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AGGLUTINATION REACTIONS OF HUMAN LEUCOCYTES.

Agglutination tests with various sera and leucocytes from 58 leukaemic patients and 61 patients without leukaemia are reported. The agglutination of white blood cells by guinea-pig serum is of limited value in the diagnosis of leukaemia, though the test may be helpful in distinguishing leukaemia from other lymphomatous disorders.Leuco-autoagglutinins were demonstrated more frequently than expected. Eleven leukaemic and six non-leukaemic sera agglutinated autologous leucocytes. White blood cell agglutinins showed no apparent relationship to maturity or numbers of circulating leucocytes or to previous blood transfusions, x-irradiation, or therapy with antimetabolites.

Agglutination↗

Rapid serotyping of groups A, B, and C meningococci by rocket-line immunoelectrophoresis and co-agglutination.

Rocket-line immunoelectrophoresis (R-LIE) with antigen containing intermediate gel, and co-agglutination utilising protein A-containing staphylococci coated with specific antibodies, were adapted for serotyping the prototypes of group B meningococci. Both were found to have the same specificity as agar gel double diffusion (AGDD) but they were more sensitive and more rapid than AGDD. R-LIE required, like AGDD, the extraction of relatively large quantities of bacteria, while the co-agglutination method, performed as a slide agglutination tests with results within a few minutes and no need of special equipment, required only a small amount of heated whole meningococcal cells. Meningococcal strains of serogroups B, C, and A from patients with carriers were serotyped and the results with all three methods were in agreement.

Agglutination Tests↗

Evaluation of Bactigen latex agglutination and Phadebact coagglutination for detection of bacterial antigens in cerebrospinal fluid.

The Bactigen latex agglutination and Phadebact coagglutination tests were evaluated for their ability to detect bacterial antigens of Haemophilus influenzae type b, Streptococcus pneumoniae (83 serotypes) and Neisseria meningitidis groups A, B, C, and Y in 214 samples of cerebrospinal fluid (CSF). Bactigen latex agglutination was more sensitive than Phadebact coagglutination: it detected 87% (59/68) of culture positive CSF specimens, whereas Phadebact detected 72% (52/72). Bactigen detected all cases of meningitis caused by S pneumoniae and H influenzae. Of the 19 specimens that were positive for N meningitidis, 74% were detected by Phadebact and only 53% by Bactigen. Gram stain results were positive for 85% of all specimens positive on culture. Bactigen was slightly more specific (97%) than Phadebact (96%). Bactigen, however, showed less specificity (81%) than Phadebact (94%) on 31 CSF specimens that were culture positive for organisms other than the test organisms. These included two CSF specimens from patients with tuberculous meningitis which gave false positive results for S pneumoniae with the Bactigen reagents. No false positive results were obtained on 104 culture negative CSF samples. Bactigen latex agglutination was superior to Phadebact coagglutination and Gram stain for the detection of S pneumoniae and H influenzae in CSF specimens from patients with bacteriologically proved meningitis.

Agglutination Tests↗

An immunoblotting procedure comprising O = 9,12 and H = d antigens as an alternative to the Widal agglutination assay.

AIMS: To compare the established Widal agglutination assay with an immunoblotting procedure. METHODS: 110 sera were used to compare the established Widal agglutination assay with an immunoblotting procedure incorporating lipopolysaccharide (LPS) (O = 9,12) and flagellar (H = d) antigens. RESULTS: Antibodies to the LPS antigens were detected in 18 sera by the Widal assay and in 37 by immunoblotting. Antibodies to the flagellar antigens were detected in 27 sera by Widal assay and in 25 by immunoblotting. CONCLUSIONS: An immunoblotting procedure incorporating O = 9,12 LPS and H = d flagellar antigens was rapid and more sensitive than the established Widal agglutination assay for providing evidence of infection with S typhi.

Agglutination Tests↗

Differential agglutination of P388 adriamycin-sensitive and P388 adriamycin-resistant leukemia cells.

Cell surface modification was studied in a subline of murine leukemia resistant to adriamycin (P388/ADR). Lectin-induced agglutination was used as a probe. Agglutination was studied using two plant lectins, wheat germ agglutinin (WGA) and Ricinus communis agglutinin-I (RCA-I). A 7-fold higher amount of WGA and 14-fold higher amount of RCA-I were required to bring about minimum agglutination of P388/S as compared to P388/ADR. The present studies clearly indicate a change in the plasma membrane of P388/ADR cells.

Agglutination↗

Agglutination of isolated platelet membranes.

Platelet membrane glycoproteins play a central role in platelet aggregation and thus in primary hemostasis. To investigate mechanisms of platelet-platelet interaction in the absence of cellular activation events, we studied immunological and functional aspects of isolated platelet membranes. Platelet membranes contained significant amounts of the inducible fibrinogen receptor, glycoprotein (GP) IIb-IIIa, which exposes conformation-dependent LIBS1 and PMI-1 epitopes in response to fibrinogen-mimetic peptides GRGDSP and HHLG-GAKQAGDV. In the presence of soluble fibrinogen, membrane-coated latex beads showed Ca(2+)-dependent agglutination that could be partially inhibited by GRGDSP but not by the biologically inactive peptide GRGESP. Thrombospondin enhanced agglutination of membrane-coated beads, which could be inhibited by polyvalent anti-thrombospondin Fab fragments and anti-thrombospondin monoclonal antibody MA-II. Mg2+ inhibited both GPIIb-IIIa- and thrombospondin-mediated agglutination of membranes in a dose-dependent manner. The results of the present study indicate that isolated platelet membranes are a useful tool to study regulation of GPIIb-IIIa- and thrombospondin-mediated platelet-platelet interaction.

Agglutination↗

Serum proteins agglutinate cilia and modify ciliary coordination.

A quantifiable assay is described in which the lateral cilia of the sea mussel Mytilus edulis are used to examine the effects of serum on ciliary function. Human sera and sera from a variety of animals have been found to induce a change in ciliary metachrony. This alteration in activity appears to be brought about by the agglutination of adjacent cilia into blocks, with the effect of increasing the wavelength; the beat frequency remains steady. Parallel studies with the rabbit cilia assay have shown the agglutination of tracheal cilia into clumps by adult bovine serum. The dilution of adult bovine serum lengthens the time before dyskinesia is first observed in both the Mytilus and rabbit assays. Fetal bovine or human cord sera do not cause ciliary dyskinesia or ciliary agglutination. The evidence presented shows that the observed ciliary dyskinesia can be induced by sera from a variety of animals and is not specific to cystic fibrosis; it also suggests that the serum immunoglobulin fraction is responsible for these effects--initial studies implicate IgM.

Adult↗

Autoimmune neutropenia in infancy due to anti-NA1 antibody: detection of antibody with immunofluorescence and agglutination test.

The sera from two patients with chronic neutropenia in infancy were examined for the presence of antineutrophil antibodies and their specificity against neutrophil antigen by using granulocyte indirect immunofluorescence test and microleukocyte agglutination test. In the microleukocyte agglutination test, the patients' sera reacted with neutrophils from their parents and normal unrelated donors having the neutrophil antigen NA1, but not with neutrophils from NA1- donors. After the absorption of patients' sera with NA1+ neutrophils, the antibody activity was completely abolished, resulting in the confirmation of the anti-NA1 antibody. In contrast, the granulocyte indirect immunofluorescence test showed positive reactions against both NA1+ and NA1- neutrophils, and the specificity for anti-NA1 was found in the results of the sera absorbed with NA1+ neutrophils. This suggested that the absorption experiment might be necessary to determine the specificity of the antibody for neutrophil antigen. Thus, we confirmed two cases with autoimmune neutropenia caused by anti-NA1 antibody. A combination of agglutination and immunofluorescence techniques would be recommended for investigation of neutrophil antibodies against the neutrophil-specific antigen.

Agglutination Tests↗

Plant protoplast agglutination and membrane-bound beta-lectins.

Plant protoplast agglutination caused by normal and immune serum and by artificial carbohydrate antigens is described. Evidence is presented that the new class of lectins, called all-beta lectins by previous authors, are present in protoplast membranes and responsible for both forms of agglutination. Some non-specific serum component and the artificial antigens are the 2 respective passive 'bridge' molecules between agglutinating protoplasts.

Agglutination↗

Plant protoplast agglutination by artificial carbohydrate antigens.

The existence of beta-lectins on protoplast surfaces is confirmed by the agglutination of protoplasts by those Yariv antigens that have sugar specificities which also interact with isolated beta-lectins. Agglutination by beta-maltosyl but not by beta-D-mannosyl Yariv antigens is used to identify some of the structural features required of the ligand for beta-lectin binding. Inhibition of agglutination by phenolic glycosides and the effect of protoplast fixation are also investigated.

Agglutination↗

Cell hybridization and cell agglutination. I. Enhancement of cell hybridization by lectins.

A great increase in hybridization frequency of cultured rodent cells was obtained when conventional cell fusion using 50% polyethylene glycol (PEG) was combined with a cell agglutination produced by plant lectins. The rate of appearance of hybrid colonies was found to be correlated with the extent of cell agglutination by lectin, as well as with cell fusion induced by subsequent PEG treatment. Phytohemagglutinin (PHA), wheat germ agglutinin, Wistaria floribunda agglutinin and concanavalin A were all active; the most effective was PHA. When parental cells in a monolayer were treated with PHA followed by PEG, the resulting hybridization frequency was very low because of markedly decreased viability, whereas the same cells in suspension yielded hybrid colonies at a higher rate. These results suggest that the enhancement of hybridization by PHA/PEG treatment was brought about by the ability of lectin to agglutinate cells.

Agglutination↗

Roles of sexual cell agglutination in yeast mass mating.

The agalpha1 mutant MAT alpha cells specifically lack the cell surface alpha-type sexual agglutination substance, which is also called alpha-agglutinin. Because the mutant cells (MATalpha agalpha1) can not form aggregates with MATa cells, MATalpha agalpha1 cells are unable to mate with MATa cells when they are co-inoculated in a liquid medium, and the mating is attenuated on solid medium. The attenuated mating ability shown in the previous studies gave us a vague idea about a physiological function of the sexual agglutinability. In order to solve the question, mating behavior of MATalpha agalpha1 cells was investigated here under conditions where the contact between MATa and MAT alpha cells is assisted by physical methods. A synthetic mutation agalpha1::URA3 was constructed and used as well as agalpha1-1 for this study to ensure the genetic defect. When a mixture of MATa and MAT alpha cells was kept on filter membrane placed on relatively dry agar medium, even agalpha1::URA3 mutant cells mated as efficiently as the wild type (AGalpha1) cells did. On filter membrane placed on moist agar medium, agalpha1 mutants mated 10-fold less efficiently than wild type cells did. The mutant cells mated 10000-time less efficiently than the wild type cells in a pellet formed by brief low speed centrifugation. In contrast, the wild type MATalpha cells mated well under all conditions tested. Under the pellet condition, a mixture of MATa and MATalpha AG alpha1 cells formed an extended and cotton-like pellet while a mixture of MATa and MATalpha agalpha1 cells formed a compact and tight pellet. These results suggest that sexual cell agglutination contributes not only to cell contact between MATa and MAT alpha cells thereby stabilizing a-alpha cell pairs, but also to construction of a uniquely organized ultra structure favorable for zygote formation and subsequent growth of diploid cells. The mating specific extended pellet formation was observed also in 4 pairs of a and alpha strains in ascosporogenous yeast genera Hansenula and Pichia.

Agglutination↗

Binding of lactoferrin to bacterial cells of the Clostridium species and their agglutination.

Cell agglutination in cell suspensions of 10 strains of Clostridium by lactoferrin (Lf) was observed by obtaining the ratio of increased absorbance (RIA) at 450 nm. The RIA values were very different among the species, being higher in the cell suspensions with bovine Lf (bLf) than in those with human Lf. The binding ability of bLf to the bacterial cells was also observed by an enzyme-linked ligand-binding assay, using the conjugate of iron-free or iron-saturated bLf with horseradish peroxidase (HRPO). The binding ability of bLf was very different among the 10 species, and showed a significant correlation with the cell agglutination of each strain. bLf formed a complex with the cells of C. perfringens, did not dissociate in 2 M NaCl or 4 M urea, but did dissociate in 1 M KSCN. These results suggest that the agglutination of cells of the Clostridium species by bLf is probably caused by the cooperative action of at least electrostatic and hydrophobic interactions between bLf and certain components of the cell surface.

Agglutination↗

[An evaluation of the direct agglutination test for the diagnosis of "mal de caderas" in horses].

The usefulness of the direct agglutination test (DA) to diagnose Mal de Caderas disease was evaluated. Forty four sera samples from two lots of horses with natural T. evansi infection (Lot 1 and Lot 2) were used. Thirteen (81.2%) of sixteen horses in which parasites were isolated gave positive agglutination titres (> or = 1:512) in the DA test. Treatment of these positive sera with 2-mercaptoethanol drops three to eight dilutions the agglutination titres in twelve samples (92%), showing the IgM nature of these antibodies. The DA test was also positive in seventeen of twenty eight horses in which parasites could not be detected. Five T. evansi infected horses, Lot three, which had high antibodies levels in the DA test, were treated with Naganol (Bayer-Germany). In four animals these antibodies were mainly IgM. In agreement with negative control for parasites, two months after treatment, IgM could not be detected while IgG antibodies remained detectable in low titres 12 months in three of the five horses. Fifty control horses sera from a T. evansi free area were AD negative. The DA and DA+2-ME are recommended as a routine method to diagnose Mal de Caderas disease in combination with parasitological diagnostic methods.

Agglutination Tests↗

Prevalence of agglutinating antibodies to Neospora caninum in raccoons, Procyon lotor.

Neospora caninum is an apicomplexan parasite that causes neonatal neuromuscular disease in dogs and abortions in cattle. Dogs are the only proven definitive host. Little is known about the prevalence of antibodies to this parasite in wildlife. Sera from 99 raccoons (Procyon lotor) were examined for agglutinating antibodies to N. caninum using the modified agglutination test employing formalin-fixed tachyzoites as antigen. Raccoons originated in Florida (n = 24, collected in 1996), New Jersey (n = 25, collected in 1993), Pennsylvania (n = 25, collected in 1999), and Massachusetts (n = 25, collected in 1993 and 1994). Ten (10%) had antibodies to N. caninum; 9 had titers of 1:50, and 1 (1%) had a titer of 1:100. The present study indicates that raccoons have minimal exposure to N. caninum. The sera were also tested for agglutinating antibodies to Toxoplasma gondii and 46 (46%) were positive; 16 had titers of 1:50, 8 had titers of 1:100, and 22 had titers of > or = 1:500.

Agglutination Tests↗

The time course of cytoadhesion, immunoglobulin binding, rosette formation, and serum-induced agglutination of Plasmodium falciparum-infected erythrocytes.

We describe morphologic characteristics of acridine orange-stained Plasmodium falciparum-infected erythrocytes and the sequential expression of several adhesion phenomena. In particular, we have studied when the adhesive and antigenic modifications appear on the infected erythrocyte surface that mediate binding to C32 melanoma cells (cytoadherence) or to erythrocytes (rosette formation) during a complete 48-hr life cycle of the parasite. The C32 melanoma cell binding started at about 12 hr and was seen during the whole life cycle with a peak around 28 hr (650 infected erythrocytes/100 C32 melanoma cells). Rosettes started to appear and immunoglobulin was found bound to the parasitized red blood cell (PRBC) somewhat later (16-20 hr). These adhesive events culminated at the mid-trophozoite/schizont stage (24-36 hr) with rosette formation and an immunoglobulin binding rate of about 50%, which decreased to about half of the peak values at the end of the life cycle. Serum-induced agglutination of the infected erythrocytes was also most extensive at 24-36 hr, but agglutination was seen with all late stage parasites, i.e., both trophozoites and schizonts at 24-48 hr of age. Taken together, adhesion to C32 melanoma cells starts prior to that of rosette formation, immunoglobulin binding, or serum-induced agglutination.

Agglutination Tests↗

Co-agglutination test for the detection of circulating antigen in amebic liver abscess.

We report here a simple and economical slide agglutination test, the co-agglutination (Co-A) test, for the detection of circulating amebic antigen in sera for the diagnosis of amebic liver abscess. Fifty serum specimens from cases of amebic liver abscess, 25 from other individuals with parasitic and miscellaneous infections, and 25 from healthy controls were tested for the presence of serum antigen by the Co-A test. Forty-five (90%) amebic liver abscess sera were found to be amebic-antigen positive by the Co-A test. None of 25 sera from healthy controls were positive for the antigen. However, false-positive results were seen with two sera from those with other parasitic and miscellaneous infection controls. These results show that the Co-A test can be used as a sensitive and specific rapid slide agglutination test for the detection of amebic antigen in the sera for diagnosis of cases of amebic liver abscess in a routine parasitology laboratory.

Agglutination Tests↗

Misidentification of Vibrio cholerae O155 isolated from imported shrimp as O serogroup O139 due to cross-agglutination with commercial O139 antisera.

Fish and shellfish products imported into Denmark are routinely analyzed for pathogenic Vibrio spp., particularly Vibrio cholerae, if products originate from subtropical or tropical areas. A V. cholerae strain that agglutinated commercial O139 antiserum but not the O1, Inaba, or Ogawa antisera was isolated from imported raw frozen shrimp. The toxigenicity of the strain was analyzed, and the results of a polymerase chain reaction showed that the V. cholerae strain did not contain the virulence genes ctx, tcpA, and zot, which are normally found in V. cholerae O1 and O139. The strain was resistant to colistin and spectinomycin. The high susceptibility of the strain to antimicrobial agents was confirmed by the lack of an SXT element, a self-transmissible, chromosomal genetic element that is normally present in 0139 strains and encodes resistance to sulfonamides, trimethoprim, and streptomycin. The strain contained two plasmids, in contrast to other O139 strains, which normally do not contain plasmids. The characteristics of the strain led to further agglutination testing with other antisera that are not commercially available, and the strain was found to agglutinate O155 antiserum in repeated testing. Manufacturers of 0139 antiserum should be aware of the closely related O antigens of the O139, O22, and O155 serogroups and should be aware that their commercial diagnostic O139 antiserum must be absorbed to remove cross-reacting agglutinins of O22 and O155 strains.

Agglutination Tests↗