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Haemolytic anaemia in previously healthy adult patients with CMV infections: report of two cases and an evaluation of subclinical haemolysis in CMV mononucleosis.

Whereas haemolytic anaemia is commonly encountered in infants and young children with cytomegalovirus (CMV) infections, it is an infrequent complication of CMV-induced infections in previously healthy adults. The data from 2 such patients are presented. One patient's Hb fell to a level of 36 g/l, and she required prednisone and blood transfusions. Her direct antihuman globulin test (DAT) was positive (IgG), and her red blood cell survival (51Cr) revealed a T 1/2 of 5 d. Both saline-agglutinating and low-molecular-weight cold agglutinins (CA) (4 degrees C) that reacted against both cord and adult cells were identified. In the second case, a moderate haemolytic anaemia (lowest Hb 87 g/l) was accompanied by negative DAT and CA studies. 20 other patients with CMV-mononucleosis were evaluated for evidence of subclinical haemolysis. Reticulocyte counts greater than 3.0% were noted in 9 of these patients. Haptoglobin values were below 0.5 g/l in 13 patients, and a positive DAT was recorded in 3/10 cases. This study documents haemolysis in many non-immunosuppressed adult patients with CMV infections. The mechanism responsible remains obscure.

Adolescent↗

48, XXYY syndrome associated with ethylenediaminetetraacetic acid (EDTA)-dependent pseudothrombocytopenia.

A 56-yr-old man with hypogonadism, gynecomastia, and mental retardation was evaluated for chromosome constitution and thrombocytopenia. Chromosomal analysis demonstrated the mosaicism of 48, XXYY and 47, XXY in the peripheral lymphocytes. Twenty out of twenty-five cells were 48, XXYY karyotype and the remaining five were 47, XXY karyotype. Thrombocytopenia was the EDTA-dependent pseudothrombocytopenia type 1 (platelet agglutination). Serological examination suggests that the platelet agglutinin belongs to IgM-kappa type. The present case exhibited both EDTA-dependent pseudothrombocytopenia and the 48, XXYY syndrome. Although this combination may have occurred purely by change, the possibility of whether or not the mosaicism of lymphocytes produces platelet agglutinin remains to be clarified.

Edetic Acid↗

Characterization, stability and in-vivo distribution of asialofetuin glycopeptide incorporating DSPC/CHOL liposomes prepared by mild cholate incubation.

In this study, a small triantennary asialoglycopeptide of fetuin (A-F2) was used as a ligand to direct liposomes to hepatocytes. A-F2 was cleaved from asialofetuin, purified, conjugated with fatty acids and incorporated into pre-formed sonicated DSPC/Chol (2:1) liposomes. A mild cholate incubation method for incorporating the A-F2 ligand on pre-formed vesicles was used. In preliminary in vivo experiments 111In3+ encapsulated in A-F2/palmityl liposomes was seen to accumulate in the liver of mice significantly faster than when encapsulated in non-ligand bearing liposomes of the same lipid composition (studied before), justifying further investigation of this system. The presence of the A-F2/fatty acid conjugate in a functional form on the vesicle surface was confirmed by their reversible agglutination in the presence of Ricinus communis agglutinin (RCA120). Effects of ligand incorporation on the vesicle size distribution, z-potential, membrane integrity and stability were monitored. The results demonstrate that highest ligand incorporation was achieved when liposomes and ligand were co-incubated in the presence of 1 mM sodium cholate. Incorporation increased with the length of the fatty acid used for A-F2 conjugation. Ligand-bearing liposomes were demonstrated to be smaller in diameter (about 30%) with a more positive z-potential in comparison to control vesicles while ligand incorporation did not influence the liposome membrane integrity. The size of the ligand-incorporating vesicles was maintained after 24 hours of incubation in isotonic buffer, proving that the vesicles do not aggregate. Although the preliminary biodistribution results may suggest that ligand bearing liposomes are accumulating in the liver, further cell culture, in vivo distribution and especially liver fractionation studies are required in order to clarify the intrahepatic localization of these liposomes and the ability to target liver hepatocytes in vivo.

Animals↗

[Immune hemolytic anemia and thrombocytopenia after peroral therapy with a penicillin derivative (phenoxymethylpenicillin)].

A previously apparently healthy male patient developed Coombs-positive haemolytic anaemia and thrombocytopenia during phenoxymethylpenicilline intake. A drug dependent IgG red cell antibody was demonstrated in patient's serum. In vitro tests with patient's serum suggested formation of complexes involving penicilline which by complement mediated mechanisms were able to agglutinate red cells in saline medium. Platelet agglutinins were demonstrated in patient's serum but their drug dependence has not been fully elucidated. The mechanism of drug mediated autoantibody synthesis might have been involved in the pathogenesis of thrombocytopenia.

Adult↗

Serologic survey for leptospirosis in Arizona beef cattle in 1981.

Sera from 1,215 beef cattle in Arizona were evaluated by leptospiral microscopic agglutination test in 1981. Over 25% had agglutinins to greater than or equal to 1 of 5 serovars of Leptospira interrogans used as antigens (canicola, grippotyphosa, hardjo, icterohaemorrhagiae, and pomona) at a titer of greater than or equal to 1:100, and 8.2% had titers of greater than or equal to 1:400 to greater than or equal to 1 serovars. The most common serovar to which reactions were detected was hardjo; agglutinins were detected at titers of greater than or equal to 1:100 in 14.3% and of greater than or equal to 1:400 in 5.5%. Cross reactions were rare at serum dilutions greater than or equal to 1:100 (2%) and extremely rare at greater than or equal to 1:400 (0.7%). Because vaccination with leptospiral bacterins is seldom practiced in Arizona beef cattle, a titer of greater than or equal to 1:100 may be useful in estimating incidence and prevalence of the disease and as an aid to diagnosis of leptospirosis.

Animals↗

Antisperm antibodies and male infertility.

Two hundred males attending a fertility clinic and 65 men who had fathered one or more children were screened for sperm antibodies using sperm agglutination and immobilisation tests. The incidence of sperm antibodies was significantly higher in the former group. Agglutinin and immobilisin were not related to sperm concentration, morphology, ejaculate volume, seminal plasma zinc, magnesium, acid phosphatase and fructose. However, sperm motility and viability were significantly reduced in the presence of sperm antibodies. There appeared to be no relationship between the presence of antisperm antibodies and Mycoplasma, aerobic or anaerobic organisms. There was a positive correlation between agglutinating and immobilising antibodies in serum and seminal plasma. This study suggests that sperm antibodies at a significant titre may lower male fertility.

Agglutination Tests↗

Immunochemical studies on lipopolysaccharide from agglutinable and non-agglutinable vibrios.

Lipopolysaccharide (LPS) prepared from Vibrio cholerae and a non-agglutinable (NAG; not agglutinable with O-group I serum according to Gardner and Venkatraman [13]) vibrio strain, isolated from a patient with cholera-like clinical symptoms, have been compared with respect to their chemical composition and immunological behavior. In addition to a significant difference in the chemical composition between the two lipopolysaccharides, the LPS from V. cholerae, unlike that from the NAG vibrio, requires prior treatment with alkali for it to be an effective antigen in the indirect hemagglutination test with sheep cells. It has been suggested that the alkali acts by removing excess O-acetyl group from LPS of agglutinable vibrios. LPS from the NAG vibrio consistently showed a lower antibody response in rabbits in terms of agglutinin and vibriocidal titer. Also, the class of agglutinin antibody elicited by LPS of the NAG vibrio was predominantly immunoglobin M, and that from V. cholerae was immunoglobulin G under comparable conditions.

Agglutination Tests↗

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↗

Sperm agglutinins in seminal plasma and serum after vasectomy: correlation between immunological and clinical findings.

The development of sperm agglutinins in serum and seminal plasma in relation to vasectomy was studied in forty-seven men by testing samples taken before vasectomy and on five occasions during the first year after vasectomy. Thirty additional patients were tested only 1 year after vasectomy. One year after vasectomy, sperm agglutinins in the serum in titres from 4 to about 4000 had developed in 62% of the entire group, while antibodies in the seminal fluid detectable by the gelatin agglutination test were present in only 4% of the group, and apart from on unusual case the titres were low here (either 4 or 8). Analysis of the modes of agglutination revealed changing patterns in several patients during the observation period, with a predominance of tail agglutinins after 1 year. In some cases, mixed agglutination was seen with serum but pure tail-to-tail agglutination with seminal plasma. The total number of spermatozoa in a pre-vasectomy ejaculate was found to be correlated with an early immune response and with the titre values after 1 year. The group of patients in whom agglutinins had developed 1 year after vasectomy were found to have significantly larger nodules at the sites of operation than those without sperm agglutinins.

Agglutinins↗

Immunoglobulin classes, secretory component, and sperm agglutinins in semen after vasovasostomy.

The immunoglobulin classes of the sperm-agglutinating antibodies in the seminal fluid and serum from four vasovasostomized men, as well as in operatively obtained testis-epididymis fluid from two of them, were investigated by specific solid-phase absorption of IgG, IgM, and IgA. In two patients the major part of the seminal agglutinins was IgA, and in one IgG, indicating that both local production and diffusion of sperm agglutinins from serum to the testis-epididymis compartment may take place. The total levels of immunoglobulins in seminal fluid generally increased after the operation, and IgM was present in two postvasovasostomy samples. This suggests a pathotopic potentiation effect of the vasectomy on the epithelial lining of the testis-epididymis compartment. Comparison of the results of solid-phase absorption with anti-IgA and anti-secretory component (SC) pointed to the presence of "free" SC in postvasovasostomy seminal fluid.

Agglutinins↗

The effect of hypothermic cardiopulmonary bypass on patients with low-titer, nonspecific cold agglutinins.

The effect of hypothermic cardiopulmonary bypass (CPB) was studied in 5 patients with strongly positive cold agglutination at 4 degrees C (experimental group) and in 10 controls. In the in vitro part of the study, the characteristics of the cold agglutinin antibodies in the experimental group included a low thermal amplitude (28 degrees C or less), a low 4 degrees C agglutination titer (1:32 or less), and nonspecificity (non-anti-I and non-anti-i). The in vivo portion of the study revealed a fall in total urine and serum free hemoglobin in both groups on going on bypass, followed by a rise for the remainder of bypass. Statistical comparison between observed and expected total free hemoglobin for both groups showed a significant rise (p less than 0.05) after bypass, thereby indicating ongoing hemolysis. No statistically significant difference between preoperative and post-operative blood urea nitrogen and creatinine levels and gross neurological status was observed in the experimental group. It was concluded that the patient with nonspecific cold agglutinins whose antibody is characterized by a low titer, a low thermal amplitude, and a lack of clinical symptomatology can undergo hypothermic CPB without increased threat of a hemolytic or vascular occlusive crisis.

Aged↗

Evaluation of a rapid, reliable, and inexpensive screening test for the serological diagnosis of human brucellosis.

In many laboratories the serological diagnosis of brucellosis is based on a tube agglutination test requiring incubation for two days and therefore causing delay in reporting of results. A slide agglutination test using an acidified, rose Bengal colored antigen that can be read after only 4 min was evaluated for its usefulness as a screening test for the presence of agglutinins in human sera. In sera of patients with culturally proven brucellosis or serological evidence for active infection the sensitivity of the slide agglutination test was 100%. Of 97 tube agglutination-negative sera none was positive in the slide test. From these results it was concluded that sera can be tested by the slide agglutination test which is quicker, easier to perform, and less expensive than the tube agglutination test. If positive, it is followed by a tube agglutination test which gives quantitative information and is necessary to follow the course of an infection.

Agglutination Tests↗

Trypanosoma lewisi: stage-specific surface antigens and exoantigens recognized by monoclonal antibodies.

The stage-specific expression of surface antigens by Trypanosoma lewisi was investigated using monoclonal antibodies directed against this parasite. Hybridomas secreting monoclonal antibodies were produced by the fusion of SP2/0-Ag 14 mouse plasmacytomas with spleen cells from rats infected previously with the Taliaferro strain of T. lewisi. Additivity enzyme-linked immunosorbent assays and indirect immunofluorescent antibody tests indicated the determinant recognized by monoclonal antibody TL40.3 (IgM) was different from those recognized by monoclonal antibodies TL40.1 (IgA), TL40.2 (IgM), and TL40.6 (IgG2 alpha). Monoclonal antibody TL40.3 agglutinated trypanosomes collected 3 days after parasite inoculation while monoclonal antibodies TL40.1, TL40.2, and TL40.6 agglutinated trypanosomes collected 6 days after inoculation. Since agglutinin titers against trypanosomes from irradiated (700 rad from a 60Co source) and nonirradiated rats were similar, expression of the antigens recognized by the monoclonal antibodies appeared to be independent of the immunological state of the host and the morphology of the parasite. The reproduction of T. lewisi in in vitro trypanostatic assays was inhibited only when the monoclonal antibodies were present in concentrations greater than or equal to those needed to agglutinate the trypanosomes. Monoclonal antibodies TL40.1 and TL40.3, but not TL40.2 and TL40.6, agglutinated erythrocytes collected later in the infection from irradiated, infected rats. None of the monoclonal antibodies agglutinated erythrocytes from nonirradiated, infected rats, from irradiated, noninfected rats or from nonirradiated, noninfected rats. This suggests that immunocompetent rats may make blocking antibodies against the exoantigens recognized by monoclonal antibodies TL40.1 and TL40.3.

Animals↗

A difference in the architecture of the surface membrane of normal and virally transformed cells.

Several tissue culture cell lines that were transformed by a tumor virus have been found to react with an agglutinin, while under identical conditions their untransformed parent cell lines did not agglutinate. Since a short treatment of the parent cell line with low concentrations of proteases exposed the same agglutinin receptor sites in a fashion indistinguishable from the transformed cells, it is proposed that both viral and chemical transformation produce changes in the architecture of the membrane, identical to those of the proteases.

Animals↗