Background
Delayed haemolytic transfusion reaction (DHTR) is mainly caused by an immune response to transfused red blood cells (RBCs). Immunized patients have a high risk of producing antibodies in ...response to further transfusion. Controlling the immune response to RBCs is therefore a major goal in sickle cell disease (SCD).
Study design
We report an observational study of eight alloimmunized SCD patients with history of severe DHTR who were treated with rituximab before a new transfusion to prevent further immunization and DHTR.
Results
Five patients showed a good clinical outcome following transfusion preceded by preemptive treatment with rituximab. The remaining patients presented mild DHTR. In all patients, the results of post‐transfusion screening tests were identical to those of pretransfusion tests; no newly formed antibodies were detected.
Conclusion
These cases suggest that rituximab prevents at least occurrence of newly formed antibodies in high responders and minimizes the risk of severe DHTR. This study confirms that DHTR is complex in SCD and does not rely only on the classical antigens/antibodies conflict. Considering potentially serious adverse effect of rituximab, this treatment should be considered cautiously, and only when transfusion is absolutely necessary in patients with history of severe DHTR linked to immunization.
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BFBNIB, DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, UILJ, UKNU, UL, UM, UPUK
1 Etablissement Français du Sang, Ile de France, Hôpital Henri Mondor, Créteil;
2 Centre de Référence des Cytopénies Auto-Immunes, Hôpital Henri Mondor, Créteil;
3 Service des Maladies Génétiques du ...Globule Rouge, Hôpital Henri Mondor Créteil;
4 Service de Médecine Interne, Hôpital Henri Mondor, Créteil;
5 Laboratoire dImmunologie, Hôpital Henri Mondor, Créteil;
6 Université de Poitiers, EA 3806, CHU de Poitiers
Correspondence: France Noizat-Pirenne, MD, PhD, Etablissement Français du Sang dIle de France, 51 Avenue du Maréchal de Lattre de Tassigny, 94000, Créteil, France. Tel: 331 56 72 76 37; Fax: 331 56 72 76 01, E-mail: france.noizat-pirenne{at}efs.sante.fr
Delayed hemolytic transfusion reaction (DHTR), a life-threatening transfusion complication in sickle cell disease (SCD), is characterized by a marked hemoglobin drop with destruction of both transfused and autologous red blood cells (RBCs) and exacerbation of SCD symptoms. One mechanism of RBCs destruction is auto-antibody production secondary to transfusion. As rituximab specifically targets circulating B cells, we thought that it could be beneficial in preventing this immune-mediated transfusion complication. We report the case of a SCD patient who previously experienced DHTR with auto-antibodies and who needed a new transfusion. DHTR recurrence was successfully prevented by rituximab administration prior transfusion, supporting the safe use of rituximab to prevent DHTR in SCD patients as a second line approach when other measures failed.
Key words: Sickle cell disease, rituximab, transfusion, auto-antibodies.
Background and Objectives Intravascular haemolytic reactions are reported in red‐cell T‐activated patients after blood transfusion. The relationship between T antigen antibodies present in normal ...plasma and these reactions remains unclear. In this study, we assessed the haemolytic activity of T antibodies in vitro in comparison with anti‐A/B antibodies.
Materials and Methods We established a haemolysis assay based on treating target red‐blood‐cells (RBCs) with 2‐aminoethylisothiouronium bromide (AET). Two hundred and seven blood donor sera were analysed for anti‐T, anti‐A/B haemolysins and anti‐T agglutinins.
Results Anti‐T haemolysins were found in 4 (1·9%) blood donor sera using a standard haemolysis method and in 174 (84%) samples using AET‐treated RBCs. Haemolysis correlated with agglutination titres (P < 10−7). With both methods, anti‐T haemolysins were much weaker than anti‐A and anti‐B haemolysins. Gradual desialylation of RBCs showed a correlation between sialic acid level as indicated by agglutination with Sambucus nigra lectin and anti‐T mediated haemolysis that was significantly increased (fold 2·4) independently of T antigen expression.
Conclusion These data indicate that, in vitro, anti‐T‐mediated haemolysis depends primarily on the degree of desialylation of target RBCs. They suggest that the haemolytic activity of T antibodies‐containing human sera is usually weak and may only become significant in the very rare setting of a profound desialylation of RBCs.
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BFBNIB, DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, UILJ, UKNU, UL, UM, UPUK
Prevention of hemolytic transfusion reactions depends upon our capacity to prevent allo-immunization and conflicts between antigens of transfused red blood cells and antibodies produced by the ...recipient. In this study, we show that to secure transfusion of sickle cell disease patients, it is necessary to take into account their immunohematologic characteristics in the organization of transfusion.
Immunohematological data of 206 chronically transfused patients have been collected as well as phenotypes of transfused units. In order to prevent allo-immunization against C and E antigens for patients typed D+C-E-c+e+ (56%), 26% of the transfused units were D-C-E-c+e+. We found that 47% of the patients had a history of allo-immunization, whereas only 15% produced an antibody the day of inclusion in the study. The non-detectable antibodies were frequently known as dangerous for transfusion. Finally, this study shows the frequency of anti-D in D+ patients and anti-C in C+ patients, pointing out the question of partial antigens.
To insure optimal transfusion safety for sickle cell disease patients, three points have to be improved: blood donation within the Afro-Caribbean community living in France, access to history of immuno-hematological data, detection of variant antigens, especially within the RH blood system.
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GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
It has been shown recently that expression of p21 is enhanced by paclitaxel. This cytotoxic compound induces mitotic spindle damage resulting in blockade of the mitotic cell cycle associated or not ...with apoptotic cell death. In the present study, we showed that, in MCF-7 cells, paclitaxel induced accumulation of p21 in cells with a G2/M DNA content, corresponding to cells either in abnormal mitosis or in an interphase-like state (decondensed chromatin) with multiple nuclei. In MCF-7 cells, the increase in p21 was subsequent to the mitotic arrest and was associated with the exit from abnormal mitosis leading to formation of cells with micronuclei. In this cell line, we noted a relationship between the elevation of p21 expression and the inhibition of p34cdc2 activity. High levels of p21 protein were also found to be associated with inactive p34cdc2/cyclin B protein complex after treatment with paclitaxel. Treatment with p21 antisense oligonucleotide partially blocked induction of p21 expression by paclitaxel and significantly reduced survival of MCF-7 cells exposed to this agent. In NIH-OVCAR-3 cells, which are deficient in basal and paclitaxel-induced p21 expression, paclitaxel led to a prolonged activation of p34cdc2 and a delayed mitotic exit associated with apoptotic cell death. These observations suggest that p21 is not required for the mitotic arrest in response to paclitaxel, but argue in favor of a role for this inhibitor in facilitating the exit from abnormal mitosis. This effectively enhances cell survival after paclitaxel-induced spindle damage.
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DOBA, EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, IZUM, KILJ, KISLJ, MFDPS, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UILJ, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Defensins have been identified as key elements of innate immunity against microbial infections. In the present study, human beta-defensin-2 (hBD-2) mRNA and peptide expression were evaluated by ...RT-PCR and Western blotting in normal human keratinocytes, in function of their stage of differentiation. In proliferating, non-differentiating keratinocytes generated in serum-free, low-calcium medium, a very low hBD-2 mRNA expression was found. A significantly higher expression was detected in high-calcium cultivated keratinocytes grown either as monolayers or as multilayers under submerged conditions. In an air-liquid interface culture of keratinocytes, allowing epidermis to be reconstructed, hBD-2 mRNA expression level was significantly higher than in the other conditions and displayed inter-individual variability as observed in native epidermis. The peptide was detected only in reconstructed epidermis. These results indicate that hBD-2 gene expression in normal human keratinocytes is dependent upon their stage of differentiation. The level of expression of hBD-1 mRNA was lower and that of hBD-3 was higher than that of hBD-2 in reconstructed epidermis. Exposure of reconstructed epidermis to bacterial lipopolysaccharide (LPS) resulted in an average 4-fold increase in hBD-2 mRNA 18 h after challenge, but not of hBD-1 and hBD-3 gene expression. These results show the selective regulation of hBD-2-encoding gene in an organotypic epidermal model, in response to LPS. They also provide evidence that in vitro reconstructed epidermis represents a useful model for studying regulation of expression of beta-defensins after skin challenge with pathogenic microorganisms in conditions as close as possible to the in vivo situation.
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NUK, OBVAL, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
La prévention des hémolyses post-transfusionnelles chez les drépanocytaires est conditionnée par notre capacité à prévenir l’allo-immunisation et les conflits antigène–anticorps. Cette étude montre ...l’importance de mettre en place une organisation transfusionnelle adaptée au profil immunohématologique particulier des drépanocytaires pour leur garantir une sécurité transfusionnelle optimale.
Les données immunohématologiques de 206 adultes drépanocytaires transfusés en dehors de l’urgence ont été analysées, ainsi que le phénotype des concentrés globulaires (CGR) délivrés. Cinquante-quatre pour cent des patients ont un phénotype D+C−E−c+e+. Pour prévenir l’allo-immunisation anti-C et anti-E, 26 % de CGR D−C−E−c+e+ ont été transfusés à ces patients D+. Quarante-sept pour cent des patients ont un historique d’allo-immunisation, alors que seulement 15 % ont une recherche d’agglutinines irrégulières positive le jour de l’inclusion, les anticorps non décelables étant souvent dangereux. Enfin, cette étude montre la fréquence importante d’anti-D chez les sujets D+ et d’anti-C chez les sujets C+, posant la question du caractère partiel des antigènes D et C.
Pour une sécurité transfusionnelle optimale des patients drépanocytaires et une meilleure adéquation entre phénotypes des receveurs et phénotypes des CGR disponibles, trois points dans l’organisation transfusionnelle doivent être améliorés : la promotion du don au sein des populations afro-antillaises doit être renforcée pour disposer de CGR D+C−E−c+e+ et éviter d’utiliser la ressource limitée de CGR D−C−E−c+e+ ; les données immunohématologiques des patients doivent être accessibles aux prescripteurs et transfuseurs, afin d’éviter les accidents immuno-hémolytiques par restimulation ; enfin, la recherche par biologie moléculaire des antigènes variants d’intérêt transfusionnel doit être mise en place chez les donneurs et receveurs afro-antillais.
Prevention of hemolytic transfusion reactions depends upon our capacity to prevent allo-immunization and conflicts between antigens of transfused red blood cells and antibodies produced by the recipient. In this study, we show that to secure transfusion of sickle cell disease patients, it is necessary to take into account their immunohematologic characteristics in the organization of transfusion.
Immunohematological data of 206 chronically transfused patients have been collected as well as phenotypes of transfused units. In order to prevent allo-immunization against C and E antigens for patients typed D+C−E−c+e+ (56%), 26% of the transfused units were D−C−E−c+e+. We found that 47% of the patients had a history of allo-immunization, whereas only 15% produced an antibody the day of inclusion in the study. The non-detectable antibodies were frequently known as dangerous for transfusion. Finally, this study shows the frequency of anti-D in D+ patients and anti-C in C+ patients, pointing out the question of partial antigens.
To insure optimal transfusion safety for sickle cell disease patients, three points have to be improved: blood donation within the Afro-Caribbean community living in France, access to history of immuno-hematological data, detection of variant antigens, especially within the RH blood system.
Full text
Available for:
GEOZS, IJS, IMTLJ, KILJ, KISLJ, NUK, OILJ, PNG, SAZU, SBCE, SBJE, UL, UM, UPCLJ, UPUK
Defensins have been identified as key elements of innate immunity against microbial infections. In the present study, human β-defensin-2 (hBD-2) mRNA and peptide expression were evaluated by RT-PCR ...and Western blotting in normal human keratinocytes, in function of their stage of differentiation. In proliferating, non-differentiating keratinocytes generated in serum-free, low-calcium medium, a very low hBD-2 mRNA expression was found. A significantly higher expression was detected in high-calcium cultivated keratinocytes grown either as monolayers or as multilayers under submerged conditions. In an air-liquid interface culture of keratinocytes, allowing epidermis to be reconstructed, hBD-2 mRNA expression level was significantly higher than in the other conditions and displayed inter-individual variability as observed in native epidermis. The peptide was detected only in reconstructed epidermis. These results indicate that hBD-2 gene expression in normal human keratinocytes is dependent upon their stage of differentiation. The level of expression of hBD-1 mRNA was lower and that of hBD-3 was higher than that of hBD-2 in reconstructed epidermis. Exposure of reconstructed epidermis to bacterial lipopolysaccharide (LPS) resulted in an average 4-fold increase in hBD-2 mRNA 18 h after challenge, but not of hBD-1 and hBD-3 gene expression. These results show the selective regulation of hBD-2-encoding gene in an organotypic epidermal model, in response to LPS. They also provide evidence thatin vitro reconstructed epidermis represents a useful model for studying regulation of expression of β-defensins after skin challenge with pathogenic microorganisms in conditions as close as possible to thein vivo situation.PUBLICATION ABSTRACT
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EMUNI, FIS, FZAB, GEOZS, GIS, IJS, IMTLJ, KILJ, KISLJ, MFDPS, NUK, OBVAL, OILJ, PNG, SAZU, SBCE, SBJE, SBMB, SBNM, UKNU, UL, UM, UPUK, VKSCE, ZAGLJ
Background: Delayed haemolytic transfusion reaction (DHTR) is mainly caused by an immune response to transfused red blood cells (RBCs). Immunized patients have a high risk of producing antibodies in ...response to further transfusion. Controlling the immune response to RBCs is therefore a major goal in sickle cell disease (SCD).Study design: We report an observational study of eight alloimmunized SCD patients with history of severe DHTR who were treated with rituximab before a new transfusion to prevent further immunization and DHTR.Results: Five patients showed a good clinical outcome following transfusion preceded by preemptive treatment with rituximab. The remaining patients presented mild DHTR. In all patients, the results of post-transfusion screening tests were identical to those of pretransfusion tests; no newly formed antibodies were detected.Conclusion: These cases suggest that rituximab prevents at least occurrence of newly formed antibodies in high responders and minimizes the risk of severe DHTR. This study confirms that DHTR is complex in SCD and does not rely only on the classical antigens/antibodies conflict. Considering potentially serious adverse effect of rituximab, this treatment should be considered cautiously, and only when transfusion is absolutely necessary in patients with history of severe DHTR linked to immunization.
Full text
Available for:
BFBNIB, DOBA, FZAB, GIS, IJS, IZUM, KILJ, NLZOH, NUK, OILJ, PILJ, PNG, SAZU, SBCE, SBMB, UILJ, UKNU, UL, UM, UPUK
Background:
Sickle cell disease (SCD) is the most common genetic disease, due to a mutation of the sixth amino acid of gene coding for the Hemoglobin (Hb) beta chain. The mutated Hb, called HbS, is ...able to polymerize which entails red blood cell (RBC) sickling, leading to chronic hemolysis and painful vaso‐occlusive crisis. Transfusion exchange is one of the therapeutic strategies used for this pathology since it allows the replacement of sickle RBCs. The quality of the transfused RBCs is essential, both to improve the transfusion yield but also to limit the pathological interactions between RBCs and endothelial cells, damaged by hemolysis. RBC concentrates can be stored up to 42 days at 4°c, however, during their hypothermic storage, the quality of RBCs decreases, mainly because of oxidative damage. Hypoxic storage technology helps to keep RBC in hypoxia, improving the concentration of ATP, 2‐3DPG, hemolysis and deformability of RBCs.
Aims:
The purpose of our study was to compare the quality of conventionally conserved RBCs with those conserved with hypoxic storage technology.
Methods:
We assessed several parameters including sickle RBC flow adhesion to HUVEC (human umbilical vein endothelial cells), previously exposed to hemolysate, to mimic endothelial damage present in patients. We also evaluated hypoxic RBC senescence parameters by flow cytometry. In that purpose, we measured intracellular calcium and reactive oxygen species (ROS) concentration as well as phosphatidylserine exposure by annexin V staining in hypoxic RBCs and control RBCs, after incubation 24 h in plasma from healthy donors and SCD patients. RBC adherence on thrombospondin (TSP) under flow was also quantified, since RBC adhesion to TSP was shown to increase during storage and since TSP concentration is elevated in sickle cell disease patients during acute chest syndrome (ACS) or at risk of ACS. In addition, we assessed hemolysis in the bags by measuring free hemoglobin concentration in the supernatant by spectroscopy.
Results:
We found a reduction in the concentration of free Hb (produced by RBC lysis) in hypoxic storage bag, a decrease in RBC senescence (ROS and calcium intracellular concentrations) in hypoxic RBCs and a reduced adherence to thrombospondin when hypoxic RBCs are incubated during 24 h in plasma from healthy donors or SCD patients. There was no difference of RBC adhesion to endothelial cells.
Summary/Conclusion:
These results suggest that hypoxic storage technology meets the safety criteria for transfusion in SCD patients and may provide significant clinical benefits.
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FZAB, GIS, IJS, KILJ, NLZOH, NUK, OILJ, SBCE, SBMB, UL, UM, UPUK
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