ALBERT

Description: Previous studies have revealed various extrinsic stimuli and factors involved in the regulation of hematopoiesis. Among these, Notch-mediated signaling has been suggested to be critically involved in this process. Herein, we show that conditional inactivation of ADAM10, a membrane-bound protease with a crucial role in Notch signaling (S2 cleavage), results in myeloproliferative disorder (MPD) highlighted by severe splenomegaly and increased populations of myeloid cells and hematopoietic stem cells. Reciprocal transfer of bone marrow cells between wild-type and ADAM10 mutant mice revealed that ADAM10 activity in both hematopoietic and nonhematopoietic cells is involved in the development of MPD. Notably, we found that MPD caused by lack of ADAM10 in nonhematopoietic cells was mediated by G-CSF, whereas MPD caused by ADAM10-deficient hematopoietic cells was not. Taken together, the present findings reveal previously undescribed nonredundant roles of cell-autonomous and non–cell-autonomous ADAM10 activity in the maintenance of hematopoiesis.

Description: Abstract 2112 Poster Board II-89 (Purpose) Asian populations have a wide variety of genetics and circumstances. Therefore, immune responses to allo-antigens may differ from those that are common among American and European populations. However, we have little information from Asian populations, and collaborative international study is required to improve the success of blood transfusions and transplantations. Therefore, we tried to analyze allo-antibodies (Abs) to erythrocyte antigens in patients from Asian countries. (Method) Forty-eight institutes, including those in Japan (29), Korea (15), Hong Kong (1), Singapore (1), Malaysia (1) and Thailand (1), participated to this cooperative study of allo-antibodies to erythrocyte antigens in Asian populations. We investigated the number of blood grouping tests, cross matching tests, and erythrocyte irregular Ab analyses as well as information of institutes including total number of bed, operations and transfusions performed. We also investigated methods adopted for screening and detecting irregular Abs to erythrocyte antigens. Furthermore, we studied the frequencies of irregular Abs to erythrocytes (D, C ,c, E, e, f, Ce, P1, M, N, S, s, Mia, Lea, Leb, Jka, Jkb, Jk3, Fya, Fyb,K, k, Kpa, Kpb, Jsa, Jsb, Dia, Dib, Lua, Lub, Xga and H), and compared them among Asian countries. The total number of independent cases included more than 866,000 patients. If a case was analyzed multiple times, we counted it as one case. Multiple antibodies detected in a patient were separately summarized. (Results) Ab screening methods adopted in these institutes included gel columns, beads columns, traditional tubes or some combination of these three. There was no difference in the methods including the erythrocyte panel adopted. Among the serum of 8,880 patients (male /female: 1 /1.55) 10,069 antibodies were detected. The numbers of these patients registered in each country are as follows; 4,222 in Japan, 2,423 in Korea, 975 in Malaysia, 622 in Thailand, 612 in Hong Kong and 26 in Singapore. On average, anti-E Ab was detected in 25.7% (min-max country, 14.3-32.2%) of patients, anti-Lea 20.9 % (3.2-30.4), anti-Mia 7.3% (0-49.4), anti-P1 6.6 % (0-10.6), anti-M 5.6 % (0-7.0), ant-c+E 4.9 % (1.6-8.5), anti-Leb 4.3 % (0-7.6), anti-D 4.0 % (0-16.2), anti-Fyb 2.2 % (0-3.7), anti-Dia 1.9 % (0-3.3), anti-Jka 1.7 % (0.8-7.7), anti-C+e 1.5 % (0-3.3), anti-Lea+Leb 1.4 % (0-5.8), anti-S 1.1 % (0-1.9), anti-c 1.1 % (0-2.3) and other Abs 9.9 %. Anti-D Ab was frequently observed in patients from Korea and Malaysia compared to Thailand, Hong Kong and Japan. Anti-Lea Ab was frequently observed in patients from Malaysia, Japan, Singapore and Korea. Anti-Dia Ab was observed in patients from Japan and Korea. Anti-Mia Ab was frequently observed in patients from Thailand and Hong Kong, followed by Malaysia and Singapore, and rarely from Japan and Korea. (Conclusion) This is the first international collaborative study on allo-immunity to erythrocyte antigens in Asian populations. There are some important differences among Asian countries with regard to the presence of anti-D, -Lea, -Dia and -Mia Abs. Further, the proportions of detected Abs, (such as anti-K, -D and -MiaAbs) differ from those reported in previous studies of patients from US and European counties (Winters et al, Hoeltge et al, Walker et al, etc). These new data will contribute to the advancement in successful blood transfusion in Asia and the world. Furthermore, the study will strengthen the network of international collaboration in future. Disclosures: No relevant conflicts of interest to declare.

Description: Abstract 3144 Poster Board III-81 Purpose Allo-immunization to erythrocytes is an important issue with regard to blood transfusions, albeit perhaps less prominent than other possible adverse effects. Several studies including pregnant and transfused cases have been conducted in US and European counties. However, a sufficient international or interracial comparative study has not been performed yet. Asian populations contain a wide variety of genetics and circumstances. They may differ in immune responses to allo-antigens that are common among American and European populations. Because no confirmed nor detail information exists for many Asian populations, collaborative international study concerning these issues will improve blood transfusions and transplantations not only in Asia but also the world. (Method) Forty-eight institutes, including those in Japan (29), Korea (15), Hong Kong (1), Singapore (1), Malaysia (1) and Thailand (1), participated in this first cooperative study of Asian population examining allo-immunity to erythrocytes. The total number of independent cases included more than 866,000 patients. Ab screening methods adopted in these institutes included gel columns, beads columns, traditional tubes or some combination of these three. If a case was tested multiple times, we counted it as one case. Multiple antibodies detected during the study in one patient were separately analyzed. Patients with unconfirmed histories were excluded from the study. We analyzed and compared the frequencies of irregular antibodies (Abs) to erythrocyte antigens between patients of different gender, patients pregnant or not, and patients with and without previous blood transfusion. Results Abs were determined in 8,880 patients (male /female: 3,528 /5,482), including 1801 /2418 from Japan, 946 /1477 from Korea, 287 /688 from Malaysia, 214 /408 from Thailand, 237 /375 from Hong Kong, and 11 /15 from Singapore. Of note, anti-D Ab was more frequently detected in females (p

Description: Backgrounds: Allo-immunization to blood cell antigens has been frequently reported among transfusion recipients with hematological diseases and cancer, as well as during pregnancy. The frequency of irregular erythrocyte antibodies (Abs) ranges from 1 to 2 % among hospitalized patients. However, the frequency varied according to the genetic diversity of the population. Several studies involving transfusion cases have been conducted in various countries including Japan (Watanabe et al, ASH 2009). However, there is currently little information about irregular erythrocyte Abs in many Asian populations. Here, we show updated data concerning variations of irregular erythrocyte Abs in patients with a history of blood transfusions between South Korea and Japan. Materials and Methods: In all, 21 institutions from South Korea and 59 from Japan participated in this study. We investigated methods employed for screening and identifying irregular erythrocyte Abs. The frequencies of irregular Abs to D, C, c, E, e, f, Ce, P1, M, N, S, s, Mia, Lea, Leb, Jka, Jkb, Jk3, Fya, Fyb, K, k, Kpa, Kpb, Jsa, Jsb, Dia, Dib, Lua, Lub, Xga and H were studied. If a case was analyzed multiple times, it was counted once. Multiple antibodies detected in the same patient were counted separately. The frequencies of irregular erythrocyte Abs on the basis of blood transfusion history were analyzed and compared between patients from South Korea and Japan. We partially selected irregular erythrocyte Abs detected in patients that were initially negative before receiving a transfusion. Among them, we compared the efficacy of detection methods between the indirect antiglobulin test (IAT) and the enzyme method at the first detection of the Abs. Results: In total, antibodies were detected in 16,438 patients (3,525 from South Korea; 12,913 from Japan). The female to male ratios of patients from the South Korean and Japanese institutions were 1.53 and 1.43, respectively. The number of patients with and without a history of previous transfusion was 1,146 and 2,100 in South Korea, 3,609 and 8,185 in Japan, respectively. Anti-E (1.4x, p

Description: Introduction: Pregnancy sometimes leads to the production of irregular erythrocyte alloantibodies (Abs). Although several investigations have been carried out in the U.S. and European countries there are few international collaborative studies on irregular erythrocyte Abs during pregnancy. Asia has a wide multiethnic population and is therefore ideal for studying diversity among ethnic groups related to transfusion medicine. However, the frequency and specificity of antigens and antibodies to erythrocytes as well as their detection is not well characterized. Therefore, the Alloimmunity to Antigen Diversity in Asian Populations Study Group was established in 2008, with the aim of studying erythrocyte alloimmunity among Asians. Here, we report similarities and differences of irregular erythrocyte Abs in pregnant patients from South Korea and Japan. Materials and Methods: Altogether, 21 institutions in South Korea and 59 in Japan participated in this study. We surveyed methods employed for screening and identifying irregular erythrocyte Abs. The frequencies of irregular Abs to D, C, c, E, e, f, Ce, P1, M, N, S, s, Mia, Lea, Leb, Jka, Jkb, Jk3, Fya, Fyb, K, k, Kpa, Kpb, Jsa, Jsb, Dia, Dib, Lua, Lub, Xga and H were investigated. If a case was analyzed multiple times, it was counted only once. Multiple antibodies detected in the same patient were counted separately. The number of patients who were pregnant at the time (pregnant patients) of the irregular erythrocyte Abs investigation was counted and compared to female patients who were not pregnant (non-pregnant female patients). Because of differences observed in anti-D Ab frequency, we investigated the number of anti-D cases in pregnant patients who received anti-D immunoglobulin, and compared it with the number of alloimmunized anti-D cases among 26 institutions in Japan from 2011 to 2013. The study was accepted with IRB in our university (#22-193, #25-252). Results: Different irregular erythrocyte Abs screening methods have been adopted at the various institutions involved in this study. The gel column method was used at 46% of institutions in South Korea and 19% of institutions in Japan, whereas the bead column method was used at 43% of institutions in South Korea and 61% of institutions in Japan. The indirect antiglobulin test (IAT) has been adopted at all institutions in both countries. The enzyme methods have been adopted at 58% of institutions in South Korea and 81% of institutions in Japan. Among more than 1.8 million patients were analyzed, antibodies were totally detected in 16,438 patients (3,525 in South Korea; 12,913 in Japan). The number of female patients was 2,113 in South Korea and 7,595 in Japan. Anti-E (1.2x, p

Description: ABO is clinically the most important blood group system in transfusion medicine and includes many variant phenotypes. To understand the molecular genetic basis of this polymorphic system, we have analyzed genomic DNAs obtained from Japanese individuals possessing variant ABO phenotypes including A2, Ax, Ael, cis-AB, Bx, and Bel. By polymerase chain reaction-single-strand conformation polymorphism (SSCP) and nucleotide sequence analyses, we identified 11 different alleles. These alleles had nucleotide sequences different from those of the previously described 13 different alleles responsible for the common ABO phenotypes. Analysis of the nucleotide sequences of the alleles responsible for those variant phenotypes showed that the amino acid residues at position 266 and 268 may be crucial for transferase specificity, whereas those at positions 214, 216, 223, 291, and 352 may be critical for the activity level. Nine of the 11 alleles, responsible for the A2, Ax, Ael, cis-AB, Bx, and Bel phenotypes, were presumed to be generated from common ABO alleles by single nucleotide mutations such as nonsynonymous substitution, deletion, or insertion. Two other alleles, responsible for the A2 and Ael phenotypes, may have originated by recombination, gene conversionlike events or accumulation of nucleotide substitutions. Our data indicate that different alleles could cause the same ABO variant phenotypes, and that these alleles do not necessarily belong to a single evolutionary lineage.

Description: Abstract 3367 Asian populations are diverse with respect to genetics, healthcare, and living circumstances. Although immune responses to erythrocyte antigens have been exhaustively characterized in various American and European populations, comparable information from Asia is lacking. Collaborative international investigations have the potential to improve transfusion and transplantation outcomes, and also advance anthropological understanding of human evolution and migration. We therefore started a comprehensive survey of alloantibodies (Abs) to erythrocyte antigens in patients from Asian countries. Here we report similaries and differences among a large number of patients in Korea and Japan. Twenty-one institutes from Korea and 38 from Japan participated in this study of alloantibodies to erythrocyte antigens. We investigated the number of blood grouping tests, cross matching tests, and erythrocyte irregular Ab analyses as well as institutional details such as the total number of beds, operations and transfusions performed. We also investigated methods employed for screening and identifying irregular Abs. Furthermore, we compared the frequencies of irregular Abs to D, C,c, E, e, f, Ce, P1, M, N, S, s, Mia, Lea, Leb, Jka, Jkb, Jk3, Fya, Fyb,K, k, Kpa, Kpb, Jsa, Jsb, Dia, Dib, Lua, Lub, Xga and H between Korea and Japan. The total number of independent cases included more than 1,135,000 patients. If a case was analyzed multiple times, it was counted once. Multiple antibodies in the same patient were separately summarized. We compared the frequencies of irregular Abs on the basis of gender, pregnancy, and transfusion history. Different methods used at different institutions did not cause statistically significant differences in results. Antibodies were detected in 10,535 patients totally, including 3,525 in Korea and 7,010 in Japan. The female to male ratios were 1.53 and 1.39, respectively. Antibody frequencies are shown in the figure. Anti-P1 and anti-Dia Abs were observed in 3.4% and 0.9% of Korean patients, respectively, versus 10.4% and 3.4% of Japanese patients. These differences were statistically significant between the two countries (p

Description: An additional decay-accelerating factor (DAF) mutation, designated as Inab phenotype in the Cromer blood group system, was recently identified in a 28-year-old Japanese woman (H.A.). The red blood cells of H.A., like those of other Inab phenotype individuals, were negative for Cromer system antigens, Cra, Tca, Dra, UMC, and IFC. The deficiency of DAF on the red blood cells of H.A. has been shown by immunoblotting with a murine monoclonal antibody to DAF. Molecular analysis has shown that H.A. is homozygous for a single nucleotide substitution, C1579→A, at the position 24 bp upstream of the 3′-end of exon 2 of the DAF gene. This substitution causes the activation of a novel cryptic splice site and results in the production of mRNA with a 26 bp deletion. The deletion introduces a reading frame shift and creates a stop codon immediately downstream of the deletion. Translation of mRNA would be terminated at the first amino acid residue of the second short consensus repeat (SCR2) domain (exon 3) of DAF. The functional domains of DAF's complement regulatory activity and the carboxy-terminal signal domains for glycosylphosphatidylinositol (GPI) anchoring are predicted to be lacking in H.A. Thus, there would be no DAF present on the cell surface.

Description: An additional decay-accelerating factor (DAF) mutation, designated as Inab phenotype in the Cromer blood group system, was recently identified in a 28-year-old Japanese woman (H.A.). The red blood cells of H.A., like those of other Inab phenotype individuals, were negative for Cromer system antigens, Cra, Tca, Dra, UMC, and IFC. The deficiency of DAF on the red blood cells of H.A. has been shown by immunoblotting with a murine monoclonal antibody to DAF. Molecular analysis has shown that H.A. is homozygous for a single nucleotide substitution, C1579→A, at the position 24 bp upstream of the 3′-end of exon 2 of the DAF gene. This substitution causes the activation of a novel cryptic splice site and results in the production of mRNA with a 26 bp deletion. The deletion introduces a reading frame shift and creates a stop codon immediately downstream of the deletion. Translation of mRNA would be terminated at the first amino acid residue of the second short consensus repeat (SCR2) domain (exon 3) of DAF. The functional domains of DAF's complement regulatory activity and the carboxy-terminal signal domains for glycosylphosphatidylinositol (GPI) anchoring are predicted to be lacking in H.A. Thus, there would be no DAF present on the cell surface.