. indica agglutinated Y. enterocolitica organisms possessing the pVY plasmid and CRMOX+ phenotype in dilutions to 1.56 micrograms/ml. In identification tests conducted parallelly agglutination solution was used in concentrations of 100 and 10 micrograms/ml. All clones of Y. enterocolitica from O3 group from cultures at 37 degrees C and with CRMOX+ phenotype possessing the pVY plasmid were agglutinated by the extract. Agglutination failed to develop in the cultures of these clones incubated at 25 degrees C. Yersinia clones not containing the pVY plasmid with CRMOX- phenotype were resistant to agglutination. The virulence plasmid was found in 44 out of 75 strains of Y. enterocolitica O3 and was identified by restriction analysis after plasmid DNA digestion with Eco RI enzyme. The obtained results agreed with those of Wauters et al. in 1995 and confirmed the opinion of these authors on the usefulness of the test with M. indica agglutinin for the identification of virulent Y. enterocolitica strains.

Agglutination Tests↗
PubMed1993

Kinetics of the autologous red cell agglutination test.

The kinetics of the autologous red cell agglutination test for detecting circulating antibodies to HIV-1 were studied. Two monoclonal anti-red blood cell antibodies (1C3/86 and 10F7MN) were used to construct Fab-peptide conjugates for the test. Both antibodies recognize glycophorin alpha on the surface of erythrocytes by immunoprecipitation or immunoblotting techniques. The number of binding sites, association and dissociation constants of 1C3/86 Fab and 10F7MN Fab' fragments were determined (n = 4.80 X 10(5) sites/erythrocyte, Ka = 0.43 X 10(7) M-1, Kd = 23 X 10(-8) M for 1C3/86, n = 4.66 X 10(5) sites/erythrocyte, Ka = 1.05 X 10(7) M-1, Kd = 9.5 X 10(-8) M for 10F7MN. The binding studies were performed under the same conditions as the autologous red blood cell agglutination test. When 0.9 microgram of anti-glycophorin Fab was added to 10 microliters of blood 0.25 microgram of 1C3/86 Fab was bound whereas 0.29 microgram for 10F7MN Fab' was bound. Antibody binding reached a plateau after 2 min and once bound did not exchange with unbound Fab over the time scale of the test. The binding of the anti-peptide antibody (cross-linking antibody) was also complete within 2 min. Addition of approximately 0.1 microgram of anti-peptide antibody gave half a maximal agglutination score. This is equivalent to 10 micrograms/ml circulating antibody. Under the agglutination test conditions, Fab-peptide conjugate was bound to 14% of available glycophorin molecules. Half maximal agglutination occurred when approximately 1.1% of the bound Fab-peptide conjugates were cross-linked. A maximum agglutination score of four occurred in the presence of 1 microgram of anti-peptide antibody equivalent to 100 micrograms/ml circulating antibody whereas an agglutination score of 1+ was elicited by only 0.32 microgram anti-peptide antibody and involved the cross-linking of approximately 160 glycophorin molecules per red cell.

Antibodies↗
PubMed1995

Human recombinant CD4 and CD4-derived synthetic peptides agglutinate immunoglobulin-coated latex particles. Evidence that residues 25-28 and 35-38 of human CD4 form two separate immunoglobulin binding sites.

It has been shown previously that amino acid residues 21-49 of the first extracellular domain of human CD4 form the core of an immunoglobulin (Ig) binding site. Synthetic peptides of human CD4 that encompass this region also bind Ig and, with higher affinity, antigen/antibody complexes. Synthetic peptides also enhance binding of both monomeric and aggregated Ig to monocytic U937 cells and Staphylococcus aureus Protein A. To better characterize the nature of the Ig binding site on CD4, we tested the ability of human recombinant CD4 (rCD4) to agglutinate polystyrene particles coated with Ig. Evidence is presented that soluble rCD4 and CD4 peptide p21-49 were capable of specific agglutination of polystyrene particles coated with polyclonal Ig of either human or sheep origin. Agglutination could be blocked by soluble human polyclonal IgG or F(ab')2 fragments. Both heparin and sulfated dextrans also inhibit agglutination, suggesting that charged residues on rCD4 played an important role in agglutination mediated by rCD4 or CD4 peptide. Similarly, aurintricarboxylic acid (ATA) also blocked agglutination of Ig-coated particles by rCD4. Agglutination mapping studies performed using truncated peptides revealed the existence of two discrete, closely related Ig binding sites (residues 25-28 and 35-38).

Amino Acid Sequence↗
PubMed1975

Role of cell membrane galactosyltransferase in concanavalin A agglutination of erythrocytes.

It has been previously observed that rabbit erythrocyte cell surface galactosyltransferase appears to play a role in concanavalin A agglutination of these erythrocytes (Podolsky et al., 1974). Further, a correlation between the occurrence or level of cell surface galactosyltransferase and concanavalin A agglutinability of other cell types has also been observed. The mechanism by which rabbit erythrocyte galactosyltransferase participates in concanavalin A agglutination has now been further defined. The enzyme was solubilized and purified. Characterization of the enzyme properties has shown them to be similar to those reported for other purified galactosyltransferases. Amino acid and carbohydrate analysis showed a high asparagine content and the presence of D-mannose. Specific alpha-mannosidase treatment of the enzyme showed that some of these D-mannose residues were terminal sugars. The purified enzyme also conferred concanavalin A agglutinability to non-agglutinable human erythrocytes. However, the ability to confer concanavalin A agglutinability was unrelated to the enzyme activity per se (as measured with fetuin acceptor) but appeared to be entirely dependent on the presence of terminal alpha-linked D-mannosyl residues in the enzyme structure. These findings suggest that the presence of terminal alpha-mannosidyl residues on cell surface glycoproteins such as galactosyltransferase may be the determining factor in agglutination of cells by concanavalin A.

Asparagine↗