Angiotensinogen, angiotensine converting enzyme and plasminogen activator inhibitor-1 gene polymorphism in chronic allograft dysfunction.

Despite dramatic improvements in first-year patient and graft survival rates, chronic allograft dysfunction (CAD) remains the leading cause of late renal allograft loss, while current immunologic strategies have little effect on this condition. The renin-angiotensin system (RAS) plays an important role in progression of chronic renal disease. It was shown that plasminogen activator inhibitor-1 (PAI-1) functions in the RAS. This study investigates the possible links between angiotensinogen (AGT M235T), angiotensin-converting enzyme (ACE) and PAI-1 genotypes with CAD. Assessments of polymorphism were performed in 127 renal allograft recipients (77 with CAD and 50 with normal renal function). Fifty healthy subjects were also considered for comparison. Genotypes were determined using polymerase chain reaction (PCR) sequence-specific primers and PCR followed by restriction fragment length polymorphism analysis. Kidney recipients with CAD had significantly higher frequencies of the TT than the recipients without CAD (P < 0.05). The transplant recipients with CAD also had significantly higher frequencies of the DD genotype than those without CAD (P < 0.05). No significant differences were observed between the allelic and genotypic distributions of PAI-1 polymorphisms. Therefore, determination of AGT M235T and ACE genotypes prior to transplantation may be useful to identify patients who are at risk for chronic renal transplant dysfunction.

Association between cyclosporine concentration and genetic polymorphisms of CYP3A5 and MDR1 during the early stage after renal transplantation.

OBJECTIVES: Cyclosporine (CsA) has a narrow therapeutic range, and its pharmacokinetic characteristics vary among individuals. It also is a substrate for cytochrome P450 (CYP) 3A and P-glycoprotein, the product of the multidrug resistance 1 (MDR1) and CYP3A5 genes. Some of the single nucleotide polymorphisms (SNPs) in these genes are associated with deficient protein expression and reduced in vivo activity. We postulated that in renal transplant recipients, these SNPs should be associated with interindividual variations in CsA pharmacokinetics. MATERIALS AND METHODS: In 88 Iranian renal transplant patients receiving CsA, CYP3A5 and MDR1 genotypes were determined by polymerase chain reaction, followed by restriction fragment length polymorphism analysis. Whole blood trough CsA concentrations were measured by radioactive immunosorbent assay. The dose-adjusted concentration (ng/mL per mg/kg/d) was calculated at 1 day (+/-2 days), 7 days, and 1 month after transplantation. RESULTS: The MDR-1 wild-type genotype (3435CC) was observed in 17 patients (19%), whereas 45 patients (51%) were heterozygous (3435CT), and 26 patients (30%) were homozygous (3435 TT) for the mutation. In the days immediately after transplantation, we found a correlation between the concentration/dose ratio and the exon 26 MDR single nucleotide polymorphisms (33.3+/-15.24 microg mg/L/kg in the CT group vs 44.1+/-28.4 microg mg/L/kg in the TT group, P=.019). This ratio was significantly higher in subjects homozygous for the mutation (3435TT). This significant difference was not seen 1 week or 1 month after transplantation. All patients had the CYP3A5*3/*3 genotype, so no differences among the CYP3A5*1/*3 genotypes were found. CONCLUSIONS: MDR-1 (3435CC) polymorphisms are associated with CsA pharmacokinetics and dose requirements in the first few days after renal transplantation. Pharmacogenetic methods could be used to help select the initial dosage and individualize immunosuppressive therapy. According to our results, the major genotype of our recipients is CYP3A5*3/*3. According to the literature, the recommended starting dosage of CsA is 9-14 mg/kg/day; however, the Iranian population has a good response with lower dosages (3-5 mg/kg/day), which may be explained by genetic differences.

Cytokine gene polymorphisms in renal transplant recipients.

OBJECTIVE: Acute rejection remains an important cause of graft loss after renal transplantation, and cytokines are key mediators in the induction and effector phases of all immune and inflammatory responses. However, the influence of gene polymorphisms on the functional immune response of transplant recipient outcomes remains controversial. MATERIALS AND METHODS: The amplification refractory mutation system polymerase chain reaction was used to detect the interleukin-10 (IL-10) (-1082 G/A), tumor necrosis factor-alpha (TNF-alpha) (-308 G/A), and interferon-gamma (IFN-gamma) (+874 T/A) single nucleotide polymorphisms in 100 of the first adult kidney recipients at our institution who were receiving cyclosporine-based immunosuppressive therapy. The diagnosis of acute rejection was based on clinical and histologic findings according to the Banff criteria. RESULTS: The results of multivariate analyses showed no significant association between episodes of acute rejection and single nucleotide polymorphisms in IL- 10, TNF-alpha genes, or dinucleotide repeat polymorphisms in the IFN-gamma gene. CONCLUSIONS: Our results demonstrate that cytokine gene polymorphisms did not influence the early outcome of kidney transplantation.

Risk of viral transmission via bone marrow progenitor cells versus umbilical cord blood hematopoietic stem cells in bone marrow transplantation.

Hematopoietic stem cell transplantation (HSCT) is the treatment of choice for children and certain adults with malignant and nonmalignant hematologic disease. Since viral infections are the major problem, this study examined those that might potentially be transmitted to HSCT recipients via bone marrow (BM) versus umbilical cord blood (UCB). BM progenitor cells, peripheral blood leukocytes, and plasma samples were collected from 30 allogenic BM donors. Umbilical cord blood hematopoietic stem cells and plasma samples were also collected from 34 UCB donors. Viral DNA extracted and purified from collected specimens was processed using nested polymerase chain reactions (PCR) to detect human parvovirus B19 (HPV B19), human herpesvirus-6 (HHV-6), varicella-zoster virus (VZV), human cytomegalovirus (HCMV), and Epstein-Barr virus (EBV). The prevalences of HCMV DNA in collected BM progenitor cells versus UCB hematopoietic stem cells were 73% versus 23%, respectively. Conversely, HHV-6 DNA was not detected in any collected specimen by simple PCR. Distribution of the other investigated virus DNAs except EBV DNA was similar in specimens collected from both groups. EBV DNA was not determined in UCB hematopoietic stem cells. The results indicate that the risk of viral transmission to BM transplant recipients via UCB hematopoietic stem cells is less than that with BM progenitor cells.

Outcome of hepatitis B and C virus infection on graft function after renal transplantation.

INTRODUCTION: Chronic liver disease resulting from hepatitis B virus (HBV) and hepatitis C virus (HCV) infections is still a major concern in kidney recipients. It is unclear whether HCV antibody status and markers of HBV infection are associated with renal dysfunction. Thus, we designed a study to investigate the incidence of HBV and HCV infection after renal transplantation and whether these infections alter graft function. METHODS: Fifty-eight patients who underwent renal transplantation participated in the study. Serum creatinine and aminotransferase levels were measured with standard automated analyzers. Anti-HCV antibodies were detected with an enzyme immunoassay, and a reverse transcriptase-polymerase chain reaction (RT-PCR) technique was used to test for HCV-RNA. Serological markers for HBV (HBsAg and anti-HBc antibody) were detected by enzyme immunoassay. All samples from patients who were seropositive for HBsAg or anti-HBc antibody were PCR-tested for HBV-DNA. A serum sample collected from living donors was tested for anti-HCV antibodies and serological markers for HBV. Serum creatinine and aminotransferase levels were also measured in living donors. RESULTS: Anti-HCV was not detected in serum samples of any cases before transplantation. However, 10 (17.2%) tested positive after transplantation. HCV-RNA was detected in 2 of the 10 patients (3.4% of all patients). None of the pretransplantation serum samples tested positive for HBsAg. However, anti-HBc antibody was identified in 8 (13.8%) of the 58 patients.. No HBV DNA was detected in serum samples of the patients with anti-HBc or HBsAg-positive. HBsAg was only detected in 1 (1.7%) recipient after transplantation. None of the 58 patients showed clinical signs or symptoms of renal dysfunction during the study period. CONCLUSION: Our data suggest that, neither HBV nor HCV infection appears to cause or contribute to renal dysfunction in the early period (1 year) after renal transplantation. Nevertheless, a long-term consequence of chronic HBV or HCV liver disease or graft loss is not impossible in renal transplant recipients.

The molecular and antigenic tissue impact of viral infections on liver transplant patients with neonatal hepatitis.

BACKGROUND: Pathogenesis of neonatal hepatitis relates to various underlying causes including viral infections. Both hepatotropic and non-hepatotropic viruses may induce liver failures in infants before birth, during delivery, or shortly after birth. OBJECTIVES: The tissue impact of HCMV, HSV, HBV, HCV, and rotavirus and adenovirus infections was evaluated in studied infants with neonatal hepatitis. METHODS: The history of viral infections was analyzed in paraffin-embedded biopsy and autopsy tissues of 22 infants with neonatal hepatitis between years 1996 and 2007, retrospectively. The tissue molecular presentation of HBV, HCV, HCMV, HSV, adenovirus, and rotavirus was evaluated by different qualitative simple and nested PCR and RT-PCR protocols. Immunohistochemistry (IHC) method was used for studying the antigenic prevalence of HSV-1, 2; HBV, HCMV and adenovirus infections. Also the laboratory liver indices of all patients with neonatal hepatitis were analyzed. RESULTS: The HBV and HSV genomes were detected in 3 (14%) of 22 infants. The rotavirus and HCV-RNA and also the HCMV-DNA were detected separately in 1 (4%) of 26 paraffin-embedded autopsy and biopsy tissues. The HBV and HSV-1 specific antigens were separately diagnosed in 1 (4%) of 26 neonatal samples by IHC protocols. Also the HSV-2 antigen was seen in 5 (23%) of 22 liver autopsy and biopsy specimens. Co-infections with HCMV, HSV, HBV, HCV, and rotavirus were detected in these infants with hepatitis. CONCLUSION: Diagnosis of single and mixed molecular and antigenic traces of HCMV, HSV, HBV, HCV and rotavirus underlines the etiologic role of these viruses in clinical pathogenesis of neonatal hepatitis.

Etiology of DNA virus infections in liver transplant recipients with neonatal hepatitis.

Neonatal hepatitis is a syndrome of symptoms associated with a history that includes any type of infectious, genetic, toxic, or metabolic causation. Various infectious agents have been implicated in hepatic inflammation in neonates including bacterial and viral pathogens, especially DNA viruses. We used molecular and antigenic methods to evaluate the role of DNA viruses, such as hepatitis type B viruses (HBV), human cytomegalovirus (HCMV), herpes simplex virus (HSV), and adenovirus, in neonatal hepatitis complications. Twenty-six paraffin-embedded biopsy and autopsy tissues obtained between 1996 and 2007 from 22 infants with neonatal hepatitis were studied retrospectively. The genome prevalence of HBV, HCMV, HSV, and adenovirus were analysed using qualitative polymerase chain reaction (PCR) protocols. The antigenic presentation of HSV-1, HSV-2, HBV, HCMV, and adenovirus were evaluated using immunohistochemistry (IHC) methods. The HCMV genome was detected separately in 1 of 22 (4.5%) paraffin-embedded autopsy and biopsy tissues. Also 3/22 (13.6%) samples were infected with HBV and HSV genomes. HBV and HSV-1 antigens were present in 1/26 (4.5%) neonatal samples and HSV-2 antigens in 5/26 (22.7%) by IHC protocols, but adenovirus and HCMV antigens were not detected among samples from infants with neonatal hepatitis. Detection of separate co-infections of HSV, HCMV, and HBV genomes in autopsy and biopsy tissues of HBV and HSV-1 or HSV-2 antigens in these patients, showed the importance of these viral infections in clinical neonatal hepatitis.

Incidence of human herpes virus-6 and human cytomegalovirus infections in donated bone marrow and umbilical cord blood hematopoietic stem cells.

This study examined the incidence of human herpes virus-6 (HHV-6) and human cytomegalovirus (HCMV) infections that are potentially transmitted to haematopoietic stem cells (HSC) transplant recipients via bone marrow (BM) or umbilical cord blood (UCB). Bone marrow progenitor cells were collected from 30 allogenic BM donors. UCB HSC were collected from 34 subjects. The extracted DNA was then processed using nested polymerase chain reaction (nPCR) technique. HCMV and HHV-6 serological status were determined by enzyme immunoassay (EIA). Nested PCR identified HCMV in 22 (73%) of 30 samples of BM progenitor cells but in only eight (23.5%) of 34 samples of UBC HSC ( P = 0.001). HHV-6 DNA was detected in 11 (36.6%) of 30 BM progenitor cells and in only one (2.9%) of 34 UBC cells ( P = 0.002). Both HHV-6 and HCMV infections were determined in nine (26.5%) of 34 bone marrow samples. The results indicate that, the risk of HCMV and HHV-6 via BM progenitor cells is higher than transmission by UCB cells ( P= 0.04).