Flow-Cytometric Analysis of HNA-2 Expression and Phenotypes Among Thai Blood Donors

BACKGROUND: Antibodies specific to human neutrophil antigen (HNA), especially HNA-2, are implicated in various conditions, including neonatal alloimmune neutropenia, febrile non-hemolytic transfusion reactions, and transfusion-related acute lung injury. The distribution of the HNA-2 phenotype frequencies in the Thai population remains unknown. This study aimed to investigate HNA-2 phenotype frequencies in Thai blood donors and to compare the relationships of sex and age with HNA-2 expression. METHODS: EDTA blood samples were collected from 220 unrelated healthy Thai blood donors, including 150 males and 70 females, with ages ranging from 20 to 57 years. Polymorphonuclear cells were isolated and stained with monoclonal antibodies clone MEM-166 and clone 2D1, which are specific to human CD177 (HNA-2) and CD45, respectively. HNA-2 expression according to sex and age was analyzed by flow cytometry. RESULTS: Among the 220 donors, HNA-2-positive and HNA-2-null-phenotype frequencies were 0.995 and 0.005, respectively. Mean antigen expression was significantly higher in women (71.01±15.46%) than in men (64.59±18.85%; P < 0.05). No significant differences in HNA-2 expression were found between different age groups. HNA-2 phenotype frequencies were similar to those in Asian, African, American, and Brazilian populations, but were significantly different from those in eastern Japanese, Korean, and French populations (P < 0.001). CONCLUSIONS: This is the first report of HNA-2 phenotype frequencies in a Thai population, and the data will be helpful in predicting the risk of HNA-2 alloimmunization and in recruiting granulocyte panel donors.

Identification of human neutrophil antibodies among volunteer blood donors in one district / 国际检验医学杂志

Objective To elucidate the distribution of human neutrophil (HNA ) antibodies among volunteer blood donors in Henan and estimate the risk rate of immune‐mediated transfusion related adverse reaction caused by HNA antibodies .Methods 156 females and 80 males were collected randomly from volunteer blood donors in Henan .All the samples′HNA antibodies were detec‐ted using LABScreen Multi kit .Results The data showed that among the 236 volunteer blood donors ,the positive rate of HNA an‐tibodies was 5 .77% (9/156)in females ,while the rates were 5 .00% (4/80)for males .Among the positive donors ,there were 7 anti‐HNA 1A ,4 anti‐HNA 4A ,1 anti‐HNA 1A/HNA 4A and anti‐HNA 1A/HNA 1C .Conclusion The frequency of HNA antibodies in females is same as the males .Our study is to elucidate the distribution of human neutrophil antibodies among volunteer blood do‐nors in Henan and estimate the risk rate of immune‐mediated transfusion related adverse reaction caused by HNA antibodies ,there‐fore it could provide theoretical basis for clinical transfusion safety .

TRALI, una amenaza enmascarada: biología y fisiopatología / TRALI, a masked menace: biology and pathophysiology

El daño pulmonar agudo ocasionado por la transfusión o TRALI (por sus siglas en inglés), definido como la aparición de un distrés respiratorio agudo en un paciente recién transfundido, pasó de ser considerado una complicación infrecuente de la terapia transfusional a ser actualmente la principal causa de mortalidad por transfusión, según sistemas de hemovigilancia de Europa y Norteamérica. Su desarrollo en forma clínicamente reconocible se atribuye a la interacción entre factores dependientes de la unidad transfundida (tipo de componente, presencia de sustancias biológicamente activas, etc.) y el estado de las respuestas celulares en el receptor. La heterogeneidad en cuanto al cuadro clínico de los pacientes afectados,la variación en el volumen infundido, el tipo de componente implicado y el tiempo desde el inicio de la transfusión hasta la aparición de los síntomas, ha hecho evolucionar la explicación a la génesis de este evento adverso, en el afán de incluir los casos sin explicación mediante las distintas hipótesis. Dos interesantes acercamientos patogénicos resultan la teoría de dos golpes y el modelo basado en el umbral que impone la relación entre los distintos factores de riesgo. La naturaleza multicausal del TRALI y el sinnúmero de variables que pueden influir en su aparición y reconocimiento, continúan haciendo de este un reto médico importante en el contexto de la medicina transfusional, donde su mejor enfoque terapéutico sigue siendo el preventivo(AU)

Transfusion-related acute lung injury (TRALI) defined as the onset of an acute respiratory distress in a recently transfused patient, has passed from been considered a rare complication of transfusion therapy to be the leading cause of transfusion-associated death, as reported by hemovigilance systems in Europe and America. In a previous paper definition, epidemiology and some clinical aspects of TRALI are reviewed. Now we focused our efforts in reviewing the incompletely understood world of its pathogenesis. Clinically recognizable TRALI´s development depends on the interaction between risk factors from both the transfused component unit (as the kind of component and substances within it) and receiver patient´s cellular response. Heterogeneity of clinical features, transfused volumes, component type and time elapsed from the beginning of transfusion to the onset of symptoms have pushed the explanations for its genesis to evolve in an effort to include as much cases as the different hypotheses allowed. Two interesting approaches to TRALI´s pathogenesis are the two hit; theory and the threshold; model imposed by risk factors interactions. The large diversity of variables and causes which can influence its onset and clinical recognition continue to make it a real challenge for clinicians, mainly within transfusion medicine, where the best therapeutic approach available is prevention(AU)

Gene Frequency of Human Neutrophil Alloantigen-3 in the Korean Population / 대한수혈학회지

BACKGROUND: In the Korean population, gene frequencies of Human Neutrophil Alloantigen-1 (HNA-1), HNA-2, HNA-4, and HNA-5 were determined using serological and genotyping methods. However, no study assessing the gene frequencies of HNA-3 among the Korean population has been conducted. The aims of this study were to report the gene frequencies of HNA-3, to estimate the risk of HNA-3a (or HNA-3b) alloimmunization, and to secure donors of granulocytes for anti-HNA-3 antibodies among the Korean population. METHODS: Genotyping of HNA-3a and HNA-3b genes of 110 healthy and unrelated Korean donors was performed using a polymerase chain reaction with sequence-specific primers. RESULTS: The observed frequencies of HNA-3a and HNA-3b were 0.695 and 0.305, respectively, among the Korean population. The HNA-3a gene frequency of Koreans was significantly lower than those of American Whites and Germans (P<0.05). The risk of HNA-3a and HNA-3b alloimmunization due to pregnancy (transfusion) was 0.065 (0.084) and 0.147 (0.250), respectively, among the Korean population. The risk of HNA-3a alloimmunization was significantly higher in the Korean population than in the German and American White populations (P<0.05). CONCLUSION: The gene frequencies of HNA-3 and the risk of HNA-3a (HNA-3b) alloimmunization due to pregnancy or transfusion among the Korean population were determined. We also identified individuals who were HNA-3a/3a or HNA-3b/3b for the granulocyte panel which could be used for anti-HNA antibody identification.

Human Neutrophil Alloantigen-3a Alloimmunization among Pregnant Korean Women / 대한수혈학회지

BACKGROUND: Alloantibodies against human neutrophil alloantigen (HNA)-3a are associated with severe and fatal transfusion related acute lung injury (TRALI). HNA-3 genotyping and HNA-3a antibody (Ab) identification are essential to diagnosis and prevention of TRALI caused by HNA-3a Ab. However there had been no laboratory for HNA-3a Ab identification in Korea. The aims of this study were to establish the HNA-3a Ab test in Korea and to estimate the incidence of HNA-3a alloimmunization among pregnant Korean women. METHODS: HNA-3a homozygotes and HNA-3b homozygotes were identified by HNA-3 genotyping. Three HNA-3a homozygotes and three HNA-3b homozygotes are included in the granulocytes panel, which consisted of 10 donors for granulocytes. Sera from 650 pregnant Korean women were tested for granulocyte Ab using a mixed passive hemagglutination assay (MPHA). When a HNA-3a Ab was detected, the woman's HNA-3 was typed to support her HNA-3a alloimmunization. RESULTS: MPHA showed positive reactions in the sera from 26 women (4.0%, 26/650). HLA Abs were detected in 18 women (2.8%, 18/650), among whom HNA Abs were identified simultaneously in 7 women. Granulocyte Abs were detected in sera from 15 women (2.3%, 15/650). The incidence of HNA-3a, HNA-1b, HNA-1a, HNA-2a, and unidentified HNA Abs among pregnant Korean women was 0.77% (5/650), 0.77% (5/650), 0.62% (4/650), 0.15 (1/650), and 0.31% (2/650), respectively. CONCLUSION: In this study, we established the HNA-3a Ab test using MPHA for diagnosis and prevention of TRALI caused by HNA-3a Ab. The incidence of HNA-3a Ab in pregnant Korean women was 0.77% (5/650).

Caracterización de los antígenos específicos del neutrófilo NA1, NA2 y SH en la población argentina / Characterization of neutrophil-specific antigens NA1, NA2 and SH in the argentinean population

Los antígenos especifícos de neutrófilos NA1 (HNA-1a), NA2 (HNA-1b) y SH (HNA-1c) son formas alotípicas del Fc gamma RIIIb y los blancos más frecuentes de los aloanticuerpos antigranulocitarios. El objetivo de este estudio fue determinar las frecuencias alélicas de los antígenos específicos de neutrófilos pertenecientes al sistema HNA-1 en donantes de sangre y amerindios de la etnia Toba de la ciudad de Rosario, Argentina. Se genotipificaron doscientos dieciocho individuos no relacionados para HNA-1a, HNA-1b y HNA-1c mediante reacción en cadena de polimerasa con cebadores secuencia específica (PCR-SSP). Las frecuencias alélicas en los donantes de sangre para HNA-1a y HNA-1b fueron 0,44 y 0,56 respectivamente y en la población amerindia Toba fueron 0,77 y 0,23 respectivamente. El alelo HNA-1c presentó una frecuencia de 0,023 en los donantes de sangre, pero no se detectó en ninguno de los individuos amerindios estudiados. Los presentes datos mostraron que las frecuencias de los alelos que codifican al sistema HNA-1 en la población mayoritaria de Rosario y en la minoritaria amerindia Toba son similares a las descriptas en europeos y otras poblaciones amerindias distantes, respectivamente.

The neutrophil-specific antigens NA1 (HNA-1a), NA2 (HNA-1b) and SH (HNA-1c) are allotypic forms of Fc gamma RIIIb and the most frequent targets of neutrophil alloantibodies. The aim of this study was to determine to gene frequencies of the neutrophil-specific antigens bolonging to the HNA-1 system in blood donors and Toba amerindians fron Rosario, Argentina. Two hundred and eighteen unrelated individual from Rosario were typed for HNA-1a, HNA-1b and HNA-1c, using polymerase chain reaction with sequence-specific primers (PCR-SSP). For the argentinean blood donors, the HNA-1a and HNA-1b gene frequencies were 0.44 and 0.56 and for the amerindians Toba were 0.77 and 0.23 respectively. The HNA-1c gene frequency in blood donors was 0.023 but the allele was absent within the amerindian individuals. The present data showed that the HNA-1 allele frequencies in the major population and the Toba amerindian from Rosario are similar to those described in European and others distant amerindians populations, respectively.

Gene Frequencies of Human Neutrophil Antigens 4a and 5a in the Korean Population / 대한진단검사의학회지

BACKGROUND: In Korean population, antigen frequencies of HNA-1a, HNA-1b, and HNA-2a are determined using a serological and genotyping method. However, no study has been done to assess the gene frequencies of HNA-4a and HNA-5a. It has been reported that the antibody against HNA-4a is associated with alloimmune neutropenia and autoimune neutropenia; however, there is no confirmed clinical report on anti-HNA-5a-related disorders. The aim of this study was to determine HNA-4a and HNA-5a gene frequencies among the Korean population. METHODS: Genotyping of HNA-4a and HNA-5a genes of 110 healthy and unrelated Korean donors was performed using a polymerase chain reaction with sequence-specific primers and an allelespecific restriction enzyme analysis. RESULTS: We found that the gene frequencies of HNA-4a and HNA-5a were 0.99 (107/110) and 0.96 (3/110), respectively, among the Korean population. But only the ones of the latter was significantly higher (P<0.01) than in the one of Caucasians. CONCLUSIONS: The gene frequencies of HNA-4a and HNA-5a were determined. We also identified an individual who was the HNA-5a-negative homozygote for the granulocyte panel that could be used for anti-HNA-5a antibody identification.

The First Case of Febrile Transfusion Reaction Caused by Anti-HNA-1b Antibody in Korea / 대한수혈학회지

Febrile nonhemolytic transfusion reaction (FNHTR) is one of the most common adverse reactions to the transfusion of blood products. It is caused by antigen-antibody reactions between patient's plasma and transfused cellular components (or the reverse) or bioactive substances such as cytokines generated by leukocytes during storage of cellular blood components. Most of the antibodies involved in FNHTR are anti-HLA antibodies. However, platelet-specific antibodies and rarely granulocyte-specific antibodies may be involved in FNHTR, We found a granulocyte antibody from a 53-year-old male with FNHTR for the first time in Korea. Fever developed during transfusion of a unit of packed RBC after partial cystectomy, and subsided after the adminstration of acetaminophen. The hemolytic transfusion reaction and sepsis were excluded after investigations. Mixed passive hemagglutination test revealed that the patient had antibody against human neutrophil antigen-1b (HNA-1b), and granulocyte antigen genotyping showed granulocyte antigen mismatches between the patient (HNA-1a homozygote) and the unit of transfused RBC (HNA-1a/HNA-1b heterozygote).