Multiple red blood cell antibodies produced by donor B lymphocytes after ABO-matched allogeneic bone marrow transplantation.

A 50-year-old man with AML[M2,t(8;21)] underwent BMT from his younger sister. At that time, he had no unexpected antibody and his blood type was O(+), CcDEe. The type of Kidd was not examined. The donor's blood type was O(+), CCDee, Jk(a+b-). One year after the BMT, the patient's blood type had changed to that of the donor's and anti-E antibody was detected. Despite the use of platelet concentrates (PCs) only, anti-c antibody was later identified. We conclude that there is a need to check red cell antibodies at regular intervals, even when using PCs only, for earlier detection of unexpected antibodies after BMT.     

Erythroid urea transporter deficiency due to novel JKnull alleles.

BACKGROUND: The Kidd blood group antigens Jka and Jkb are encoded by the red blood cell (RBC) urea transporter gene. Homozygosity for silent JK alleles results in the rare Jk(a-b-) phenotype. To date, seven JKnull alleles have been identified, and of these, two are more frequent in the Polynesians and Finns. This study reports the identification of other JKnull alleles in Jk(a-b-) individuals of different ethnic or geographic origins. STUDY DESIGN AND METHODS: Nine Jk(a-b-) samples and a sample from a Jk(a-b+) mother of a Jk(a+b-) baby were investigated. Polymerase chain reaction amplification and sequence analysis of the JK gene was performed. Western blotting and urea lysis were used to confirm Jk(a-b-) RBCs. RESULTS: Four novel alleles were identified: two different nonsense mutations, 202C>T (Gln68Stop) and 723delA (Ile262Stop) were identified on otherwise consensus JK*1 and JK*2 alleles, respectively. A missense mutation, 956C>T (Thr319Met), was identified in a JK*1 allele from an African-American and a JK*2 allele in two people of subcontinental Indian descent. Immunoblotting and urea lysis confirmed absence of JK glycoprotein in RBC membranes from a sample carrying the 956C>T mutation. Other previously described JKnull mutations were found in samples of origins other than in which they were first identified. CONCLUSION: The molecular bases of the Jk(a-b-) phenotype are diverse and this is the first report of JKnull alleles in individuals of African and subcontinental Indian descent. Although rare, these alleles should be taken into consideration when planning genotyping strategies for blood donors and patients.     

The Kidd minus-minus phenotype in a Caucasian

This is the reported example in a Caucasian with the phenotype Jk(a-b-). A strongly reacting antibody was present which was of a single specificity and which could be completely absorbed by either Jk(a+b-) or Jk(a-b+) cells. Since no compatible donors could be found, autotransfusion was done. Family study failed to reveal any other members with the Jk(a-b-) phenotype. The question of a third allele at the Kidd locus is considered.     

深圳地区汉族无偿献血人群Kidd血型系统基因多态性的分子遗传学研究

目的 探讨深圳地区汉族无偿献血人群Kidd血型系统基因多态性的分子遗传学背景.方法 采用简单随机抽样法选择2015年1月至2015年12月于深圳市血液中心无偿献血的74例汉族献血者为研究对象.采用红细胞尿素溶血试验及常规血清学方法鉴定献血者的Kidd血型表型.采用PCR扩增JK基因第4～11外显子及部分内含子片段,并且采用直接测序法对PCR扩增产物进行序列分析.结果 ①74例献血者的Kidd血型表型分别为Jk(a+b+)37例,Jk(a+b-)16例,Jk(a-b+)21例,未筛查出稀有Kidd血型表型Jk(a-b-).②74例献血者的DNA样本均成功扩增JK基因第4～5外显子、第6外显子、第7外显子、第8～9外显子、第10外显子、第11外显子区域.③DNA序列分析结果显示,37例Jk(a+b+)表型献血者DNA样本中,JK基因存在第9外显子c.838G＞A和第7内含子c.664-68C＞T杂合突变.21例Jk(a-b+)表型献血者DNA样本中,JK基因为c.838AA和c.664-68CC纯合型,而16例Jk(a+b-)表型献血者DNA样本中,JK基因为c.838GG和c.664-68TT纯合型.74例献血者DNA样本中,JK基因均为第8内含子c.811+84TT纯合型.结论 深圳地区汉族献血人群Kidd血型系统中,除存在JK基因第9外显子c.838G＞A(p.Asp280Asn)突变以外,还存在第7内含子c.664-68C＞T碱基置换,该突变可能与JKa/JKb基因多态性相关.并且JK基因第8内含子均为c.811+84TT纯合型,其在中国人群中属于常见的基因型.     

The Kidd (JK) Blood Group System

The Kidd blood group system was discovered in 1951 and is composed of 2 antithetical antigens, Jk^a and Jk^b, along with a third high-incidence antigen, Jk3. The Jk3 antigen is expressed in all individuals except those with the rare Kidd-null phenotype. Four Kidd phenotypes are therefore possible: Jk(a+b−), Jk(a−b+), Jk(a+b+), and Jk(a−b−). The glycoprotein carrying the Kidd antigens is a 43-kDa, 389-amino acid protein with 10 membrane-spanning domains which functions as a urea transporter on endothelial cells of the renal vasa recta as well as erythrocytes. The HUT11/UT-B/JK (SLC14A1) gene encoding this glycoprotein is located on chromosome 18q12-q21. The Jk^a and Jk^b antigens are the result of a single-nucleotide polymorphism present at nucleotide 838 resulting in an aspartate or asparagine amino acid at position 280, respectively. The Kidd blood group can create several difficult transfusion situations. Besides the typical acute hemolytic transfusion reactions common to all clinically relevant blood group antigens, the Kidd antigens are notorious for causing delayed hemolytic transfusion reactions due to the strong anamnestic response exhibited by antibodies directed against Kidd antigens. The Kidd-null phenotype is extremely rare in most ethnic groups, but is clinically significant due to the ability of those with the Kidd-null phenotype to produce antibodies directed against the high-incidence Jk3 antigen. Anti-Jk3 antibodies behave in concordance with anti-Jk^a or anti-Jk^b possessing the capability to cause both acute and delayed hemolytic reactions. Antibodies against any of the 3 Kidd antigens can also be a cause of hemolytic disease of the fetus and newborn, although this is generally mild. In this review, we will outline the makeup of the Kidd system from its historical discovery to the details of the Kidd gene and glycoprotein, and then discuss the practical aspects of Kidd antibodies and transfusion reactions with an extended focus on the Kidd-null phenotype. We will end with a brief discussion of the donor aspects related to the screening and supply management of blood from donors with the rare Jk(a−b−) phenotype.     

Further evidence of heterogeneity within the Kidd blood group system

Serological studies on blood of selected individuals show that red blood cells from individuals genetically JkaJk and JkbJk give single dose agglutination reactions with anti-Jka and anti-Jkb respectively, but react as strongly with anti-JkaJkb as do cells from random individuals. A Jk(a+b-) Oriental person has been found whose red blood cells give a single dose reaction with anti-Jka, but react more weakly than do cells from random people in tests with anti-JkaJkb.The results support a conclusion that the antigen recognized by anti-JkaJkb is a distinct and separate antigen of the Kidd blood group system.     

Acute hemolytic anemia associated with a chlorpropamide-induced apparent auto-anti-Jka

A patient with acute hemolytic anemia and a positive direct antiglobulin test was found to be Jk(a + b +) with anti-Jka in her serum. For 2 weeks prior to admission, the patient had taken chlorpropamide, a hypoglycemic agent. The drug was discontinued upon the diagnosis of hemolytic anemia, and the hemoglobin concentration gradually increased. When chlorpropamide was added to the patient's serum in vitro, it enhanced the reactivity of the anti-Jka, and 40 days posttransfusion, the serum would only react with Jk(a+) red cells when chlorpropamide was present. These findings suggest that a chlorpropamide-dependent antibody with Jka specificity had formed. We do not know why the antibody induced by chlorpropamide reacted preferentially with Jk(a+) red cells.     

DNA-based typing of blood groups for the management of multiply-transfused sickle cell disease patients

BACKGROUND: The usefulness of DNA genotyping for RBC antigens as a tool for the management of multiply-transfused patients with sickle cell disease (SCD) to overcome the limitations of hemagglutination assays was evaluated. STUDY DESIGN AND METHODS: Blood samples from 40 multiply-transfused SCD patients were studied by hemagglutination and by PCR-RFLP for antigens or genes in the Rh (D, C/c, E/e), Kell, Kidd, and Duffy systems. RESULTS: Discrepancies were found between hemaglutination and DNA typing test results in six patients: two were discrepant in Rh typing (one was D- by hemagglutination and RhD by DNA, and one was E+e- and RhEe by DNA), two were discrepant in Duffy typing [both were Fy(a+b-) and Fy(b)/Fy(b) by DNA], and four were discrepant in Kidd typing [Jk(a+b+) and Jk(b)/Jk(b) by DNA; two of these samples were also discrepant in Duffy]. Stored segments from blood units that had been recently transfused to these six recipients were phenotyped, confirming that the transfused RBCs were the source of the discrepancy between genotype and phenotype. CONCLUSION: DNA typing of blood groups by PCR-RFLP in peripheral blood WBCs contributes to the management of transfusions in SCD patients by allowing a more accurate selection of donor units.     

上海地区部分人群Jk(a-b-)、Di~b-、Wr~b-、K_0、Ena-、Tja-、Ge-稀有血型筛选

目的　了解上海地区人群中部分稀有血型的分布 ,以解决临床稀有血型用血问题。方法　采用 2M尿素对受检红细胞进行Kidd血型系统的Jk(a -b - )表型筛选 ;利用稀有单克隆、多克隆抗血清 ,对Diego血型系统的Dib-、Wrb- ,Kell血型系统的K、K0 ,MNS血型系统的Ena - ,P血型系统的Tja - ,Miltinberger血型系统的Murf,Gerbich血型系统的Ge -进行筛选。筛选方法采用试管法间接抗人球蛋白试验 (IAT) ,U型 96孔微量板IAT和盐水直接离心法试验。结果　在 6 4 5人中筛选到Di(a +b - ) 1例 ,90 0人中筛选到Mur(+) 6例 ,10 0 5 7人中筛选到K(+) 7例 ,4 84 0 0人中筛选到Jk(a -b - ) 2例 ,而在 4 0 0 0人的筛选中未发现Wrb-、K0 、Ge -和Ena -表型。结论　上海地区人群中Dib-、Mur+、Jk(a-b - )的红细胞表型明显高于白种人和黑人。     

Blood group genotyping facilitates transfusion of beta-thalassemia patients

We evaluated the usefulness of blood group genotyping as a supplement to hemagglutination to determine the red blood cell (RBC) antigen profile of polytransfused patients with beta-thalassemia. We selected 10 alloimmunized patients who were receiving antigen-matched RBCs based on phenotype, and had clinical evidence of delayed hemolytic transfusion reaction. DNA was prepared from blood samples and RH E/e, K1/K2, FY A/FY B, and JK A/JK B alleles were determined by PCR-RFLP. RH D/non-D was determined according to the PCR product size associated with the RHD gene sequence in intron 4 and exon 10/3'UTR. RH C/c was tested by multiplex PCR. The phenotypes and genotypes of nine of the 10 samples were discrepant. Five of the discrepancies occurred in the Rh system. One sample was phenotyped as Rhcc and genotyped as RH C/C, and two samples were phenotyped as RhCc and genotyped as RH C/C. Two other samples were phenotyped as RhEe and genotyped as RH e/e. Three samples had discrepancies in the Kidd system with phenotype Jk(a+b+) and were genotyped as homozygous for JK B. One sample had a discrepancy in the Duffy system: it was phenotyped as Fy(a+b-) and homozygous for FY B. Genotyping was very important in determining the true blood groups of many polytransfused patients with beta-thalassemia, and it assisted in the identification of suspected alloantibodies and the selection of antigen-negative RBCs for transfusion.     

Jk(a−b−) red blood cells resist urea lysis

Falsely high automated platelet counts in a patient with aplastic anemia were found to be due to increased resistance of the red blood cells to urea lysis. The patient's blood group Jk(a-b-). Further investigation revealed that this phenomenon occurred with all of eight bloods of the phenotype Jk(a-b-) but not with red blood cells of other phenotypes tested. We therefore report an association of a rare blood group phenotype with unusual red blood cell behavior in vitro.     

Genomic characterization of the kidd blood group gene:different molecular basis of the Jk(a-b-) phenotype in Polynesians and Finns

BACKGROUND: The clinically important Kidd (JK) blood group antigens are carried by the urea transporter in red cells. The rare Jk(a-b-) phenotype can be caused by homozygosity at the JK locus for a silent allele, JK: This phenotype has been recorded in many ethnic groups, but it is most abundant among people originating from the Polynesian Islands and Finland. The molecular basis for Jk(a-b-) is unknown in these populations. STUDY DESIGN AND METHODS: Blood samples from individuals of Swedish, Polynesian, and Finnish origin were collected and characterized by routine JK blood group serology and JK genotyping. Genomic DNA covering the exons and intervening introns of the JK gene coding region was amplified by polymerase chain reaction, and fragments were directly sequenced. RESULTS: Exon and partial intron sequences in the coding region of the JK gene were determined. Finnish and Polynesian Jk alleles were analyzed; the only deviations from consensus were a splice-site mutation (G-->A) in Polynesians, causing skipping of exon 6, and a T871C substitution predicted to disrupt a potential N-glyco-sylation motif (NSS-->NSP) in Finns. Methods for rapid detection of silent Jk alleles were developed for clinical application. CONCLUSION: Polynesians and Finns have two different molecular alterations in their Jk alleles, both of which can now be determined by polymerase chain reaction.     

流式细胞术应用于Kidd血型Jka抗原检测的建立

目的:建立流式细胞术鉴定Kidd血型Jka抗原的实验方法,并验证其准确性。方法:随机选取96(人)份无偿献血者血液样本,将红细胞与IgG性质抗-Jka混合孵育,随后结合荧光素Alexa Fluor 647标记的羊抗人IgG二抗,采用流式细胞术检测样本红细胞表面Jka抗原。通过流式细胞术获得每个样品的荧光直方图。利用血清学凝集试验比较衡量流式细胞术检测Jka抗原强度的准确性,利用PCR-SSP与基因测序分型验证流式细胞术检测Jka抗原阴阳性的正确性。结果:流式细胞术对于红细胞Jka抗原强度的检测与血清学鉴定结果相一致,血清学凝集强度越高,流式细胞术检测荧光强度也越高,Jka抗原强度也越高。流式细胞术检测灵敏度优于血清学方法,Jka抗原阳性包括弱阳性样本与阴性样本的荧光强度差别很大,极易区分。全部样本流式细胞术检测结果与基因分型结果完全一致,证实了流式细胞术的准确性。结论:成功建立了流式细胞术鉴定红细胞Kidd血型Jka抗原的方法,灵敏度优于传统的血清学方法,可以准确区分不同强度的Jka血型抗原。     

应用血清学和分子生物学方法筛选浙江汉族人群中部分稀有血型

本研究探讨浙江汉族人群部分红细胞血型系统稀有表型的分布情况。利用血清学技术或分子生物学方法分别筛选H系统H-、MNS系统GPA-和s-、Rh系统Rhnull、Rhmod、D--、CCDEE和CCdEE、Gerbich系统GPC-、I系统i+、Lutheran系统Lub-、Kell系统k-和Jsb-、Duffy系统Fya-、Ok系统Oka-、Diego系统Dib-。利用尿素溶血试验筛选Kidd系统Jk(a-b-)表型。结果表明:1 618例献血者中检出1例Di(a+b-),1 007例献血者检出3例Fy(a-b+),633例Rh阴性献血者检出1例CCdEE。大规模筛选中未发现Jk(a-b-)、H-、GPA-、s-、GPC-、成人i+、Lub-、k-、Jsb-、Lub-和Oka-稀有血型。结论:在献血人群中发现Di(a+b-)、Fy(a-b+)、CCdEE稀有表型,提供了浙江汉族人群部分红细胞稀有血型的分布数据。     

Methods for the detection of Jk heterozygotes: interpretations and applications

The lytic properties of red cells from Jk(a+,b+), Jk(a-, b+) and Jk(a+b-) (normal), Jk(a-b-), and obligate Jk heterozygotes were studied. The Jk(a-b-) cells did not hemolyze for at least 15 minutes in either 2 M urea or methylurea, whereas normal cells were completely hemolyzed in 2 minutes. Red cells from Jk heterozygotes demonstrated intermediate levels of hemolysis when compared to normal and Jk(a-b-) red cells. In addition, these cells had less than 10 percent hemolysis when suspended in 2 M methylurea prepared in 0.4 percent phosphate-buffered saline (pH 7.2). This method may be an alternative to hemagglutination titration scoring for the detection of Jk, especially in paternity cases.     

An apparent anti-Jka reacting only in the presence of methyl esters of hydroxybenzoic acid

An apparent anti-Jka, reacting only in the presence of methyl esters of hydroxybenzoic acid (HBA), was detected in an individual with Jk (a+) red blood cells. The antibody was first detected when a commercial low- ionic-strength-solution preparation containing the preservative methyl paraben (a methyl ester of hydroxybenzoic acid) was used in compatibility testing. Negative reactions were obtained when regular saline, albumin, and enzyme technics were employed. The positive indirect antiglobulin tests were due to cell-bound complement; no IgG was detected. The patient had no clinical or hematologic evidence of hemolytic anemia and was transfused with Jk(a+) red blood cells with no ill effects. It is postulated that immune complexes formed between the antibody and the chemical may result in a site complementary to some aspect of the Jka antigen. When this complex binds to Jk(a+) cells, complement is activated and some complement components remain on the cells. We do not know why only Jk(a+) cells were involved in these reactions.     

Anti-Jka, -C, and -E in a single patient, initially demonstrable only by the manual hexadimethrine bromide (Polybrene) test, with incompatibilities confirmed by 51Cr-labeled red cell studies

Published reports have confirmed the superior sensitivity of the manual hexadimethrine bromide (Polybrene) test (MPT) for demonstrating many alloantibodies in vitro; however, the clinical significance of alloantibodies demonstrable exclusively by MPT has not been shown conclusively. A patient with macroglobulinemia experienced chills, fever, hemoglobinemia, and hemoglobinuria following the transfusion of 1 unit of red cells (RBCs) shown to be compatible by the low-ionic-strength antiglobulin (LIS-AG) method. Serologic investigation was negative. Intravascular hemolysis occurred with a second "compatible" unit. Serologic studies were again negative by LIS-AG and ficin-AG methods, but revealed anti-Jka by MPT. Both donors were Jk(a+b-), and 51Cr studies of the second donor's RBCs revealed a t1/2 of less than 30 minutes, with marked intravascular hemolysis. A LIS-AG-compatible Jk(a-) unit was transfused uneventfully, but with no rise in hematocrit. MPT next revealed anti-C; subsequent 51Cr studies with the Jk(a-), Cc donor's RBCs showed a 51Cr t1/2 of 100 minutes with slight intravascular lysis. Four transfusions of Jk(a-), C- blood were uneventful, but 5 days later the patient's hemoglobin declined. The following day, anti-E was demonstrable exclusively by MPT. 51Cr-labeled Jk(a-), C-, E- RBCs had normal 24-hour survival. The patient's hemoglobin rose to 11 g per dl following transfusions of Jk(a-), C-, E- RBCs, and he was discharged. In vitro studies employing the patient's purified IgM paraprotein revealed no interference with alloantibody binding or detection.     

Erythroid urea transporter deficiency due to novel JKnull alleles

BACKGROUND: The Kidd blood group antigens Jk^a and Jk^b are encoded by the red blood cell (RBC) urea transporter gene. Homozygosity for silent JK alleles results in the rare Jk(a–b–) phenotype. To date, seven JK ^null alleles have been identified, and of these, two are more frequent in the Polynesians and Finns. This study reports the identification of other JK ^null alleles in Jk(a–b–) individuals of different ethnic or geographic origins.
STUDY DESIGN AND METHODS: Nine Jk(a–b–) samples and a sample from a Jk(a–b+) mother of a Jk(a+b–) baby were investigated. Polymerase chain reaction amplification and sequence analysis of the JK gene was performed. Western blotting and urea lysis were used to confirm Jk(a–b–) RBCs.
RESULTS: Four novel alleles were identified: two different nonsense mutations, 202C>T (Gln68Stop) and 723delA (Ile262Stop) were identified on otherwise consensus JK * 1 and JK * 2 alleles, respectively. A missense mutation, 956C>T (Thr319Met), was identified in a JK * 1 allele from an African-American and a JK * 2 allele in two people of subcontinental Indian descent. Immunoblotting and urea lysis confirmed absence of JK glycoprotein in RBC membranes from a sample carrying the 956C>T mutation. Other previously described JK ^null mutations were found in samples of origins other than in which they were first identified.
CONCLUSION: The molecular bases of the Jk(a–b–) phenotype are diverse and this is the first report of JK ^null alleles in individuals of African and subcontinental Indian descent. Although rare, these alleles should be taken into consideration when planning genotyping strategies for blood donors and patients.     

Assessment of apparent non-maternity using individual specific gene probes

In a case of disputed paternity mother and child showed a genetic incompatibility in the Kidd system of the erythrocytes: mother Jk(a-b+), child Jk (a+b-). For the first time the RFLP technique solved this problem, using four different Jeffreys' highly polymorphic single locus DNA probes MS 1, MS 31, g 3 and MS 43. We found that mother and child possessed one identical gene observed in each of the four probes. The confidence of somatic genuineness of this mother-child pair is very strong: p = 10(9). In addition, the above-mentioned four DNA probes showed identical results in 12 cases of paternity exclusion and 18 nonexclusions found by 23 conventional hereditary systems. Furthermore, in five cases of bone marrow transplantation success of this therapy was proven by the RFLP technique, which differentiates between recipient genes and donor genes. Our investigations on hereditary so far comprise five families with 10 children and one family over four generations.