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1.  Prevalence and specificities of red cell alloantibodies among blood recipients in the Malaysian state of Kelantan 
Background:
Red blood cell (RBC) alloantibodies may be formed following exposure to RBC antigens. In most cases, the alloimmunization develops during pregnancy or from previous blood transfusions. The RBC antigens and their alloantibodies vary among different human populations and ethnic groups, and they do have a clinical significance for their adverse immunological reactions.
Aims:
This study aimed at studying the prevalence of RBC alloantibodies at the Blood Transfusion Unit of Hospital Raja Perempuan Zainab II in Kota Bharu, Malaysia.
Patients and Methods:
A cross-sectional study was performed utilizing data obtained in the years 2007 and 2008. Data of antibody screening tests from 5719 patients were examined.
Results and Discussion:
The overall prevalence of alloimmunization was 65 (1.13%). The majority of these had a single alloantibody (76.9%), whereas the remaining 23.1% had multiple antibodies. The anti-E antibody comprised the most common alloantibody (24.6%) followed by the anti-Lewis (a) antibodies (18.5%) and the anti-M antibody (13.8%). There were more female recipients than males.
Conclusions:
It was concluded that the findings of this work have been comparable with other published works, and that the main factors associated with alloantibody formation were multiple transfusions and pregnancies. The study also emphasizes the necessity for carrying out immunohematology studies prior to every blood transfusion especially in cases that require multiple transfusions for a long period of time such as in thalassemia patients.
doi:10.4103/0973-6247.75997
PMCID: PMC3082716  PMID: 21572715
RBC antigens; alloantibodies; blood transfusion
2.  Frequency & specificity of RBC alloantibodies in patients due for surgery in Iran 
Background & objectives:
Red blood cell alloimmunization is common in patients receiving multiple blood transfusions. Since the probability of repeat transfusion increases with longer life expectancy, it is important to study to which extent alloimmunized patients with a history of transfusion are prone to form alloantibodies after transfusion events. The aim of this study was to retrospectively analyze the alloimmunization against RBCs among transfused patients who were to undergo elective surgery in Tehran, Iran.
Methods:
A total of 3092 occasionally transfused patients, who were to undergo elective surgery, in four hospitals in Tehran were included in the study. For patients with alloantibodies, the data about sex, date of birth, history of transfusion, surgery, abortion and alloantibody specificity were collected.
Results:
Clinically significant alloantibodies were found in 30 patients. The presence of positive antibodies in the patients for whom cross-match had been done was one per cent. Most of them had surgery history or transfusion record during the preceding year. The three most frequent alloantibodies were anti-K (23.53%), anti- E (20.59%) and anti-c (17.56%).
Interpretation & conclusions:
The most common clinically significant alloantibodies identified in men and women were anti-K and anti-E, respectively. The most common causes of alloimmunization for men were surgery history and transfusion record and for women pregnancy.
PMCID: PMC3788212  PMID: 24056603
Alloimmuization; RBC antigens and antibodies; serological testing; transfusion-surgery
3.  Evaluation of Alloimmunization Rate and Necessity of Blood Type and Screening Test among Patients Candidate for Elective Surgery 
Introduction
Alloimmunization is a reaction of the immune system to foreign antigens. For prevention of alloantibody formation, performing of type and screen test is necessary on a patient’s blood specimen as part of pre-transfusion testing.
Materials and method
In this cross-sectional study, type and screen test done for 1420 patients with elective surgery for detection of alloantibody in Imam Khomeini hospital in Ardabil.
Results
Prevalence of alloantibody in this population was 0.92% (13 patients) and 99.2% (1407 patients) showed no alloantibody in their serum. The most prevalent alloantibody was anti-K, anti-E and anti-c. No significant relationship observed between sex and alloimmunization rate.
Conclusion
Performance of type and screen test play an important role in reducing the rate of alloimmunization, and also, could reduce the demands for blood reservation in hospital blood banks.
PMCID: PMC3913156  PMID: 24505544
Alloimmunization; Type and screen test; Elective surgery
4.  Detection of alloimmunization to ensure safer transfusion practice 
Background:
Serological safety is an integral part of overall safety for blood banks. Emphasis is on the use of routinue Red Blood Cell (RBC) antibody screen test, at set time intervals, to reduce risks related to alloantibodies. Also emphasis is on importance of issuing antigen negative blood to alloantibody positive patients. Effect of using leucodepleted blood on the rate of alloimmunization is highlighted. The concept of provision of phenotypically matched blood is suggested.
Materials and Methods:
Antibody screen test is important to select appropriate blood for transfusion. Repeat antibody screen testing, except if time interval between the earlier and subsequent transfusion was less than 72 hours, followed by antibody identification, if required, was performed in patients being treated with repeat multiple blood transfusions. Between February 2008 and June 2009, repeat samples of 306 multi-transfused patients were analyzed. Search for irregular antibodies and reading of results was conducted using RBC panels (three-cell panel of Column Agglutination Technology (CAT) and two cell panel of the Solid Phase Red Cell Adherence Technology (SPRCAT). Specificities of antibodies were investigated using appropriate panels, 11 cell panel of CAT and 16 cell panel of SPRCA. These technologies, detecting agglutination in columns and reactions in solid phase, evaluate the attachment of irregular incomplete antibody to antigen in the first phase of immunological reaction more directly and hence improve the reading of agglutination. Three to four log leuco reduced red blood cells were transfused to patients in the study using blood collection bags with integral filters.
Results:
Alloimmunization rate of 4.24% was detected from 306 multiply transfused patients tested and followed up. The Transfusion therapy may become significantly complicated.
Conclusion:
Red cell antibody screening and identification and subsequent issue of antigen negative blood have a significant role in improving blood safety. Centers that have incorporated antibody screen test and identification have ensured safe transfusion. Identified patients should be flagged in a database and information shared. Such patients can be given carry-on cards and educated about the names of the identified antibodies. Full red cell phenotyping of individuals, patients and donors, can be feasibility.
doi:10.4103/0973-6247.115577
PMCID: PMC3757774  PMID: 24014944
Antibody screen Test; antibody identification; cell panel
5.  Immune regulation in chronically transfused allo-antibody responder and nonresponder patients with sickle cell disease and β-thalassemia major 
American journal of hematology  2011;86(12):1001-1006.
Red blood cell alloimmunization is a major complication of transfusion therapy. Host immune markers that can predict antibody responders remain poorly described. As regulatory T cells (Tregs) play a role in alloimmunization in mouse models, we analyzed the Treg compartment of a cohort of chronically transfused patients with sickle cell disease (SCD, n = 22) and β-thalassemia major (n = 8) with and without alloantibodies. We found reduced Treg activity in alloantibody responders compared with nonresponders as seen in mice. Higher circulating anti-inflammatory IL-10 levels and lower IFN-γ levels were detected in non-alloimmunized SCD patients. Stimulated sorted CD4+ cells from half of the alloimmunized patients had increased frequency of IL-4 expression compared with nonresponders, indicating a skewed T helper (Th) 2 humoral immune response in a subgroup of antibody responders. All patients had increased Th17 responses, suggesting an underlying inflammatory state. Although small, our study indicates an altered immunoregulatory state in alloantibody responders which may help future identification of potential molecular risk factors for alloimmunization.
doi:10.1002/ajh.22167
PMCID: PMC3618679  PMID: 21953592
6.  Alloantibodies and Australia antigen after open heart surgery 
Journal of Clinical Pathology  1974;27(1):45-49.
The development of irregular serum antibodies (alloantibodies) following massive transfusion was studied in 144 patients who had undergone cardiac valve replacement. An overall incidence of alloimmunization of 8·3% was found. Of the 15 antibodies detected nine were Rhesus anti-E and four anti-Kell. The incidence of anti-E formation in Rhesus E-negative patients was 11·4%; the corresponding figure for anti-Kell was 3·2%. The results suggest that the risk of allo-immunization is directly related to the volume of blood transfused, and Rhesus E-negative persons appear to be at particular risk.
Australia (Au) antigen investigations were also carried out in 102 of these cases. Three patients were Au antigen positive, one of them developing acute hepatitis. In each case no Au antigen could be detected in the donor blood that was used.
It is suggested that tests for alloantibodies and for the Australia antigen should become part of the routine follow-up of any patient receiving massive transfusion.
PMCID: PMC477984  PMID: 4206877
7.  Conservative management of Beta-thalassemia major cases in the sub-division level hospital of rural West Bengal, India 
Background:
The ideal management of thalassemia involves a multidisciplinary therapeutic team approach and should be preferably done at a comprehensive thalassemia care center with all sorts of specialists and the backup of a well-equipped blood bank. However, in developing country like ours, these facilities are not available in rural set up. So, a situation where conservative therapy with regular blood transfusion is the only choice left to innumerable thalassemic children.
Objective:
To evaluate the existing conservative management protocol of Beta-thalassemia major patients in the setup of a subdivision level Government Hospital of rural West Bengal, India.
Materials and Methods:
The study was performed between December 2009 and December 2011. Beta-thalassemia major patients, registered in blood bank for moderate transfusion regimen, were taken in study. All the patients were screened for Transfusion Transmittable Infections at the time of registration and thereafter periodically every six months. Iron chelation therapy was given simultaneously with transfusion at a dose of 20 to 40 mg/kg/day for six days. The patients were advised to follow up with chelation therapy at home by daily infusion with a goal of maintaining serum ferritin level below 1000 ng/ml. Over this long period of study, the patients were periodically evaluated for complications.
Results:
The average blood requirement (ml/kg/year) in 1-5 years, 6-10 years, and 11-15 years were 110, 150, and 180, respectively. Incidence of Hepatitis C Virus infection in 1-5 years and 6-10 years were 1.75% and 2.08%, respectively. It is well seen that serum ferritin level increase with ascending age as does the blood consumption.
Conclusion:
Conservative management may be the best alternative and at times the only hope for patients in developing country like ours. However, in order to decrease the disease load, steps need to be taken to introduce preventive measures.
doi:10.4103/0976-9668.107269
PMCID: PMC3633258  PMID: 23633844
Beta-thalassemia major; blood transfusion; conservative management; iron chelation; prevention program
8.  Alloimmunization screening after transfusion of red blood cells in a prospective study 
Background
Several irregular red blood cell alloantibodies, produced by alloimmunization of antigens in transfusions or pregnancies, have clinical importance because they cause hemolysis in the fetus and newborn and in transfused patients.
Objective
a prospective analysis of patients treated by the surgical and clinical emergency services of Hospital de Clínicas of the Universidade Federal do Triângulo Mineiro (HC/UFTM), Brazil was performed to correlate alloimmunization to clinical and epidemiological data.
Methods
Blood samples of 143 patients with initial negative antibody screening were collected at intervals for up to 15 months after the transfusion of packed red blood cells. Samples were submitted to irregular antibody testing and, when positive, to the identification and serial titration of alloantibodies. The Fisher Exact test and Odds Ratio were employed to compare proportions.
Results
Fifteen (10.49%) patients produced antibodies within six months of transfusion. However, for 60% of these individuals, the titers decreased and disappeared by 15 months after transfusion. Anti-K antibodies and alloantibodies against antigens of the Rh system were the most common; the highest titer was 1:32 (anti-K). There was an evident correlation with the number of transfusions.
Conclusions
Given the high incidence of clinically important red blood cell alloantibodies in patients transfused in surgical and clinical emergency services, we suggest that phenotyping and pre-transfusion compatibilization for C, c, E, e (Rh system) and K (Kell system) antigens should be extended to all patients with programmed surgeries or acute clinical events that do not need emergency transfusions.
doi:10.5581/1516-8484.20120051
PMCID: PMC3459635  PMID: 23049421
Blood transfusion; Blood group antigens; Hemolysis; Immunophenotyping; Emergencies
9.  HLA Alloimmunization Is Associated With RBC Antibodies in Multiply Transfused Patients With Sickle Cell Disease 
Pediatric blood & cancer  2010;54(4):552-558.
Background
Alloimmunization to minor red blood cell (RBC) antigens occurs commonly in sickle cell disease (SCD). Patients with alloimmunization demonstrate increased risk for new alloantibody formation with subsequent transfusion. Alloimmunization to human leukocyte antigens (HLA) can occur with RBC transfusion and may result in graft rejection during stem cell or organ transplantation. The prevalence and risk factors for HLA alloimmunization in multiply transfused pediatric SCD patients are unknown.
Procedure
A cross-sectional study of HLA alloimmunization in SCD patients aged 3–21 years with a history of ≥3 RBC transfusions was performed to test the hypothesis that HLA alloimmunization is associated with RBC alloimmunization. Antibodies to class I and class II HLA were measured by Flow Panel Reactive Antibody (FlowPRA®).
Results
Seventy-three SCD patients (30 with RBC antibodies) were tested. HLA antibodies were detected in 25/73 (34%) patients; class I HLA antibodies occurred in 24/73 (33%) and class II HLA antibodies occurred in 3 (4%). Among patients with RBC antibodies, 16/30 (53%) had HLA antibodies, while 9/43 (21%) patients without RBC antibodies had HLA antibodies (OR 4.32 [1.6–12.1]). In a multivariate analysis, antibodies to RBC antigens were an independent predictor of HLA alloimmunization (P =0.041). The association of RBC and HLA immunization was strongest among patients with no history of chronic transfusion therapy.
Conclusions
This analysis is the first description of HLA alloimmunization in pediatric SCD patients who receive primarily leukoreduced RBC transfusions and demonstrates that HLA alloimmunization tendency is associated with antibodies to RBC antigens.
doi:10.1002/pbc.22327
PMCID: PMC3722881  PMID: 19890898
alloimmunization; human leukocyte antigen (HLA); sickle cell disease; transfusion
10.  Red cell alloimmunization and autoantibodies in Egyptian transfusion-dependent thalassaemia patients 
Introduction
The objective of this study was to explore the frequency of red cell alloantibodies and autoantibodies among β-thalassaemia patients who received regular transfusions.
Material and methods
This study included 501 patients with β-thalassaemia. This work planned to study the presence of alloantibodies and autoantibodies to different red cell antigens in multitransfused thalassaemia patients using the ID. Card micro typing system.
Results
Of a total of 501 β-thalassaemia patients included in the study, 11.3% of patients developed alloantibodies; 9.7% of these alloantibodies were clinically significant. The most common alloantibodies were anti-K, anti-E and anti-C. The rate of incidence of these alloantibodies was 3.9%, 3.3% and 1.7% respectively. Autoantibodies occurred in 28.8% of the patients and 22.1% of these antibodies were typed IgG. There was a significant association between splenectomy with alloimmunization and autoantibody formation (p = 0.03, p = 0.001 respectively). There was no significant association between alloantibody, autoantibody formation and number of transfused packed red cells.
Conclusions
Alloimmunization to minor erythrocyte antigens and erythrocyte autoantibodies of variable clinical significance are frequent findings in transfused β-thalassaemia patients. There is an association between absence of the spleen and the presence of alloimmunization and autoantibody formation.
doi:10.5114/aoms.2010.14473
PMCID: PMC3284076  PMID: 22371805
β-thalassaemia; alloimmunization; autoantibodies
11.  Blood transfusion among thalassemia patients: A single Egyptian center experience 
Background:
Although red cell transfusions are lifesavers for patients with thalassemia, they are responsible for a series of complications and expose the patients to a variety of risks.
Material and Methods:
This cross-sectional study included 464 Egyptian beta(β) thalassemia major patients whose age ranged between 10 months and 31 years (mean 10.2 ± 6.6 years). All patients were subjected to thorough history taking with special emphasis on blood transfusions regarding rate of blood transfusion, type of received blood, and history of previous transfusion reactions in addition to type of chelation and compliance to iron chelation therapy and history of diabetes. Serum ferritin and pretransfusion hemoglobin assessment were done for all patients.
Results:
The mean pretransfusion hemoglobin level was 5.7 ± 1.16 g/dl. Allergic reactions were observed in 3.9% of the patients during the period of the study, while the history of previous allergic reaction was given by 72% of the patients. Deferiprone showed better compliance (58.6%) than deferoxamine (26.3%). The prevalence of diabetes was 10.1% among the studied group. On comparing diabetics to nondiabetics, serum ferritin, transfusion intervals, and age were statistically higher among diabetics (P<0.001).
Conclusion:
Lower pretransfusion hemoglobin and high rate of prevalence of diabetes, in addition to better compliance to deferiprone than deferoxamine, were detected among the patients.
doi:10.4103/0973-6247.106728
PMCID: PMC3613658  PMID: 23559762
β-thalassemia; blood transfusion; allergic transfusion reactions; Iron chelators
12.  Frequency of Sensory Neural Hearing Loss in Major Beta-Thalassemias in Southern Iran 
Iranian Journal of Pediatrics  2010;20(3):308-312.
Objective
The thalassemias are among the most common genetic disorders worldwide, occurring more frequently in the Mediterranean region. The aim of this study was to determined frequency of sensory-neural hearing loss in major ß- thalassemias transfusion dependent patients in south of Iran.
Methods
A cross sectional study on 308 cases of major beta-thalassemia patients referring to Thalassemia Center of Shiraz University of Medical Sciences between 2006–2007 years. The diagnosis of ß- thalassemia major was based on clinical history, complete blood count and hemoglobine electrophoresis. Clinical data such as serum ferritin level, deferoxamine (DFO) dose, mean daily doses of DFO (mg/kg) and audiometric variables was recorded.
Findings
Out of 308 cases, 283 (96.5%) had normal hearing and 10 (3.5%) sensorineural hearing loss. There was no statically significant difference between two groups regarding mean age, weight, age at the first blood transfusion, age at the first DFO infusion.
Conclusion
We found the lowest incidence of sensorineural hearing loss in a large population of patients suffered from major thalassemia who received DFO. We show that DFO is not ototoxic at a low dose. When considering all related literature, as a whole there has been much critical misrepresentation about DFO ototoxicity.
PMCID: PMC3446050  PMID: 23056722
Thalassemia; Sensorineural Hearing Loss; Blood Transfusion; Deferoxamine; Ferritin
13.  Prevalence of Anti HCV Infection in Patients with Beta-Thalassemia in Isfahan-Iran 
Objectives:
Hepatitis C virus (HCV) is the major cause of post-transfusion hepatitis infection (PTH). Patients with thalassemia major are at high risk of hepatitis C due to the blood transfusion from donors infected by HCV. The aim of this study was to detect the prevalence of anti-HCV antibodies and risk factors in multitransfused thalassemic patients in Isfahan-Iran to establish more preventive strategies.
Methods:
This study was conducted to assess the patients with beta-thalassemia in Isfahan hospitals during 1996-2011 for HCV infection. A structured interview questionnaire was developed by the trained researcher to collect the demographic and risk factors. Statistical analysis was done by Chi-square test, Mann-Withney and multiple logistic regressions using SPSS software, version 15.
Results:
466 patients with major thalassemia participated in this study. The mean age of patients was 17.46 ± 8.3. Two hundred and seventy (58.3%) and 193 (41.7%) of participants were male and female, respectively. The prevalence of HCV was estimated 8% among thalassemia patients. History of surgery, history of dental procedure, number of units transfused per month, number of transfusion per month and duration of transfusion had significant association with HCV seropositivity in univariate analysis. There were no statistical significant risk factors for HCV seropositivity in multiple logistic regression models.
Conclusions:
Our findings revealed that blood transfusion was the main risk factors for HCV infection among beta-thalassemic patients. Therefore, more blood donor screening programs and effective screening techniques are needed to prevent transmission of HCV infection among beta-thalassemic patients.
PMCID: PMC3399295  PMID: 22826753
Beta-thalassemia; HCV infection; Iran
14.  Frequencies of maternal red blood cell alloantibodies in Port Harcourt, Nigeria 
Background:
Alloantibodies of clinical importance can cause transfusion reactions or hemolytic disease of the fetus and newborn (HDFN). The frequencies of these antibodies have not been reported in our locality.
Aims:
To determine the frequency of occurrence of alloantibodies among pregnant women in Port Harcourt, Nigeria.
Settings and Design:
This is a prospective study, which was carried out in the Braithwaite Memorial Specialist Hospital, Port Harcourt, Nigeria.
Materials and Methods:
Screening and identification of red blood cell alloantibodies was done on the sera of 500 pregnant women using the DiaMed, DiaCell, and DiaPanel reagents (Cressier, Switzerland). ABO and Rh blood groups were done using antisera bought from Biotec (Ipswich, UK).
Results:
Alloantibodies were identified in the serum of 17 of the 500 (3.4%) pregnant women. The specificity of the antibodies was as follows: anti-C 6 (1.2%), anti-E 3 (0.6%), anti-Jsb 3 (0.6%), and anti-K 5 (1.0%). No anti-D was identified despite 8.6% of the study population being Rhesus D (Rh D) negative. The distribution of the antibodies was found to be independent of the blood groups of the participants (χ2 = 4.050, P = 0.670). Blood group O constituted the highest percentage (48.0%).
Conclusion:
This study has identified the presence of non-Rh D antibodies to the proportion of 3.4%. Rh D antibody was absent in this population irrespective of the relatively high percentage of Rh D negative women. There is a need to determine the actual risk these antibodies may pose to the antenatal women and to include antibody screening and identification in routine antenatal care.
doi:10.4103/0973-6247.75987
PMCID: PMC3082715  PMID: 21572714
Alloantibodies; frequencies; non-Rh D antibodies; HDFN; Nigeria
15.  Frequency of RBC Alloantibodies in Chinese Surgical Patients 
Background
The aim of this study was to evaluate the frequency of red blood cell (RBC) alloantibodies in surgical patients.
Methods
Blood samples of 18,980 Chinese surgical patients were collected and tested between January 2009 and September 2010. For patients with RBC alloantibodies, sequences of antibodies were identified with the DiaMed Asia antibody screening system. Data regarding sex, age, transfusion history, pregnancy history, and alloantibody specificity were collected.
Results
39 alloantibodies were identified from 18,980 samples, yielding a prevalence of 0.21%. The most frequently identified alloantibodies were Rhesus system antibodies (28/39, 71.8%), including anti-E (17/39; 1 anti-E accompanied by anti-Fyb), anti-c (5/39), anti-cE (2/39), anti-Cw (1/39), anti-D (1/39), and anti-e (1/39). Other antibodies included anti-M (5/39), anti-Lea (2/39), anti-Leb (1/39), anti-K+S (1/39), anti-JKa (1/39), and anti-JSa (1/39). The frequency of alloantibodies was greater in females than in males (31 vs. 8).
Conclusion
The results show a higher prevalence of RBC alloantibodies in females than in males. Anti-E was the most common alloantibody identified in this Chinese surgical population and was also more frequent in females compared to males.
doi:10.1159/000339811
PMCID: PMC3434327  PMID: 22969699
Red blood cells; Alloantibody; Surgery
16.  The First Korean Case Report of Anti-Gerbich 
Annals of Laboratory Medicine  2012;32(6):442-444.
In this study, we report the first Korean case of an anti-Gerbich (Ge) alloantibody to a high-incidence antigen that belongs to the Ge blood group system. The alloantibody was detected in a middle-aged Korean woman who did not have a history of transfusion. Her blood type was B+, and findings from the antibody screening test revealed 1+ reactivity in all panels except the autocontrol. The cross-matching test showed incompatible results with all 5 packed red blood cells. Additional blood type antigen and antibody tests confirmed the anti-Ge alloantibody. While rare, cases of hemolytic transfusion reaction or hemolytic disease in newborns due to anti-Ge have been recently reported in the literature. Therefore, additional further studies on alloantibodies to high-incidence antigens, including anti-Ge, are necessary in the future.
doi:10.3343/alm.2012.32.6.442
PMCID: PMC3486941  PMID: 23130346
Ge; Blood group antigens; Transfusion
17.  Prevention of red cell alloimmunization by CD25 regulatory T cells in mouse models 
American journal of hematology  2007;82(8):691-696.
Transfusion therapy is currently an effective therapeutic intervention in a number of diseases, including sickle cell disease. However, its use is complicated by a high incidence of red blood cell (RBC) alloimmunization in the transfusion recipients. The identification of T regulatory cells (Tregs) among the CD4+CD25+ T cell subset as key regulators of peripheral tolerance in mice as well as humans has opened an exciting era in the prevention and treatment of autoimmune disease and for improving organ transplantation. However, their potential in inducing transfusion tolerance remains to be explored. We used red cells from mice transgenic for human glycophorin A blood group antigen as donor cells and transfused wild-type mice to induce alloantibodies, as an experimental system to study RBC alloimmunization. We found that depletion with anti-CD25 enhanced the alloantibody production, indicating that CD25 Tregs play an important role in regulation of alloantibody responses. More importantly, adoptive transfer of purified population of CD4+CD25+ but not CD4+CD25– cells from naïve mice prevented the induction of IgG and IgM alloantibody production in transfusion recipients, with a concomitant reduction in activated splenic B cells and macrophages. Similarly, adoptive transfer of purified populations of CD4+CD25+ cells from naïve mice into naïve syngeneic recipients inhibited the anti-Ig response to rat RBCs in the recipients but transfer of control CD4+CD25– cells did not. Altogether, our results demonstrate that Tregs participate in the control of trans-fusion-associated RBC alloantibody responses, opening up the possibility that Treg immunotherapy may be exploited for suppressing transfusion immunization events.
doi:10.1002/ajh.20959
PMCID: PMC3607942  PMID: 17492644
18.  Prevalence, specificity and risk of red blood cell alloantibodies among hospitalised Hubei Han Chinese patients 
Blood Transfusion  2014;12(1):56-60.
Background
The prevalence, specificity and risk of red blood cell alloantibodies vary widely among different geographic areas, races, and diseases and according to different methods of study, but no data are available on the Chinese Han population, who were investigated in the present study.
Materials and methods
Antibody screening was conducted among 42,517 hospitalised Hubei Han Chinese individuals using column agglutination technology. Samples that were positive in antibody screening were subjected to antibody identification by the tube test. Clinical data, including gender, age, race, transfusion history and records of alloantibody detection, transfusion reactions or haemolytic disease of the newborn, were collected to analyse the prevalence and specificity of alloantibodies and complications associated with them.
Results
A total of 212 patients with alloantibodies were identified among 42,517 patients, yielding a prevalence of 0.50% in this study. Significantly different prevalence rates were observed according to age and sex. The most frequently identified alloantibodies were anti-E (87/212, 41.0%), anti-D (45/212, 21.2%), anti-M (41/212, 19.3%) and a combination of anti-E and anti-c (13/212, 6.1%). Haemolytic disease was observed in 13 infants with anti-D, three infants with anti-E and one infant with anti-Fya alloantibodies. Delayed haemolytic transfusion reactions occurred in four patients with alloantibodies.
Discussion
In hospitalised Hubei Han Chinese individuals, the overall prevalence of alloantibodies was 0.50%, with anti-E, anti-D and anti-M being the most frequently identified alloantibodies. These results indicate that anti-D and anti-E alloantibodies were major risk factors for haemolytic disease of the newborn or delayed haemolytic transfusion reactions in this study population.
doi:10.2450/2013.0013-13
PMCID: PMC3926729  PMID: 24333071
RBC alloantibodies; antibody prevalence; antibody specificity; Han Chinese
19.  Transfusion Transmitted Hepatitis: Where Do We Stand Now? A One Center Study in Upper Egypt 
Hepatitis Monthly  2012;12(4):286-291.
Background
Despite progress made in the prevention of transfusion-transmitted infections (TTI) over the last few years, they continue to be a problem in many parts of the world, particularly in multitransfused patients.
Objectives
The aim of this study was to estimate the prevalence of hepatitis B virus (HBV), hepatitis C virus (HCV), and to evaluate the screening and vaccination program among our cohort of multitransfused children from Qena, Upper Egypt.
Patients and Methods
One-hundred children suffering from diseases requiring repeated blood transfusions were included in the study. They were classified into group 1, which included 67 children with thalassemia, and group 2, which included 33 children with hemophilia. Screening for hepatitis B surface antigen, hepatitis B surface antibody, hepatitis B core antibody and antibody to HCV was done using a second-generation enzyme-linked immunosorbent assay technique.
Results
Only 12% of all patients were either acutely or chronically infected with HBV. 46% were immune due to previous vaccination, whereas 39% of patients were not protected from HBV infection. HCV antibodies were positive in 45% of cases. Seventy-eight patients had a complete hepatitis B vaccination in the form of three doses as documented by birth certificate. Thirty-six patients mentioned history suggestive of hepatitis. The prevalence of the studied hepatitis markers was similar in both the thalassemia and hemophilia groups of children.
Conclusions
Transfusion-transmitted hepatitis is still a major problem for multitransfused children in Egypt. More effort is required to reduce the infection rate through proper screening of blood and blood products, strict emphasis on receiving the vaccine, regular follow-up for those children with a hepatitis B antibody titer, and providing booster doses for those in need.
doi:10.5812/hepatmon.852
PMCID: PMC3360939  PMID: 22690237
Child; Hepatitis; Transfusion; Egypt
20.  Determination of Hepatitis C Genotypes and the Viral Titer Distribution in Children and Adolescents with Major Thalassemia 
Iranian Journal of Pediatrics  2010;20(1):75-81.
Objective
Hepatitis C virus (HCV) is an etiological agent responsible for occurrence of post-transfusion hepatitis in thalassemic patients. This study identified hepatitis C genotypes in pediatric and adolescent thalassemic patients and their correlation with age, blood transfusion, HCV RNA viral titer and liver function.
Methods
This study considers cross-sectional data from the Center for Thalassemia in Zahedan (Iran) carried out between August 2005 and September 2007. Twenty multitransfused patients suffering from β-thalassemia major and chronic HCV infection (13 males, 7 females) were included in the study. Patients were considered eligible for the study if they were seropositive for HCV RNA polymerase chain reaction (PCR) before initiation of evaluation. Blood sample was taken for HCV genotype and viral titer as well as biochemical markers. Type specific primer and real-time RT-PCR HCV were used for determination of viral genotype and HCV-RNA titer.
Findings
There was a significant positive correlation between serum HCV RNA titer and genotypes (P<0001). Serum HCV RNA levels were found higher in genotype 3a than in others. The most prevalent genotype in thalassemic patients was genotype 3a (40%) followed by 1b (25%), unclassified (20%) and la (15%). There was no meaningful relationship between genotype, Alanine aminotranferease, ferritin and alkaline phosphatase. Age, serum HCV RNA titer and number of transfusions were the only significant factors associated with genotypes (P<015, P<0.0001 and P<0.001 respectively).
Conclusion
This study showed that HCV genotype and viral titer are related to the number of blood transfusions received by thalassemic patients. Screening donated blood in blood banks would prevent the occurrence of hepatitis C in this high-risk group.
PMCID: PMC3445996  PMID: 23056686
Hepatitis C; Virus Titer; Viral Load; Liver Function Tests; Thalassemia; Iran
21.  A Newly Recognized Blood Group in Domestic Shorthair Cats: The Mik Red Cell Antigen 
Background
Naturally occurring alloantibodies produced against A and B red cell antigens in cats can cause acute hemolytic transfusion reactions. Blood incompatibilities, unrelated to the AB blood group system, have also been suspected after blood transfusions through routine crossmatch testing or as a result of hemolytic transfusion reactions.
Hypothesis
Incompatible crossmatch results among AB compatible cats signify the presence of a naturally occurring alloantibody against a newly identified blood antigen in a group of previously never transfused blood donor cats. The associated alloantibody is clinically important based upon a hemolytic transfusion reaction after inadvertent transfusion of red cells expressing this red cell antigen in a feline renal transplant recipient that lacks this red cell antigen.
Methods
Blood donor and nonblood donor cats were evaluated for the presence of auto- and alloantibodies using direct antiglobulin and crossmatch tests, respectively, and were blood typed for AB blood group status. Both standard tube and novel gel column techniques were used.
Results
Plasma from 3 of 65 cats and 1 feline renal transplant recipient caused incompatible crossmatch test results with AB compatible erythrocytes indicating these cats formed an alloantibody against a red cell antigen they lack, termed Mik. The 3 donors and the renal transplant recipient were crossmatch-compatible with one another. Tube and gel column crossmatch test results were similar.
Conclusions and Clinical Importance
The absence of this novel Mik red cell antigen can be associated with naturally occurring anti-Mik alloantibodies and can elicit an acute hemolytic transfusion reaction after an AB-matched blood transfusion.
PMCID: PMC3132507  PMID: 17427390
Blood type; Cat; Gel column crossmatch; Hemolytic transfusion reaction; Renal transplant
22.  Therapeutic Value of Combined Therapy with Deferasirox and Silymarin on Iron Overload in Children with Beta Thalassemia 
Background
Beta thalassemia is an inherited hemoglobin disorder resulting in a severe, chronic anemia requiring life-long blood transfusion that induces iron overload. Silymarin is a flavonoid complex isolated from Silybin marianum with a strong antioxidant activity, inducing an hepatoprotective action, and probably, a protective effect on iron overload. The aim of this work was to determine the silymarin value in improving iron chelation in thalassemic patients with iron overload treated with Deferasirox.
Patients and Methods
This study was conducted on 40 children with beta thalassemia major under follow-up at Hematology Unit, Pediatric Department, Tanta University Hospital with serum ferritin level more than 1000 ng/ml and was divided into two groups. Group IA: Received oral Deferasirox (Exjade) and silymarin for 6 months. Group IB: Received oral Deferasirox (Exjade) and placebo for 6 months and 20 healthy children serving as a control group in the period between April 2011 and August 2012 and was performed after approval from research ethical committee center in Tanta University Hospital and obtaining an informed written parental consent from all participants in this study.
Results
Serum ferritin levels were markedly decreased in group IA cases compared with group IB (P= 0.001).
Conclusion
From this study we concluded that, silymarin in combination with Exjade can be safely used in the treatment of iron-loaded thalassemic patients as it showed good iron chelation with no sign of toxicity.
Recommendations
We recommend extensive multicenter studies in a large number of patients with longer duration of follow-up and more advanced techniques of assessment of iron status in order to clarify the exact role of silymarin in reducing iron overload in children with beta thalassemia.
doi:10.4084/MJHID.2013.065
PMCID: PMC3867224  PMID: 24363880
23.  Patient-to-Patient Transmission of Hepatitis C at Iranian Thalassemia Centers Shown by Genetic Characterization of Viral Strains 
Hepatitis Monthly  2013;13(1):e7699.
Background
Hepatitis C is prevalent among thalassemia patients in Iran. It is mainly transfusion mediated, in particular among patients treated before 1996 when blood screening was introduced.
Objectives
The current study aimed to investigate why patients still seroconvert to anti-HCV in Iranian thalassemia centers.
Patients and Methods
During 2006-2007 sera were sampled from 217 anti-HCV positive thalassemia patients at nine thalassemia centers in Tehran and Amol city, where 34 (16%) patients had been infected after 1996. The HCV subtype could be determined by sequencing and phylogenetic analysis of partial NS5B and/or 5׳NCR-core region in 130 strains.
Results
1a (53%) was predominant followed by 3a (30%), 1b (15%), and one strain each of 2k, 3k and 4a. Phylogenetic analysis revealed 19 clades with up to five strains diverging with less than six nucleotides from each other within subtypes 1a and 3a. Strains in seven clades were from nine patients infected between 1999 and 2005 and similar to strains from eight patients infected before 1996, indicating ongoing transmission at the centers. Further epidemiological investigation revealed that 28 patients infected with strains within the same clade had frequently been transfused at the same shift sitting on the same bed. An additional eight patients with related strains had frequently been transfused simultaneously in the same room.
Conclusions
The results suggest nosocomial transmission at these thalassemia centers both before and after the introduction of blood screening. Further training of staff and strict adherence to preventive measures are thus essential to reduce the incidence of new HCV infections.
doi:10.5812/hepatmon.7699
PMCID: PMC3622054  PMID: 23585766
Hepatitis C; Thalassemia; Iran
24.  Alloimmunization to transfused HOD RBCs is not increased in mice with sickle cell disease 
Transfusion  2011;52(2):231-240.
Background
Increased rates of RBC alloimmunization in patients with sickle cell disease may be due to transfusion frequency, genetic predisposition, or immune dysregulation. To test the hypothesis that sickle cell pathophysiology influences RBC alloimmunization, we utilized two transgenic mouse models of sickle cell disease.
Study Design and Methods
Transgenic sickle mice, which express human α and βS globin, were transfused with fresh or 14-day stored RBCs containing the HOD (hen egg lysozyme, ovalbumin, and human Duffyb) antigen; some recipients were inflamed with poly (I:C) prior to transfusion. Anti-HOD alloantibody responses were subsequently measured by ELISA and flow crossmatch; a cohort of recipients had post-transfusion serum cytokines measured by bead array.
Results
Both Berkeley and Townes homozygous (SS) and heterozygous (AS) mice had similar rates and magnitude of anti-HOD RBC alloimmunization following fresh HOD RBC transfusion compared with control animals; under no tested condition did homozygous SS recipients make higher levels of alloantibodies than control animals. Unexpectedly, homozygous SS recipients had blunted cytokine responses and lower levels of anti-HOD alloantibodies following transfusion of 14-day stored RBCs, compared with control animals.
Conclusions
In sum, homozygous βS expression and the ensuing disease state are not alone sufficient to enhance RBC alloimmunization to transfused HOD RBCs in 2 distinct humanized murine models of sickle cell disease under the conditions examined. These data suggest other factors may contribute to the high rates of RBC alloimmunization observed in humans with sickle cell disease.
doi:10.1111/j.1537-2995.2011.03255.x
PMCID: PMC3218203  PMID: 21790627
red blood cells (sickle cell disease); alloimmunization; transfusion medicine
25.  β-Thalassemia: HiJAKing Ineffective Erythropoiesis and Iron Overload 
Advances in Hematology  2010;2010:938640.
β-thalassemia encompasses a group of monogenic diseases that have in common defective synthesis of β-globin. The defects involved are extremely heterogeneous and give rise to a large phenotypic spectrum, with patients that are almost asymptomatic to cases in which regular blood transfusions are required to sustain life. As a result of the inefficient synthesis of β-globin, the patients suffer from chronic anemia due to a process called ineffective erythropoiesis (IE). The sequelae of IE lead to extramedullary hematopoiesis (EMH) with massive splenomegaly and dramatic iron overload, which in turn is responsible for many of the secondary pathologies observed in thalassemic patients. The processes are intimately linked such that an ideal therapeutic approach should address all of the complications. Although β-thalassemia is one of the first monogenic diseases to be described and represents a global health problem, only recently has the scientific community started to focus on the real molecular mechanisms that underlie this disease, opening new and exciting therapeutic perspectives for thalassemic patients worldwide.
doi:10.1155/2010/938640
PMCID: PMC2873658  PMID: 20508726

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