C1-inhibitor treatment in patients with severe complement-mediated autoimmune hemolytic anemia.

Complement-mediated (CM) autoimmune hemolytic anemia (AIHA) is characterized by the destruction of red blood cells (RBCs) by autoantibodies that activate the classical complement pathway. These antibodies also reduce transfusion efficacy via the lysis of donor RBCs. Because C1-inhibitor (C1-INH) is an endogenous regulator of the classical complement pathway, we hypothesized that peritransfusional C1-INH in patients with severe CM-AIHA reduces complement activation and hemolysis, and thus enhances RBC transfusion efficacy. We conducted a prospective, single-center, phase 2, open-label trial (EudraCT2012-003710-13). Patients with confirmed CM-AIHA and indication for the transfusion of 2 RBC units were eligible for inclusion. Four IV C1-INH doses (6000, 3000, 2000, and 1000 U) were administered with 12-hour intervals around RBC transfusion. Serial blood samples were analyzed for hemolytic activity, RBC opsonization, complement activation, and inflammation markers. Ten patients were included in the study. C1-INH administration increased plasma C1-INH antigen and activity, peaking at 48 hours after the first dose and accompanied by a significant reduction of RBC C3d deposition. Hemoglobin levels increased briefly after transfusion but returned to baseline within 48 hours. Overall, markers of hemolysis, inflammation, and complement activation remained unchanged. Five grade 3 and 1 grade 4 adverse event occurred but were considered unrelated to the study medication. In conclusion, peritransfusional C1-INH temporarily reduced complement activation. However, C1-INH failed to halt hemolytic activity in severe transfusion-dependent-CM-AIHA. We cannot exclude that posttransfusional hemolytic activity would have been even higher without C1-INH. The potential of complement inhibition on transfusion efficacy in severe CM-AIHA remains to be determined.

Reduction of anti-K-mediated hemolytic disease of newborns after the introduction of a matched transfusion policy: A nation-wide policy change evaluation study in the Netherlands.

BACKGROUND: During pregnancy, maternal red blood cell (RBC) antibodies can lead to life-threatening fetal hemolysis and anemia. Women can become immunized by a pregnancy or an unmatched transfusion. Our aim was to quantify the effect of a nationwide K-matched transfusion policy for women of childbearing age potential to prevent K-immunization in pregnancy. STUDY DESIGN AND METHODS: In this nation-wide policy change evaluation study we determined the occurrence of RBC antibodies before and after introduction of a K-matched transfusion policy and evaluated the cause K alloimmunization 10 years after introduction of this measure. K-matched transfusion for females under 45 years of age is advised in the Dutch transfusion guideline since 2004. We used laboratory data from pregnancies with RBC antibodies identified in the period 1999-2018 obtained as part of a population-based screening program in the Netherlands. RESULTS: Tests of 36 286 pregnancies produced a positive antibody screening result which concerned anti-K in 1550 pregnancies. The occurrence of anti-K decreased from 67.9 to 20.2 per 100 000 pregnancies. The relative risk reduction was 0.70 which largely exceeded the relative risk reduction of 0.27 for antibodies against RBC antigens for which no preventive matching is required. The number of pregnancies at risk for anti-K-mediated disease decreased from 9.7 to 4.2 per 100 000 pregnancies. CONCLUSIONS: A K-matched transfusion policy is associated with a major decrease in a number of pregnant women with anti-K and pregnancies at risk for anti-K-mediated disease. A relatively simple measure is now shown to impact prevention of hemolytic disease in the fetus and newborn.

Plasma thrombopoietin levels as additional tool in clinical management of thrombocytopenic neonates.

Plasma thrombopoietin (Tpo) levels distinguish thrombocytopenia resulting from increased platelet destruction or decreased platelet production. We investigated whether measuring plasma Tpo levels in thrombocytopenic newborns is of diagnostic value to establish the underlying mechanism of thrombocytopenia.Tpo levels were measured with in-house developed ELISA in samples referred to our center because of thrombocytopenia noticed in the first 10 days of life. Clinical data were collected.Plasma Tpo levels <128 AU/ml were found in the majority (92%) of 121 newborns with immune-mediated thrombocytopenia (n = 104) and thrombocytopenia due to bacterial infections (n = 7); increased plasma Tpo levels (≥128 AU/ml) were found in thrombocytopenic newborns with severe asphyxia (n = 24). Highly increased plasma Tpo levels (>200 AU/ml) were found in thrombocytopenic neonates with congenital viral infections (n = 22) or amegakaryocytosis (n = 6). A plasma Tpo level <128 AU/ml excludes (negative predictive value 96%, 95% CI 90-99) severe asphyxia, congenital viral infections and amegakaryocytosis as the cause for thrombocytopenia in newborns.Increased plasma Tpo levels indicate that thrombocytopenia in newborns, as a result of various nonimmune disorders, is often caused by (temporary) bone marrow suppression/failure. Measurement of plasma Tpo levels provides the clinician with an additional tool to decide on the differential diagnosis, the necessity for subsequent diagnostics and treatment in neonates with thrombocytopenia.

A national Transfusion Register of Irregular Antibodies and Cross (X)-match Problems: TRIX, a 10-year analysis.

BACKGROUND: The Transfusion Register of Irregular Antibodies and Cross-match Problems (TRIX) is a unique national database in the Netherlands that was launched in 2007. Transfusion laboratories register the presence of irregular RBC alloantibodies for their patients and can consult the database for information that is relevant for pretransfusion testing, unknown in their own laboratory information system. STUDY DESIGN AND METHODS: Data from the TRIX database 10 years after implementation have been analyzed to demonstrate the added value of TRIX for transfusion practice. TRIX antibody registration, antibody disappearance likelihood, and differences between men and women have been analyzed and evaluated. RESULTS: In the 10-year period 2007 to 2016, a total of 80,164 alloantibodies have been identified and registered in 62,110 individuals. Of the antibodies, 81% were reported in women and 19% in men (female:male, 4.3:1). Rh (DCcEe and Cw ), K, Fya , and Jka antibodies account for 65.6% of all antibody registrations. M and Lewis antibodies account for 18.6% of all antibodies. Antibody disappearance likelihood is relatively high for the clinically relevant antibodies directed against Jkb , s, Fyb , and e. Antibodies directed against D, Fya , and K have a relatively low antibody disappearance likelihood. CONCLUSION: TRIX is a unique and useful tool for transfusion laboratories for timely and up-to-date information on the presence of erythrocyte antibodies, which improves pretransfusion testing and compatible blood selection. TRIX also provides macro data on the prevalence of individual antibodies and antibody disappearance likelihoods that can be used for developing blood type matching strategies for patient groups at risk. © 2019 AABB.

[Ceftriaxone-induced immune haemolytic anaemia and multi-organ failure]. / Hemolyse en multiorgaanfalen door ceftriaxon.

BACKGROUND: Drug-induced immune haemolytic anaemia (DIIHA) is caused by various drugs or their metabolites. Cephalosporins are associated with haemolytic anaemia but multi-organ failure is rarely described. CASE DESCRIPTION: We report the case of a 57-year-old female who was diagnosed with neuroborreliosis and treated with ceftriaxone. The patient developed severe DIIHA. Massive intravascular haemolysis led to shock and acute renal failure, necessitating mechanical ventilation and dialysis. Treatment with ceftriaxone was discontinued and glucocorticoids were prescribed. The patient recovered slowly but fully. CONCLUSION: Ceftriaxone-induced immune haemolytic anaemia is a rare but potentially fatal condition.

Postnatal outcome in neonates with severe Rhesus c compared to rhesus D hemolytic disease.

BACKGROUND: Neonates with Rhesus c (Rh c) hemolytic disease of the fetus and newborn (HDFN) are often managed in the same way as neonates with Rhesus D (Rh D) HDFN, although evidence to support this policy is limited. The objective of this study was to evaluate neonatal outcome in severe Rh c HDFN compared to Rh D HDFN. STUDY DESIGN AND METHODS: A retrospective study of (near-)term neonates with severe Rh c (n = 22) and Rh D HDFN (n = 103; without additional antibodies) admitted to the Leiden University Medical Center between January 2000 and October 2011 was conducted. The need for intrauterine transfusions (IUTs), phototherapy, exchange transfusions (ETs), and top-up transfusions up to 3 months of age were recorded and compared between both groups. RESULTS: Although there was a trend for a slightly more severe antenatal course for Rh D HDFN reflected by an earlier need for and higher number of IUTs (median [interquartile range], 2 [1.5-4] vs. 2 [1-2] in Rh c HDFN; p = 0.070), no significant differences were found for the postnatal course between Rh c and Rh D group in days of phototherapy (mean, Days 4.8 and 4.6, respectively; p = 0.569), need for ET (50% vs. 44%, respectively; p = 0.589), and top-up transfusions (62% vs. 78%, respectively; p = 0.128). CONCLUSION: Postnatal outcome in neonates with severe Rh c HDFN is similar compared to neonates with severe Rh D hemolytic disease in terms of days of phototherapy, need for ET, and need for top-up transfusions. These results justify a similar postnatal management of neonates with Rh D and Rh c HDFN.

Molecular analysis of the York antigen of the Knops blood group system.

BACKGROUND: Antigens of the Knops blood group system are present on complement component (3b/4b) receptor 1 (CR1/CD35), which is a transmembrane glycoprotein encoded by the CR1 gene. Eight of the nine known antigens of this system are linked to polymorphisms in Exon 29. The molecular background of one antigen, York (Yk(a)), has not yet been described. STUDY DESIGN AND METHODS: We aimed to identify a polymorphism associated with the absence of Yk(a) to enable molecular typing. Yk(a)-negative individuals were identified by serologic typing. Their CR1 gene was partially sequenced and compared to that of Yk(a)-positive individuals. Loss of Yk(a) antigen was investigated by expressing the SCR22/23 domain of both wild-type and mutated CR1 as a GPI-linked protein on HEK293 cells. RESULTS: We observed that absence of the Yk(a) antigen is caused by a mutation in Exon 26 of the CR1 gene. This 4223C>T mutation results in a 1408T>M change at the protein level. Ten of 117 donors (8.5%) were homozygous TT, confirming the Caucasian frequency of 8% Yk(a)-negative individuals. Serologically, these TT donors showed a Yk(a)-negative phenotype, while CC/CT individuals were Yk(a)-positive. While the Yk(a) antigen was present on HEK293 cells expressing wild-type constructs, cells expressing the 4223C>T variant were Yk(a) negative. CONCLUSION: We identified a 4223C>T sequence variation in the CR1 gene causing absence of the Yk(a) antigen of the Knops blood group system. With this finding, all polymorphisms of the known Knops blood group antigens have been revealed, enabling molecular testing to contribute to red blood cell alloantibody identification procedures.

Blood group does not correlate with disease severity in patients with Fabry disease (alpha-galactosidase A deficiency).

Blood groups B and P1 are substrates for the lysosomal enzyme alpha-galactosidase A. Therefore, patients with alpha-Gal A deficiency and blood groups B or P1 may exhibit more severe disease. In 48 Fabry patients distribution of blood group was not different from that in the Dutch population. No patient had blood group B. Clinical symptoms did not differ between bloodgroup P1 or P2 patients. We conclude that blood groups B and P1 are not overrepresented in Dutch Fabry patients. Blood group P1 is not correlated with more severe disease and cannot be considered a significant risk factor.

Plasma thrombopoietin levels in patients with chronic renal failure.

INTRODUCTION: Thrombopoietin (Tpo) is the most important regulator of thrombocytopoiesis. The main sites of Tpo production are the liver and the kidney produce Tpo. In the current study, the influence of renal failure on overall Tpo production was evaluated. MATERIALS AND METHODS: Tpo levels were measured in 23 patients on hemodialysis (HD) and 16 patients on chronic ambulatory peritoneal dialysis (CAPD). Plasma glycocalicin (GC) levels and platelet counts were measured as parameters of platelet mass and platelet turnover. RESULTS: Platelet counts were significantly lower in the HD group, both before 207+/-98 x 10(9)/l (P<0.001) and after hemodialysis 202+/-102 x 10(9)/l (P<0.001) when compared to healthy controls, 293+/-79 x 10(9)/l. No significant difference was found between platelet counts in patients on CAPD and healthy donors. Mean plasma Tpo levels of HD patients were higher both before 23+/-18 AU/ml (P<0.0001) and after dialysis 25+/-26 AU/ml (P<0.0001), as compared to Tpo levels in healthy controls (11+/-8 AU/ml). Patients on CAPD had significantly higher Tpo concentrations, 29+/-25 AU/ml than healthy controls (P<0.0001). There was no difference in Tpo level between the HD and CAPD group. No correlation was found between Tpo concentration and platelet count, hematocrit, creatinine or uremia levels. The GC concentration was significantly higher in HD patients and CAPD patients when compared to healthy controls. There was no correlation between GC and Tpo level or platelet count. CONCLUSION: These results confirm the increased platelet turnover in patients with chronic renal failure. Moreover this study shows that the kidney does not seem to play a major role in the overall Tpo production in the body.

Three parameters, plasma thrombopoietin levels, plasma glycocalicin levels and megakaryocyte culture, distinguish between different causes of congenital thrombocytopenia.

Fourteen children with congenital thrombocytopenia were analysed in order to unravel the mechanisms underlying their thrombocytopenia and to evaluate the value of new laboratory tests, namely measurement of plasma thrombopoietin (Tpo) and glycocalicin (GC) levels and analysis of megakaryocytopoiesis in vitro. Three groups of patients were included. The first group (n = 6) was diagnosed with congenital amegakaryocytic thrombocytopenia. They had no megakaryocytes in the bone marrow, three out of four patients showed no megakaryocyte formation in vitro, and all had high Tpo and low GC levels. Mutations in the thrombopoietin receptor gene, c-mpl, were the cause. The second group of patients (n = 3) had normal Tpo and severely decreased GC levels. In bone marrow, normal to increased numbers of atypical, dysmature megakaryocytes were present. In vitro megakaryocyte formation was quantitatively normal. A defect in final megakaryocyte maturation and subsequent (pro-)platelets may be the cause of the thrombocytopenia. The patients in the third group (n = 5) had Wiskott-Aldrich syndrome (WAS). They had normal Tpo and GC levels and normal megakaryocyte formation both in vivo and in vitro. This corresponded with the generally accepted hypothesis that thrombocytopenia in WAS is due to increased platelet turnover. In conclusion, different causes of congenital thrombocytopenia can be distinguished using three parameters: Tpo and GC plasma levels and in vitro analysis of megakaryocytopoiesis. Therefore, these parameters may be helpful in early diagnosis of different forms of congenital thrombocytopenia.