The challenging blood cross-matching and resolution for multiple myeloma patients in different disease stages / 中国输血杂志

【Objective】 To explore the challenging blood cross-matching and resolution for multiple myeloma (MM) patients in different disease stages. 【Methods】 For a patient who was first diagnosed as MM and scheduled for blood transfusion, his blood was cross matched with donors’ blood by microcolumn gel method and tube test. When the major side of cross-matching was agglutinated, the patient’s plasma was cross matched with donors’ red blood cell (RBC) by polybrene test, then plasma dilution cross matched with donors’ RBC by microcolumn gel method. For a patient who was diagnosed as recurrent refractory MM and scheduled for blood transfusion, his blood was cross matched with donors’ blood by microcolumn gel method. 【Results】 1) Case 1 was a first-visit outpatient. The major side of microcolumn cross-match test was agglutinated with the shape of fine line. The result of tube method also showed agglutination of major sides, and the rouleaux were detected by the microscopy. Then polybrene method and microcolumn gel method (after plasma diluted) were applied for cross-matching again with the above two donors’ blood and showed compatibility. 2) Case 2 was a recurrent refractory MM patient. The major and minor sides of microcolumn cross-match test were both agglutinated with the shape of granular. The patient was treated with anti-CD38 monoclonal antibody. The RBCs, after treated with dithiothreitol (DTT) was used to cross match with patient plasma by microcolumn test, and the result was compatible. 【Conclusion】 Polybrene method and microcolumn gel method after plasma diluted are suitable for blood cross-matching of newly diagnosed MM patients, also for those treated with CD38 monoclonal antibody, as the drug interference with cross-matching can be eliminated by DTT.

Removal of serological interference in the treatment of multiple myeloma with CD38 monoclonal antibody using polybrene method / 中国输血杂志

【Objective】 To investigate the clinical feasibility of polybrene method to remove serological interference in patients receiving anti-CD38 monoclonal antibody for the treatment of multiple myeloma, and its detection performance of alloantibodies. 【Methods】 For patients receiving anti-CD38 monoclonal antibody for the treatment of multiple myeloma, unexpected antibody screening and cross-matching blood test were performed by polybrene method. 【Results】 The polybrene method can remove the interference of anti-CD38 monoclonal antibodies on blood group serology; both methods can effectively remove anti-CD38 monoclonal antibody to detect anti-E, anti-D, anti-Fya and anti-S antibodies.The titers of anti-D, anti-E, anti-Fya and anti-S alloantibodies, yielded by enhanced polybrene method, were higher than those of the polybrene method.Seven patients received K-antigen-negative blood transfusion without any adverse reactions to blood transfusion. 【Conclusion】 For the treatment of multiple myeloma using CD38 monoclonal antibody, the polybrene method can quickly and effectively remove the interference of daratumumab with blood group serology.

Daratumomab interference with pre-transfusion examination of patients with multiple myeloma / 中国输血杂志

【Objective】 To solve daratumomab interference with blood compatibility testing in multiple myeloma (MM) patients treated by daratumomab(DARA). 【Methods】 The irregular antibodies screening before and after the DARA treatment, and the major side crossmatch via coombs' test and polybrene method, respectively, were performed to resolve the nonspecific interference in a MM patient’s cross-matching test, produce by DARA. 【Results】 The initial panreactivity on the major side with agglutination (3+ ~4+ ), produce by DARA, was overcome by dithiothreitol (DTT) treatment, and turner out to be none agglutination. Otherwise, DARA had no effect on the crossmatch using polybrene method. 【Conclusion】 Antibody screening and identification should be conducted before DARA treatment in MM patients, and DARA interference with blood compatibility testing can be resolved by DTT treatment or the crossmatch using polybrene method.

Interference of CD38 monoclonal antibody in blood compatibility testing and its countermeasures: A general consensus among experts / 中国输血杂志

With continuous discovery of tumor immune targets and continuous changes in antibody research and development technology, antibody drugs are becoming more and more widely used in clinical practice. However, some targets are not only expressed on tumor cells, but also on red blood cells. Therefore, the clinical application of antibodies against the corresponding targets may interfere with the detection of blood transfusion compatibility, resulting in difficulty in blood matching or delay of blood transfusion. This consensus summarizes the current solutions for the interference of CD38 monoclonal antibody (CD38 mAb) in transfusion compatibility testing. After analyzing the advantages and disadvantages of different methods, polybrene and sulfhydryl reducing agents [dithiothreitol (DTT) or 2-mercaptoethanol (2-Me)], as a solution for CD38 mAb interference in blood compatibility testing, are recommended for Chinese patients, so as to eliminate blood transfusion interference produce by CD38 mAb and further provide a pre-transfusion workflow for clinicians and technicians in Department of Blood Transfusion.

-D- phenotype screening by polybrene method / 中国输血杂志

【Objective】 To develop a novel screening reagent for -D- phenotype preliminary screening based on the difference in RhD antigen expression level of -D- phenotype and normal RhD phenotype. 【Methods】 RhD antigen expression of -D-phenotype and Rh D-- gene carrier were detected by flow cytometry. By adjusting the concentration of polybrene in the screening system, the red blood cells with high RhD antigen expression level agglutinated, and the preliminary screening of the -D-phenotype and its gene carriers was realized. 【Results】 According to the quantitative results of immunofluorescence intensity (MFI) analysis by flow cytometry, the expression level of RhD antigen in -D- phenotype cells (284 360±16 698, n=3) was about 3 times normal RhD positive cells (98 642±35 908, n=9)(P<0.01), while RhD antigen expression level of RhD-- gene carrier (181 109±39 455, n=4) was about 2 times normal RhD positive cells(P<0.01). RhD antigen expression (144 538±227 445, n=7) of the positive cells screened by 15 μL 3% fresh red blood cell suspension and screening system 35 μL (1 μL IgG anti-D, 29 μL polybrene polybrene, and 5 μL low ionic strength solution) was about 1.5 times normal RhD positive cells. 【Conclusion】 The polybrene preliminary screening system, which can be used for high-throughput screening of -D- phenotype, is a reliable technical method for frequency study of this phenotype.

Estudio de la desialización producida por larvas musculares de Trichinella spiralis / Desialylation study produced by muscle larvae of Trichinella spiralis

Trichinella spiralis es agente causal de una zoonosis endémica en Argentina. El objetivo fue estudiar la desialización eritrocitaria producida por larvas musculares (LM) de T. spiralis. Se trabajó con concentrados de LM, incubados en partes iguales con glóbulos rojos (GR) Grupo O (37 °C) durante 3 horas (con y sin agitación controlada) durante 150 minutos, a intervalos de 30 minutos, para estudiar el curso de la desialización en el tiempo. Los GR controles fueron incubados de la misma manera, con igual volumen de solución fisiológica. Se aplicó el método de titulación de la agregación, calculando título y coeficiente de puntuación total (CexpST). Se encontró que los eritrocitos incubados con LM presentaron mayor agregación que los controles. El valor medio de CexpST con agitación (0,43) fue significativamente menor que cuando los GR no se agitaron (0,72). El estudio de la desialización eritrocitaria en el tiempo mostró que el título de los GR control disminuyó significativamente a los 90 minutos en 5/10 repeticiones y a los 150 minutos en 9/10. El aumento del tiempo de incubación produjo el incremento de la desialización excepto a los 120 y 150 minutos donde no existieron diferencias significativas en el valor de CexpST. La experiencia realizada in vitro, sugeriría que en la infección in vivo, las LM podrían captar el ácido siálico a partir de los residuos sializados presentes en la célula muscular.

Trichinella spiralis is the cause of an endemic zoonosis in Argentina. The objective was to study the erythrocyte desialylation by T. spiralis muscle larvae (ML) applying an aggregation titulation method. We worked with ML concentrates, which were incubated with an equal volume of O Group erythrocytes (RBCs) at 37° C for 3 hours, (with and without controlled agitation) and for 150 minutes, taking samples at 30 minutes intervals to study the course in time of the desialylation. RBCs control were incubated with an equal volume of physiological saline solution. Aggregation titulation method was applied and the title and total score coefficient (TSexpC) were calculated. The results showed that erythrocytes incubated with ML showed more aggregation than controls. The average TSexpC with agitation (0.43) was significantly lower than when erythrocytes were not stirred (0.72). The course in time of the erythrocyte desialylation showed that the RBCs contol title decreased significantly at 90 minutes in 5/10 repetitions and at 150 minutes in 9/10. Increasing the incubation time produced an increase in erythrocyte desialylation, except at the 120 and 150 minutes interval where no significant differences in TSexpC values were found. The in vitro experience would suggest that in cases of in vivo infection, ML could capture sialic acid from sialylate residues present in the muscle cell.

Application of 3 kinds of method of blood crossmatching in neonatal blood transfusion security / 国际检验医学杂志

Objective To investigate the application values of 3 kinds of crossmatching method saline medium method,polybrene test and microcolumn gel test in the neonatal blood transfusion security.Methods 174 newborns needing blood transfusion were simultaneously performed the isotype crossmatching by the saline method,polybrene test and microcolumn gel test.The irregular antibody and the direct antiglobulin test were routinely conducted,the samples with positive irregular antibody were further per-formed the antibody specificity identification.Results 14 cases (8.05%)of crossmatching incompatibility were found by the saline method,1 case (0.57%)by the polybrene test and 62 cases (35.63%)by the microcolumn gel test;among which,31 cases were secondary side crossmatching incompatibility caused by positive direct antiglobulin test,1 was the incompatibility of 3 methods caused by anti-D antibodies.The difference between the polybrene test and the microcolumn gel test was of statistical significance (P <0.05).Conclusion The blood crossmatching by the microcolumn gel test in blood transfusion of newborns requires quite few amount of blood sample,is simple to operate and easy to be standardized.With results clearly and easily measured and sensitivity higher than that of polybrene test,its result is clear and easy to be judged,its sensitivity is higher than that of the polybrene test, which has the important significance for ensuring the neonatal blood transfusion safety.

Efecto producido por Ascaris lumbricoides sobre la carga superficial eritrocitaria utilizando el Método de Polibrene / Effect of Ascaris lumbricoides on the erythrocyte superficial charge using Polybrene Method

Los ácidos siálicos (AS) contenidos en glucoproteínas y glucolípidos participan en diversas funciones biológicas y su presencia en el eritrocito tiene importancia hemorreológica y hemodinámica. Se considera que los AS intervendrían en la interacción parásito-hospedador. El objetivo de este trabajo fue estudiar el efecto producido por A. lumbricoides sobre la carga superficial eritrocitaria utilizando el Método de Polibrene. Se trabajó con 51 extractos de parásito adulto ([EA]) y con 4 concentrados larvales ([CLAL]: 2000/ 1000/ 500/ 250 larvas/ mL). El método fue aplicado en eritrocitos no tratados (Control) y tratados con [EA]/ los 4 [CLAL] de forma simultánea. El tratamiento con las larvas se realizó en 18 experiencias. Los eritrocitos tratados con el 37, 25% de los [EA] presentaron menor agregación en relación al Control. Los análisis estadísticos mostraron que este efecto no se relacionó con la concentración proteica del [EA]. La disminución de agregado en el tratamiento con [CLAL] fue dependiente de la concentración larvaria y se relacionó a concentraciones = a 500 larvas/ mL. Se observó que la fijación [EA]/ [CLAL] al eritrocito no era fuerte. Los resultados demostraron que A. lumbricoides secuestra AS eritrocitario. Esta captación podría influir en la patología y / o participar en la evasión de la respuesta inmune del hospedador.

Sialic acids (SA) contained in glycoproteins and glycolipids participate in various biological functions; besides, its presence on the erythrocyte surface has both hemodynamical and hemorheological importance. SA are thought to intervene in the parasite-host interaction. The aim of this work was to study the effect of Ascaris lumbricoides on the erythrocyte superficial charge by using Polybrene Method. Work was carried out on 51 adult parasite extracts ([AE]) and 4 (larvae) larvae concentrates ([ALLC]: 2000 / 1000 / 500 / 250 larvae/ mL). The method was applied to non-treated (Control) and treated red cells with [AE] / all 4 [ALLC] simultaneously. Larvae´ treatment was conducted over 18 experiences. Erythrocytes treated with 37. 25% of the [AE] had lower aggregation than the Control. The statistical analysis showed that this effect was not related to the protein concentration of [AE]. The aggregate decrease in the treatment with [ALLC] was dependent on larvae concentration and it was related to larvae concentrations = 500 larvae/ mL. It was observed that the [AE]/ [ALLC] fixation to the erythrocyte was not strong. The results have shown that A. lumbricoides sequestrates SA erythrocyte. This capture could affect the pathology and/or participate in the escape of the host's immune response.

Comparison of four methods for determination of IgG red blood cell antibodies / 第三军医大学学报

Objective To compare the specificity, sensitivity, titers, and rapidity of four methods including papain technique, anti-globulin technique, polybrene test, and micro-column gel test for determination of IgG red blood cell antibodies. Methods Twelve kinds of IgG red blood cell antibodies such as anti-D, anti-E, anti-C, anti-c, anti-e, anti-Jk a, anti-Jk b, anti-Fy a, anti-Fy b, anti-k, anti-S, and anti-s were checked by the four methods. Results Seven kinds of IgG red blood cell antibodies including anti-D, anti-E, anti-C, anti-c, anti-e, anti-Jk a, and anti-Jk b were detected using papain technique (7/12). All of the 12 kinds of IgG antibodies were detected by anti-globulin technique (12/12). Eleven kinds of IgG red blood cell antibodies except anti-k were examined with polybrene test (11/12) and all the antibodies were also determinated by micro-column gel test (12/12). The titers of the antibodies determination suggested that papain technique was less sensitive than other three methods, while the micro-column gel test was more sensitive than other three methods in examination of all the antibodies. The lasting time of four techniques were: papain technique 45 min, anti-human globulin technique 60 min, polybrene test 5 min, and micro-column gel test 30 min. Conclusion Papain technique has some limitation in determination of IgG antibodies and anti-globulin technique is complicated because of long period incubation and multiple wash of red blood cells. Polybrene test is the most simple and convenient technique for determination of IgG antibodies. Micro-column gel test is the most sensitive method in determination of IgG antibodies.