Secretor status of blood group O mothers is associated with development of ABO haemolytic disease in the newborn.

BACKGROUND AND OBJECTIVES: Identification of antibody characteristics and genetics underlying the development of maternal anti-A/B linked to inducing haemolytic disease of the foetus and newborn could contribute to the development of screening methods predicting pregnancies at risk with high diagnostic accuracy. MATERIALS AND METHODS: We examined 73 samples from mothers to 37 newborns with haemolysis (cases) and 36 without (controls). The secretor status was determined by genotyping a single nucleotide polymorphism in FUT2, rs601338 (c.428G>A). RESULTS: We found a significant association between secretor mothers and newborns developing haemolysis (p = 0.028). However, stratifying by the newborn's blood group, the association was found only in secretor mothers to blood group B newborns (p = 0.032). In fact, only secretor mothers were found in this group. By including antibody data from a previous study, we found higher median semi-quantitative levels of IgG1 and IgG3 among secretor mothers than non-secretor mothers to newborns with and without haemolysis. CONCLUSION: We found that the maternal secretor status is associated with the production of anti-A/B, pathogenic to ABO-incompatible newborns. We suggest that secretors experience hyper-immunizing events more frequently than non-secretors, leading to the production of pathogenic ABO antibodies, especially anti-B.

ABO haemolytic disease of the newborn: Improved prediction by novel integration of causative and protective factors in newborn and mother.

BACKGROUND AND OBJECTIVES: Prediction of haemolytic disease of the foetus and newborn (HDFN) caused by maternal anti-A/-B enables timely therapy, thereby preventing the development of kernicterus spectrum disorder. However, previous efforts to establish accurate prediction methods have been only modestly successful. MATERIALS AND METHODS: In a case-control study, we examined 76 samples from mothers and 76 samples from their newborns; 38 with and 38 without haemolysis. The IgG subclass profile of maternal anti-A and anti-B was determined by flow cytometry. Samples from newborns were genetically analysed for the A2 subgroup, secretor and FcγRIIa receptor alleles. RESULTS: Surprisingly, we found a correlation between the newborn secretor allele and haemolysis (p = 0.034). No correlation was found for FcγRIIa alleles. The A2 subgroup was found only in newborns without haemolysis. Unexpectedly, different reaction patterns were found for maternal anti-A and anti-B; consequently, the results were treated separately. For the prediction of haemolysis in A-newborns, the maternal IgG1 subclass determination resulted in an accuracy of 83% at birth. For B-newborns, an accuracy of 91% was achieved by the maternal IgG2 subclass determination. CONCLUSION: We improved the prediction of ABO-HDFN by characterizing maternal anti-A and anti-B by flow cytometry and we presented genetic traits in newborns with correlation to haemolysis. We propose a new understanding of A- and B-substances as immunogens that enhance the maternal immune response and protect the newborn, and we suggest that the development of ABO-HDFN is different when caused by maternal anti-A compared to maternal anti-B.

Prediction of ABO hemolytic disease of the newborn using pre- and perinatal quantification of maternal anti-A/anti-B IgG titer.

BACKGROUND: Hemolysis in fetus/newborns is often caused by maternal antibodies. There are currently no established screening procedures for maternal ABO antibodies harmful to fetus/newborn. We investigated the clinical significance, and predictive value of maternal anti-A/B titer for hyperbilirubinemia in ABO-incompatible newborns. METHODS: We conducted a case-control study of blood group O mothers and their ABO-compatible (O) vs. -incompatible (A/B) newborns receiving phototherapy, and of ABO-incompatible newborns receiving phototherapy vs. no phototherapy. Newborn data and treatment modalities were recorded, and total serum bilirubin and hemoglobin were measured. Maternal anti-A/B immunoglobulin-γ (IgG) titers were measured prenatally and perinatally, and negative and positive predictive values (NPV, PPV) were calculated to assess the risk of developing hyperbilirubinemia requiring phototherapy. RESULTS: We found a significantly higher maternal IgG antibody titer in the case group (p < 0.001). Maternal anti-A/B titers at first trimester had modest predictive values: NPV = 0.82 and PPV = 0.65 for neonatal hyperbilirubinemia; titers at birth improved the predictive values: NPV = 0.93 and PPV = 0.73. Newborn hemoglobin was significantly lower in incompatibles compared to compatibles (p = 0.034). Furthermore, increased anti-A/B IgG production during pregnancy was associated with hyperbilirubinemia and hemolysis in incompatible newborns. CONCLUSIONS: There was a significant association between maternal anti-A/B IgG titer and hyperbilirubinemia requiring treatment. IMPACT: Maternal anti-A/B IgG titer in the first trimester and at birth is predictive of hemolytic disease of the ABO-incompatible newborn. Increased IgG anti-A/B production throughout pregnancy in mothers to ABO-incompatible newborns developing hyperbilirubinemia contrasts a constant or reduced production in mothers to newborns not developing hyperbilirubinemia. Screening tools available in most immunohematology laboratories can identify clinically important IgG anti-A/B. Use of maternal samples taken at birth yielded NPV = 0.93 and PPV = 0.73.

Thirty-three-day storage of dithiothreitol-treated red blood cells used to eliminate daratumumab interference in serological testing.

BACKGROUND AND OBJECTIVES: Daratumumab binds CD38 on red blood cells causing interference with indirect antiglobulin tests. Dithiothreitol is used to eliminate interference allowing detection of alloantibodies. Haemolysis is observed during storage of dithiothreitol-treated antibody identification panel cells. The objective of this study was to develop a modified method for dithiothreitol treatment to reduce the haemolysis during 33 days of storage and still be able to eliminate daratumumab interference. MATERIALS AND METHODS: Panel cells were treated with various volumes of 0·2 m dithiothreitol supplied by various manufacturers. Haemolysis Index of dithiothreitol-treated and untreated panel cells was measured and compared on days 1, 15 and 33. Antibody screening tests with dithiothreitol-treated screening cells were performed on samples from 15 daratumumab-treated patients (dose 16 mg/kg) and 34 patients with known alloantibodies. Antibody identifications with dithiothreitol-treated panel cells were performed on seven additional known alloantibodies. RESULTS: Dithiothreitol treatment with a ratio of 30:25 (red blood cells:dithiothreitol) showed the same degree of haemolysis as with untreated panel cells. Daratumumab interference was eliminated in all 15 samples from daratumumab-treated patients. Twenty-six of 34 alloantibodies were detected, and all seven additional alloantibodies were identified using the modified dithiothreitol treatment. Eight alloantibodies within the Kell system were negative. No decrease in the reaction strength was observed during the 33-day storage period. CONCLUSION: The modified dithiothreitol method was able to reduce haemolysis during storage and to detect and identify alloantibodies in the presence of daratumumab.

Pneumatic tube transport of blood samples affects global hemostasis and platelet function assays.

INTRODUCTION: Pneumatic tube systems (PTS) are frequently used for rapid and cost-effective transportation of blood samples to the clinical laboratory. The impact of PTS transport on platelet function measured by the Multiplate system and global hemostasis measured by the TEG 5000 was evaluated. METHODS: Paired samples from healthy adult individuals were obtained at two study sites: Rigshospitalet (RH) and Nordsjaellands Hospital (NOH). One sample was transported by PTS and one manually (non-PTS). Platelet function was assessed by platelet aggregation (Multiplate) and global hemostasis was assessed by a variety of thrombelastography (TEG) assays. Multiplate (n = 39) and TEG (n = 32) analysis was performed at site RH, and Multiplate (n = 28) analysis was performed at site NOH. RESULTS: A significant higher agonist-induced platelet aggregation was found for PTS samples compared to manual transport at site NOH (P < .02, all agonists). No significant difference was found at site RH (P > .05, all agonists). For Kaolin TEG, samples transported by PTS showed a significant lower R-time and higher Angle (P < .001). No significant differences in MA and LY30 was found (P > .05). ACT of RapidTEG was significantly reduced (P = .001) and MA of Functional Fibrinogen TEG was significantly increased (P < .001) after PTS transport. No significant impact of PTS was observed for TEG assays with heparinase (P > .05). CONCLUSIONS: Depending on the type of PTS, transportation by PTS affected platelet aggregation measured by Multiplate. Furthermore, PTS alters TEG parameters possibly reflecting coagulation factors. Clinical laboratories should evaluate the effect of the local PTS on Multiplate and TEG results.