Prokaryotic expression, polyclonal antibody preparation, spatio-temporal expression profile and functional analysis of c-Myc of Helicoverpa armigera (Lepidoptera: Noctuidae) / 生物工程学报

c-Myc protein encoded by c-Myc (cellular-myelocytomatosis viral oncogene) gene regulates the related gene expression through the Wnt/β-catenin signaling pathway, and has received extensive attention in recent years. The purpose of this study was to express Helicoverpa armigera c-Myc gene (Ha-c-Myc) by using prokaryotic expression system, prepare the polyclonal antibody, examine the spatio-temporal expression profile of Ha-c-Myc, and investigate the possible function of Ha-c-Myc in regulating H. armigera sterol carrier protein-2 (SCP-2) gene expression. The Ha-c-Myc gene was amplified by PCR and cloned into a prokaryotic expression plasmid pET-32a(+). The recombinant plasmid pET-32a-Ha-c-Myc was transformed into Escherichia coli BL21. IPTG was used to induce the expression of the recombinant protein. Protein was purified by Ni2+-NTA column and used to immunize New Zealand rabbits for preparing the polyclonal antibody. The Ha-c-Myc expression levels in different developmental stages (egg, larva, prepupa, pupa, and adult) of H. armigera and different tissues (midgut, fat body, head, and epidermis) of the prepupa were determined by real-time quantitative reverse transcription PCR (qRT-PCR). Ha-c-Myc siRNA was synthesized and transfected into H. armigera Ha cells. The relative mRNA levels of Ha-c-Myc and HaSCP-2 in Ha cells were detected by qRT-PCR. Results showed that the pET-32a-Ha-c-Myc recombinant plasmid was constructed. The soluble Ha-c-Myc protein of about 65 kDa was expressed in E. coli. The polyclonal antibody was prepared. Western blotting analysis suggested that the antibody had high specificity. Enzyme linked immunosorbent assay (ELISA) showed that the titer of the antibody was high. Ha-c-Myc gene expressed at all developmental stages, with high levels in the early and late instars of larva, and the prepupal stage. Tissue expression profiles revealed that Ha-c-Myc expressed in various tissues of prepupa, with high expression level in the midgut, but low levels in the epidermis and fat body. RNAi results showed that the knockdown of Ha-c-Myc expression significantly affected transcription of HaSCP-2, leading to a 50% reduction in HaSCP-2 mRNA expression level. In conclusion, the Ha-c-Myc was expressed through a prokaryotic expression system, and the polyclonal anti-Ha-c-Myc antibody was obtained. Ha-c-Myc may promote the expression of HaSCP-2 and play an important role in the lipid metabolism of H. armigera. These results may facilitate further study on the potential role and function mechanism of Ha-c-Myc in H. armigera and provide experimental data for exploring new targets of green pesticides.

Correlation Analysis of Hemolytic Transfusion Reaction Induced by Low Titer Antibody / 中国实验血液学杂志

OBJECTIVE@#To establish the diagnostic process of low titer blood group antibody in the occurrence of adverse reactions of hemolytic transfusion.@*METHODS@#Acid elusion test, enzyme method and PEG method were used for antibody identification. Combined with the patient's clinical symptoms and relevant inspection indexes, the irregular antibodies leading to hemolysis were detected.@*RESULTS@#The patient's irregular antibody screening was positive, and it was determined that there was anti-Lea antibody in the serum. After the transfusion reaction, the low titer anti-E antibody was detected by enhanced test. The patient's Rh typing was Ccee, while the transfused red blood cells were ccEE. The new and old samples of the patient were matched with the transfused red blood cells by PEG method, and the major were incompatible. The evidence of hemolytic transfusion reaction was found.@*CONCLUSION@#Antibodies with low titer in serum are not easy to be detected, which often lead to severe hemolytic transfusion reaction.

Retrospective Analysis of Irregular Antibodies Causing Hemolytic Disease of the Fetus and Newborn in Jiangxi Province / 中国实验血液学杂志

OBJECTIVE@#To analyze the characteristics of antibody-specific distribution, laboratory detection results of hemolytic disease of the fetus and neonatal(HDFN) caused by irregular blood group antibodies other than ABO, and its correlation with the clinical situation.@*METHODS@#The non-ABO-HDFN cases in our hospital from October 2012 to December 2021 were selected as the research objects, and the cases diagnosed with ABO-HDFN in the same period were randomly selected as the control group, and the data of antibody specific distribution, total bilirubin, direct antibodies, maternal history, age of the children, the presence or absence of combined ABO-HDFN, and whether to exchange/transfuse blood were retrospectively analyzed. The characteristics of non-ABO-HDFN in Jiangxi province were analyzed.@*RESULTS@#The detection rate of non-ABO-HDFN in Jiangxi province increased. Among 187 non ABO-HDFN cases, the highest percentage of Rh-HDFN was detected (94.6%). Compared with the control group of ABO-HDFN, the non-ABO-HDFN had higher mean integral value of direct antibody, higher peak total bilirubin, and longer duration. Anti-M-HDFN may have severe disease but the direct antibody weak positive/negative, it was easy missed in clinical and delayed the treatment. There is no correlation between the specificity of irregular antibodies, the sex of the child, the mother's previous childbirth history, the presence or absence of combined ABO-HDFN and the need for blood exchange/transfusion(P>0.05).@*CONCLUSION@#The irregular antibodies of causing non ABO-HDFN in Jiangxi area are mainly Rh blood group system, followed by MNS blood group system. Understanding the characteristics of HDFN disease, serological features and the correlation with clinical indexes will help to detect and treat non ABO-HDFN in time and reduce the risk of complications.

Analysis of Irregular Blood Group Antibody Distribution and Blood Transfusion Efficacy in Patients with Malignant Tumor / 中国实验血液学杂志

OBJECTIVE@#To investigate the distribution of irregular blood group antibodies in patients with malignant tumors, and to analyze the relationship between it and efficacy of blood transfusion in patients.@*METHODS@#5 600 patients with malignant tumors treated in Shanxi Bethune Hospital from January 2019 to December 2021 were selected as the research subjects. All patients received blood transfusion, and cross matching test was conducted before blood transfusion, irregular antibody results of patients were tested; the irregular distribution of blood group antibodies was observed, and the relationship between it and efficacy of blood transfusion in patients was analyzed.@*RESULTS@#Among 5 600 patients with malignant tumors, 96 cases were positive for irregular antibody, and the positive rate was 1.71%; the main blood group systems involved in the irregular antibody positive of 96 patients with malignant tumors were RH, MNSs and Duffy system, among which Rh blood group was the most common, and the proportion of anti-E was the highest; among the malignant tumor patients with positive blood group irregular antibody, the proportion of female was higher than that of male; the proportion of patients aged >60 years was the highest, followed by patients aged >40 and ≤50 years, and the proportion of patients aged 18-30 years was the lowest; the patients with positive blood group irregular antibody were mainly in blood system (including lymphoma), digestive system, reproductive and urinary system; the positive rate of irregular antibody of patients in the ineffective group was higher than that of patients in the effective group, the difference was statistically significant (P<0.05). Logistic regression analysis results showed that, irregular antibody positive was a risk factor for ineffective blood transfusion in patients with malignant tumor (OR>1, P<0.05).@*CONCLUSION@#The irregular blood group antibody positive of patients with malignant tumor are mostly female, and the proportion of patients aged >60 is the highest, which is mainly distributed in malignant tumors of blood system, digestive system and urogenital system, and the positive blood group irregular antibody is related to the efficacy of blood transfusion in patients.

Hemofilia A: una enfermedad huérfana / Hemophilia A: a rare disease

La hemofilia A es una enfermedad hereditaria ligada al cromosoma X, causada por mutaciones en el gen F8 del factor VIII de la coagulación. Se considera una enfermedad huérfana, ya que su prevalencia es baja, de 26,6 por cada 100.000 nacidos vivos de sexo masculino. Los pacientes con hemofilia A tienen fases de inicio y amplificación de la coagulación relativamente normales y son capaces de formar el tapón plaquetario inicial en el lugar de la hemorragia, pero debido a la deficiencia del factor VIII, son incapaces de generar una cantidad de trombina en la superficie de las plaquetas, que sea suficiente para estabilizar el coágulo de fibrina. En un paciente masculino con hemorragias inusuales debe descartarse un trastorno de coagulación tipo hemofilia A, y se debe solicitar un recuento de plaquetas y un tiempo de protrombina (TP), los cuales usualmente son normales, y un tiempo de tromboplastina parcial activado (TPT) que se presenta prolongado. Para el diagnóstico diferencial con otras coagulopatías se realiza la medición de factores de coagulación, y pruebas de corrección cuando existe la sospecha de un inhibidor o de una hemofilia adquirida. Los pacientes afectados pueden presentar formas leves, moderadas o severas de la enfermedad, según el nivel plasmático del factor. En Colombia y en el mundo, la hemofilia fue reconocida como una enfermedad huérfana que representa un problema de salud pública, debido a su proceso de atención altamente especializado, que incrementa los costos asociados con la asistencia sanitaria, y afecta la calidad de vida de los pacientes y de aquellos que los rodean, además de que representa un reto diagnóstico que requiere constante actualización, para que pueda ser tratada de manera efectiva

Hemophilia A is an X-linked inherited disease caused by mutations in the coagulation factor VIII F8 gene. It is considered a rare disease, as its prevalence is 26.6 per 100,000 live male births. Patients with hemophilia A have a relatively normal coagulation onset and amplification phases, and are able to form the initial platelet plug at the site of hemorrhage; but due to factor VIII deficiency, they are unable to generate a sufficient amount of thrombin on the platelet surface to stabilize the fibrin clot. In a male patient with unusual bleeding, a hemophilia A-type coagulation disorder should be ruled out, and blood tests such as a platelet count and prothrombin time (PT), which are usually normal, and an activated partial thromboplastin time (APTT), which is prolonged, should be requested immediately. For differential diagnosis with other coagulopathies, measurement of coagulation factors and correction tests are performed when there is suspicion of an inhibitor or acquired hemophilia. Affected patients may present mild, moderate or severe forms of the disease, depending on the plasma level of the factor. In Colombia and worldwide, hemophilia was recognized as a rare disease that represents a public health problem due to its highly specialized care, which increases the costs associated with health care, and affects the quality of life of patients and those around them, as well as representing a diagnostic challenge that requires constant updating, so that it can be treated effectively

Erythrocyte phenotyping for feline AB system in domestic cats from the Ilhéus-Itabuna microregion, Bahia, Brazil / Fenotipagem eritrocitária para sistema AB felino em gatos domésticos da microrregão Ilhéus-Itabuna, Bahia, Brasil

This study aimed to determine the erythrocyte phenotypes of the feline AB system and to check the presence of antigens other than those present in the feline AB system in domestic cats from Ilhéus-Itabuna microregion, Bahia, Brazil. Three-hundred feline blood samples were collected at the Veterinary Hospital of the "Universidade Estadual de Santa Cruz" (UESC) and in home visits to perform blood phenotyping using the tube-method testing. The reverse phenotyping was made between cats that tested phenotype B with blood samples of cats that tested phenotype A to confirm the blood phenotype B. The cross-tested among cats with phenotype A was made in order to verify the presence of different antigens of AB system in this blood phenotype. The results underwent macroscopic and microscopic analyses. Among the 300 animals tested, regarding breed, 290 were mixed-breed cats and among the remaining ten, five were Persians, four Siamese, and one Angora. 297 (99%) presented with phenotype A (including all the breeding cats) and three (1%) with phenotype B, and all this cats were mixed-breed cats. None (0%) of the cats showed the phenotype AB. All phenotype B bloods reacted to reverse phenotyping with phenotype A, confirming the phenotype B of these cats. All phenotype A bloods were compatible among each other, so no further erythrocyte antigens were detected through this test. The mother of one of the phenotype B cats was identified and had phenotype A, demonstrating phenotype A parents with phenotype B offspring. This finding indicates heterozygosis in the studied population. This data enable to conclude that the studied population presented different erythrocyte phenotypes, subsequently highlighting the importance of conducting phenotype analyses in these animals before performing blood transfusion to avoid serious hemolytic complications associated with incompatibility.(AU)

O objetivo deste estudo foi determinar a frequência dos fenótipos eritrocitários do sistema AB felino e verificar a presença de outros antígenos, não pertencentes ao sistema AB felino, em gatos domésticos das cidades de Ilhéus e Itabuna, Bahia, Brasil. Amostras de sangue de 300 gatos foram coletadas no Hospital Veterinário da Universidade Estadual de Santa Cruz (UESC) e em visitas domiciliares para realizar a fenotipagem sanguínea usando o método de tubo. A fenotipagem reversa foi realizada em gatos que testaram o fenótipo B com amostras que testaram o fenótipo A, para confirmação do fenótipo sanguíneo. O teste cruzado foi realizado entre gatos do fenótipo A, para pesquisar a presença de diferentes antígenos do sistema AB dentro desse fenótipo sanguíneo. Os resultados foram submetidos a análises macroscópicas e microscópicas. Dos 300 animais testados, 110 eram machos e 190 fêmeas, e suas idades variaram de cinco meses à 15 anos. Sobre as raças, 290 eram gatos sem raça definida e dos 10 restantes, cinco eram Persas, quatro eram Siameses e um Angorá. 297 (99%) apresentaram fenótipo A (incluindo todos os gatos de raça) e três (1%) tiveram fenótipo B, sendo todos esses gatos sem raça definida. Nenhum (0%) dos gatos apresentou fenótipo AB. Todos os sangues com fenótipo B reagiram na fenotipagem reversa com o fenótipo A, confirmando o fenótipo B desses gatos. Todos os sangues com fenótipo A foram compatíveis entre si, portanto nenhum antígeno eritrocitário adicional foi detectado através deste teste. A genitora de um dos gatos com fenótipo B, foi encontrada e a mesma possuía fenótipo A, demonstrando pais com fenótipo A e cria com fenótipo B. Esse achado indica heterozigose na população estudada. Esses dados levam à conclusão de que diferentes fenótipos eritrocitários estão presentes na população estudada e destacam a importância da realização de testes fenotípicos nesses animais antes dos procedimentos de transfusão, a fim de evitar complicações hemolíticas graves decorrentes do envolvimento de animais incompatíveis.(AU)

Clinical Application of Screening Cell Combination Method in the Prediction of Red Blood Cell Alloantibody / 中国实验血液学杂志

OBJECTIVE@#To explore the clinical application of screening cell combination method in the prediction of red blood cell alloantibody, so as to provide basis for clinical diagnosis.@*METHODS@#From October 2018 to April 2020, 9 680 samples were screened with automatic blood group instrument, 79 patients with positive alloantibodies were identified by 4 sets of screening cells from different manufacturers (referred to as combined method). At the same time, cell panel Panocell-16 was used for comparative analysis. Meanwhile, the combined method was also used to identify the antibodies of 20 samples from National Center for Clinical Laboratories external quality assessment (EQA) in China and 12 samples from WHO EQA.@*RESULTS@#The 79 alloantibodies included anti-Mia antibody (7 cases), anti-M antibody (13 cases), anti-Le@*CONCLUSION@#The combined method can identify the alloantibodies of red blood cells in Chinese population. The screening cells can be used for screening of irregular antibodies without wasting reagents at the same time.

Retrospective Analysis of Serological Characteristics and Distribution of Patients with Mimicking Antibodies / 中国实验血液学杂志

OBJECTIVE@#To understand the characteristics of patients with mimicking specificity autoantibodies through the analysis of the causes of autoantibodies, specificity of antibodies, strategy of blood transfusion, effect of transfusion and distribution of antibodies in China and abroad.@*METHODS@#A total of 23 patients who applied for blood in our hospital from January 2017 to June 2019 were identified as mimicking specificity autoantibodies by antibody identification or absorption-elution test. The causes of mimicking specificity autoantibodies, antibody specificity, blood transfusion strategy and blood transfusion effect were analyzed. The relevant articles on antibodies published in China and abroad were summarized and sorted out, and the distribution of antibodies was analyzed.@*RESULTS@#All the 23 patients with mimicking specificity autoantibodies were Rh blood group system antibodies, of which mimicking anti-Ce autoantibodies were the most common (34.8%), followed by mimicking anti-e autoantibodies (26.1%), mimicking anti-D autoantibodies (21.7%), mimicking anti-C autoantibodies (8.7%) and mimicking anti-E autoantibodies (8.7%). Except for 2 cases with suspected history of blood transfusion, the other 21 cases had a history of blood transfusion / pregnancy. The most common cause of mimicking autoantibodies was drug, followed by infection and autoimmune diseases. The hemoglobin (Hb) of pretransfusion in the blood transfusion group was (48.4±23.9) g/L, which was significantly lower than (86.0±38.9) g/L in the non-transfusion group (P<0.01). Except for 2 cases who could not evaluate the effect of blood transfusion, the effective rate of transfusion was 100%. According to the retrospective statistics of 32 related articles published in China and abroad, the most type of mimicking antibodies were in Rh blood group system, accounting for 79.28%, among which anti-E was the main part of all mimicking autoantibodies, accounting for 21.95%. The following ones were in Kidd system MNSs system, and Kell system.@*CONCLUSION@#Combined with the clinical symptoms and the degree of difficulty of blood matching, the best strategy of blood transfusion should be selected to ensure the safety of blood transfusion.

Early stage of antibody-mediated rejection after lung transplantation： A case report and literature review / 中南大学学报(医学版)

Antibody-mediated rejection (AMR) is a rare and serious complication after lung transplantation, with no characteristic of pathological manifestation, no systematic standard treatment, and the poor efficacy and prognosis. We reported a case of early AMR after lung transplantation and the relevant literature has been reviewed. A male patient presented with symptoms of cold 99 days after transplantation and resolved after symptomatic treatment. He admitted to the hospital 14 days later because of a sudden dyspnea and fever. Anti-bacteria, anti-fungi, anti-virus, and anti-pneumocystis carinii treatment were ineffective, and a dose of 1 000 mg methylprednisolone did not work too. The patient's condition deteriorated rapidly and tracheal intubation was done to maintain breathing. Serum panel reactive antibody and donor specific antibody showed postive in humen leukocyte antigen (HLA) II antibody. Pathological examination after transbronchial transplantation lung biopsy showed acute rejection. Clinical AMR was diagnosed combined the donor-specific antibody with the pathological result. The patient was functionally recovered after combined treatment with thymoglobuline, rituximab, plasmapheresis, and immunoglobulin. No chronic lung allograft dysfunction was found after 3 years follow up. We should alert the occurrence of AMR in lung transplantation recipient who admitted to hospital with a sudden dyspnea and fever while showed no effect after common anti-infection and anti-rejection treatment. Transbronchial transplantation lung biopsy and the presence of serum donor-specific antibody are helpful to the diagnosis. The treatment should be preemptive and a comprehensive approach should be adopted.

Establishment of Quantitative SPR Assay for Antibodies Against Human Platelet Antigen-1a / 中国实验血液学杂志

OBJECTIVE@#To establish quantitative surface plasmon resonance (SPR) assay for antibodies against human platelet antigen-1a (HPA-1a).@*METHODS@#Recombinant protein was fixed on the chip surface by amino coupling method. SPR assay was used to detect the standard antibodies against HPA-1a at different conceatration. The optimal experimental parameters were determined, and standard curves were constructed with linear regression. Moreover, the sensitivity, specificity, accuracy and precision of the assay were evaluated.@*RESULTS@#The quantitative SPR assay for HPA-1a antibodies was established. The determination ranges were 0-20 IU, with accuracy (recovery rate) was 97.75%-103.08%. The intra-assay precision [coefficients of variation (CV)] was 3.53%-4.29%, and the inter-assay precision (CV) was 2.08%-4.40%. For specificity test, several kinds of monoclonal and human antibodies against platelet membrane protein were tested and no positive result was observed.@*CONCLUSION@#The established quantitative SPR assay for HPA-1a antibodies shows good sensitivity, specificity, accuracy and precision, and this rapid and simple method provides a new reference method for scientific research and clinical antibody detection.

Frequência de anticorpos irregulares identificados em pacientes atendidos em um hemonúcleo no sudoeste do Paraná no ano de 2017 / Frequency of irregular antibodies identified in patients treated at the hemotherapy service in southwest of Paraná in 2017

Quando um indivíduo é exposto a antígenos eritrocitários não próprios, ocorre uma resposta imunológica, que leva à produção de anticorpos irregulares voltados contra esses antígenos. Esse processo é conhecido como aloimunização eritrocitária e acontece em decorrência de transfusões de sangue ou gestações incompatíveis. Na medicina transfusional a pesquisa de anticorpos irregulares é fundamental, pois a falha na detecção de um aloanticorpo pode provocar reações transfusionais, aloimunizações, anemias hemolíticas autoimunes e doença hemolítica perinatal. Este estudo tem por objetivo analisar a frequência de anticorpos irregulares de pacientes atendidos no Hemocentro Regional de Francisco Beltrão, Paraná, no ano de 2017. Os dados foram coletados a partir da revisão de registros em arquivos do Laboratório de Imunohematologia do Hemonúcleo. Foram avaliados dados de 49 protocolos de pacientes que apresentaram dificuldades transfusionais no ano de 2017. Dentre os pesquisados, 37 pacientes (75,5%) apresentaram anticorpos irregulares. Dentre os anticorpos anti-eritrocitários observados neste estudo, evidenciou-se a presença de doze pacientes com anti-D (27,2%), seis pacientes com anti-K (13,6%), quatro pacientes com anti-C (9,0%) e em seis pacientes (13,6%) foi observada a presença de autoanticorpos. Este estudo indica que, nos pacientes transfundidos, os anticorpos mais frequentes foram os aloanticorpos Anti-D do Sistema Rh, provavelmente devido ao seu alto grau de imunogenicidade. A prevalência desses anticorpos é semelhante a vários estudos encontrados na literatura.

When an individual is exposed to not-self red blood cell antigens, an immune response occurs, which leads to the production of irregular antibodies directed against these antigens. This process is known as erythrocyte alloimmunization and occurs as a result of blood transfusions or incompatible pregnancies. In transfusion medicine, the search for irregular antibodies is essential, since failure to detect an alloantibody can cause transfusion reactions, alloimmunizations, autoimmune hemolytic anemias, and perinatal hemolytic disease. This study aims at analyzing the frequency of irregular antibodies of patients seen at the Regional Blood Center of Francisco Beltrão, Paraná, in 2017. The data were collected from the review of records in files of the Immunohematology Laboratory of Hemonúcleo. Data from 49 protocols of patients who had transfusion difficulties in 2017 were evaluated. Among those surveyed, 37 patients (75.5%) had irregular antibodies. Among the anti-erythrocyte antibodies observed in this study, the presence of twelve patients with anti-D (27.2%), six patients with anti-K (13.6%), four patients with anti-C (9.0 %), and in six patients (13.6%) with the presence of autoantibodies were observed. This study indicates that, in transfused patients, the most frequent antibodies were the Rh System Anti-D alloantibodies, probably due to their high degree of immunogenicity. The prevalence of these antibodies is similar to several studies found in the literature.

Seguimiento inmunológico después de trasplante renal: una propuesta para la práctica clínica en Colombia / Immunological follow-up of patients with renal transplants: A proposal for clinical practice in Colombia

El daño del injerto es un proceso multifactorial que se inicia tempranamente después de la mayoría de los trasplantes de donantes sin HLA idéntico. Puede deberse a las comorbilidades del receptor, al estado del donante, al tiempo de isquemia, y al fenómeno de isquemia y reperfusión, entre otros, condiciones que inducen factores metabólicos e inmunológicos que finalmente desembocan en la disfunción del injerto. Sin embargo, entre el momento del trasplante y la aparición de los signos y síntomas existe un periodo que puede tardar semanas o años. Por ello, después del trasplante renal, es importante hacer un seguimiento racional que incluya la evaluación clínica y permita anticiparse al daño inmunológico del injerto. En este ensayo se propone un algoritmo de seguimiento del injerto renal después del trasplante.

Graft damage is a process that starts at the moment of transplantation, due to comorbidities of receptor, donor status, ischemia time, ischemia-reperfusion phenomenon, among others, those induce metabolic and immune factors that ultimately trigger clinical manifestations of graft dysfunction. However, the preclinical progression between the time of transplantation and the appearance of signs and symptoms of graft damage can take weeks to years. Therefore, the implementation of rational monitoring approaches during the posttransplantation period is critical and should include not only the clinical follow-up but also anticipate immunological graft damage. In the present essay, we propose an immunological monitoring algorithm for the post-renal transplantation period.

Efectividad de inducción de tolerancia inmune en niños con hemofilia A y aloanticuerpos neutralizantes / Immunotolerance induction effectivity in hemophilia A children and neutralizing alloantibodies

Resumen: Introducción: El desarrollo de aloanticuerpos neutralizantes anti-factor VIII en hemofilia A es la complicación más seria relacionada al tratamiento. La inducción de tolerancia inmune (ITI) o inmunotolerancia es el único tratamiento que erradica inhibidores, permitiendo utilizar nuevamente factor VIII para el tratamiento o profilaxis de eventos hemorrágicos. Objetivo: reportar la experiencia en niños sometidos a inmunotolerancia en la red pública del país. Pacientes y Método: Análisis retrospectivo y descriptivo de 13 niños con Hemofilia A severa e inhibidores persistentes de alto título, que recibieron ITI y seguimiento completo. Se utilizó concentrado de FVIII plasmático en dosis de 70-180 UI/Kg/diarias, definiendo éxito como la negativización del inhibidor y recu peración de la vida media del FVIII. Resultados expresados en media (rango). Resultados: En 13 pacientes se identificó el inhibidor, a una edad de 17,6 meses (2-48), tras 35,2 días (9-112) de exposición a FVIII. Once pacientes (84,6%) recuperaron la vida media del FVIII, tras 49,6 meses (26-70) de tratamiento. En los pacientes que respondieron, el título del inhibidor se negativizó en 7,3 meses (1-20). Conclusiones: En niños con hemofilia A e inhibidores persistentes de alto título, la ITI tiene un elevado éxito. Dado que el tiempo de respuesta es variable, la inmunotolerancia debe ser personalizada.

Abstract: Introduction: The development of anti-factor VIII neutralizing antibodies in hemophilia A is the most severe com plication related to treatment. Immune tolerance induction (ITI) is the only known treatment for eradicating inhibitors. A successful ITI allows using factor VIII (FVIII) again for the treatment or prophylaxis of hemorrhagic events. Objective: To report the experience of pediatric patients who underwent ITI in the country's public health care network. Patients and Method: Retrospective and descriptive analysis of 13 pediatric patients with severe Hemophilia A and high-titer inhibitors persis tence who underwent ITI and complete follow-up. Plasma-derived FVIII concentrate was used at 70 180 IU/kg/day doses. The success of the treatment is defined by achieving a negative titer and a half life recovery of the FVIII. The results were expressed in median (range). Results: In 13 patients, the inhibitor was identified at an average age of 17.6 months, after 35.2 days of exposure to the FVIII. 11 patients (84.6%) recovered the half-life of FVIII after 49.6 months of treatment. In the patients who responded to treatment, the inhibitor titer was negative at 6 months on average. Conclusions: ITI is the treatment of choice for patients with hemophilia A and inhibitors persistence. ITI must be perso nalized since the time response is variable in each patient.

The rare holley antibody associated with a severe hemolytic transfusion reaction: the importance of this antibody identification to find a compatible blood unit / Raro anticorpo associado à reação transfusional hemolítica grave: a importância de sua identificação para encontrar uma unidade sanguínea compatível

ABSTRACT The correct identification of erythrocyte antibodies is fundamental for the searching for compatible blood and haemolytic transfusion reactions prevention. Antibodies against antigens of high prevalence are difficult to identify because of the rarity of their occurrence and unavailability of negative red cells for confirmation. We report a case of 46-years-old woman, diagnosed with hemoglobinopathy, and who had symptomatic fall in hemoglobin levels (5.3g/dL) after blood transfusion suggestive of transfusion reaction. The patient's blood type was O RhD-positive. Irregular antibody screening was positive and demonstrated a panreaction against all erythrocytes tested, but this result was not reactive with dithiothreitol. Using negative red cells for antigens of high prevalence of our inventory we could identify in the serum of the same erythrocytes an anti-Holley antibody associated with anti-E. Molecular analysis confirmed that the patient was negative for E and Holley antigens. The crossmath with compatible units confirmed the results. Holley is a high prevalence antigen of the Dombrock blood system whose negative phenotype is extremely rare in all populations and is associated with hemolytic transfusion reactions. This is an antibody that is difficult to identify because laboratories need to have experience in solving complex cases, and have available a large stock of rare sera and erythrocytes, as well other tools such as enzymes, thiol reagents and molecular tests. The correct identification of a rare antibody is initial and mandatory for searching of compatible donors, and to guarantee a satisfactory transfusional support.

RESUMO A correta identificação dos anticorpos eritrocitários é fundamental na busca de sangue compatível e na prevenção das reações transfusionais hemolíticas. Anticorpos contra antígenos de alta prevalência são de difícil identificação, devido à raridade de sua ocorrência e à indisponibilidade de hemácias negativas para sua confirmação. Apresentamos aqui o caso de uma paciente do sexo feminino, 46 anos, com diagnóstico de hemoglobinopatia, que apresentou queda sintomática dos níveis de hemoglobina (5,3g/dL) após transfusão sanguínea, sugestiva de reação transfusional. O tipo sanguíneo da paciente era O RhD-positivo. A pesquisa de anticorpos irregulares foi positiva, demonstrando panreação contra todos os eritrócitos testados, mas não reativo ao ditiotreitol. Utilizando hemácias selecionadas negativas para antígenos de alta prevalência do nosso inventário, foi possível identificar no soro da mesma um anticorpo anti-Holley associado a um anti-E. A análise molecular confirmou que a paciente era negativa para os antígenos E e Holley, e as provas de compatibilidade com unidades fenotipadas confirmaram os resultados. Holley é um antígeno de alta prevalência do sistema sanguíneo Dombrock, cujo fenótipo negativo é extremamente raro em todas as populações e está associado a reações transfusionais hemolíticas. Trata-se de anticorpo de difícil identificação, pois os laboratórios precisam ter experiência na resolução de casos complexos, grande estoque de soros e eritrócitos raros, além de outras ferramentas, como enzimas, reagentes tiol e testes moleculares. A identificação correta de um anticorpo raro é inicial e obrigatória para a busca de doadores compatíveis, garantindo um suporte transfusional satisfatório.

Analysis of the Irregular Antibodies of the Patients and Its Clinical Significance / 中国实验血液学杂志

OBJECTIVE@#To retrospectively analyze the identification results of irregular antibodies, to clarify the distribution features and to explore the relation of alloantibodies and autoantibodies with the immunized history of patients and disease kinds.@*METHODS@#49 820 patients who applied for red blood transfusion during Sep 1st 2017 to Sep 1st 2018 were selected. All the specimens were screened for the antibody by microcolumn gel antiglobulin technique, which then were identified for irregular antibody.@*RESULTS@#Antibodies were found in 861 (1.73%) of all 49 820 transfused samples. The alloimmunization history of the patients with antibodies was significantly different between male and female (χ=18.54，P＜0.01). The alloantibody was the most common, accounting for 59.50% in all of the antibodies. Warm autoantibody, anti-E, anti-M, anti-cE and anti-Ce accounted for 68.5% of the antibodies. The blood group of Rh, MNS and Lewis were responsible for 92.40% of alloantibody, especially anti-E accounted for the largest percentage(38.60%) of alloantibody. Patients with alloantiboies experienced much more the alloimmunization and transfusion history (χ=20.13，P＜0.01；χ=5.40，P＜0.05) . The distribution of auto and alloantibody was very significantly different among the ddifferent isease (χ=51.8，P＜0.01), Hematopathy, solid tumor and osteoarthropathy were often associated with alloantibody, otherwise, autoantibodies often occurred in hematopathy and autoimmune disease.@*CONCLUSION@#The most important factor that results in antibody-screening positive is alloantibody, in which anti-E antibody from Rh blood group system in most common.

Anti-PP₁P(k) (Tj(a)) Antibody in a Korean Female Patient with p Phenotype Confirmed by Genotyping

Extended Red Blood Cell Phenotype Matching Is Dependent on Ethnicity and Specificity of RBC Alloantibodies

No abstract available.

Caracterización de donantes de sangre con rastreo de anticuerpos irregulares positivo en Montería, Colombia 2012-2015 / Blood donors characterization with positive irregular antibodies in Montería, Colombia 2012- 2015

Introducción: Los anticuerpos irregulares corresponden a aquellos distintos a los anticuerpos naturales anti-A o anti-B, los cuales pueden aparecer en respuesta a la exposición a un antígeno eritrocitario extraño (transfusión o trasplante) o por incompatibilidad materno-fetal. Objetivo: Caracterizar a los donantes con rastreo de anticuerpos irregulares positivo de un banco de sangre de Montería, Colombia, durante el periodo 2012-2015. Métodos: Estudio transversal y retrospectivo, con fuente de información secundaria, basada en los resultados del rastreo de anticuerpos en los donantes de un banco de sangre de Montería, Colombia, entre los años 2012 y 2015. La población estuvo conformada por todos los donantes voluntarios registrados en el tiempo del estudio (35 248 donantes), a quienes se les realizó rastreo de anticuerpos. Como muestra, se seleccionaron todos los casos que tuvieron resultados positivos (71 donantes). Los datos fueron organizados en tablas y analizados en el software SPSS 21.0, Microsoft Excel y en Epidat versión 3.1. Resultados: El 0,2 por ciento de la población presentó un rastreo de anticuerpos positivo con un intervalo de confianza entre 0,15 y 0,25 por ciento. Los anticuerpos irregulares fueron más frecuentes en los hombres y en donantes O Rh positivo. Se encontraron Ac irregulares con 13 especificidades diferentes, con predomino de anti-M, anti-Lea, anti-D y anti-E y porcentajes respectivos de 27,78 por ciento, 20,83 por ciento, 9,72 por ciento y 8,33 por ciento. El 50 por ciento de los donantes tenía 30,5 años o menos, el 49,3 por ciento había donado previamente y el 9,9 por ciento recibió al menos una transfusión en algún momento de su vida. Conclusión: La frecuencia de donantes con rastreo de anticuerpos irregulares positivo fue baja, el sexo masculino presentó mayor porcentaje, se detectó principalmente en el grupo sanguíneo O y dentro de los anticuerpos irregulares, anti-M presentó una mayor frecuencia(AU)

Introduction: Irregular antibodies correspond to those other than natural anti-A or anti-B antibodies, which may appear in response to exposure to a foreign erythrocyte antigen (transfusion or transplantation) or due to maternal-fetal incompatibility. Objective: To characterize the donors with positive irregular antibody screening of a blood bank in Monteria, Colombia during the period 2012-2015. Methods: Cross-sectional and retrospective study, with secondary information source, based on the results of the antibody screening in donors of a blood bank in Monteria, Colombia from 2012 to 2015. The population consisted of all voluntary donors registered in the study time (35 248 donors), who were screened for antibodies. As a sample, all cases that had positive results (71 donors) were selected. The data was organized in tables and analyzed in the software SPSS 21.0, Microsoft Excel and in Epidat version 3.1. Results: 0.2 percent of the population presented a positive antibody screen with a confidence interval between 0.15 and 0.25 percent Irregular antibodies were more frequent in men and in O Rh positive donors. Thirteen types of irregular antibodies were found, with predominance of anti-M, anti-Lea, anti-D and anti-E and respective percentages of 27.78 percent, 20.83 percent, 9.72 percent and 8.33 percent. 50 percent of the donors were 30.5 years old or less, 49.3 percent had previously donated and 9.9 percent received at least one transfusion at some point in their lives. Conclusion: The frequency of donors with irregular positive antibody screening was low, the male sex had a higher percentage, it was detected mainly in blood group O and within the irregular antibodies, anti-M showed a higher frequency(AU)

Screening and Identification of Blood Group Alloantibody by Surface Plasmon Resonance Technique and Its Preliminary Application / 中国实验血液学杂志

OBJECTIVE@#To investigate the feasibilily of screening and identifying the red blood cell type alloantibodies by means of surface plasman resonance(SPR) technique so as to provide a new method for detecting the transfusion compatibility of red blood cells.@*METHODS@#The RBC antigens for screening the alloantibody were fixed on the SPR chip surface by means of amino coupling method; the analysis conditions of SPR chip were optimized and then the control serum with RBC blood group antibody positive was detected; the performance of SPR chip for detection of serum was analysed; the consistance of rusults detected by SPR technique and microcolum agglutination for clinieal samples of 129 thalasstmia patients with history of lone-term blood transfusion were compared; at the same time, the blood group amtibodies in 7 patients with blood group antibody positive were identified before blood transfusion by using SPR chip so as to select the RBC antigen compatible blood for transfusion; and the efficacy of RBC transfusion was followed up and evaluated.@*RESULTS@#The repeatability, sensitivity and specificity of SPR chip technique for detecting the blood group alloantibodies all were better. The SPR technique and microcolumn agglutination method were not significant different for screening blood group alloantibodies (χ2 = 0.333, P＞0.05), and the overall consistency was 97.2%; the results of SPR technique in 7 patients with positive blood group antibodies were as follows: 3 cases with anti-E, 1 case anti-M, 1 case anti-C, 1 case anti-Jka and 1 case autoantibody, which were consistent with the results of microcolumn agglutination tests, and the compatible red blood cells were selected for transfusion, of which the infusion of 6 cases was effective. In only 1 case the infusion was ineffective because of autoantibody.@*CONCLUSION@#For screening and identification of blood group alloantibodies, the performance of SPR chip technique is equivalent to the micro-column agglutination, but the procedure of SPR technique is simpler, faster and high-throughput and label-free, which can meet the basic requirements for rapid screening and identification of blood group alloantibodies before transfusion of red blood cells.