[ first experience of stem cell labeling in iran using 111In- oxine]

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 1x106MSCs] were mixed, and suspended with 50 micro Ci 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 micro Ci/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

HLA class II allele and haplotype frequencies in Iranian patients with leukemia

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

HLA class II allele and haplotype frequencies in Iranian patients with acute myelogenous leukemia and control group

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

Comparison of functional competence of umbilical cord and adult peripheral blood dendritic cells in allogenic mixed leukocyte reaction

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] wereenriched as HLA-DR[+] cells, lacking the CDS, CDllb, 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 Tlymphocytes 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

Cytokine gene polymorphisms in iranian patients with beta-thalassemia major

beta-thalassemia as a hereditary disease is defined as defective synthesis of beta -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 beta -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 Thl and Th2 cytokines in Iranian patients with beta -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-y, G alleles at position -238 TNF-oc and 166 IL-2, and C allele at position -590 IL-4. TGF- beta haplotype TG/TG increased whereas TGF- beta 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

Th1 and Th2 cytokine gene polymorphism in Iranian patients with chronic myelogenous leukemia

Bakground: lt has been hypothesized that genetic factors other than histocompatibility disparity may play a role in predisposition to developing Chronic Myelogenous Leukemia [CML]. In this regard, Thl and Th2 cytokines and their gene polymorphism seems to be important. Overall expression and secretion of cytokines is dependent, at least in part, on genetic polymorphism [nucleotide variations] within the promoter region or other regulatory sequences of cytokine genes. The majority of polymorphisms described are single nucleotide polymorphism [SNPs]. The objective of this study was to analyze the genetic profile of Thl and Th2 cytokines in 30 Iranian patients with CML and 40 healthy subjects

Methods: In the patients and control subjects, the allelic and genotype frequencies were determined for the cytokine genes. All typing were performed by PCR-SSP assay. Allele and genotype frequencies were calculated and compared with those of normal controls

Results: The results showed that the most frequent alleles in our patients were TGF-3 TG/TG, IL-4 T at position -1089, C at position -590, T at position -33 and IL-10 A at position -1082. Whereas the following alleles - TGF-3 CG/CG and IL-10 C at position -592 - were seen in much lower frequencies

Conclusion: In conclusion, it could be suggested that the frequency of high producing TGF-3 alleles and low producing IL-4 and IL-10 alleles in the CML patients is higher than the normal subjects