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Kunhai Wu,1 Lufei Chen,1 Huifang Huang2 1Blood Transfusion Department, Fujian Provincial Maternity and Children’s Hospital, Fuzhou, Fujian, People’s Republic of China; 2Intensive Care Unit, Fujian Provincial Maternity and Children’s Hospital, Fuzhou, Fujian, People’s Republic of ChinaCorrespondence: Huifang HuangIntensive Care Unit, Fujian Provincial Maternity and Children’s Hospital, Fuzhou, Fujian, People’s Republic of ChinaTel +86 13906913057Fax +86 591-87555474Email 190596715@qq.comPurpose: The study aimed to analyse the clinical effect of homotype ET method and compatible ET method in neonates with hyperbilirubinemia caused by non-blood-group antibodies (non-blood-group antibodies refers to due to cause other than blood-group antibodies) as well as to provide relevant reference materials for clinical diagnosis and treatment in Fuzhou city.Patients and Methods: Between January 2011 and December 2020, a total of 125 neonates, who were admitted to the Sentinel hospital of Fuzhou city, Fujian Provincial Maternity and Children’s Hospital for hyperbilirubinemia and treated with ET therapy were selected. They were divided into homotype ET (n=32) and compatibility ET (n=93) treatment groups. We analysed changes in serum total bilirubin (STB), serum indirect bilirubin (SIB), and platelet count before and after ET; retrospectively analysed the neonates’ clinical data; and compared findings between the 2 groups.Results: The homotype ET group and the compatible ET group showed higher STB, SIB, and platelet levels before ET and decreased levels of the same after ET; the difference was statistically significant (P< 0.001). Compared with the compatible ET group, the decreases in STB, SIB, and platelet count in the homotype ET group showed no significant difference (P> 0.05).Conclusion: ET therapy is the most timely and effective treatment method for severe hyperbilirubinemia in neonates clinically, and the effects of homotype and compatible ET are similar.Keywords: hyperbilirubinemia, exchange transfusion, neonate, homotype, compatibility

ZusammenfassungEine positive Verträglichkeitsprobe bei scheinbar negativem Antikörperstatus ist ein seltenes Ereignis, das jedoch häufig eiliger Aufklärung bedarf. Zu den Ursachen gehören Verwechslungen von Proben oder Patienten, Fehlbestimmungen, neu nachweisbare irreguläre Alloantikörper, Antikörper gegen niedrigfrequente Antigene, gegen Reagenzienbestandteile oder gegen das Merkmal A1. Ein standardisierter Arbeitsfluss hilft, die Ursachen rationell und schnell aufzuklären und so eine sichere Versorgung des Patienten zu ermöglichen.

Blood group O or B recipients wait longer for a kidney transplant. We studied the distribution of anti-ABO blood group antibody titres in patients awaiting a kidney transplant, and modelled the effect of altering the UK National Kidney Allocation Scheme to allow for patients with 'LOW' titres (≤1:8, ≤3 dilutions) to receive a deceased donor ABOi (ddABOi) transplant. In a prospective study of 239 adult patients on the waiting list for a transplant in 2 UK centres, ABO-antibody titres (anti-A and anti-B) were measured. Based on the proportions of 'LOW' anti-A or anti-B antibodies, four simulations were performed to model the current allocation rules compared with variations allowing ddABOi allocation under various conditions of blood group, HLA matching, and waiting time. The simulations permitting ddABOi resulted in more blood group B recipients being transplanted, with median waiting time reduced for this group of recipients, and more equitable waiting times across blood groups. Additionally, permitting ddABOi resulted in greater numbers of 000MM allocations overall in compatible transplants under modelled conditions. Changing allocation in the UK to permit ddABOi in patients with 'LOW' titres would not change the total number of transplants, but redistributes allocation more equitably amongst blood groups, altering waiting times accordingly.

Colorectal cancer (CRC) is a common cause of cancer-related deaths largely due to CRC liver metastasis (CRLM). Identification of targetable mechanisms continues and includes investigations into the role of inflammatory pathways. Of interest, MAPK is aberrantly expressed in CRC patients, yet the activation status is not defined. The present study assessed p38&gamma, activation in CRC patients, cancer cells, and tissues of cotton top tamarin (CTT) and common marmoset (CM). The primate world is an overlooked resource as colitis-CRC-prone CTT are usually inure to liver metastasis while CM develop colitis but not CRC. The results demonstrate that p38&gamma, protein and phosphorylation levels are significantly increased in CRC patients compared to normal subjects and CTT. Furthermore, p38&gamma, phosphorylation is significantly elevated in human CRC cells and hepatoblastoma cells but not in CM colon. Additionally, carcinoembryonic antigen (CEA) and biliary glycoprotein (BGP) are induced in the CRC patients that showed p38&gamma, phosphorylation. Inhibition of p38 MAPK in CRC cells showed a significant decline in cell growth with no effect on apoptosis or BGP level. Overall, p38&gamma, is activated in CRC tumorigenesis and likely involves CEA antigens during CRLM in humans but not in the CTT or CM, that rarely develop CRLM.

In this chapter, the characteristics of an antibody, ABO blood group antibodies, and the detection of blood group antibodies are discussed. ABO blood group antibodies, the most important antibodies in the human body, have many common features with general antibodies in terms of basic functions and biological characteristics. Clearance of ABO blood group antibodies (mainly IgG and IgM) is the key to ABO-incompatible (ABOi) transplantation. Antibody removal technology includes plasma exchange technology, immuno-adsorption technology, and plasma double filtration technology.

The presence of multiple alloantibodies or an antibody to a high-prevalance antigen in a patient sample can pose challenges in antibody identification. The pattern of reactivity seen on an antibody panel may show various strengths of reactivity by different methods of testing or same strength of reactivity at one or more phases of testing. To ensure proper identification, multiple investigative tools may be used. We review one of these methods-inhibition by soluble substances-which has become an expansion of our toolbox within the past 10 years. Alloantibodies can be inhibited using specific soluble substances. These soluble substances occur naturally in various fluids or can be manufactured. When a patient sample contains multiple antibodies, clinically significant or not, inhibition of one may help determine specificities of others. Specific inhibition of a particular antibody will also help to confirm its presence. The presence of multiple alloantibodies or an antibody to a high-prevalance antigen in a patient sample can pose challenges in antibody identification. The pattern of reactivity seen on an antibody panel may show various strengths of reactivity by different methods of testing or same strength of reactivity at one or more phases of testing. To ensure proper identification, multiple investigative tools may be used. We review one of these methods—inhibition by soluble substances—which has become an expansion of our toolbox within the past 10 years. Alloantibodies can be inhibited using specific soluble substances. These soluble substances occur naturally in various fluids or can be manufactured. When a patient sample contains multiple antibodies, clinically significant or not, inhibition of one may help determine specificities of others. Specific inhibition of a particular antibody will also help to confirm its presence.

The Hemolytic Disease of the Fetus and Newborn (HDFN), if untreated, can have severe consequences on baby’s health, eventually causing death. Prevention and diagnosis of pregnancies at risk of HDFN consists in quantitative analysis in hospital every 2-4 weeks of blood group antibodies in maternal serum. The most important antibodies to be monitored are anti- Rhesus D (or anti-RhD) ones. Nowadays several portable systems for different healthcare applications have been developed towards decentralization of diagnosis thanks to the enormous advances in electrochemical biosensing. Home-monitoring of pregnancies at risk of HDFN could significantly improve patients’ life quality. Here we demonstrate for the first time label-free quantitative detection of anti-RhD antibodies by means of electrochemical impedance spectroscopy (EIS), suitable for integration within a complete portable device. The specificity of the sensor was also proved.

BACKGROUND: Patients transfused at more than one health care facility face safety risks, because their transfusion record is fragmented. Blood group antibodies documented at one facility may be unknown to others. Because many antibodies are evanescent, access to prior antibody records is important for preventing incompatible transfusions and delayed hemolytic reactions. The study goal was to quantify multisite transfusion activity and its impact on antibody record accuracy. STUDY DESIGN AND METHODS: Patients (n = 100) undergoing hospital transfusion testing were surveyed to determine the locations and dates of any prior trans

BACKGROUND: Concurrent alloantibodies are defined as two or more blood group (BG) antibodies coexisting in a given patient. These antibodies are significant because they can present major problems in compatibility testing. The goals of this study were to determine the properties of concurrent BG antibodies. STUDY DESIGN AND METHODS: The transfusion records of 18,750 patients at a Veterans Affairs medical center were reviewed to identify alloimmunized individuals. The following data were collected on patients making concurrent antibodies: antibody specificities, time of first detection, whether the antibodies disappeared over time, and, if so, their time of disappearance. RESULTS: Multiple alloimmunization occurred in 21.7% (96/443) of alloimmunized patients, constituting 39.9% (230/577) of all antibodies. The rate at which an antibody was concurrent with another antibody varied by antigenic specificity (p

A commercially prepared antibody screening system, Selectogen®, is now available in Denmark. A total of 291 selected sera underwent extended antibody studies by means of the Selectogen procedure and the routine methods of the blood grouping laboratory in Arhus. One hundred and thirty three sera contained irregular antibodies which could be analyzed in detail with respect to sensitivity of the methods used. The comparison showed the value of the two-stage papain technique in studies aimed at revealing the presence of weak irregular antibodies. The Selectogen albumin technique was not found to be a satisfactory substitute for the enzyme technique. Good results were obtained by using the Selectogen albumin indirect Coombs technique, but even that could not replace the enzyme technique, particularly not in the demonstration of weak Rhesus or Lewis antibodies.

A survey is presented of 4168 sera containing irregular blood-group antibodies. The antibodies were found during 309,335 antibody screenings and 135,402 cross matchings of blood samples from about 200,000 individuals. About 50 per cent of the antibodies were anti-D, sometimes together with other antibodies in Rh negatives (D neg), 8 per cent were Rh antibodies in Rh positives (D pos) and many of these antibodies were detectable only by a two-stage enzyme technique and not by AHG-technique. Almost 15 per cent of the antibodies belonged to the Lewis-system. There was a significantly increased number in females of the fertile age, and there was a significant decrease in the number of group O individuals among the Lewis-immunized persons. The low number of group O individuals indicate a hitherto unrecognized relationship between the ABO- and Lewis-systems. Altogether 2.1 per cent of the individuals described possessed one or more irregular antibodies. The importance is discussed of avoiding complications from these antibodies during blood transfusions. It is concluded that a reliable cross-match should include a two-stage enzyme technique as well as the usual saline and AHG tests. It is emphasized that the antibody screening is an efficient prophylactic measure, particularly for smaller hospitals, where complicated cross-matchings are impractical.

Removal of blood group antibodies against the donor organ prior to ABO-incompatible transplantation can prevent episodes of hyperacute rejection. We are developing a specific antibody filter (SAF) device consisting of immobilized synthetic Atrisaccharide antigens conjugated to polyacrylamide (Atri-PAA) to selectively remove anti-A antibodies directly from whole blood. In this study, we evaluated eight anti-A IgM monoclonal antibodies (mAbs) using Enzyme-Linked Immunosorbent Assay (ELISA) to determine their specificity for binding to Atri-PAA. Five of the eight mAbs met our criteria for specificity by binding to Atri-PAA with at least five times greater affinity compared to the negative controls. These selected mAbs will be studied for their binding characteristics to Atri-PAA which will aid in the development of the SAF. The study of kinetics of antibody removal and quantification of antibody removal will be used in our mathematical model to maximize the antibody removal rate and binding capacity of the SAF.

Alloantibodies to blood group antigens are produced because there are differences between the blood group antigens on recipient and donor red blood cells. Historically, and currently, blood group antibodies and antigens are detected by haemagglutination, which requires appropriate reagents and considerable skill. Over the past two decades there has been an astounding pace of growth in the field of molecular biology techniques and even more recently in the understanding of the basis of many blood group antigens and phenotypes. Thus, we are now able to determine blood group polymorphisms and predict blood group antigen expression using molecular approaches. This article reviews molecular approaches to identify donors with rare blood types, either due to the absence of a high prevalence antigen or an absence of a combination of antigens.

Purpose: The aim of this study is to investigate the mechanism of long-term acceptance of ABO-incompatible renal allograft. Methods: Thirty-four ABO-incompatible kidney transplant recipients with blood group O were entered in this study. Eighteen and 16 recipients received a blood group B (B-to-O) and A1 (A1-to-O) allograft, respectively. Sixteen (O-to-O) recipients were control subjects. The titers of blood group antibodies were measured before pre-conditioning and following transplantation. Blood group antigens were stained in 32 graft biopsy specimen. Results: In 18 (B-to-O) and 16 (A1-to-O) recipients, only anti-B and anti-A1 antibodies, respectively, were significantly suppressed compared with those in 16 (O-to-O) more than 1 year post-transplantation. In eight (B-to-O) and seven (A1-to-O) recipients, only anti-B and anti-A1 antibodies, respectively, were significantly suppressed 1 year post-transplantation, compared with those pre-conditioning. Blood group antigens were stained with same intensity and pattern in any biopsy specimen under all conditions. There was no significant deposition of IgG and IgM in any biopsy specimen. These suggested that production of anti-donor blood group antibodies appeared to be specifically suppressed in long-term stable ABO-incompatible kidney transplant recipients. Conclusion: Accommodation of ABO-incompatible kidney transplantation could be explained by donor-specific blood group antibody suppression.

SUMMARY: Objectives: To review the incidence and clinical features of intravenous immunoglobulin (IVIg)-induced haemolysis. Background: Haemolysis can be a severe complication of IVIg administration. It is due to the passive transfer of blood group antibodies and may result in significant anaemia and renal failure. Methods: We report a case of severe IVIg-induced haemolysis; review the data reported to vigilance groups (The Medicines and Healthcare Products Regulatory Agency, European Union Drug Regulatory Authorities, Food and Drug Administration and the Canada Vigilance Centre) between January 1998 and May 2012; and systematically review IVIg-induced haemolysis case reports (between January 1948 and January 2013). Results: Nine hundred-twenty five cases of IVIg-induced haemolysis were identified from a review of cases reported to vigilance groups; 62 case reports were included in the systematic review. The majority of these were due to administration of doses of at least 2gkg-1 of IVIg (97%). IVIg-induced haemolysis was reported most commonly for patients with blood group A (65%) or AB (26%). One case report noted that in two patients with IVIg-induced haemolysis both received IVIg from the same batch. Conclusion: We make the following recommendations for the management of suspected cases of IVIg-induced haemolysis: Stop IVIg infusion and perform tests for haemolysis. Check titres of anti-blood group antibodies in IVIg. Provide supportive management for patient with fluid and/or red blood cell transfusions if necessary. Consider quarantine of the IVIg batch if found to be high titre for anti-A/B. Report reaction to regulatory/vigilance body. © 2013 British Blood Transfusion Society.

This study was aimed to review and establish the practice of exchange transfusion (ET) with reconstituted blood in neonates and to observe fall of bilirubin and its comparison with related studies. Twenty-five neonates diagnosed as hemolytic disease of newborn (HDN) were selected for this study, in which exchange transfusion was carried out as one of the treatments for hyperbilirubinemia. Out of the 25 cases, 15 were of Rhesus (Rh) HDN, while ABO and other blood groups constituted 6 and 4 HDN cases respectively. First, the neonates′ and mother′s blood samples were subjected to relevant investigations. After that, for neonates having Rh HDN, O Rh negative cells suspended in AB plasma were given, O Rh positive cells suspended in AB plasma were given to ABO HDN; and O positive cells, which were indirect Coomb′s cross-matched compatible with neonates′ and mother′s serum / plasma, suspended in AB plasma were given to the neonates having HDN because of other blood group antibodies. The exchange transfusion (ET) was carried out taking all aseptic precautions by Push-Pull technique with double-volume exchange transfusion method. The average post-exchange fall in serum indirect bilirubin was (52.01%) in all 25 cases, which was found to be more significant than the previous studies. Looking into the superiority of the exchange transfusion in HDN by reconstituted blood, the reconstituted blood can be modified and supplied as per the requirement and conditions.

We report the use of bioactive paper for typing of secondary human blood groups. Our recent work on using bioactive paper for human blood typing has led to the discovery of a new method for identifying haemagglutination of red blood cells. The primary human blood groups, i.e., ABO and RhD groups, have been successfully typed with this method. Clinically, however, many secondary blood groups can also cause fatal blood transfusion accidents, despite the fact that the haemagglutination reactions of secondary blood groups are generally weaker than those of the primary blood groups. We describe the design of a user-friendly sensor for rapid typing of secondary blood groups using bioactive paper. We also present mechanistic insights into interactions between secondary blood group antibodies and red blood cells obtained using confocal microscopy. Haemagglutination patterns under different conditions are revealed for optimization of the assay conditions.

The sensitivity of the low ionic strength solution antiglobulin test (LISS-AGT), polyethylene glycol antiglobulin test (PEG-AGT), low ionic strength solution solid-phase anti-globulin test (LISS-SPAT), gel low ionic strength solution antiglobulin test (GEL-LISS), and gel papain test (GEL-PAP) was compared in titration studies of 460 sera containing identified IgG alloantibodies. The GEL-PAP was 100% sensitive to detect Rh antibodies, whereas the PEG-AGT was the most sensitive to detect Kell, Duffy, Kidd, Ss, and rare blood group antibodies. The better performance of PEG-AGT was especially obvious with Kell, Duffy, and Ss antibodies (S = 100%). When the sensitivity of the LISS-AGT, PEG-AGT, GEL-LISS, and GEL-PAP was evaluated in different routines, the GEL-LISS showed to be more sensitive than PEG-AGT in the detection of clinically significant antibodies. These discrepant results showed that the performance of a technique may change when it is applied as a routine. © 1996 Wiley-Liss, Inc.

Introduction: Patients with hematological and oncological malignancies are among the most frequent recipients of chronic red blood cell (RBC) transfusion therapy. One of the risks associated with chronic RBC exposure is the development of blood group antibodies. However, there are few extensive studies of alloimmunization rates associated with hematological or oncological malignancies in adult patients despite the frequency with which they are transfused. As such, the aims of this study were: 1) to determine the alloimmunization rate associated with malignant conditions in general, 2) to establish and compare alloimmunization rates for individuals with different forms of hematological and oncological neoplasia, and 3) to determine if any oncological disorder(s) appeared to predispose individuals to alloimmunization such that antibody mitigation measures could be employed for these diagnoses. Methods: Patients included were those undergoing type and screen testing at the study site (n=18,750 unique, active subjects in our facility's transfusion record database). For each of these subjects, the electronic medical record was retrospectively reviewed to determine if a history of a hematologic or oncologic malignancy was present. Disorders grouped as 'hematologic malignancies' in this study included: acute leukemia (myeloid or lymphoid), mast cell diseases, multiple myeloma, myelodysplastic syndromes, myeloproliferative disorders, and non-Hodgkin lymphoma (T- or B-cell). 'Non-hematologic' cancers in this study were broadly grouped as follows: bladder, colon, head/neck, lung, prostate, skin, soft tissue/sarcoma. If a patient had a history of any of the above neoplastic diseases, further inclusion in the study required: 1) that the patient undergo at least one RBC unit transfusion after their diagnosis was established and 2) that at least one follow-up antibody screen test be performed after their first RBC transfusion post-diagnosis. When these additional inclusion criteria were met, the following data were collected for each subject: malignancy type, race/ethnicity, gender, and the number and specificity of antibodies detected. Ratio data were compared using the chi-square test with Yates' correction for continuity; P values Results: A total of 69 patients became alloimmunized after their diagnosis of a malignancy; these patients were overwhelmingly male (66/69; 95.7%) and most identified as white, non-Hispanic (50/69; 72.5%). The alloimmunization rate among patients with any malignancy diagnosis was 1.5% (69/4687), which was significantly lower than the general, historical alloimmunization rate at our facility (443/18750; 2.4%; P=0.0019). Of total transfused patients with a malignancy, most were diagnosed with a non-hematological disorder(4371/4687; 93.3%) and such malignancies were associated with an alloimmunization rate of 1.3% (55/4371) following transfusion. By comparison, the alloimmunization rate for transfused patients with hematological malignancies was 2.8% (14/501), which was significantly higher (P=0.011). When alloantibody development was analyzed on a disorder-by-disorder basis, the five highest alloimmunization rates were seen in: myelodysplastic syndromes (8/71; 11.2%), soft tissue cancers (3/50; 6.0%), acute leukemia (1/34; 2.9%), renal cancer (3/113; 2.7%), and myeloproliferative disorders (2/75; 2.7%). Alloimmunization rates under 2% were observed for all other hematological and non-hematological malignancies encountered in our patient population. Conclusions: RBC alloimmunization rates in transfused patients with malignancy were significantly lower than historical controls, suggesting that the immunosuppressive nature of these disorders and/or their associated therapies may play a role in limiting blood group antibody development. Hematologic disorders were associated with higher alloimmunization rates than non-hematologic malignancies, which may reflect a larger transfusion burden in these illnesses (a factor to be evaluated in future analyses). Overall, only myelodysplastic syndromes and soft tissue malignancies were associated with antibody formation rates substantially higher than historical controls. Therefore, consideration for prophylactic antigen matching to prevent alloimmunization may be warranted for these conditions. Disclosures No relevant conflicts of interest to declare.

Seven monoclonal antibodies (MAbs) selected from a large panel of human IgG-specific murine MAbs were characterized serologically and studied for their ability to cooperate in routine antihuman globulin agglutination tests. In binding inhibition experiments, three of these MAbs were shown to bind simultaneously to immobilized human IgG molecules. Cooperation among these MAbs increased significantly the capacity of the individual MAbs to agglutinate red cells sensitized with weak blood group antibodies. These results demonstrate the usefulness of selected MAb blending for the preparation of potent antihuman IgG reagents from murine monoclonal antibodies.

The Red Cross Blood Centre of Madrid, Spain, is part of the regional organization for blood and plasma donation and typing in Madrid Autonomous Community. To increase the number of patients we serve and identify rare antigens that may be specific to our multiracial population, we implemented the BioArray HEA BeadChip™ system. Herein we describe our experience with the training, installation, and implementation of the HEA BeadChip™ system. We provide information about the blood group antibodies responsible for acute and delayed hemolytic reactions. Future objectives for use are described.

Background and Objectives The aim of the study was to evaluate, in an international collaboration, three lyophilized intravenous immunoglobulin (IVIG) preparations for their suitability to standardize and control haemagglutination testing for anti-A and anti-B in IVIG products. Materials and Methods Twenty-three laboratories tested candidate IVIG reference reagents consisting of a Positive control (07/306), a Negative control (07/308), and a specifically formulated Limit preparation (07/310) to define the maximum (e.g. pharmacopoeial) limits of anti-A and anti-B in IVIG products, where limits are applicable. Laboratories performed direct haemagglutination using papain-treated erythrocytes and/or indirect antiglobulin tests. Results For both methods, there was up to 16-fold variation in anti-A and anti-B titres, although there was good agreement over a two-fold titre range for anti-A and anti-B between laboratories for both 07/306 and 07/310 using the direct method. Comparative titration data for 07/306 and 07/310 indicated that the use of a ‘Limit’ reference reagent would facilitate identification of higher titre batches when the direct haemagglutination method is used. Conclusions The establishment of preparations 07/306, 07/308 and 07/310 as reference reagents by the World Health Organization will facilitate global standardization of haemagglutination tests for anti-A and anti-B, ensure that such tests are sufficiently sensitive and specific, and facilitate identification of batches that exceed maximum recommended levels of anti-A and anti-B. The Commission of the European Pharmacopoeia and the United States Food and Drug Administration have adopted the same reference reagents including the maximal specifications defined by preparation 07/310.

The concentrations of anti-A and anti-B IgM and IgG antibodies have been studied in the serum of a patient with blood group AB who received a type A donor liver. A newly developed ABO-ELISA was used for this purpose and the values were compared to hemagglutination titers. During the postoperative study period over 8 weeks, the anti-A and anti-B levels showed a higher fluctuation than was measured in preoperative samples. Thus, in this AB-type patient, anti-A IgM varied 10-fold, anti-A IgG 20-fold and anti-B IgG 16-fold. Peak values corresponded to rejection episodes. Immunoactivation in the patient was further documented by the presence of abnormally high levels of soluble interleukin-2 receptors (slL-2R) in serum samples. The study shows that monitoring of anti-A/B antibodies may represent a further criterion to follow-up transplanted patients during the critical postoperative graft acceptance period.

Antibody identification is one of the last areas of laboratory medicine to embrace computerization. This is due in part to the complex nature of antibody identification. Until recently, computer programs written to assist with antibody identification have been slow and cumbersome. However, technological advances in computer hardware have greatly improved the response time for these applications. Still, blood bank technologists sometimes shun assistance from the computer because they consider antibody investigation as an intellectually stimulating aspect of their job that they do not wish to turn over to a computer. Computer programs for antibody investigation are probably most useful for technologists who perform antibody investigations infrequently. They can also be invaluable for experienced technologists performing complex investigations. When integrated with a computerized blood bank record-keeping system, antibody investigation software can improve storage and retrieval of test results. Database applications for retrieving rare cells and sera offer the potential to improve the productivity and capability of any antibody investigation laboratory. And word processing applications greatly enhance the potential for generating meaningful reports of test results. Immunohematology 1993;9:53–55.

A micro enzyme-linked immunosorbent assay (ELISA), using lyophilized stroma as carrier of red blood cell antigens, which stays stable longer than usual, using intact erythrocytes, was developed for determination of blood-group antibodies in the ABO, Kell and Lewis-systems. Stroma being fixed on microtiter plates was incubated with antisera and peroxidase-conjugated anti-human globulin. The transformation of the substrate added was determined photometrically. A binding of antibodies to the stroma could be demonstrated up to an antibody dilution of 1:1024 for the ABO-system, of 1:512 for the Kell-system and of 1:64 for the Lewis-system. By standardization of this method the quantitative determination of antibodies becomes possible without being restricted by the limited stability of intact erythrocytes.

Current textbooks for transfusion medicine state that anti-A and/or anti-B (anti-A/B) agglutination titers--and thus the respective antibody concentrations--reach their maximum in individuals 5 to 10 years old and then gradually decline with the increasing age of the individual. This statement is largely based on a study by Thomsen and Kettel that dates to 1929. In the present article, ABO antibodies in sera of 175 healthy persons aged 61 to 97 years, as well as sera of 170 newborn infants and children aged 0 to 17 years, were analyzed. Microhemagglutination tests were performed with all sera and complemented by ABO enzyme-linked immunosorbent assays to measure the immunoglobulin class (IgM, IgG, and IgA) of the anti-A/B. As in a previous study using sera of persons aged 20 to 67 years, individual differences exceeded age-related changes for all variables. Median values of IgG and IgA anti-A/B were elevated in elderly persons of blood group O, whereas no significant changes were observed in other variables. In particular, the decrease in agglutination titers with the increasing age of the individuals was far less pronounced than previously described; even in sera of persons aged 90 to 97 years, median agglutination titers of 128 were found. Results in the sera of children confirm previously reported data that agglutination titers and IgM anti-A/B reached adult levels at the age of 5 to 10 years.

The presence of residual donor red cells in transfused patients impairs the ability to obtain a reliable phenotype for subsequent allogeneic blood transfusions. Post transfusion phenotyping is particularly important in transfused patients developing multiple or weak blood group antibodies. However, separation of autologous and transfused red cells from recently transfused patients is necessary for the proper identification of any suspected red cell alloantibody or autoantibody. The standard technique for the separation of autologous from transfused red cells involves microhematocrit centrifugation with phytate ester solution, a laborious and time consuming method. We have examined the effectiveness of a simplified microhematocrit technique, omitting the use of esters, to separate autologous and transfused cells by performing Rhesus (Rh) and Kell (K) blood group phenotyping, both manually and by automation, pre and post transfusion. The Rh phenotype and K type were recorded on pre- and post-transfusion peripheral blood samples from 100 newly transfused operating theatre patients, both manually and by automation using the DiaMed Classic ID-Gel Station and Diaclon ’Rh subgroups+K’ gel cards. On post transfusion blood samples, separation of autologous from transfused cells was performed using the Haematospin 1300, based on the difference in the specific gravity between reticulocytes and mature red cells where the autologous cells concentrate in the top layer of the microcapillary tube. This separation was performed on blood samples taken three days post transfusion, based upon the normal reticulocyte maturation time of three days. On all samples tested, there was 100% correlation between manual and automated ID-GEL Station results. On Rh phenotyping of post transfusion blood samples prior to cell separation, 46 of the 100 patients had a mixed field (Mf) reaction. Repeat Rh phenotyping of day three post transfusion samples, following separation of autologous cells using the microhematocrit centrifugation technique, showed complete correlation with pre transfusion Rh phenotyping results in all but two cases, where a Mf reaction persisted. These two cases had had massive blood transfusions (40 and 36 units of red cells respectively) compared to the amount of red cells (mean 3.3 units, range 1–15) transfused to the other 98 patients. One week after initial Rh phenotyping, allowing sufficient time for autologous reticulocyte recovery in these two patients, repeat Rh phenotyping of separated autologous cells correlated with that of the pre transfusion results. 6 of the 100 patients were K positive on pre transfusion testing. Repeat testing of post transfusion blood samples prior to cell separation showed a Mf reaction for the K antigen in two of these 6 cases, suggesting transfusion of K antigen negative red cell units. Once cell separation was complete, both patients were confirmed as K positive. In conclusion, the simplified microhematocrit centrifugation technique and automated ID-Gel station are reliable, sensitive and less hazardous in providing an accurate Rh phenotype and K type on patient blood samples taken three days post transfusion and could be easily performed in the standard blood transfusion laboratory. The only limitation to this technique is an inability to separate autologous and transfused cells from patients who have undergone massive transfusions within a short space of time.

Anti-A and anti-B blood group antibodies in the cynomolgus monkey sera were serologically characterized. 2-Mercaptoethanol (2-ME) inactivated these antibodies of both group-A and group-B sera. Furthermore, soluble human ABH substances neutralized these antibody activities. However, these activities were not inhibited by anti-human IgG or IgA serum but by anti-human IgM serum. By gel filtration on Sephacryl S-300, the anti-A and anti-B antibodies reacting readily in saline media were detected in the fractions corresponding to the IgM antibodies. Thus, anti-A and anti-B antibodies in group-B and group-A sera were judged to be mainly of IgM. In contrast to group-A and group-B sera, group-O serum contained IgG antibodies of high titers besides IgM antibodies. Neither 2-ME nor anti-human IgM serum affected these antibody activities. On the basis of the results of the inhibition test using anti-human immunoglobulin sera as well as of gel filtration, group-O sera were divided into the following two categories, the one having only IgG reacting well in the Coombs method and the one having both IgM and IgG.

A two-channel Auto Analyzer II system (AAII) was tested for routine screening of blood group antibodies in 8, 980 sera during past 18 months. A low ionic strength polybrene method (LISP) was employed to detect IgG antibody.Manual conventional method was tested in parallel and compared with this method. AAII was not outstanding with respected to detection of some naturally occurring antibodies such as anti-Lewis, anti-P1. For the purpose of preferential detection of antibodies in the P and Lewis systems, we used commercial panel red cells instead of pooled red cells made in our laboratory.On the contrary, weak immune antibodies missed in the manual method were well detected by AAII. These results suggest that this system is ideally suitable to the detection of IgG antibodies which appears early after transfusion.we consider that the automated LISP method is very useful in a laboratory engaged in the detection and identification of red cell antibodies.

Blood banks face significant problems supplying blood units compatible with erythrocyte antigens other than ABO and rhesus D. In this study the number of such units was evaluated between 1973 and 1982, and it was found that 1.5% of the total number of blood units were compatible with other antigens than ABO and rhesus D. Based on data of the number of units requested for each such antigen and the frequency of the antigen in the European population, a logistical system providing sufficient numbers of ‘antibody-compatible’ blood units is proposed.

P blood group system antibodies are encountered frequently in clinical transfusion practice. The authors describe an improved method for removing P antibodies from patient specimens without dilution. Solid- phase synthetic P antigens were used to neutralize serums containing P blood group system antibodies; 26 alloantibodies with specificity other than P were not inhibited. The synthetic P antigens are also used to characterize the heterogeneous immunochemical fine specificity of antibodies to P1 and P + P1 + Pk.

Immunologic injury is an important contributing factor in failure of aortic valve allografts. The etiology of this immune reaction is unknown, but blood group antibodies have been postulated. In 46 patients, ABO and rhesus blood group compatibility bore no relationship between the success or failure or aortic valve allotransplantation. Therefore, circulating blood group antibodies do not contribute to valve failure, and recipients do not have to receive aortic valve allografts matched for ABO or rhesus compatibility.