NOR polyagglutination and Sta glycophorin in one family: relation of NOR polyagglutination to terminal alpha-galactose residues and abnormal glycolipids.

BACKGROUND: This report describes the characterization of polyagglutinable red cells (RBCs), identified in two generations of a Polish family. CASE REPORT: Untreated and modified RBCs of the proposita (TS) were tested by serologic methods, using human sera, antibodies, lectins, and inhibitors of agglutination. Moreover, glycophorins were characterized by sodium docecyl sulfate-polyacrylamide gel electrophoresis and Western blotting, and glycolipids were purified, fractionated by thin-layer chromatography, and detected with Ricinus communis agglutinin I (RCA-I, specific for galactose residues) and Griffonia simplicifolia IB4 lectin (GSL-IB4, specific for Gal alpha1-3Gal- structure). Some of the experiments were also performed on RBCs of members of TS's family. RESULTS: Polyagglutination, found in four members of TS's family, was identified as the second case of an earlier described NOR polyagglutination. The polyagglutination was decreased by treating the RBCs with alpha-galactosidase and was inhibited by a neutral glycolipid fraction from NOR+ RBCs. Detection of neutral glycolipids of TS's RBCs on the thin-layer plate by RCA-I and GSL-IB4 revealed the presence of components that were not detectable in control RBCs. Moreover, Western blotting of RBC membranes from five family members with glycophorin monoclonal antibodies and agglutination assays with anti-St(a) and anti-Dantu sera identified the presence of St(a) glycophorin in four members of the family, two of whom were NOR+ and two NOR-. CONCLUSION: Our results showed that two rare features of TS's RBCs, NOR polyagglutination and St(a) glycophorin, are inherited independently, and that NOR+ RBCs contain neutral glycolipids with an abnormal oligosaccharide structure, most likely terminated with alpha-galactosyl residues.

The value of gel test and ELAT in autoimmune haemolytic anaemia.

Three methods for detection of warm type IgG autoantibody were evaluated using 400 blood samples from 147 patients suspected of autoimmune haemolytic anaemia (AIHA). Three direct antiglobulin techniques (DAT) were used: conventional tube DAT, gel-DAT by micromethod and gel-DAT enzyme linked antiglobulin test (ELAT). Eluate examinations confirmed the presence of autoantibodies on red cells. These tests were compared directly using 126 selected blood samples from 85 patients with IgG molecules on their red cells detected by the gel test. In 106 of these samples, collected from 65 patients with clinical symptoms of AIHA, the presence of autoantibody was confirmed by acid elution. The ELAT was positive in 100 samples (94%), 87 samples for tube DAT (82%). The ELAT as well as the tube DAT was negative in 20 samples with non-reactive eluates by gel test. The gel-DAT was therefore not fully specific and detected IgG on red cells of patients with hypergammaglobulinaemia. However, due its higher sensitivity it proved useful as a screening test. The ELAT allowed changes in the number of IgG molecules per red cell to be monitored quantitatively. Both methods play a part in the diagnosis and monitoring patients with warm type IgG auto-antibody.

Enzyme-linked antiglobulin test for the evaluation of the amount of IgG autoantibodies on red blood cells.

An enzyme-linked antiglobulin test (ELAT) was adapted to the assessment of IgG autoantibodies on red cells. Due to the worked out standard curve and the performance of the test on microplates, ELAT became a simple and inexpensive technique. It can be used for the detection of IgG within the range from 70 to 37,500 molecules per red cell. This is a simple quantitative method that can be used in immunohaematological studies of red cells.

Active immunization of children with leukemias and lymphomas against infection by hepatitis B virus.

Engerix B vaccine was administered to 54 children with leukemias and lymphomas aged from 2 to 15 years. In 36 cases chemotherapy was completely stopped, and 18 cases were receiving maintenance treatment. Engerix B was given at 0, 1, 2 and 6 months in a dose of 20 micrograms to children < 10 years, and 40 micrograms to older patients. The effectiveness of active immunization was demonstrated after complete therapy cessation in 88% of cases. The levels of antibodies determined 1 year after primary vaccination remained high, and in most of the vaccinated children they were > 1000 mIU/mL. In children vaccinated in the course of maintenance treatment the levels of antibodies did not give sufficient protection against infection.

[Screening for risk factors in agricultural schools of 5 Austrian federal counties ]. / Risikofaktorscreening in landwirtschaftlichen Schulen aus 5 österreichischen Bundesländern.

While screening activities have already been performed in a wide range in adults in Austria, extensive investigations in children and juveniles have so far only been done in a very limited number. In 1094 out of 1732 juveniles attending agricultural schools all lipid parameters were fully documented and thus could be evaluated. On an average, the total cholesterol (169 mg/dl) was too high; the worst findings for lipid parameters cholesterol, HDL-cholesterol, triglycerides, apolipoprotein-A1 were found in pupils of Burgenland. The cholesterol/HDL-cholesterol ratio in Burgenland was significantly (p < 0.01) higher than in Lower Austria. Female participants had a higher total cholesterol, HDL-cholesterol, and apolipoprotein-A1, but lower triglycerides and also lower blood pressure. Juveniles with a positive family history showed lower total cholesterol, but higher triglycerides as well as HDL-cholesterol. In heavy smokers, but also in occasional smokers, a lower total cholesterol, but decreased HDL-cholesterol and apolipoprotein-A1 and increased triglycerides were found. These findings underline the high prevalence of hypercholesterolemia in juveniles justifying screening activities and possibly therapeutic intervention at this young age.

[Evaluation of screening and complementary tests for anti-HCV antibodies]. / Ocena testów przegladowych i uzupelniajacych, sluzacych do wykrywania przeciwcial anty-HCV.

Screening for antibodies to hepatitis C virus (HCV) in blood donors by second generation tests was started on May 1991. On July 1992 obligatory testing of every blood or plasma donation was implemented. The incidence of anti-HCV evaluated in the first period (236.590 sera) was 1.4%. In the second period (296,573 sera) the incidence dropped to 0.9%. 489 sera repeatedly positive in screening were examined by a complementary test, 4-RIBA. Compatible positive results were obtained in 72.8% of the sera. 9.4% of the sera were negative and 17.8% gave indeterminate results. Reactivity was most frequently (95.7%) encountered to the structural core HCV peptide. The value of 4-RIBA was discussed. In conclusion, it was pointed out, that blood donors deferral should be based on repeated screening.

[Prevention of hepatitis B by passive immunization of children with chronic hematologic disease immunoglobulin]. / Zapobieganie metoda biernej immunizacji zakazeniu wirusem zapalenia watroby typu B u dzieci chorych na przewlekle choroby krwi.

In order to prevent the hepatitis virus B (HBV) infection, in 66 children aged from 6 months to 14 years with acute and chronic leukaemias, Hodgkin's and non-Hodgkin's lymphomas, testicular tumours, and aplastic anaemia a specific immunoglobulin was used intravenously (Hepatect made by Biotest). Hepatect was given every month to children with proliferative diseases throughout the whole time of intensive chemotherapy, and to children with aplastic anaemias during the administration of antilymphocytic globulin, prednisone, and Anapoln. Fourteen children were excluded from the analysis due to lack of systematic follow-up. Among 52 studied children, in most cases considerable fluctuations were observed of antibody concentration, the maximal values of which were of 150 mIU/ml. In 35 children, with the exception of sporadic falls, the anti-HBs antibody level remained level was noted, in two cases the presence of antibodies was revealed only sporadically. One of these children was infected with HBV. In all, three children were infected (5.76% of all children in the studied group). Perhaps the use of higher doses of Hepatect and its more frequent administration in children showing low anti-HBs level after routine doses might reduce further the incidence of infection.

[Prediction of the severity of fetal erythroblastosis using the erythrocyte phagocytosis test and evaluating anti-Rh antibody titers and their levels by autoanalysis]. / Przewidywanie ciezkosci choroby hemolitycznej noworodków za pomoca testu erytrofagocytozy, miana przeciwcial anty-Rh i ich stezenia ocenianego w autoanalizatorze.

Erythrophagocytosis++ test was performed in 45 pregnant Rh-negative women together with routine anti-Rh antibodies assay with indirect antiglobulin test and their concentration determined in an autoanalyser. It was found that the degree of ++phagocytosis of the standard erythrocytes immunized with maternal anti-Rh antibodies by the monocytes from healthy subjects is an important predictive factor enabling to foresee a severity of hemolytic disease in newborn babies. ++Phagocytosis over 50% predicts a severe course of the disease whereas under 20%--a mild form of the serologic conflict.

Correlation of monocyte-monolayer assay results, number of erythrocyte-bound IgG molecules, and IgG subclass composition in the study of red cell alloantibodies other than D.

Comparisons have been made between the serological and immunological characteristics of 42 blood group alloantibodies (other than D) covering twelve systems using a monocyte-monolayer assay (MMA), a radiometric antiglobulin test for antibody binding and IgG subclass determinations. The results of the MMA correlated well with the level of IgG molecules bound on incompatible cells, and the highest levels in both cases were associated with the presence of the IgG3 subclass. However, limited clinical data shows that, while in general the MMA clearly identifies the clinically significant antibodies, the correlation with the degree of clinical outcome is less well defined, and in some instances other factors may be operating to ameliorate the in vivo effect of the antibody.

Analysis of immune response to red blood cell antigens in multitransfused patients with different diseases.

The rate of alloimmunization to red blood cell antigens in 1502 multitransfused patients, mainly with blood disorders, was analyzed in a retrospective study. The overall incidence of alloantibodies was 5.7%. Three groups of patients were identified with different potential for antibody production. The lowest probability (1.8%) of alloimmunization was found in the group of patients with lymphoproliferative syndromes, acute myeloid leukaemia and burn disease. The highest probability (33.4%) of immune response to red blood cell antigens was found in patients with AIHA, liver cirrhosis and myelodysplastic syndrome. In the group of patients with chronic myeloid leukaemia, pancytopenias, anaemias of various origin and aplastic anaemia the probability of alloimmunization ranged from 5.7% to 13.6%. A possible role of genetic-factors and immune competence status in post-transfusion alloimmunization is briefly discussed.

First example of By phenotype.

In ABO blood grouping red cells of a young healthy woman behaved like O, but in her serum anti-B was lacking. By absorption-elution technique the very weak B activity was revealed. The saliva contained B and H substances, although the B to H ratio was significantly reduced, when compared to the normal B and Bm secretors. In the serum B transferase activity was not demonstrated. The presented results were comparable to those described in persons with Ay phenotype. Therefore, this weak B variant can be classified into By category.

Alloantibody anti-Gerbich: blood transfusion problems and family studies.

In a patient with myelodysplastic syndrom complement fixing IgG1 anti-Ge three weeks after Ge positive serologically compatible blood transfusion was found. The patient and one of his children were Ge-1, -2, -3. Red cells were elliptocytic. The patient's nonconsanguineous wife and other family members were Ge positive. In the red cell membranes of Ge negative persons glycophorin C was lacking. Abnormal glycophorin was present in the red cell membranes of all family members. Anti-Ge appeared to be of no clinical significance. Transfusion of serologically incompatible Ge positive blood provided beneficial effect. After this transfusion anti-Ge disappeared from the serum and was not detected at the whole follow-up, although the patient received Ge positive blood several times. Immunological tolerance towards Ge antigen is suggested.

[Serological markers of hepatitis B virus and cytomegalovirus in patients with hemophilia]. / Serologiczne znaczniki wirusa zapalenia watroby typu B i wirusa cytomegalii u chorych na hemofilie.

In 100 adult patients with severe haemophilia A (78 patients) and B (22 patients) sera were screened for the presence of serological markers of hepatitis B virus (HBV) and of cytomegalovirus (CMV) and liver function tests were performed which included measurement of serum aminotransferase AST and ALT activities, total bilirubin concentration and plasma levels of factor VII and X. In all the patients at least one out of five determined HBV markers (HBsAg. HBeAg, anti-HBs, anti-HBc and anti-HBe) was detected. HBsAg was found in 10% of the patients, and its prevalence in haemophiliacs B was higher than than observed in haemophiliacs A (22.7% and 6.4%, respectively). HBsAg appeared more frequently in patients receiving factor VIII concentrates (16.7%) than in those treated with cryoprecipitate (4.5%). Anti-CMV antibody was detected in sera of 98% of the patients. In 1/3 samples of cryoprecipitate anti-HBc or anti-HBs were present, and in the half of samples anti-CMV occurred. Abnormal liver function tests indicating chronic hepatitis or liver cirrhosis were obtained in 8 patients. Raised ALT activity which could suggest chronic infection with non-A, non-B virus occurred in 6 cases. The present study indicates that haemophiliacs frequently transfused with plasma products are at high risk for viral infections leading to liver dysfunction.

Auto-anti-D blocking D determinants on red cells in pregnancy.

Positive direct antiglobulin test due to IgG1 autoantibody of anti-D specificity on red cells of an apparently healthy pregnant woman was found. At the period of the highest autoantibody activity D epitope were completely blocked and even by special methods used for Rh typing in patients with AIHA the detection of D antigen was not possible. Free auto-anti-D caused benign HDN in the infant. The autoantibody production was transient and stopped after delivery. D variant included into category V on the red cells of the patient under study and her two siblings was recognized. This finding, confirmed by in vitro study, gives the explanation of the complete blocking of D epitopes by anti-D autoantibody.