TET2, DNMT3A, IDH1, and JAK2 Mutation in Myeloproliferative Neoplasms in southern Iran.

Background: Five epigenetic regulator mutations are considered in myeloproliferative neoplasms (MPN) that have prognostic and therapeutic values. Objective: We aimed to evaluate these mutations in MPNs among the Iranian population. Methods: We selected 5 mutations in 4 epigenetic regulatory genes [TET2, DNMT3A, IDH1 (rs147001633&rs121913499), and JAK2)] and evaluated 130 patients with MPNs including 78 Philadelphia chromosome negative (49 ETs, 20 PVs, and 9 PMFs) and 52 Philadelphia chromosome-positive patients as well as 51 healthy controls. Results: Eight patients (6.5%) carried the DNMT3A mutation, 35 (27%) were positive for TET2 mutation and 64 (49.3%) had the JAK2V617F mutation. In the healthy controls, 16 (31.4%) cases had the TET2 mutation (15 Heterozygote + 1 Homozygote) and one had heterozygote JAK2 mutation. There was no statistically significant difference between patient groups for any of these mutations, except for JAK2. The JAK2 mutation rate was 18 (90%), 25 (51%), 7 (77.8%), 14 (26.9%) in polycythemia vera, essential thrombocythemia, primary myelofibrosis, and chronic myelocytic leukemia, respectively. Patients aged 60 and older were more likely to carry the TET2 mutation (23% vs. 39% in younger and older than 60 years old individuals, p=0.025). IDH1 was not detected at all and PV had the highest TET2 mutation 7(35%). Two PMF patients had a history of bone marrow transplantation that were negative for IDH1and DNMT3A and one was positive for TET2 mutation. Conclusion: In the normal Iranian population, the heterozygote form of TET2 mutation is significant, especially in the elderly. No association was found between JAK2 and TET2 mutations. Both of them are more prevalent in the age group of 60 years and older. DNMT3A mutation has a low prevalence and occurs in both positive and negative MPNs.

Importance of miR-UL-148D Expression Pattern in Cytomegalovirus Infected Transplant Patients.

Background: MicroRNAs (miRNAs) are endogenous, 18-22 nucleotide non-coding RNA molecules. Human cytomegalovirus (HCMV) is a ubiquitous and particular herpes virus that encodes miRNAs, which increases gradually in the presence of infection. One of the important viral miRNAs is HCMV-miRUL-148D, which plays a role in establishing and maintaining viral latency. Objective: The current study aimed to evaluate the expression levels of HCMV-miRUL-148D in active and inactive HCMV infected transplant patient groups compared to healthy individuals. Methods: Total RNA was extracted from blood samples of 60 solid organ transplant patients and 30healthy controls. In-house SYBR Green Real-Time PCR evaluated the expression levels of studied miRNAand gene. Results: The expression level of the UL-148D gene was significantly higher in the active HCMV infectedpatients (p=0.001) compared to other groups. While the miRUL-148D expression level significantly increased in the inactive HCMV-infected patients (p<0.001) compared to other groups. Conclusion: Increased miRUL-148D expression level in the inactive HCMV-infected transplant patients indicates the potential role of this miRUL-148D as a biomarker of the HCMV latent stage.

Association between Interleukin-21, 23 and 27 Expression and Protein Level with Cytomegalovirus Infection in Liver Transplant Recipients.

BACKGROUND: Cytokines have regulatory crosstalk with CMV infection leading to manage of post-liver transplantation virus-related outcomes. OBJECTIVE: To investigate the link between IL-21, IL-23 and IL-27 mRNA and protein level with active CMV infection, which was evaluated in reactivated and non-reactivated liver transplant recipients. METHODS: Two groups of liver transplant recipients were enrolled in this study-54 without and 15 with active CMV infection. 3 EDTA-treated blood samples were taken on day 1, 4, and 7 post-liver transplantation. Plasma and buffy coats of all samples were separated. All samples were analyzed for CMV reactivation using antigenemia technique. The separated plasma of positive samples was used for viral DNA extraction and protein evaluation. For evaluating the mRNA expression level by real-time PCR, RNA extraction and cDNA synthesis were done for all samples. Also, the protein level of studied genes was estimated by ELISA. RESULTS: The expression level of IL-21, IL-23A and IL-27A cytokine genes was increased in CMV reactivated liver transplant recipients in comparison with CMV non-reactivated ones; IL-27A expression pattern was significant (p=0.001) at all sampling times. IL-21 significantly increased on the 2nd and 3rd (p=0.028 and 0.01, respectively) sampling days in CMV reactivated compared with non-reactivated patients. The expression level of IL-23A cytokine significantly increased on the 3rd (p=0.017) sampling day in CMV reactivated compared with non-reactivated liver transplant recipients. CONCLUSION: Elevation in the expression level of IL-21, IL-23A and IL-27A mRNA and protein level in CMV reactivated patients emphasized on the antiviral role of these cytokines in CMV reactivated liver transplant recipients.

Dysregulated Expression of CD28 and CTLA-4 Molecules in Patients with Acute Myeloid Leukemia and Possible Association with Development of Graft versus Host Disease after Hematopoietic Stem Cell Transplantation.

BACKGROUND: Dysregulated expression of co-stimulatory molecules is one of the immune escape mechanisms employed in hematologic malignancies like acute myeloid leukemia (AML). OBJECTIVES: To evaluate the expression of the CD28 and CTLA-4 molecules in 62 adults with de novo AML and its correlation with the development of acute graft vs host disease (GVHD) after hematopoietic stem-cell transplantation. METHODS: The relative expression of CD28 and CTLA-4 was measured by quantitative SYBR Green real-time PCR method in a group of patients and controls as well as different risk groups (high, intermediate and favorite risk), M3 vs non-M3 and GVHD vs non-GVHD patients. RESULTS: The mRNA expression of CD28 (7.9-fold) and CTLA-4 (5.7-fold) was significantly increased in AML patients compared with healthy controls (p=0.006 and 0.02, respectively). Although the mean expression of both CD28 and CTLA-4 was increased in high-risk group compared with low-risk and intermediate-risk groups, the difference was not statistically significant. Also, the mean expression of the CTLA-4, but not CD28, was significantly higher in M3 patients compared with non-M3 ones (p<0.001). The expression of CD28 was upregulated in GVHD patients, while the expression of CTLA-4 was slightly lower in GVHD patients compared with non-GVHD patients, though the difference was not statistically significant. There was no significant correlation between the expression of CD28 and CTLA-4 and laboratory parameters like white blood cells and platelets counts, and hemoglobin and lactate dehydrogenase level in AML patients. CONCLUSIONS: CD28 and CTLA-4 molecules are aberrantly expressed in peripheral blood leukocytes of AML patients and might contribute to the development of aGVHD after hematopoietic stem cell transplantation.

Genetic variation of TNF-α and IL-10, IL-12, IL-17 genes and association with torque teno virus infection post hematopoietic stem cell transplantation.

Little is known about the role of genetic variation in the genes for cytokines and susceptibility to viral infection especially torque teno virus (TTV) following allogeneic hematopoietic stem cell transplantation. In this study, the association between interleukin-12, interleukin-17, interleukin-10 (IL-12,-17,-10) and tumor necrosis factor-α (TNF-α) polymorphisms was evaluated in patients with TTV infection who underwent allogeneic hematopoietic stem cell transplantation from South of Iran. The single nucleotide polymorphisms in the cytokine genes including IL-12 (-1188A/C), IL-17 (-197G/A), IL-10 (-1082G/A, -819C/T and -592C/A) and TNF-α (-308 G/A) were analyzed by PCR-RFLP methods. While our results did not show any association between IL-17, IL-12 and IL-10 (-819C/T and -1082G/A) polymorphisms and TTV infection status, heterozygote genotype of IL-10 (-592C/A) had direct correlation with TTV infection and A allele of TNF-α (-308G/A) showed a protective effect against TTV infection (P = 0.05 and P = 0.025, respectively). Within the group of patients who experienced acute graft-versus-host disease, the AA genotype and the A allele of IL-17 (-197 G/A) were significantly higher in non-infected patients compared to infected ones (P = 0.024 and P = 0.057, respectively). It was also observed that among infected patients, the GG genotype of IL-17 and AA genotype of TNF-α were significantly increased in hematopoietic stem cell transplanted patients with low grade (grade I+II) acute graft-versus-host disease compared to high grade (grade III and IV) disease (P = 0.056 and P = 0.056, respectively). Taken together, genetic variation of IL-10 (-592C/A) and TNF-α (-308G/A) genes might be associated with susceptibility to TTV infection post hematopoietic stem cell transplantation. Keywords: TNF-α; interleukins; torque teno virus (TTV); hematopoietic stem cell transplantation (HSCT); graft versus host disease (GvHD).

Association of IL-12 and TNF-α Polymorphisms with Graft-Versus-Host Disease in Allogeneic Hematopoietic Stem Cell Transplant Recipients.

BACKGROUND: Cytokines are important factors determining the outcome of transplantation. The host ability in cytokine production may be affected by cytokine genes polymorphisms. OBJECTIVE: To investigate the effect of IL-12 and TNF-α gene polymorphisms on outcome of hematopoietic stem cell transplantation. METHODS: 90 bone marrow transplant recipients were included in this study. 30 (33%) of 90 recipients experienced graft-versus-host disease (GVHD). IL-12 and TNF-α gene polymorphisms were evaluated by PCR-RFLP and ARMS-PCR method, respectively. RESULTS: No significant difference in the distribution of IL-12 (rs3212227 +1188 A/C) and TNF-α (rs 1800629 -308 G/A) genotypes and alleles was observed between those with and without GVHD. There was no significant association between the distribution of genotypes and the recipient sex. CONCLUSION: IL-12 (rs3212227 +1188 A/C) and TNF-α (rs 1800629-308 G/A) genotypes and alleles were not risk factors for development of GVHD.

Interaction between Herpes Virus Infections and IL10 and Risk of Bone Marrow Suppression.

BACKGROUND: Syndrome of transient bone marrow suppression may result from various extra-hematological diseases, such as immunological deregulations, and viral infectious diseases secondarily affecting the function of hematopoietic stem cells. OBJECTIVE: To evaluate the pathogenic role of herpes viruses and their contraction with IL10 cytokine gene polymorphism, which can impair hematopoiesis in patients with transient bone marrow suppression. METHODS: In a cross-sectional study 30 patients who admitted to Namazi Hospital, affiliated to Shiraz University of Medical Sciences, with transient bone marrow suppression were recruited. Diagnosis of the transient bone marrow suppression was made by expert hematologists. A control group consisting of 100 healthy unrelated individuals was also included. One EDTA-treated blood sample was collected from each studied patients and plasma was isolated. The molecular prevalence of cytomegalovirus and HHV8 evaluated was evaluated using real-time and nested PCR protocols, respectively. The SNPs of the IL10 (rs 1800896-1082G/A) cytokine gene was evaluated by PCR-RFLP method. RESULTS: Cytomegalovirus and HHV8 infections were found in 2 and 3 of studied patients with transient bone marrow suppression. Significant higher frequency of IL10 G allele and GG genotype were found in HHV8-infected patients comparing to uninfected ones. Higher frequencies of A allele and AG and AA genotypes of IL10 were found in cytomegalovirus-uninfected patients comparing to infected ones, respectively. The significant higher frequencies of IL10 AA and AG genotypes were found in controls compared to bone marrow suppressed patients. CONCLUSION: IL10 genetic polymorphism might have determinative role in resistance to the cytomegalovirus, especially HHV8 infections, in patients with bone marrow suppression. Focus in new interaction between HHV8 infection and IL10 genetics in bone marrow suppressed patients should be completed by the analysis of the anti-herpes virus immunity in future studies.

MMP-2 gene expression at mRNA level in HBV and HCV infected patients.

Matrix metalloproteinases (MMPs) family play a determinative role in the development of liver fibrosis, metastasis, unregulated angiogenesis, and tumor growth. In this study the possible association between the MMP-2 gene expression level and risk of chronic hepatitis B virus (HBV) or hepatitis C virus (HCV) infections were evaluated in liver transplanted patients. Formalin fixed paraffin embedded (FFPE) liver tissue samples were collected from 225 transplant patients between years 2012 and 2016. The presence of HBV and HCV infections were analyzed in patients studied using molecular and immunologic diagnostic protocols according to the instructions of the manufacturers. Patients were divided to HBV, HCV, and HBV with HCV co-infected groups. A healthy control group was also included. For the quantitative analysis of MMP-2 mRNA gene expression an in-house-SYBR Green Real-Time PCR method was performed. The level of MMP-2 mRNA expression showed a significant increase in all studied viral hepatitis infected patient groups in comparing with healthy controls. The MMP-2 gene expression level increased in HBV infected patients when compared with HCV and HBV with HCV co-infected patients, but not significantly. Results showed a significant increase in MMP-2 expression level in all viral hepatitis single and coinfected liver transplanted patients when compared with the controls and also in HBV infected patients when compered with other viral infected ones, need to confirm in further completed studies.

Association between IRF1 Gene Expression and Liver Enzymes in HBV-infected Liver Transplant Recipients with and without Experience of Rejection.

BACKGROUND: Liver function indices and anti-viral immune regulatory markers can both improve graft outcomes, which lead to better post-transplantation management and increase the possibility of surveillance in liver transplant recipients with chronic hepatitis B virus (HBV) infection. OBJECTIVE: To determine the association between the interferon regulatory factor 1 (IRF1) mRNA levels and liver enzymes in HBV-infected liver transplant recipients with and without experience of rejection. METHODS: A total of 46 chronic HBV-infected patients who had undergone liver transplant surgery was divided into 2 groups of recipients "with rejection" and "without rejection.". Blood samples were collected form each patient on days 1, 4, and 7 post-transplantation. A SYBER GREEN real-time PCR was used to evaluate the expression level of IRF1 in liver recipients. Liver enzyme activities were also measured in all patients. RESULTS: The expression of IRF1 in the patients with rejection was up-regulated at all 3 follow-up days compared with those without rejection. The serum levels of ALT and AST were more than normal levels at 3 follow-up times in both study groups. Significant differences were found in IRF1 gene expression levels and also serum ALT levels between those with and without rejection after 7 days post-transplantation. CONCLUSION: The IRF1 expression and serum ALT levels were increased significantly in patient with rejection compared to those without rejection. IRF1, an inflammatory factor, may also intensify induction of inflammatory pathways in engrafted liver and promote liver inflammation and injuries leading to liver enzymes elevation in patients with graft rejection.

MMP-2 gene expression at mRNA level in HBV and HCV infected patients

@#Matrix metalloproteinases (MMPs) family play a determinative role in the development of liver fibrosis, metastasis, unregulated angiogenesis, and tumor growth. In this study the possible association between the MMP-2 gene expression level and risk of chronic hepatitis B virus (HBV) or hepatitis C virus (HCV) infections were evaluated in liver transplanted patients. Formalin fixed paraffin embedded (FFPE) liver tissue samples were collected from 225 transplant patients between years 2012 and 2016. The presence of HBV and HCV infections were analyzed in patients studied using molecular and immunologic diagnostic protocols according to the instructions of the manufacturers. Patients were divided to HBV, HCV, and HBV with HCV co-infected groups. A healthy control group was also included. For the quantitative analysis of MMP-2 mRNA gene expression an in-house-SYBR Green Real-Time PCR method was performed. The level of MMP-2 mRNA expression showed a significant increase in all studied viral hepatitis infected patient groups in comparing with healthy controls. The MMP-2 gene expression level increased in HBV infected patients when compared with HCV and HBV with HCV co-infected patients, but not significantly. Results showed a significant increase in MMP-2 expression level in all viral hepatitis single and coinfected liver transplanted patients when compared with the controls and also in HBV infected patients when compered with other viral infected ones, need to confirm in further completed studies.

Improving the potency of DNA vaccine encoding HIV-1 Nef antigen using two endogenous adjuvants in mouse model.

BACKGROUND: DNA immunization can induce long-term immune responses, which are required to design an effective HIV vaccine. It was shown that antigen-expressing plasmids can increase the protective immunity against infectious diseases such as: influenza and malaria. However, DNA-based immunizations have poor immunogenicity, thus the use of potent immunoadjuvants can enhance their potency. METHODS: In the current study, preparation of the recombinant HIV-1 Nef, Gp96 and HMGB1 DNA constructs was performed in bacterial system. Then, the immunogenicity of DNA construct harboring HIV-1 Nef gene (pcDNA-Nef) was studied using two endogenous adjuvants (pcDNA-HMGB1 and pcDNA-Gp96) in BALB/c mouse model. RESULTS: Our data showed that co-injection of pcDNA-Nef with pcDNA-HMGB1 effectively raised both humoral and cell-mediated immune responses in mice as compared to pcDNA-Nef adjuvanted with pcDNA-gp96. Indeed, co-immunization of HIV-1 Nef DNA with HMGB1 DNA significantly induced high levels of IgG2a and IFN-γ directed toward Th1 responses and also cytotoxic T lymphocytes (CTLs) activity in comparison with other immunized groups. CONCLUSION: These findings suggest that the full length of HMGB1 gene could be a more efficient adjuvant for improvement of therapeutic HIV DNA-based immunization compared to the full length of gp96 gene (Tab. 1, Fig. 3, Ref. 58).

Interleukin-28B rs12979860 C/T Polymorphism and Acute Cellular Rejection after Liver Transplantation.

BACKGROUND: Interleukin-28 (IL-28B) rs12979860 C/T polymorphism is a known predictor of sustained virological response after antiviral treatment in hepatitis C. IL-28B affects the innate immune system as well as intrahepatic expression level of interferon-stimulated genes. OBJECTIVE: To investigate the effect of recipient IL-28B polymorphism on occurrence of acute rejection after liver transplantation. METHODS: 140 liver allograft recipients were selected. Acute rejection episodes were recorded in 39 patients (AR group); the remaining had normal graft function (non-AR group). 70 normal subjects were also studied as the control group. The IL-28B rs12979860 was genotyped through PCR-RFLP method. RESULTS: No significant difference was found between AR and non-AR groups in terms of genotype and allele frequency. However, the CC genotype was significantly (p<0.001) more frequent in patients than in the control group; the C allele variants increased the risk of end-stage liver disease (OR: 2.60). CONCLUSION: Liver damage in association with the carriage of IL-28B C allele is associated with a higher likelihood of developing cirrhosis.

Theoretical and Instrumental Studies of the Competitive Interaction Between Aromatic α-Aminobisphosphonates with DNA Using Binding Probes.

Fluorescence spectroscopy, UV-visible absorption spectroscopy, circular dichroism (CD) spectroscopy, viscometry, cyclic voltammetry (CV), and differential pulse voltammetry (DPV) were applied to investigate the competitive interaction of DNA with two aromatic α-aminobisphosphonates and neutral red dye (NR, intercalator) and Hoechst (Ho, groove binder) as spectroscopic probes, in a Tris-hydrogen chloride buffer solution (pH 7.4). The principal component analysis (PCA) was applied to determine the number of chemical components presented in complexation equilibrium of DNA with the aromatic α-aminobisphosphonates (B1 and B2). The spectroscopic and voltammetric studies showed that the groove binding mode of interaction is predominant in the solution containing DNA and α-aminobisphosphonates. Furthermore, the results indicated that α-aminobisphosphonate with the lengthy N-alkyl chains had a stronger interaction. The PCA and theoretical quantum mechanical and molecular mechanic methods were also utilized to determine the structure of DNA with the two α-aminobisphosphonates (B1 and B2).

Association of TLR4 polymorphisms and polyomavirus BK infection in liver transplant patients.

Toll-like receptors (TLRs) may have a role in orchestrating the immune responses against polyomavirus BKV and also may influence liver transplant outcomes. However, the clinical relevance of this experimental observation has not been examined. Improving knowledge regarding details of genetic source of TLR polymorphisms can promote new therapeutic strategies to inhibit virus related clinical disorders in post-liver transplantation. Therefore, the Asp299Gly and Thr399Ile TLR4 polymorphisms were evaluated in liver transplanted patients with and without polyomavirus BK infection. In a cross sectional study, 144 liver transplant patients received allograft at Transplant Center of Namazi Hospital affiliated to Shiraz University of Medical Sciences who were recruited between years: 2014- 2015. Patients were followed for the graft outcome and acute rejection episode(s) and divided into two groups based on experiencing acute rejection or not. The genomic DNA of polyomavirus BK was diagnosed in studied patient using qualitative nested- PCR technique. Analysis of TLR4 gene polymorphisms were analyzed using PCR-RFLP protocols. The polyomavirus BK infection was found in 15 of 144 (10.4%) liver transplanted patients. A total of 14 of 15 (93.3%) and all of polyomavirus BK infected patients have been shown to be homozygous wild type AA genotype of TLR4-Asp299Gly (A896G) and CC genotype of TLR4- Thr399Ile (C1196T) polymorphisms. Homozygous mutant GG genotype of Asp299Gly (A896G) was found in 3 (2.1%) of the studied patients. Homozygous mutated TT genotype of Thr399Ile (C1196T) was found in only 5 (3.5%) of the liver recipients. There were no significant differences between homozygous wild type genotypes of studied TLR4 SNPs for liver transplant patients with or without polyomavirus BK infections. Significant association was also not found between homozygous mutated genotype of TLR4 SNPs for patients experiencing rejection episodes. However further completed studies on larger population and with longer follow-up are needed to confirm these results.

CXCL9 expression and polyomavirus BK infectivity in renal transplant patients with nephropathy.

Polyomavirus BK is an important risk factor for nephropathy and renal lose after kidney transplantation. CXCL9 is a key immunoregulatory molecule which participates in stimulation and migration of immune cells to the infected sites. Thus, the main aim of this study was to evaluate the expression levels of CXCL9 mRNA and serum levels in the infected polyomavirus BK infected renal transplant patients with and without nephropathy compared with healthy controls. This cross sectional study was performed on three studied groups including: polyomavirus BK infected vs. non-infected renal transplant patients with nephropathy and healthy controls. The mRNA and serum levels of CXCL9 were evaluated on the studied patient and control samples using an in-house comparative real time PCR and ELISA methods, respectively. The mRNA expression and serum levels of CXCL9 were both increased in polyomavirus BK infected compared with non-infected renal transplant patients and also in comparing with healthy controls. This upregulation was significant in the serum level in polyomavirus BK infected vs. non-infected patients and also in comparing with controls. According to these results, polyomavirus BK can induce renal complications via stimulation of inflammatory biomarkers like chemokine. Confirmation of the increasing of the expression and production of CXCL9 as a pro-inflammatory chemokine in renal transplanted polyomavirus BK infected patients with nephropathy need to confirm in further completed studies with longer follow-up.

Generation of Islet-like Cell Aggregates from Human Adipose Tissue-derived Stem Cells by Lentiviral Overexpression of PDX-1.

BACKGROUND: Pancreatic duodenal homeobox1 (PDX-1) is a transcription factor that is important in regulating pancreas development and maintaining ß-cell function. ß-cell replacement is an effective approach for the treatment of type 1 diabetes. Human adipose-mesenchymal stem cells (hAMSCs) are the ideal population cells for differentiating into insulin-producing cells. OBJECTIVE: To determine if islet-like cell aggregates production could be generated from hAMSCs by lentiviral overexpression of PDX-1. METHODS: After isolation of hAMSCs, characteristics of these cells were identified by flow-cytometic analysis and multilineage differentiation studies. PDX-1 gene delivered into hAMSCs through lentiviral vector for differentiating hAMSCs into insulin-producing cells (IPCs) at the utilized protocol for 14 days. Characteristics of IPCs were evaluated by immunocytofluorescence, dithizone staining, and quantitative reverse transcription PCR. In response to high glucose medium, insulin release was detected by chemiluminescence enzyme immunoassay. RESULTS: The islet-like cell aggregates appeared about 10 days after introduction of PDX-1 into hAMSCs. PDX-1 induced its own expression (auto-induction), a number of islet-related genes such as Ngn3, Nkx2-2, and insulin. The insulin-positive cells were detected in the PDX-1 transduced cells. In response to glucose challenge test, secretion of insulin hormone in the medium with high glucose concentration significantly increased in the PDX-1-transduced cells related to medium with low glucose concentration. CONCLUSION: Introduction of lentiviral PDX-1 significantly induces hAMSCs to differentiate into islet-like cell aggregates, which may provide a source of adipose stem cells-derived insulin-producing cells for cell replacement therapy in type 1 diabetes.

Frequency of Polyomavirus BK Infection in Kidney Transplant Patients Suspected to Nephropathy.

BACKGROUND: Polyomavirus BK is a major cause of nephropathy in immunosuppressed transplanted patients. Non-invasive diagnostic protocols such as molecular detection of polyomavirus BK replication are a useful strategy to predict BK virus-associated nephropathy (BKVAN). OBJECTIVE: To determine the prevalence of polyomavirus BK infection among kidney transplant patients suspected to have BKVAN. METHODS: In a cross-sectional study 108 kidney transplanted patients whose laboratory and clinical presentation were in favor of nephropathy between 2010 and 2012, were enrolled for analysis. Polyomavirus BK replication was evaluated in plasma and tissue samples of studied patients using a quantitative real-time PCR. Active cytomegalovirus infection was analyzed in studied patients using antigenemia method. A possible association between polyomavirus BK infection with clinical and laboratory risk factors of BKVAN were evaluated. RESULTS: The polyomavirus BK replication was found in 17 (15.7%) of 108 of plasma and 9 (11%) of 82 tissue samples in kidney transplanted patients. Cytomegalovirus co-infection was found in 3 of 17 and 3 of 9 plasma and tissue samples in polyomavirus BK infected patients, respectively. Significant associations were found between polyomavirus BK infection with tubulointerstitial nephritis and acute cellular rejection, as important pathologic findings of BKVAN. CONCLUSION: Diagnosis of single and co-infection of polyomavirus BK infection in plasma samples is a useful assay to evaluate the risk of BKVAN in kidney transplant patients. Established threshold values for studied viral infections have beneficial use in screening of kidney transplant patients at risk of BKVAN, need to confirm and standardized in completed further studies.

Polymorphism of the IL-18 and CD40 genes and Liver Transplant Outcome in Iranian Patients.

BACKGROUND: Cytokines and co-stimulatory molecules are important factors determining the outcome of transplantation. OBJECTIVE: To investigate the effect of IL-18 and CD40 gene polymorphisms on the outcome of liver transplantation. METHODS: 150 liver transplant recipients were included in this study. Alleles and genotypes frequencies for IL-18 (rs1946519) and CD40 (rs1883832) were determined in 28 acutely rejected (AR group) and 122 non-acutely rejected (non-AR group) liver transplant recipients. IL-18 and CD40 gene polymorphisms were evaluated by PCR-RFLP methods. RESULTS: There were no significant associations between IL-18 and CD40 polymorphism with acute rejection in liver transplant patients. IL-18TT and TG genotypes had a significant association with rejection in women compared to men. After grouping the liver recipients according to living vs cadaver donors, a significant association was found between CC genotype of CD40 and rejection in male living donor recipients. IL-18 TG genotype had a significant association with rejection in female cadaver donor recipients. CONCLUSION: There is no correlation between all genotype and alleles of IL-80 and CD40 polymorphism and the outcome of liver transplantation. However, gender and type of donor affect the correlation between all genotype and alleles of IL-18 and CD40, and the outcome of liver transplantation.

Differential Expression of Insulin-Like Growth Factor-I Receptor on Human Bone Marrow-Derived Mesenchymal Stem Cells Induced by Tumor Necrosis Factor-α.

BACKGROUND: Cell-based therapy has been implicated in the treatment of liver diseases. Mesenchymal stem cells from various sources such as bone marrow are available. These cells are one of the major candidates in cell therapy. The production of insulin-like growth factor-I increases in the regenerating organ. The insulin-like growth factor-I in liver regeneration is effective after binding to insulin-like growth factor-I receptor. OBJECTIVE: To test our hypothesis that tumor necrosis factor-α can stimulate mesenchymal stem cells to express insulin-like growth factor-I receptor. METHODS: Bone marrow was aspirated from normal human donor after taking informed consent. Cells were isolated and cultured. Identification of cells was done by flowcytometry and functional tests. The fourth passage cells were treated with tumor necrosis factor-α at two doses of 1 and 10 ng/mL, and incubated for 2, 10, 24, and 48 hours. Insulin-like growth factor-I receptor gene expression was studied using real-time polymerase chain reaction. RESULTS: Flowcytometry showed that the human bone marrow mesenchymal stem cells were positive for CD90 and negative for CD45 and CD80. The insulin-like growth factor-I receptor gene expression was increased in tumor necrosis factor-α treated in comparison with untreated cells. CONCLUSION: Treatment of human bone marrow-derived mesenchymal stem cells with tumor necrosis factor-α increases gene expression of insulin-like growth factor-I receptor. This finding may be used for increasing the effectiveness of stem cell therapy in those with acute hepatic failure.

The molecular prevalence of viral infections in transplant candidates with bone marrow suppression, shiraz, southern iran, 2010.

BACKGROUND: Transient bone marrow suppression, characterized by acute inability of the bone marrow to produce circulating blood cells, may strongly relate to the pathogenesis of some viral infections. OBJECTIVE: To study the prevalence of some DNA and RNA viruses in patients with transient bone marrow suppression. METHODS: EDTA-treated blood samples were collected from 27 patients with clinically- and laboratory-confirmed transient bone marrow suppression. The genomic DNA of hepatitis B virus, adenovirus, polyomavirus BK, and parvovirus B19, and genomic RNA of hepatitis C and G viruses were extracted and amplified by sensitive and specific in-house simple and nested PCR and RT-PCR protocols, respectively. The risk factors that might be related to the studied viral infections were analyzed. RESULTS: Hepatitis B virus infection was diagnosed in 9 (33%) of 27 patients; adenovirus infection in 2 (7%); and parvovirus B19 infection in 7 (26%) of 27 patients. The genomic DNA of polyomovirus BK was not detected in any patients. Both hepatitis C and G viruses were found in 3 (11%) of 27 patients. CONCLUSION: Diagnosis of the high prevalence of hepatitis B virus, and parvovirus B19 in patients with transient bone marrow suppression, reflects the importance of these viral infections in introducing bone marrow suppression. This hypothesis should be confirmed in further studies.