Browsing by Author "Ramzi, Mani"

Now showing 1 - 4 of 4

Diagnosis and Monitoring of Human Cytomegalovirus Infection in Bone Marrow Transplant Recipients by Quantitative Competitive PCR
(Başkent Üniversitesi, 2006-06) Ziyaeyan, Mazyar; Sabahi, Farzaneh; Alborzi, Abdolvahab; Mahboudi, Fereidoun; Kazemnejad, Anooshirvan; Ramzi, Mani; Moravej, Ali; Jaberi, Marjan Mojtahed
Objectives: Human cytomegalovirus (HCMV) is a common cause of infection worldwide. Severe cytomegalovirus disease is usually observed in immunodeficient individuals such as bone marrow transplant (BMT) or AIDS patients. In these patients, proof of viral presence is not enough for making clinical decisions; one must report the quantity of virus or viral load in appropriate clinical specimens to demonstrate the relationship between disease severity and HCMV infection. The goal of this study was to use quantitative competitive polymerase chain reaction (PCR) to determine HCMV viral load in 26 BMT recipients. Materials and Methods: Peripheral blood was collected weekly for 100 days from 26 BMT recipients. Qualitative and quantitative competitive PCRs on 105 mononuclear cells were performed for each patient. The same tests were performed once for each of 26 donors. In addition, the anti-HCMV humoral response was detected by performing IgM and IgG ELISAs in donors and recipients prior to transplantation. Results: Of 26 BMT donors and recipients, 25 and 26 were IgG positive, and 2 and 6 had HCMV-specific IgM antibodies, respectively. From 313 total clinical specimens tested, 255 had positive qualitative PCR results. Results of quantitative PCR on the same specimens demonstrated that in 14 patients, viral copy number per 105 cells had increased, pointing toward HCMV reactivation. In others, changes in viral copy number were mostly around 100/105 cells, with an upper limit of 300/105 cells. Conclusions: Owing to the high prevalence of cytomegalovirus in our country, the chance of viral reactivation and HCMV infection/disease upon transplantation must be seriously considered. Therefore, use of quantitative PCR in PCR-positive patients is highly recommended to demonstrate active infection that may lead to HCMV disease during the posttransplant period. This also could help physicians begin pre-emptive therapy that would be for a shorter treatment period and provide for better outcomes in infected BMT patients.
Interleukin-10 Gene Polymorphism in Bone Marrow Transplant Recipients
(Başkent Üniversitesi, 2008-03) Azarpira, Negar; Geramizadeh, Bita; Darai, Masumeh; Aghdaie, Mahdokht Hossein; Ramzi, Mani
Objectives: Graft-versus-host disease is the main complication after hematopoietic stem cell transplant, occurring even after donor and recipient human leukocyte antigen matching, apparently because of donor/recipient minor histocompatibility antigen mismatches and cytokine polymorphisms. Interleukin-10 suppresses several activities of the immune response by inhibiting T helper 1 and T helper 2 cells. These properties suggest that interleukin-10 could act as a suppressive mediator and prevent graft-versus-host disease. This study evaluates the association between the interleukin-10 promoter gene polymorphism and transplant outcomes among 18 recipients of cytokine-mobilized peripheral blood stem cells from human leukocyte antigen-matched sibling donors. Materials and Methods: We analyzed 3 single-nucleotide polymorphisms in the proximal region of the interleukin-10 promoter gene (-1082/-819/-592) by the amplification refractory mutation system and polymerase chain reaction-restriction fragment length polymorphism methods. Eighteen donors and their recipients who had undergone an allogeneic peripheral blood stem cell transplant at the Bone Marrow Transplant Center in Nemazi Hospital (Shiraz, Southern Iran) between September 2005 and September 2006 were enrolled. Results: The GCC haplotype (1082*G/819*C/592*C) was predominant in both the donor and the recipient, but no significant correlations were present between the GCC haplotype in either the donor or the recipient and the risk of acute graft-versus-host disease (P = .56). Conclusions: The interleukin-10 promoter gene polymorphism was found not to be associated with acute graft-versus-host disease in patients after an allogeneic peripheral blood stem cell transplant from human leukocyte antigen-matched sibling donors. Additional studies with larger samples are necessary to further define the influence of interleukin-10 on the immune response after bone marrow transplant.
Methylenetetrahydrofolate Reductase C677T Genotypes and Clinical Outcome Following Hematopoietic Cell Transplant
(Başkent Üniversitesi, 2007-12) Azarpira, Negar; Geramizadeh, Bita; Darai, Masumeh; Aghdaie, Mahdokht Hossein; Ramzi, Mani
Objective: Methotrexate may be used as a prophylactic agent against graft-versus-host disease in hematopoietic cell transplant. The drug exerts its effect on folate metabolism; 5,10-methylenetetrahydrofolate reductase is a critical enzyme involved in this cycle and is related to the toxicity of methotrexate. Methods: We examined the association of a single nucleotide polymorphism at position 677 in the 5,10-methylenetetrahydrofolate reductase gene and the clinical outcomes of patients treated with allogeneic hematopoietic cell transplant. Genotyping of 5,10-methylenetetrahydrofolate reductase was performed by polymerase chain reaction-restriction fragment length polymorphism on 30 patients receiving hematopoietic cell transplant and their HLA-matched related donors. Patients were given a short course of methotrexate as prophylaxis to prevent graft-versus-host disease. Results: Donors and recipients who carried a 677T allele showed mildly higher total bilirubin, aspartic transaminase, and alanine transaminase levels, but these increases above the normal values were not statistically significant (P > .05). The platelet recovery to 20 000/µL and granulocyte recovery to 500/µL were slower for patients who carried a 677T allele, but these correlations also were not statistically significant. The 5,10-methylenetetrahydrofolate reductase genotypes of neither the donors nor the recipients had any effect on the incidence of acute graft-versus-host disease. Conclusions: No association was observed between the C677T polymorphism and the outcome parameters for any of the different genotypes studied here. Additional studies with larger samples are necessary to further elucidate the influence of 5,10-methylenetetrahydrofolate reductase genotyping on clinical outcomes of patients treated with hematopoietic cell transplant who receive methotrexate.
Quantification of Human Cytomegalovirus DNA by a New Capture Hybrid Polymerase Chain Reaction Enzyme-Linked Immunosorbent Assay in Plasma and Peripheral Blood Mononuclear Cells of Bone Marrow Transplant Recipients
(Başkent Üniversitesi, 2008-12) Ziyaeyan, Mazyar; Kadivar, Mohammad; Pourabbas, Bahman; Mahboudi, Fereidoun; Ramzi, Mani; Alborzi, Abdolvahab; Sabahi, Farzaneh
Objectives: Quantitative monitoring of human cytomegalovirus infections is helpful in determining appropriate antiviral management in patients who receive bone marrow transplants. We sought to design and evaluate a new cytomegalovirus capture hybrid polymerase chain reaction enzyme-linked immunosorbent assay (PCR-ELISA) in plasma and peripheral blood mononuclear cells to monitor cytomegalovirus infection in bone marrow transplant recipients. Patients and Methods: Twenty-six patients who received allogeneic bone marrow transplants, including 17 male patients and 9 female patients (9 adults, 17 children), were enrolled in this study. A total of 313 consecutive whole blood specimens, before and from 7 to 120 days after transplant, was evaluated in the study. A newly designed biotinylated probe-mediated quantitative competitive PCR-ELISA test was used to determine cytomegalovirus load in specimens of peripheral blood mononuclear cells and plasma. Results: All 26 patients were cytomegalovirus seropositive before transplant. Capture hybrid PCR-ELISA of peripheral blood mononuclear cells detected cytomegalovirus DNA in 287 of 313 specimens (91.7%) even in cases with no active cytomegalovirus infection. in plasma, cytomegalovirus DNA was detected in 114 of 313 specimens (36.4%). Increasing titers of cytomegalovirus DNA were detected in 14 of 26 patients (53.8%). Conclusions: The quantitative capture hybrid PCR-ELISA was able to diagnose and monitor cytomegalovirus infection in patients who received bone marrow transplants. Detection of cytomegalovirus DNA in plasma was more predictive of the onset of cytomegalovirus-related clinical symptoms, compared to detection in peripheral blood mononuclear cells.