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Introduction: Haemolytic Disease of the Foetus and Newborn (HDFN) is widely known to be caused by the presence of irregular Red Blood Cell (RBC) antibodies in pregnant females. To prevent HDFN, it is crucial to identify these antibodies in pregnant women. Aim: To evaluate the prevalence of irregular red cell antibodies in pregnant women in a tertiary care hospital. Materials and Methods: A cross-sectional retrospective study was conducted in the Department of Transfusion Medicine at Indraprastha Apollo New Delhi, from November 2009 to December 2017. A total of 8,217 prenatal females were examined for irregular red cell antibodies. Antibody screening and blood grouping were performed. The Capture-R ready screen was used to confirm screen-positive tests. Adsorption, elution, and other advanced techniques were utilised as needed. Data was collected from patient records and entered into a Microsoft excel spreadsheet. The statistical analysis was conducted using Statistical Package for Social Sciences (SPSS) software version 14.0 (USA). Statistical tests, including the Chi-squared and Fisher’s exact test, were performed, with a p-value less than 0.05 considered statistically significant. Results: Out of the 8,217 prenatal females examined, 105 (1.27%) had positive screening results with the four cell panel. Red cell alloimmunisation was observed in groups that were RhD negative (n=93/105; 88.57%) and positive (n=12/105; 11.42%). The most prevalent alloantibody among RhD negative individuals was alloantibody anti-D (n= 83/93; 89.24%), followed by alloantibody anti C+ anti D (n=13/93; 13.97%). Anti-D was the most prevalent alloantibody overall (n=83/105; 79.04%). Conclusion: Alloimmunisation rates were notably high among women lacking the Rh D antigen. Individuals with a history of adverse obstetric outcomes showed a statistically significant association with alloimmunisation. Therefore, it is recommended to screen such individuals for alloantibodies to enable early detection and improve the management of HDFN.

Objective: Despite an increase in the rate of successful live donor renal transplantation done annually, the number of potential recipients with acceptable donors is relegated to the ever-expanding cadaver-donor waiting list due to sensitization to human leukocyte antigen (HLA) antibodies. If not sufficiently suppressed, these preformed HLA antibodies can trigger antimicrobial resistance (AMR) and early graft loss. To ameliorate this situation, various desensitization treatments are administered to provide a survival benefit to highly sensitized patients. Method: One hundred and six patients in the time frame of January 2017 to March 2019 were included in the study group. The desensitization protocol included therapeutic plasma exchange and administration of low-dose intravenous immunoglobulin (100 mg/kg per therapeutic plasma exchange (TPE) session) to highly sensitized patients (treatment group) who subsequently underwent renal transplantation after negative pre-transplant Centers for Disease Control and Prevention Luminex crossmatch (CDC/LumXM). We compared graft survival rates between the group undergoing desensitization (treatment group) and matched control group of patients that underwent HLA-compatible transplantation. Results: In the treatment group, Kaplan-Meier analysis estimates an average rate of patient graft survival of 95.2% at 3 years post-transplant, as compared with the rate of 86.9% in the same time frame for the control-matched group (p < 0.05 for both comparisons). Conclusion: Desensitization treatment with TPE before live donor renal transplantation in the case of patients with HLA sensitization provides better survival benefits along with monitoring for donor-specific antibodies (DSAs) and other infections, rather than waiting for a compatible organ donor. The data lays out evidence that desensitization treatments can assist overcome HLA incompatibility barriers in live donor renal transplantation.

Background and Objectives: The anti-A and anti-B are naturally occurring antibodies in the human body but alloantibodies may be formed in individuals who lacks that corresponding red cell antigen after any sensitising event like blood transfusion or pregnancy. The aim of this study was to find the incidence of evanescenct RBC alloantibodies that may not be detectable at the time of pre-transfusion screening. Methods: This was a retrospective observational study from January 2017 to May 2019 in a tertiary care centre from North India. The data on antibody screening and identification retrieved from the departmental records. The patients were screened for red blood cell antibody and those with a positive screening test were further investigated for antibody identification. Antibody screening was performed by solid phase method using Immucor Capture-R Ready Screen (4 cell). Confirmed positive samples were investigated with advance methods of Antibody ID using Reagent panel Red Blood Cells Capture-R Ready ID (16 Cell) in Automated Analyzer Iris or Neo (Immucor Inc. Nocross, GA, USA). As and when required other advance techniques of adsorption and elution, etc were also performed too. Results: The total number of patient screened during this period were 8071. Amongst them 312 were positive for antibody screening (3.8%), of those 216 (62.9%) cases had alloantibodies and 96 (30.7%) cases had autoantibodies. Among the 216 cases of alloantibody, 147 (68.1%) cases had single alloantibody and 69 (31.9%) cases had multiple alloantibodies. Amongst the patients having single alloantibody, the prevalence was highest for Anti-D 54 (36.7%), followed by Anti-E 27 (18.3%), Anti-C 8 (5.4%), Anti-c 7 (4.7%), Anti-K 9 (6.1%), Anti-Fyb 11 (7.4%), Anti-M 18 (12.2%), Anti-N 4 (2.7%), Anti-Fyb 3 (2%), Anti-Lea 3 (2%), Anti-Cw 2 (1.3%) and Anti-Mia 1 (0.6%). Of the 96 patients with autoantibody 72 had warm autoantibody, 10 had cold autoantibody and there were 8 cases of mixed warm and cold autoantibodies. There were 6 patients with warm autoantibody with underlying alloantibody. Conclusion: In conclusion, the high prevalence of Anti-Rh (Anti-D, E), Anti-M, Anti-Fyb antibodies in patient requiring transfusion reiterates the importance of stringent antibody screening and identification as an essential step in preventing delayed hemolytic transfusion reaction.

INTRODUCTION: Autoimmune hemolytic anemia is characterized by increased red cell destruction and/or decreased red cell survival due to autoantibodies directed against self-antigens on red cells. Since autoantibodies react with self and nonself red blood cells (RBCs), they tend to mask the underlying clinically significant alloantibodies and many a times mimic a specific pattern like alloantibodies. MATERIALS AND METHODS: We discuss three immune hematological cases of warm autoantibodies. Antibody screening was performed by solid-phase red cell adherence (SPRCA) technique on a fully automated platform NEO Iris (Immucor Inc., USA). In case of a positive antibody screen, antibody identification was performed using SPRCA, NEO Iris (Immucor Inc., USA). Alloadsorption for adsorbing the autoantibodies was done using in-house prepared allogenic packed RBCs – R1R1, R2R2, and rr. RESULTS: All cases had warm autoantibody with a broad specificity against self-Rh antigens. Anti “C” and Anti “e” antibodies were identified in case 1 and autoanti “e” antibody in cases 2 and 3. Case 3 had underlying alloanti “E” along with autoanti “e” which posed a transfusion challenge. CONCLUSION: Our case series highlights the importance of detecting the nature of the antibody whether it is alloantibody or autoantibody with antigen specificity. This would help in selecting appropriate antigen negative blood units for transfusion purpose.

BACKGROUND: Transfusion of ABO-compatible single donor platelets (SDP) is preferable for better outcomes over group switchover SDP. The use of SDP containing ABO-incompatible plasma is associated with a risk of allergic and acute hemolytic transfusion reactions. Moreover, high titer O group donors SDP impose a further threat to patient safety. Platelet additive solution (PAS) is used worldwide for the storage of platelets which reduces plasma volume available in SDP. SSP + (Macopharma) is one such PAS which can provide improved availability, logistical management, decrease wastage, and improvement in patient safety. The aim of this study was to assess the feasibility of using PAS to obtain low titer SDP units which can be utilized across a larger patient population and to study quality control parameters of these units. MATERIALS AND METHODS: The study was performed in the department of Transfusion Medicine from June 2017 to January 2018 after clearance from the Institutional Review Board. The study design comprised two cohorts (A and B). In cohort A, the temporal trend of in-vitro changes in the quality parameters was tested and analyzed for PAS modified and unmodified products on days 1, 5 and 7. In cohort B, the original plasma from the SDP donors of all blood group donors except the AB group was tested for antibody titers before (prepreparation) and after modification (postpreparation) by PAS. RESULTS: In cohort A, in the control group, there was a significant change in the mean platelet volume, potassium, and bicarbonate levels from day 1 to day 7, whereas no significant change in the biochemical parameters was noted in the study group where PAS was used. In cohort B, on comparing the anti-A and anti-B, before and after modification of SDP with PAS, there was a significant reduction in the median titers across all the groups studied. CONCLUSION: PAS added SDP is an efficient strategy to reduce the ABO-antibody levels significantly. PAS added SDP also helps in the better inventory management of available groups.

INTRODUCTION: Therapeutic plasma exchange has been widely employed by clinicians for removal of the toxic constituents from plasma by filtration of whole blood and subsequent removal of plasma and reinfusion of cellular components along with a replacement fluid. It has become an accepted therapeutic modality in paediatric patients for numerous indications including but not limited to renal transplant, haemolytic uremic syndrome and Guillain Barre Syndrome. But, data on safety and efficacy are mainly derived from studies in the adult population with very limited data available in the paediatric age group. However, it is technically challenging in children due to their small circulating volume. This study discusses the clinical indications, efficacy, and safety of therapeutic plasma exchange in paediatric population. METHOD: We retrospectively reviewed the data of children (up to 18 years of age) who underwent TPE between January 2017 and March 2019 at our Hospital. Main features of the TPE procedures i.e. frequency of TPE, site of vascular access, type of replacement fluid used, instrument used, plasma volume processed, priming of the circuit, adverse events if any and outcome of the patients were analysed. RESULTS AND CONCLUSION: A total of 114 procedures were performed on these 24 patients. Fifteen patients with Category I indication showed good clinical outcome in terms of attainment of target ABO titre and/or decrease in the donor specific antibody. TPE is an effective therapeutic option in selected paediatric disorders. Our series of data on TPE procedures from paediatric perspective has shown safety and efficacy of the therapy.

BACKGROUND: HELLP (hemolysis, elevated liver enzymes, low platelet count) syndrome occurs in about 0.5%–0.9% of all pregnancies, but its prevalence is higher in patients with severe preeclampsia, accounting for a substantial maternal and perinatal morbidity and mortality. According to the latest American Society for Apheresis guidelines, Therapeutic plasma exchange (TPE) performed for postpartum cases and antepartum HELLP syndrome cases fall in Categories III and IV, respectively. MATERIALS AND METHODS: Retrospective analysis was done at our tertiary care center from January 2014 to June 2019 for patients diagnosed with HELLP syndrome. Clinical data for age, gestational age at the time of diagnosis, type of delivery, outcome of pregnancy, history of preeclampsia /eclampsia, hemoglobin levels, AST, ALT, LDH, platelet counts, prothrombin time, activated partial thromboplastin time, international normalised ratio, complete blood count, was obtained from patients' electronic medical records. The TPE was initiated within 24 hrs of diagnosis. All TPE was done on Spectra Optia apheresis system (Terumo BCT, Inc, USA). Statistical testing was conducted with the statistical package for the social science system version SPSS 20.0 and R-3.2.0. Continuous variables were expressed as mean±SD and were compared between Pre and Post TPE records of patients by using the paired T test. RESULTS: Nine patients fulfilled the criteria of HELLP syndrome. Seven (77.8%) were diagnosed in the postpartum period and 2 (22.2%) during the second trimester. Out of the total nine patients, two patients (22.2%) recovered completely and were discharged on day 15 ± 7 days, whereas 4 (44.4%) patients were discharged on day 21 ± 7 days with the advice of hemodialysis. Two (22.2%) patients had an intrauterine death and were discharged 3–4 days after the demise. In all these patients (except one), the TPE was initiated within 24 h of the diagnosis. A significant increase in platelet count and decrease in the lactate dehydrogenase levels (P < 0.05) was observed post TPE. CONCLUSION: Our data showed that TPE improved the treatment outcome in patients with HELLP syndrome despite being a Category III and IV indication among postpartum and antenatal females, respectively. However, a timely diagnosis and management are of paramount importance for a favorable outcome. TPE needs to be performed within 24 h of the diagnosis postdelivery when the patient is not responsive to the usual therapies, especially in class I HELLP syndrome.

Granulocyte transfusion (GTx) is an efficient and compelling treatment option for patients with neutropenia following hematopoietic stem cell transplant. The donor pool for granulocyte harvest is limited to close friends and family members and the donors accepted are often of the same ABO Rh type. We report a case of ABO-incompatible prophylactic GTx, in a case of acute myeloblastic leukemia. Postcollection processing of the granulocyte product was done to reduce the red blood cell volume to

Background and Aim: Renal transplantation (RT) is the most successful and ideal renal replacement therapy for end-stage renal disease patients. Renal allograft rejection has always been one of the major barriers in successful RT. Our aim was to report the role of therapeutic plasma exchange (TPE) in acute humoral rejection (AHR) patients who underwent live-related RT (LRRT) and their renal allograft outcome at our center. Materials and Methods: A prospective observational study was conducted from July 1, 2014, to December 31, 2016. Patients with biopsy-proven AHR and treated with TPE along with other lines of treatment after undergoing LRRT were included in the study. ABO-incompatible individuals, pediatric patients, and patients undergoing second transplants were excluded from the study. Clinical history, donor and graft details, management, and patient and graft survival were noted. Results: Of the 1608 patients who underwent LRRT, 49 (37 males, 76%; 12 females, 24%; mean age 39.5 ± 13.3 years) had biopsy-proven AHR (3.04%) and were treated with TPE. A total of 281 TPEs were performed with an average of 5.7 TPE/patient (range 2–12). Of the 49 patients, 38 patients (78%) with favorable response underwent 213 (75.8%) TPEs (average of 5.6 TPE/patient; range: 2–12), whereas 11 patients (22%) with unfavorable response underwent 68 (24.2%) TPEs (average of 6.2 TPE/patient; range: 3–8). Blood urea (P = 0.012) and serum creatinine (P = 0.038) levels at the time of rejection were significant predictors of response to TPE therapy. The average length of stay in our study population was 33 ± 22 days. Six months posttransplant, the patient and graft survival were 93.3% and 89.5%, whereas at 12 months, they were 89.3% and 81.5%, respectively. Conclusion: TPE is a safe and effective adjunct therapy for treating AHR patients.

Background and Objectives: Red cell exchange (RCE) is a type of therapeutic apheresis procedure aimed at removing the deformed red blood cells of a patient and replacing them with those obtained from healthy donors. It is a commonly used but yet underutilized procedure for the treatment of hemoglobinopathies such as sickle cell disease (SCD) (acute or chronic), and other diseases and conditions such as ABO-incompatible hematopoietic stem cell transplantation, severe erythrocytosis, hereditary hemochromatosis, severe infection with intra-erythrocytic parasites such as malaria, babesiosis or cases of a drug overdose, poisoning and to perform envenomation. The objective of the study is to study and understand the efficacy of RCE in the management of patients in various clinical scenarios. Methods: This is a retrospective study carried out at a tertiary care hospital in North India over a period of 4 years (2016–2020). All the RCE were performed on Spectra Optia apheresis (System 1 P0 4552 (Terumo BCT, Lakewood, CO, USA) which comes with a built-in software (Version 11) for performing RCE. Results: A total of 8 exchanges were performed in five patients. Three of these patients had undergone the procedure twice. Two out of the five patients were found to be antibody screen positive. Corresponding antigen-negative units of packed red cells were issued for these patients. The targeted and final fraction of red cell remaining values which were kept at ≤30% was achieved in all cases. Conclusion: RCE is a safe and effective therapeutic treatment modality for patients of SCD. It is a simple and relatively safe treatment modality that provides immediate relief in acute complications and also prevents the risk of long-term complications such as iron overload and iron-induced organ damage.

There are occasions when tests performed before considering a patient for transplant are ambiguous and require further workup. One such condition is the presence of a positive virtual crossmatch (VXm) (anti-human leukocyte antigen [HLA-A]*26: 01 antibody in this case) with a negative complement-dependent cytotoxicity, Luminex, and flow crossmatch. To ascertain the nature of the antibody, the beads used in single-antigen bead assay (SAB) were treated by acid to denature the antigens and retested with the control and test sample. The mean fluorescence intensities (MFIs) from the patient sera with acid-treated beads increased considerably as compared to the regularly untreated SAB indicating additional antigen epitopes become available by the denaturation process. The MFIs of the antibodies from that of the control sera were reduced to half on testing with the acid-treated SAB assay, indicating that HLA antigen HLA-A*26 was susceptible to acid treatment. Therefore, results of VXm should be interpreted with caution.

CD38 is a disulfide-linked molecule present on red blood cells (RBCs) and daratumumab; an anti-CD38 monoclonal antibody is a novel agent for treating multiple myeloma patients. It also binds to the RBC along with the plasma cells in concern, creating a menace in the immunohematology workups and requires the use of dithiothreitol-treated cells to rule out its interference. Appropriate and timely communication with the clinicians about the patient history goes a long way in solving complex looking immunohematology workups.

The human leukocyte antigen (HLA) matching plays an important role in determining the clinical outcome of renal transplantation. The development of donor specific antibodies (DSA) against HLA is associated with antibody mediated allograft tissue injury, poor outcome and rejection. The DQ-DSA develops in a denovo pattern and its unfavorable impact on renal transplantation has not yet been widely reported. We investigated the clinical significance of DQ-DSA in a patient diagnosed with hypertension, CKD stage V on maintenance hemodialysis (MHD) for second renal transplant. The histocompatibility workup before the first transplant included low resolution HLA-A, B, DR typing of both patient and donor. HLA type of the patient was HLA-A*29, 68, HLAB*44, 44, DRB1*07, 11. HLA type of the donor was HLA-A*03, 68, HLA-B*39, 44, DRB1*07, 10 with a 3/6 match. The HLA antibody screen and complement dependent cytotoxicity crossmatch (CDC) were found to be negative. No therapeutic plasma exchanges (TPE) were done during stay and post-transplant the patient was on triple immunosuppressant therapy. After four years the patient was diagnosed with recurrent membranoproliferative glomerulonephritis and second renal transplant was planned, therefore, histocompatibility workup was initiated. HLA antibody screen was found to be positive for HLA class II. Initially only HLA-A, B, DR typing was performed and that too only low resolution, further, high resolution HLA typing was done for HLA-DR and DQ to rule out if these antibodies are de-novo DQ/DR DSA. We analyzed that the patient had developed de-novo DSA against HLA-DRB1*10:01 (DR10), MFI-2374 and DQB1*06:01 (DQ6), MFI-15315. This study suggests the role of DQ antibodies in determining the graft survival and to highlight the need of HLA DQ typing as a routine of the diagnostic work-up in a solid organ transplant.

ABO incompatibility and preformed antibodies against the human leukocyte antigen (HLA) are two impermissible barriers to a successful renal transplantation, especially in highly sensitized patient population. With the availability of effective desensitization regimens, good patient and graft outcomes have been reported. As transfusion medicine specialists we report our experience, where patient presented with dual histocompatibility barriers i.e. ABO incompatibility along with preformed donor-specific antibodies (DSA) and negative complement dependent lymphocytotoxicity (CDC) HLA crossmatch. The desensitization strategy followed for our patient included rituximab (375 mg/m2), bortezomib (1.3 mg/m2) and eleven pre-transplant therapeutic plasma exchange (TPE) followed by intravenous immunoglobulin (100 mg/kg per TPE session). Anti-B titer of 1:1 and negative Luminex crossmatch (LumXm) class II DSA (less than 1000 mean fluorescence intensity; MFI), was achieved prior to renal transplantation. Fifteen months post-transplant, patient is doing well with serum creatinine level of 0.8 mg/dL with repeat LumXm class II DSA negative (891 MFI). The desensitization regimen followed proved to be effective in our case.

Background: Alloantibodies may be detected in blood donors who have either been transfused previously or female donors with previous obstetric events. These antibodies can occasionally cause severe transfusion reaction, if a large amount of plasma or whole blood is transfused, as in massive transfusions and pediatric patients. Aims: The present study aims to assess the prevalence of red cell antibodies in healthy blood donors at a tertiary care hospital-based blood bank in India. Materials And Methods: A total of 82,153 donor samples were screened for irregular red cell antibodies between January 2012 and December 2015 at the Department of Transfusion Medicine, Indraprastha Apollo Hospitals, New Delhi. Antibody screening was performed by solid phase method using Immucor Capture–R ready screen (pooled cells) on fully automated immunohematology analyzer Galileo Neo (Immucor Inc., Norcross, GA, USA). Positive tests were further confirmed using Capture-R ready screen (4 cell panel). Advanced investigations to identify the antibody/ies were performed on confirmed positive samples. Antibody identification was conducted using various cell panels (Immucor Capture-R Ready-ID, Panocell-10, Ficin Treated). An advanced technique such as adsorption and elution was performed as per requirement. Results: Screening with pooled cells and 4 cell panel was positive in 227 donors (0.27%), 150 of these donors had autoantibodies, 1 had autoantibodies with underlying alloantibody anti-Jka (0.001%), and 76 had alloantibodies (0.09%) alone in their plasma. Anti-M was the most common antibody (43 donors) identified, followed by anti-D (21 donors). Anti-N was detected in 4; anti-Jka, anti-C, and anti-E in two donors each followed by anti-P1 and anti-Leb in 1 donor. Conclusion: Antibodies against red cells can be present in healthy donors detection of which is important in providing safe blood to the patient. The prevalence of red blood cell antibody in healthy donors in this study was found to be 0.27%, while the prevalence of alloantibodies was 0.09%. The majority of alloantibodies were anti-M (56.57%) and anti-D (27.63%).

Transplant recipients are always at a risk of developing anti-human leukocyte antigen (HLA) antibodies due to prior sensitizing events such as blood transfusions, multiple pregnancies, or transplantation. Unexpected positive outcomes can be seen in complement dependent cytotoxicity (CDC) based assays due to underlying autoimmune disorders or pharmacological treatment (rituximab/intravenous immunoglobulin/anti-thymocyte globulin administration), therefore, limiting its value. CDC based assay results strongly depend on the vitality of the donor lymphocytes, highlighting another major limitation of this assay. Thus, as an alternative approach, solid phase based crossmatch assays were introduced which function independently of the cell quality and have higher sensitivity and specificity in detecting anti-HLA antibodies. We describe a case where the patient awaiting renal transplantation from living related donor was evaluated by pretransplant histocompatibility testing for the detection of anti-HLA antibodies. The histocompatibility testing revealed positive CDC anti-human globulin and flow crossmatch along with negative Luminex based assays (HLA antibody screen, luminex crossmatch, and luminex single bead assay). Detailed histocompatibility workup revealed immunoglobulin G autoantibodies which were complement activating and lympocytoxic in nature.

Background: Blood transfusion of contaminated components is a potential source of sepsis by a wide range of known and unknown pathogens. Collection mechanism and storage conditions of platelets make them vulnerable for bacterial contamination. Several interventions aim to reduce the transfusion of contaminated platelet units; however, data suggest that contaminated platelet transfusion remains very common. Aim: A pathogen inactivation system, “INTERCEPT”, to inactivate bacteria in deliberately contaminated platelet units was implemented and evaluated. Materials and Methods: Five single-donor platelets (SDP) and five random donor platelets (RDP) were prepared after prior consent of donors. Both SDP and RDP units were deliberately contaminated by stable stock ATCC Staphylococcus aureus and Escherichia coli, respectively, with a known concentration of stock culture. Control samples were taken from the infected units and bacterial concentrations were quantified. The units were treated for pathogen inactivation with the INTERCEPT (Cerus Corporation, Concord, CA) Blood system for platelets (Amotosalen/UVA), as per the manufacturer's instructions for use. Post illumination, test samples were analyzed for any bacterial growth. Results: Post-illumination test samples did not result in any bacterial growth. A complete reduction of >6 log10S. aureus in SDP units and >6 log10Escherichia coli in RDP units was achieved. Conclusion: The INTERCEPT system has been shown to be very effective in our study for bacterial inactivation. Implementation of INTERCEPT may be used as a mitigation against any potential bacterial contamination in platelet components.

Introduction: Our study presents an analysis of the trends of ABO antibody titers and the TPE (Therapeutic Plasma Exchange) procedures required pre and post ABO incompatible renal transplant. Materials and Methods: Twenty nine patients underwent ABO incompatible renal transplant during the study period.The ABO antibody titers were done using the tube technique and titer reported was the dilution at which 1+ reaction was observed. The baseline titers of anti-A and anti-B antibodies were determined. The titer targeted was ≤8. Patients were subjected to 1 plasma volume exchange with 5% albumin and 2 units of AB group FFP (Fresh Frozen Plasma) in each sitting. TPE procedures post-transplant were decided on the basis of rising antibody titer with/ without graft dysfunction. Results: The average number of TPE procedures required was 4-5 procedures/patient in the pretransplant and 2-3/patient in the post-transplant period. An average titer reduction of 1 serial dilution/procedure was noted for Anti-A and 1.1/procedure for Anti-B. Number of procedures required to reach the target titer was not significantly different for Anti-A and Anti-B (P = 0.98). Outcome of the transplant did not differ significantly by reducing titers to a level less than 8 (P = 0.32). The difference in the Anti-A and Anti-B titers at 14th day post-transplant was found to be clinically significant (P = 0.042). Conclusion: With an average of 4-5 TPE procedures pretransplant and 2-3 TPE procedures post transplants, ABO incompatible renal transplantations can be successfully accomplished.

Patients presenting with hyperleukocytosis secondary to acute leukemia, with total leukocyte count or blast count more than 100,000/μL are often considered for leukapheresis, especially if clinical signs of leukostasis are present. Leukostasis is often associated with high morbidity and mortality in patients with leukemic processes. The main goal of management of hyperleukocytosis and/or leukostasis is to reduce the blast count before initiation of chemotherapy. Leukapheresis is often used prophylactically to prevent leukostasis or to provide symptomatic relief. We, as transfusion medicine specialists, present our experience of doing therapeutic leukapheresis in patients presenting with hyperleukocytosis with or without presenting features of leukostasis.

Background: Red cell alloimmunization is an acknowledged complication of blood transfusion. Current transfusion practices for thalassemia do not cater to this risk. Serological phenotyping is usually not reliable in these cases unless performed before the first transfusion. Under such circumstances, molecular blood grouping is an effective alternative. Aim: To perform molecular blood group genotyping in chronically transfused thalassemia patients and assess the risk of antigenic exposure and incidence of alloimmunization with current transfusion protocols. Materials and Methods: Molecular blood group genotyping was performed for 47 chronically transfused thalassemia patients. Their 1-year transfusion records were retrieved to assess the antigenic exposure and the frequency thereof. Results: Of 47 patients, 6 were already alloimmunized (3 with anti-E and 3 with anti-K) and were receiving the corresponding antigen negative units. We observed that random selection of ABO and Rh D matched units resulted in 57.7% ±8.26% chance of Rh and Kell phenotype matching also. Forty-four patients had received one or more antigenic exposures at least once. The 6 already alloimmunized patients were further exposed to antigens other than the ones they were immunized to. During the study period, only one patient developed an alloantibody, anti-E with exposure to antigens C (92%) and/or E (32%) at each transfusion. Conclusion: Several factors apart from mere antigen exposure may influence the development of alloimmunization as most of our patients received antigenic exposures but not alloimmunized. Our data provide an impetus for future large-scale studies to understand the development of alloimmunization in such patients.

As the incidence of end-stage renal disease (ESRD) is rapidly increasing, the demand for dialysis and transplantation has dramatically increased, which has led to concerns about the availability and equitable allocation of kidneys for transplantation. The distribution of HLA-A, B and DR alleles in 148 renal transplant recipients and 191 live related prospective donors from 2009 to 2010 were analyzed. Allele frequencies and haplotype frequencies were calculated in recipients and donors. The prospective donors were further analyzed on the basis of their relationship to the patients and according to the sex ratio. A significant female preponderance was noted in the prospective donor population, most of whom were either siblings or parents of the recipients. On the contrary, the recipient population predominantly comprised of males. The most frequent HLA-A, HLA-B, HLA-DRB1 alleles in renal transplant patients were HLA-AFNx0111, AFNx0102, AFNx0101, AFNx0124; HLA-BFNx0135, BFNx0140, BFNx0144, BFNx0115, BFNx0152, and HLA-DRB1FNx0115, DRB1FNx0107, DRB1FNx0113, DRB1FNx0111 respectively. The most frequent HLA-A, HLA-B, HLA-DRB1 alleles in prospective donors were HLA-AFNx0102, AFNx0111, AFNx0133, AFNx0124; HLA-BFNx0135, BFNx0144, BFNx0140, BFNx0115 and HLA-DRB1FNx0115, DRB1FNx0107, DRB1FNx0111, DRB1FNx0113 respectively. AFNx0111-BFNx0135, AFNx0102-DRB1FNx0115, BFNx0140-DRB1FNx0115 were the most common HLA A-B , HLA A-DR, HLA B-DR haplotypes respectively in renal transplant patients, whereas, AFNx0111-BFNx0135, AFNx0111-DRB1FNx0115, BFNx0144-DRB1FNx0107 were the most common haplotypes in renal donors. In three locus haplotype, HLA-AFNx0102-BFNx0140-DRB1FNx0115 was the most frequent haplotype in patients, whereas, in prospective renal donors HLA-AFNx0133-BFNx0144-DRB1FNx0107 was the most frequent haplotype.

Introduction. The MIC (MHC class I chain-related) genes are a group of nonclassical MHC genes, located in the MHC class 1 region of chromosome 6. The aim of the present study was to find the prevalence of MHC class 1 chain-related (MICA) alloantibodies in patients undergoing live-related donor renal transplantation and its role in short-term graft survival. The role of blood transfusion in the formation of these antibodies was also studied. Materials and Methods. Pretransplant samples of patients undergoing renal allograft transplantation were tested for anti-MICA antibodies. Association of various demographics, HLA-A + B + DRB1 mismatches, anti-HLA antibody screen, and anti-MICA antibodies was assessed using Pearson’s chi-square test. Results. Out of 646 serum samples, 94 (14.6%) were positive and 552 (85.4%) were negative for anti-MICA antibodies. Patients with anti-MICA antibody had a graft survival 89.3% as compared to 94.7% in patients without anti-MICA antibody (P0.05). Conclusion. Preformed MICA antibodies independently increase the risk of kidney rejection and therefore recommend that guidelines should be formed for mandatory testing of these antibodies prior to renal transplant.

Background: Transfusion of blood components and age of transfused packed red cells (PRCs) are independent risk factors for morbidity and mortality in cardiac surgeries. Materials and Methods: We retrospectively examined data of patients undergoing cardiac surgery at our institute from January 1, 2012 to September 30, 2012. Details of transfusion (autologous and allogenic), postoperative length of stay (PLOS), postoperative complications were recorded along with other relevant details. The analysis was done in two stages, in the first both transfused and nontransfused individuals and in the second only transfused individuals were considered. Age of transfused red cells as a cause of morbidity was analyzed only in the second stage. Results: Of the 762 patients included in the study, 613 (80.4%) were males and 149 (19.6%) were females. Multivariate analysis revealed that factors like the number and age of transfused PRCs and age of the patient had significant bearing upon the morbidity. Morbidity was significantly higher in the patients transfused with allogenic PRCs when compared with the patients not receiving any transfusion irrespective of the age of transfused PRCs. Transfusion of PRC of over 21 days was associated with higher postoperative complications, but not with in-hospital mortality. Conclusion: In patients undergoing cardiac surgery, allogenic blood transfusion increases morbidity. The age of PRCs transfused has a significant bearing on morbidity, but not on in-hospital mortality. Blood transfusion services will therefore have to weigh the risks and benefits of providing blood older than 21 days in cardiac surgeries.

Context: The clinically significant antibodies are those active at 37°C and/or by the indirect antiglobulin test. Most of the published literature refers to antibodies of Lewis blood group system to be insignificant, whereas antibodies to M and N blood groups are associated with variable clinical significance. Aims: The aim of this study is to find the frequency and clinical significance of antibodies to M, N and Lewis blood group systems. Settings and Design: The study was carried out retrospectively from January 2009 to December 2012. Materials and Methods: Antibody screening was performed by solid phase red cell adherence (SPRCA) technique using four cell screening panel on a fully automated platform GALILEO (Immucor Inc. USA). In case of a positive antibody screen, antibody identification was performed using SPRCA (GALILEO, Immucor Inc. USA). Results: A total of 49,077 red cell antibody screens were performed and a total of 427 identifications of red cell antibodies were carried out. A total of 304 specific antibodies were detected: 8.22% of antibodies were of anti-M specificity and 2.96% were of anti-N specificity. Majority (84%) of anti-M and 77.78% of anti-N were of Immunoglobulin G (IgG) class reacting at 37°C. 1.31% of the antibodies were directed against Lewis system antigens of which 0.65% were anti-Lea and 0.65% were anti-Leb. Half of the Lewis system antibodies, i.e., 1 each of anti-Lea and anti-Leb were of IgG class. Conclusion: Our study highlights the importance of detecting the thermal amplitude of antibodies with variable clinical significance especially if both IgG and IgM types of antibodies are associated with it so as to establish their clinical significance.

Background: Patients with thalassemia major are largely transfusion dependent and are thus exposed to a variety of risks such as transmission of infectious diseases, iron overload and alloimmunization. This study was performed to determine the prevalence of human immune deficiency virus (HIV), hepatitis B virus (HBV), hepatitis C virus (HCV) and red cell antibodies among multiple-transfused thalassemic patients in and around the national capital region. Materials and Methods: The Department of Transfusion Medicine, Indraprastha Apollo Hospitals, conducted this study in collaboration with the National Thalassemia Welfare Society over a period of 1 year starting February2011. Blood samples from the patients were tested for blood group, red cell alloantibody/ies, anti-HIV, anti-HCV and hepatitis B surface antigen (HBsAg) by ELISA and for the respective viral ribonucleic acid (RNA) or deoxyribonucleic acid (DNA) by nucleic acid testing (NAT). Results: A total of 462 thalassemics which consists of 290 males and 172 females were tested. The overall alloimmunization rate was 4.1% and anti-Kell was the most common antibody identified. Thirteen cases (2.8%) were positive for HBsAg by ELISA, 107 (23.1%) were reactive for anti HCV and 11 (2.38%) for anti HIV antibodies. Further screening and discriminatory assays by NAT confirmed the presence of HBV DNA in 11 cases, HIV RNA in 7 cases and HCV RNA in 48 cases. Conclusion: In spite of advances in Immunohematology and infectious marker testing in recent years, the rates of alloimmunization and infectious marker positivity remains high among multiply transfused patients like thalassemics. Provision of safe and adequate blood supply to these patients is a key to improving their quality-of-life and longevity.

Context: Extensive bleeding associated with liver transplantation is a major challenge faced by transplant surgeons, worldwide. Aims: To evaluate the blood component consumption and determine preoperative factors that predict the same in living donor liver transplantation (LDLT). Settings and Design: This prospective study was performed for a 1 year period, from March 2010 to February 2011. Materials and Methods: Intra- and postoperative utilization of blood components in 152 patients undergoing LDLT was evaluated and preoperative patient parameters like age, gender, height, weight, disease etiology, hemoglobin (Hb), hematocrit (Hct), platelet count (Plt), total leukocyte count (TLC), activated partial thromboplastin time (aPTT), international normalized ratio (INR), serum bilirubin (T. bilirubin), total proteins (T. proteins), albumin to globulin ratio (A/G ratio), serum creatinine (S. creatinine), blood urea (B. urea), and serum electrolytes were assessed to determine their predictive values. Univariate and stepwise discriminant analysis identified those factors, which could predict the consumption of each blood component. Results: The average utilization of packed red cells (PRCs), cryoprecipitates (cryo), apheresis platelets, and fresh frozen plasma was 8.48 units, 2.19 units, 0.93 units, and 2,025 ml, respectively. Disease etiology and blood component consumption were significantly correlated. Separate prediction models which could predict consumption of each blood component in intra and postoperative phase of LDLT were derived from among the preoperative Hb, Hct, model for end-stage liver disease (MELD) score, body surface area (BSA), Plt, T. proteins, S. creatinine, B. urea, INR, and serum sodium and chloride. Conclusions: Preoperative variables can effectively predict the blood component requirements during liver transplantation, thereby allowing blood transfusion services in being better prepared for surgical procedure.

Background : In India, the incidence of breast cancer has increased in the urban population, with 1 in every 22 women diagnosed with breast cancer. It is important to know the HER2/neu gene status for a better prognostication of these patients. Aim : The aim of this study was to compare the efficacy of fluorescence in situ hybridization (FISH) and immunohistochemistry (IHC) for determining HER2/neu alteration in breast carcinoma. Materials and Methods : A total of 188 histologically proven breast carcinoma cases between the years 2007 and 2011 were retrospectively analyzed on the paraffin tissue sections by both IHC and FISH techniques. FISH for HER2/neu gene amplification was performed on cases where the IHC status was already known and the results were compared. Results : A total of 64 (30%) patients were found to be amplified and the remaining 124 (65.9%) cases were found to be unamplified through FISH. Patients observed with 3+ reading on IHC were later confirmed as unamplified in 29.5% cases through FISH. Conclusion : It has been confirmed with the present study that IHC is a prudent first-step technique to screen tissue samples for HER2/neu gene status, but should be supplemented with the FISH technique especially in equivocal cases.

Background: Until lately, anti-HBc antibodies were considered an effective marker for occult Hepatitis B virus (HBV) infection and have served their role in improving blood safety. But, with the development of advanced tests for HBV DNA detection, the role of anti-HBc in this regard stands uncertain. Materials and Methods: Anti-HBc and HBsAg ELISA and ID-NAT tests were run in parallel on donor blood samples between April 1, 2006 and December 31, 2010 at the Department of Transfusion Medicine, Indraprastha Apollo Hospitals, New Delhi. A positive ID-NAT was followed by Discriminatory NAT assay. Results: A total of 94 247 samples were tested with a total core positivity rate of 10.22%. We identified nearly 9.17% of donors who were reactive for anti-HBc and negative for HBsAg and HBV DNA. These are the donors who are potentially non-infectious and may be returned to the donor pool. Conclusion: Although anti HBc testing has a definite role in improving blood safety, centers that have incorporated NAT testing may not derive any additional benefit by performing anti-HBc testing, especially in resource-limited countries like ours.

Introduction: The Procleix Ultrio Plusassay is a new-generation qualitative in vitro nucleic acid amplification test used to screen for human immunodeficiency virus type 1 (HIV-1) RNA, hepatitis C virus (HCV) RNA and hepatitis B virus (HBV) DNA in blood donors. This study was performed to compare the Procleix Ultrio assay with the new-generation Procleix Ultrio Plus Nucleic Acid Test (NAT) assays. Materials and Methods: Ten thousand three hundred and two donor samples were run in parallel for ID NAT using the Procleix Ultrio and the Procleix Ultrio Plus assay. Simultaneously, enzyme-linked immunosorbent assay testing was performed on an EVOLIS Walk away System for HIV, HCV, HBsAg and anti-HBc. Reactive samples were confirmed using polymerase chain reaction. Results: In the 10,302 samples tested during the study period, we identified 15 NAT yields, and all these revealed HBV DNA in the discriminatory assays. Eight of these were exclusive yields from the Ultrio Plus assay and the remaining seven cases were determined as HBV NAT yield, both by the Procleix Ultrio as well as the Ultrio Plus assays, i.e. "Combined" yields. No HCV or HIV 1 yields were detected during the study period by either of two assays. Conclusion: With an overall yield rate of 1 in 687 and an exclusive yield rate of 1 in 1287, the Procleix Ultrio Plus assay proved to be highly sensitive in detecting occult HBV infections.