Abstract

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 Background and Objectives

  The aim of the present study was to evaluate red blood cell chimerism after bone marrow transplantation by flow cytometry.

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 Materials and Methods

  In order to perform this assay, FITC labeled antibodies against blood groups ABH, Rh, Kell, Duffy, Kidd, MNS were used.14 hematologic patients under BMT were selected for this study. The required sample was 5 ml peripheral blood that is collected in tubes containing EDTA. At first, donor and recipients red cells phenotypes were identified with the use of both agglutination and flow cytometry methods then, on post-transplantation days of 15, 30 and 60, only blood samples of the recipients were analyzed by flow cytometry for the antigens differing from donors to recipients. Antibody screening test and titration of ABH Isohemagglutinins were performed on recipients' plasma samples and then repeated on post- transplantation day of 60.

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 Results

  After BMT, red cell chimerism was detected in all 14 patients (in 9 patients on post-transplantation day of 15 and in 5 patients on day of 30). Antibodies against minor blood groups and Rh blood group were not detected at all. The occurrence of chimerism was not inhibited by ABO incompatibility of donors and recipients but in patients who were ABH incompatible with their donors, ABH isohemagglutinins titer following transplantation decreased. Although the presence of isohemagglutinins did not prevent chimerism but it seems these antibodies by attaching to their related antigens on chimeric red cells membrane prevented corresponding antigen detection.

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 Conclusions

  Now by using flow cytometry, red cell phenotyping is applicable and reticulocyte analysis is much easier to perform so that chimerism can be detected in patients who have recently experienced blood transfusion. Moreover, through further evaluation of red cell chimerism and detection of recipient autologous red cells, disease relapse can be predicted.

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 Key words: Bone Marrow Transplantation (BMT), Chimerism, Red cells, Blood group antigens, Hematological disorders

Abstract

Background and Objectives

HTLV is a member of Retroviridae family whose infection can lead to leukemia. Since in most regions of Iran (like Bushehr) donated blood units are not tested for HTLV infection, multiple transfusion patients are at high risk. In this study, we evaluated serological prevalence of the infection among Busherian patients.

Materials and Methods

This was a descriptive study in which samples of all Busherian multi-transfused patients were taken based on dispersion of thalassemic and hemodialysis hospital wards. Blood samples for detection of HTLV I/II infection markers were tested by ELISA method and for  confirmation of positive ELISA results western blot technique was used. Data were analyzed by using SPSS software and Chi-square.

Results

Out of 642 Busherian multi-transfused patients including 455 (70.78%) thalassemic, 86(13.3%) hemophilic and 101 (15.7%) hemodialysis patients, 41 all thalassemic were ELISA positive out of whom 22(53.7%) were male and 19(46.3%) female. Infection in 14 cases was confirmed by western blot technique (p=0.77). Statistical analysis did not show any significant correlation between western blot results and gender (p=0.56). Neither was there any significant difference between western blot test results across different age ranges (p=0.67). The significant point in this research was the detection of a thalassemic patient with dual infection of HTLV type I and II.

Conclusions

Detection of HTLV infection among multi-transfused population indicated that such an infection may exist among blood donors of Bushehr.

Key words:  Transfusion HTLV I/II, Thalassemia, Hemophilia

  Abstract

 Background and Objectives

  The co-existence of recipient’s and donor’s hematopoietic systems after allogeneic marrow transplantation is called mixed chimerism. Chimerism analysis provides a national method of assessing the ability of different conditioning regimens, graft-versus-host disease (GVHD), prophylactic regimens, and cellular therapy to promote engraftment.

  Materials and Methods

  The association of mixed chimerism with acute graft-versus-host disease (GVHD), disease recurrence, survival, and relapse free survival was investigated in 91 patients 12 and 79 of whom underwent either bone or peripheral blood HLA-identical marrow transplantation respectively. Chimerism was assessed using multiplex amplification of shorty tandem repeats (STR-PCR). Cases included thalassemics (19 subjects), AML (29), ALL (20), CML (18) and others (5). Median age was 21 (age range of 3-50). There were 38 females (41.8%) and 53 males (58.2%). Conditioning was busulfan plus cyclophosphamide in 34 patients, busulfan plus fludarabin in 51 patients and busulfan plus fludarabin plus anti-thymocyte globulin in 6 patients. Median time of follow up was 13 months. Data was analyzed using SPSS statistical software.

 Results

  On day 30 after transplantation, mixed chimerism (MC) was observed in 15 patients (16.5%), complete donor chimerism (CC) in 72 patients (79%), and no chimerism in 4 patients. The incidence of acute GVHD was significantly lower in mixed chimeras than in complete chimeras (p=0.01) but there was no significant difference in acute GVHD grade (I, II vs. III, IV) between two groups. The incidence of relapse and overall survival were 17.6% and 88.9% respectively showing no significant difference between MC and CC. Relapse free survival was 80.2% and not significantly different between two groups.

 Conclusions

  Despite some previous reports, we found no significant difference in survival and relapse rate between MC and CC.

 Key words: Allogenic, Bone Marrow Transplantation , Chimerism, PCR

  Abstract

 Background and Objectives

 GVHD occurs in 20–40% of recipients of HLA-matched sibling donor grafts. A number of studies suggest that polymorphism in cytokine genes influence susceptibility to post-BMT complications. One of the cytokine gene family (IL-1RN) encodes IL-1 receptor antagonist (IL-1Ra), an anti-inflammatory molecule that competes for receptor binding with IL-1α and IL-1β. The overall contribution of IL-1 to the proinflammatory response depends on the balance between these three molecules. In this study, we decided to evaluate the polymorphisms of IL-1Ra gene that might influence the outcome of BMT.

 Materials and Methods

 In this cohort study, patients' (n = 175) and donors' (n = 175) DNA from a total of 350 human leucocyte antigen (HLA)-identical sibling allogeneic BMTs were analysed. IL-1Ra gene polymorphism was studied by PCR-VNTR method. PCR products were then visualized by electrophoresis in 2% agarose gel. Then, the correlation between donor and recipient genotype and GVHD grade for their respective transplant was assessed.

 Results

 The frequency of the aGVHD grades was divided to grade 0 (38.3%), grade I (17.7%), grade II (27.4%), grade III (13.7%), grade IV (2.9%). Correlation between donor and recipient genotype and GVHD grade for their respective transplant was assessed. We observed no significant correlation between the IL-1Ra polymorphism and incidence of aGVHD, although possession of the allele 2 in the individual genotype was associated with less severe acute GVHD.

 Conclusions

 The observed allele frequencies were different from those previously reported. None of the polymorphisms showed association with the presence of acute GVHD. However, presence of allele 2 showed association with aGVHD. If the recipient possesed allele 2, the probability of aGVHD was 46% and in the absence of allele 2, it was 62%. Recipient age, donor age, recipient disease particularly thalassemia were the most remarkable risk factors for acute GVHD.

 Key words : Bone marrow transplantation, Genotype, Graft Versus Host Disease

  Abstract

 Background and Objectives

 Drug resistance is a common cause of treatment failure in malignant disorders especially hematological malignancies. Overexpression of P-glycoprotein (pgp) is the most common cause of drug resistance. Studies on the importance of pgp are controversial that is attributed to the lack of a standard method for pgp assay and the lack of consistency with clinical data. The aim of this study is Multiple drug resistance (MDR) evaluation according to pgp expression in leukemic cells at the beginning of treatment, and prediction of responses to treatment, relapse or death.

 Materials and Methods

 Between June 2001 and May 2006, we studied 185 leukemic patients. Diagnosis was performed on each case via morphology, based on immunotypes, and by cytogenetic method pgp expression was also evaluated by flowcytometry. For pgp evaluation, we collected peripheral blood or bone marrow samples in EDTA. Then, patients went under treatment by standard protocols and their responses to treatment were observed. Patient follow ups for response to treatment, complete remission, resistance to induction, relapse or death were conducted. Finally, clinical data were compared with laboratory results. Mann-Whitney, kruscal-valis, kaplan-meier, and chi-square tests were used for data analysis.

 Results

 From 185 patients with the median age of 28.5 years (within the age range of 11-76), 128 patients showed AML and 57 ALL. 62% were male and 38% female. Pgp expression showed no significant correlation with age, sex, type of leukemia, and white blood cells count. There was no significant association between pgp expression and AML subtype. Two-year Overall Survival (OS) and Disease Free Survival (DFS) were 63% and 42% respectively. DFS showed significant correlation with pgp in ALL and non-M3 subtype of AML (P=0.024).

 Conclusions

 Expression of pgp would be a prognostic factor for relapse prediction and might be important in treatment planning.

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 Key words : Multiple drug resistance, P-Glycoprotein, Acute myelocytic leukemia, Acute lymphocytic leukemia