kamran alimoghadam

Allogeneic Hematopoietic Stem Cell Transplantation (HSCT) is a curative treatment option for many patients with Acute Myeloid Leukemia (AML); however, it can lead to complications of Graft-Versus-Host-Disease (GVHD) which can affect the quality of life and overall survival. The aim of this study was to assess the effects of both acute and chronic GVHD on survival rate in patients with AML who received HSCT. In a longitudinal study, 587 patients with AML who underwent bone marrow transplantation in Tehran-Iran between1991 and 2011 were recruited. All patient records were analyzed for the occurrence of adverse events including acute and chronic GVHD and leukemia relapse. Data were analyzed using Log-rank, Kaplan-Meier, Univariate and Multivariate Cox Regression models. The five-year overall survival (OS) was found to be 71.9% (95% CI: 67.40-76.41). Also there was a significant relationship between cGVHD and OS (P=0.001, HR = 0.476, 95%). Hazard of death in these patients was less than those who did not experience an occurrence of cGVHD and aGVHD (HR= 0.629, P= 0.078). A significant relationship between cGVHD and relapse was observed (P< 0.001) indicating that patients who developed cGVHD experienced a better survival rate. A significant relationship was also found between overall survival and aGVHD grade (P< 0.001). Hazard of death (HD) for cGVHD and relapse variables were estimated to be 0.554 and 3.869. This study is one of the largest studies (regarding the number of participants) done to date in the Middle East with quite a long duration (20 years). cGVHD appears to have a positive influence on survival rate in patients with AML who received HSCT. It is recommended that further studies investigate the underlying reason or mechanisms behind this.

The aim of the present clinical trial was to investigate the efficacy of autologous bone marrow mesenchymal stem cells (BM-MSCs) in glycemic control of diabetic patients without using any immunosuppressive drugs over a nine-month period. Twenty-three patients with T1DM, at 5 to 30 years of age and in both sexes, participated in this study. This trial consisted of two phases; in the end of the first phase (three month after the transplantation), if the patient still needed exogenous insulin to control his/her glycemic state, the second phase of study was performed. In both phases, 100 milliliter of mixed mesenchymal stem cells and normal saline containing 2×10⁶ autologous cells/kg for each patient was delivered to patients through cubital vein. All patients were evaluated at 1, 3, 6 and 9 months after the procedure. Twenty-one patients underwent a second injection. Nine patients (39%) responded positively and 14 patients (61%) responded negatively based on their HbA1c levels and insulin requirements in both injections. Two patients became insulin-free during two rounds of injections. In responder patients, mean levels of C-peptide and HbA1c as well as prescribed insulin dosage significantly decreased compared to baseline measures (P=0.002, P=0.007 and P<0.001). In the second phase, responder patients did not show significant reduction in C-peptide levels compared to the baseline of the second phase. Mean levels of HbA1c and prescribed insulin dosage significantly decreased in comparison to the beginning of the study (P<0.05). Transplantation of BM-MSC can be viewed as a promising, simple, safe, and efficient therapeutic modality for T1DM.

The aim of the present clinical trial was to investigate the efficacy of autologous bone marrow mesenchymal stem cells (BM-MSCs) in glycemic control of diabetic patients without using any immunosuppressive drugs over a nine-month period. Twenty-three patients with T1DM, at 5 to 30 years of age and in both sexes, participated in this study. This trial consisted of two phases; in the end of the first phase (three month after the transplantation), if the patient still needed exogenous insulin to control his/her glycemic state, the second phase of study was performed. In both phases, 100 milliliter of mixed mesenchymal stem cells and normal saline containing 2×10⁶ autologous cells/kg for each patient was delivered to patients through cubital vein. All patients were evaluated at 1, 3, 6 and 9 months after the procedure. Twenty-one patients underwent a second injection. Nine patients (39%) responded positively and 14 patients (61%) responded negatively based on their HbA1c levels and insulin requirements in both injections. Two patients became insulin-free during two rounds of injections. In responder patients, mean levels of C-peptide and HbA1c as well as prescribed insulin dosage significantly decreased compared to baseline measures (P=0.002, P=0.007 and P<0.001). In the second phase, responder patients did not show significant reduction in C-peptide levels compared to the baseline of the second phase. Mean levels of HbA1c and prescribed insulin dosage significantly decreased in comparison to the beginning of the study (P<0.05). Transplantation of BM-MSC can be viewed as a promising, simple, safe, and efficient therapeutic modality for T1DM.

Semi-Markov multi-state models are very important to describe regression and progression in chronic diseases and cancers. Purpose of this research was to determine the prognostic factors for survival after acute leukemia using multi-state models.

In this descriptive longitudinal research, a total of 507 acute leukemia patients (206 acute lymphocyte leukemia, (ALL), 301 acute myeloid leukemia (AML)) at Shariati Hospital, Tehran, Iran were selected. The median of follow up time was 1.5 year. A multi-state model with four states, (state 1: bone marrow transplantation, state 2: chronic graft verses host disease( cGvHD), state 3: platelet recovery, state 4: death, (absorb state) ) were defined. Weibull distribution was considered as transition time between states. Data was analyzed using R Ver 2.13 software.

With increasing sojourn time in state 1, hazard of cGVHD increased, the effects of some covariates were not constant during disease time for example after platelet recovery, death hazard of acute lymphoblastic leukemia patients was 2.15 times of acute myeloid leukemia patients. There was a negative correlation between sojourn time in state 1 and state 2(r = -0.17, P<0.001).

Semi-Markov multi-state models are useful models to accommodate multiple events and time dependent covariates.

Arsenic has been used as an effective medicine in the treatment of severe promyelocytic leukemia in recurrence and resistance cases toward ATRA. In this study, it has been used as induction and maintenance therapy after remission.Method & Material: We studied 31 patients diagnosed by APL. Arsenic was started at a dose of 0.15 mg/kg (daily) until patient’s bone marrow remission.(1) We started Arsenic as a consolidation therapy after 28 days rest and then we continued the treatment with Arsenic each 3-4 months during 2 years for 14 days. 4 patients died (12.9%) during the first 15 days of treatment. 27 patients (87%) went into remission. 2 patients refused the continuation of treatment regardless the remission. 25 patients received a long term treatment. The disease of 3 out of 25 patients recurred during follow -up period.1 patient died during the treatment after recurrence and 3 others given ATRA & Arsenic went into remission. Now, It has been past 2 months since the end of their remission.The recurrence appeared in the form of full involvement of thoraco-lumbar which was observed as an extensive tumor on MRI and was found to cover the mentioned area.1 patient faced CNS fungal infection during neutropenia period and then recovered after operation and proper treatment; however his vision was severely damaged. As 4 patients faced leukocytosis over 1000/000ml, we were obliged to discontinue arsenic for 3-4 days and chemotherapy by Danurobicine was prescribed for 2 days.The patient’s follow-up and the median survival time were 54 and 48 months, respectively. The overall survival was 80.6%. Arsenic as the first line therapy for APL is an effective treatment. Consistent long- term therapy with intervals will reduce the risk of disease recurrence.

Invasive aspergillosis is a major cause of morbidity and mortality in hematologic malignant patients which have received intensive cytotoxic therapy or undergone blood and marrow transplantation (BMT). Early clinical and radiological diagnosis of IA is almost impossible and so mortality is very high. The main purpose of our study was to assay the diagnostic value of serum galactomannan (a fungal antigen found in the sera of infected patients) level in early diagnosis of invasive aspergillosis in patients with hematologic malignancies and BMT.Method and materials: During 2009and2010, 70 adult patients with neutropenic fever of unknown origin in Shariati and Imam Khomeini centers of cancer and BMT were tested for galactomannan serum level on the 7th day of fever or before starting antifungal treatment. The OD index of 0.5 or more was taken as positive result. Tissue biopsy from lung or sinus was performed in 8 patients and there were 7 positive results for aspergillus species. Galactomannan EIA test result was positive in 26 cases (37.1%). There were 7 cases of proven IA with positive tissue (lung or sinus) biopsy and 20 cases of probable IA. Overall, 27 cases were positive for IA (gold standard= proven IA+pobable IA). The study yields a sensitivity and specificity of 77.8% and 88.4% for this test, respectively. Positive and negative predictive value of the test with confidence interval of 95% was 80.8 % (60-92.7%) and 86.4 %(72.0-94.3%), respectively. Considering these results, galactomannan EIA test results must be interpreted cautiously as an alternative test to prove IA, and it doesn’t eliminate the need for other diagnostic criteria such as clinical symptoms, biopsy, imaging, etc. However, the test is substantially helpful in recognizing true cases of the disease.

At present, the only curative treatment for β-thalassemia major is allogenic bone marrow transplantation accompanied with considerable mortality and morbidity in class III β-thalassemia. Regarding few case reports on successful non-myeloablative stem cell transplantation in class III β-thalassemic cases, we evaluated the effectiveness of this type of allogenic stem cell transplantation, considering less toxic non-myeloablative conditioning regimen. In this prospective study in Shariati Hospital bone marrow transplantation center during 2001-3, 13 class III β-thalassemia patients (on the basis of history and physical examination and liver biopsy) were transplanted with peripheral blood and bone marrow stem cells from their HLA-identical siblings. Non-myeloablative conditioning regimen included fludarabine; busulan; antithymocyte globulin. Graft versus host disease (GVHD) prophylactic regimen was cyclosporin and metothrexate. In the case of the declining chimerism, the patients were treated with donor lymphocyte infusions (DLI). The conditioning regimen was tolerated well without any considerable toxicity in hematologic, gastrointestinal and pulmonary systems. Five (38.5%) patients had acute and 2(15.4%) had chronic GVHD. Two patients died after transplantation. While two cases had a thalassemia-free survival. Although associated with high graft failure and the recurrence of disease, non-myeloablative stem cell transplantation may be used as a curative, less toxic post-transplantation treatment for class III ß-thalassemia.

In order to achieve the best therapeutic strategy with which to treat multiple myeloma (MM), it is necessary to understand the molecular mechanisms of myeloma cell growth. In addition to genetic defects, the bone marrow microenvironment (BMM) plays a primary role in MM pathophysiology. For many years, interleukin-6 (IL-6) was considered to be a central growth factor in myeloma cells. However, recent evidence indicates that increasing numbers of cytokines enhance myeloma cell growth in BMM. In this study, we investigated the possibility that leptin could be a member of cytokine network that supports myeloma cells.

The expression of leptin and its receptors in cell lines, bone marrow (BM) and mononuclear peripheral blood (PB) were analysed by RT-PCR. Additionally, leptin serum levels were analysed by ELISA and the effect of leptin on growth of cell lines by cell culture.According to our results,leptin and its receptors were expressed in cell lines, myeloma BM and PB mononuclear cells (MNC). However the PB mononuclear cells of treated myeloma patients did not express leptin and differed in the expression of leptin receptors. Acute lymphoid leukemia, B-blasts (B-ALL) did not express leptin and its receptors. Untreated myeloma and B-ALL patients had high and low levels of leptin in their serum, respectively. Recombinant exogenous leptin enhanced the growth of RPMI8866 but did not affect the growth of U66 and Jurkat.Regarding this study and the findings of other studies, leptin may play a role in MM pathophysiology. Therefore leptin and its receptors might be analysed as a therapeutic target for myeloma BMM.

Acute myeloblastic leukemia (AML) is the most common form of acute leukemia in adults. One of the major problems in this disease is the emergence of leukemic blast cells that are resistant to anticancer drugs. This phenomenon is termed multidrug resistance (MDR). One cause of the MDR is the expression of the MDR1 gene and its product, P-glycoprotein (Pgp). In this study, we tried to inhibit the MDR phenotype with MDR1/mRNA/Pgp in the leukemic cells using short interfering RNA (siRNA) and two nano particles as non-viral vectors. The Pgp expressing cell line was established from a parental K562 cell line with increasing concentration of doxorubicin and named KDI/20. In order to reverse the MDR phenotype due to pgp expression, siRNA against MDR1/mRNA was synthesized. siRNA was used on the KDI/20 cells in combination with two nano particles as non-viral vectors: (1) Fugene 6 transfection reagent (cationic lipid) and (2) polyethylenimine (cationic polymer). The effect of these complexes was assessed at the cellular level by flowcytometry (for Pgp detection) and molecular level by Real Time - PCR.The result showed 79% reduction in Pgp by siRNA /Fugene 6 and 86% reduction in the Pgp by siRNA/PEI at the cellular level in flowcytometry. Also 62% reduction in the MDR1/mRNA by siRNA /Fugene 6 and 74% reduction in the MDR1/mRNA by siRNA /PEI at the molecular level by Real Time – PCR. siRNA could be an efficient method in order to post transcriptional gene silencing and there were no significant differences between two nano particles in gene silencing in this study.

The field of stem cell biology and regenerative medicine is rapidly moving toward translation to clinical practice. Stem cell therapy seems to be a new treatment option for some diseases. So, tracking the distribution of stem cells is crucial to their therapeutic use. Based on this fact we labeled human mesenchymal stem cells (MSCs) with 111In- oxine for the first time in our country. The aim of this study was to investigate the possibility of stem cell labeling in Iran. In addition the researchers assessed the cell viability, specific activity and labeling efficiency after labeling. After culturing mesenchymal stem cells (MSCs), for radio-labeling, the sample (which contained 1×106MSCs) were mixed, and suspended with 50 µCi 111In-oxine, and then incubated for 20 min at the room temperature. The cells were then washed with normal saline twice to remove the free 111In. The labeling efficiency, specific activity and cell viability was 70.6%, 31.70 µCi/106 and 100%, respectively. It seems that, this method is practical and easily applicable with acceptable efficiency and specific activity in our laboratory settings using pharmaceutical produced in Iran.

Previous studies demonstrated significant differences in a number of HLA allele frequencies in leukemia patients and normal subjects. In this study, we have analyzed HLA class II alleles and haplotypes in 110 leukemia patients (60 acute myelogenous leukemia “AML”, 50 chronic myelogenous leukemia”CML”) and 180 unrelated normal subjects. Blood samples were collected from all of the patients and control subjects. DNA was extracted by salting out method and HLA typing was performed using PCR-SSP method. Significant positive association with AML was obtained for HLA-DRB1*11allele (35% vs. 24.7%, P=0.033). Two alleles including HLA-DRB4 and –DQB1*0303 were significantly less frequent in AML patients than in controls. HLA-DQB1*0303 allele was never observed in CML patients compared with allele frequency in controls (4.2%). According to haplotype analysis, HLA-DRB1*0101/DQA1*0104/-DQB1*0501 frequencies were significantly higher and –DRB1*16/-DQA1*01021/-DQB1*0501 frequencies were significantly lower in CML patients than in controls. In conclusion it is suggested that HLA-DRB1*16 allele and HLA-DRB1*15/-DQA1*0103/-DQB1*06011 and –DRB1*16/-DQA1*01021/-DQB1*0501 haplotypes predispose individuals to AML and HLA-DRB4 allele predispose to CML. Future studies are needed to confirm these results and establish the role of these associations in AML and CML.

Previous studies have demonstrated some significant differences in HLA allele frequencies in leukemic patients and normal subjects. We have analyzed HLA class II alleles and haplotypes in 60 Iranian patients with acute myelogenous leukemia (AML) and 180 unrelated normal subjects. Blood samples were collected after obtaining informed consents. From the patients and control DNA extraction and HLA typing were performed using PCR-SSP method. Significant positive association with the disease was found for HLA-DRB1*11 allele (35% vs. 24.7%, p=0.033). Two alleles including HLA-DRB4 and –DQB1*0303 were found to be significantly decreased in patients compared to controls. Regarding haplotype analysis, no significant association was found between case and control groups. It is suggested that HLA-DRB1*11 allele plays as a presumptive predisposing factor while the HLA-DRB4 and –DQB1*0303 alleles are suggested as protective genetic factors against acute myelogenous leukemia. Larger studies are needed to confirm and establish the role of these associations with acute myelogenous leukemia.

HLA compatibility between transplant donor and recipient is one of themajor determinants of transplant outcome. To determine HLA class I by PCR- Sequence-Specific Oligonucleotide Probe (PCR-SSOP) in cord blood donors. Genomic DNA of 142 cord blood samples registered at the Cord Blood Bank of Iran at Hematology، Oncology، and Bone Marrow Transplantation Research Center، was prepared and HLA class I was determined by the PCR-SSOP. A total of 284 HLA-A alleles was identified of which A*02 and A*24 were the most common. Among 284 HLA-B and HLA-C alleles، B*35، B*51، Cw*4 and Cw*12 were the most frequent alleles in the studied population. Amplification of HLA loci with PCR-SSOP has proved to be a reliable method for HLA-A، -B and -C genotyping.

Many investigators have used xenogeneic, especially murine stromal cells and fetal calf serum to maintain and expand human stem cells. The proliferation and expansion of human hematopoietic stem cells in ex vivo culture were examined with the goal of generating a suitable protocol for expanding hematopoietic stem cells for patient transplantation. Using primary fetal liver cells, we established a serum-free culture system to expand human primitive stem/progenitors cells. Non-enriched cord blood CD34+ cells were cultured on a monolayer of mouse primary fetal liver cells in the presence of trombopoietin, flt3/flk2 ligand, and/or stem cell factor, IL-6 and IL-3 under serum-free conditions. After 1 or 2 weeks of culture, cells were examined for clonogenic progenitors and percentage of CD34+ CD38- cells. In the presence of trombopoietin, flt3/flk2 ligand, and stem cell factor, fetal liver cells supported expansion of CD34+ cells more than 10 to 20 fold. In addition, colony forming unit-cell assay was expanded more than 5- and 10-fold after 1 and 2 weeks of culture, respectively. These results strongly suggest that fetal liver cells may be a suitable feeder layer for expansion of hematopoietic progenitors from umbilical cord blood in vitro. Iran. Biomed.

Dendritic cells (DCs) are the most potent stimulators of primary T cell responses and play a key role in immune reactions after stem cell transplantation. Very little is known about the cord blood (CB) dendritic cells and their potential involvement in the low incidence and lower severity of acute graft-versus-host disease after CB transplantation. The aim of this study was the isolation of cord blood and peripheral blood dendritic cells and comparison of their functional competence and determination of their probable role in graft versus host disease after stem cell transplantation. In this study, fresh peripheral blood DCs (PBDCs) were enriched as HLA-DR+ cells, lacking the CD3, CD11b, CD14, CD16, CD19 and CD56, using immunomagnetic bead depletion. For cord blood dendritic cells (CBDCs) enrichment CD34+ and CD66b+ cells were needed to be depleted too. Immunomagnetically enriched PB/CB dendritic cells were co-cultured with adult T lymphocytes and cell proliferation was measured by 3H-thymidine incorporation. Results showed that CBDCs were significantly poor stimulators of the mixed leukocyte reaction as compared with PBDCs (P< 0.05). The demonstrated impairment of CBDCs function could be of importance in interpretation of the low incidence and milder severity of graft-versus-host disease (GVHD) in umbilical CB transplantation compared with peripheral blood or bone marrow stem cell transplantation.

-thalassemia as a hereditary disease is defined as defective synthesis of -globin chains, resulting in erythropoiesis abnormalities and severe anemia. Different studies have shown that cytokines and cytokine gene polymorphisms play a major role in the pathogenesis of -thalassemia. Single nucleotide polymorphisms (SNPs) within the promoter region or other regulatory sequences of cytokine genes lead to overall production of cytokines. To analyze the genetic profile of Th1 and Th2 cytokines in Iranian patients with -thalassemia major. Allelic and genotype frequencies of cytokine genes were determined in 30 thalassemia patients and 40 healthy subjects using PCR-SSP assay. Allele and genotype frequencies were calculated and compared with those of normal controls. The results of our study show a significant decrease in A allele at position UTR 5644 IFN-, G alleles at position -238 TNF- and 166 IL-2, and C allele at position -590 IL-4. TGFhaplotype TG/TG increased whereas TGF- haplotype CG/CG and IL-10 haplotype GCC/ACC decreased significantly in all patients. Data of this investigation suggest that variations among cytokine gene polymorphisms may contribute to the disease susceptibility. A finding which needs to be fairly clarified in other ethnic groups.