Cytomegalovirus infection in recipients of related and unrelated donor bone marrow transplants: no evidence of increased incidence in patients receiving unrelated donor grafts

Recent reports suggest an increased incidence of cytomegalovirus (CMV) infection in recipients of unrelated donor (UD) bone marrow transplantation (BMT).
In this study we have collated the incidence of CMV infection and disease in sequential UD ( n  = 119) and related donor (RD; n  = 79) BMT performed in a single institution over a 7-year period. Low-risk patients (CMV seronegative recipient and donor) accounted for 51% of UD BMT ( n  = 61) and 62% of RD BMT ( n  = 49), with CMV excretion documented in one RD BMT only. The remaining high-risk patients received identical prophylaxis regimens with aciclovir and intravenous immunoglobulin (IVIG). Two groups consisting of 58 UD BMT (median age 9.0 years, range 0.7–45.3 years) and 30 RD BMT (median age 13.6 years, range 1.6–47.6 years) were analysed. CMV reactivation/re-infection was documented in 15 UD BMT (26%) and 10 RD BMT (33%) ( P  = 0.72), and CMV disease in four UD BMT (8%) and four RD BMT (13%) ( P  = 0.533). In this series the risk of CMV excretion and disease following UD BMT was similar to that following RD BMT.     

Decreased serum level of antibody against human cytomegalovirus in patients with Behçet’s disease

Behçets disease (BD) is a connective tissue disorder characterized by recurrent orogenital ulcer, uveitis, and skin lesions. Recurrent aphthous ulcer is associated with human cytomegalovirus (HCMV). To investigate the possible role of HCMV in BD, we measured the titers of IgG, IgM, and IgA anti-HCMV antibodies in 73 Korean patients with BD, 50 with scleroderma, 70 with systemic lupus erythematosus, and 50 from healthy controls by indirect immunofluorescent staining. The titer of IgG anti-HCMV antibody was significantly lower in patients with BD than in controls (geometric mean 3115.4 vs 9687.6, P=0.0001 by Wilcoxons rank sum test), as was the titer of IgA anti-HCMV antibody (geometric mean 1.9 vs 15.7, P=0.0001, Wilcoxons rank sum test). In conclusion, we found significantly lower antibody responses to HCMV in patients with BD.     

Effective inhibition in animals of viral pathogenesis by a ribozyme derived from RNase P catalytic RNA

A functional RNase P ribozyme (M1GS RNA) was constructed to target the overlapping mRNA region of two murine cytomegalovirus (MCMV) capsid proteins essential for viral replication: the assembly protein (mAP) and M80. The customized ribozyme efficiently cleaved the target mRNA sequence in vitro. Moreover, 80% reduction in the expression of mAP and M80 and a 2,000-fold reduction in viral growth were observed in cells expressing the ribozyme. In contrast, there was no significant reduction in viral gene expression and growth in cells that either did not express the ribozyme or produced a “disabled” ribozyme carrying mutations that abolished its catalytic activity. When the ribozyme-expressing constructs were delivered into MCMV-infected SCID mice via a modified “hydrodynamic transfection” procedure, expression of ribozymes was observed in the livers and spleens. Compared with the control animals that did not receive any M1GS constructs or received the disabled ribozyme construct, animals receiving the functional ribozyme construct exhibited a significant reduction of viral gene expression and infection. Viral titers in the spleens, livers, lungs, and salivary glands of the functional ribozyme-treated SCID mice at 21 days after infection were 200- to 2,000-fold lower than those in the control animals. Moreover, survival of the infected animals significantly improved upon receiving the functional ribozyme construct. Our study examines the use of M1GS ribozymes for inhibition of gene expression in animals and demonstrates the utility of RNase P ribozymes for gene targeting applications in vivo.     

Serological evaluation of the role of cytomegalovirus in the pathogenesis of IDDM: a prospective study

Summary To study the possible temporal association between primary cytomegalovirus infection and the appearance of islet cell autoantibodies or the development of insulin-dependent diabetes mellitus (IDDM) cytomegalovirus antibodies were analysed from follow-up sera of 46 initially non-diabetic siblings of diabetic children who either manifested clinical IDDM (22 siblings) or turned islet cell antibody positive (24 siblings) during the prospective observation (mean follow-up time 2.9 years). Secondly, cytomegalovirus antibodies were analysed during pregnancy in 96 mothers whose child presented with IDDM before the age of 7 years and in 96 control mothers who gave birth to a non-diabetic child. Thirdly, a case-control series including 90 newly-diagnosed young children with IDDM and their 90 control subjects was analysed. No seroconversions were found in cytomegalovirus antibodies during the follow-up of the 46 siblings indicating no temporal association with islet cell antibody seroconversion or manifestation of clinical diabetes. During the follow-up 17 (37%) siblings were constantly seronegative and 29 (63%) seropositive for cytomegalovirus IgG and there was no difference between islet cell antibody positive and negative siblings. Cytomegalovirus IgG and IgM were not different in pregnant mothers who gave birth to a subsequently diabetic child compared to control mothers, or in newly-diagnosed diabetic children compared to control children. Cytomegalovirus IgA was higher in newly-diagnosed diabetic children than in control children (p<0.005). This difference disappeared when only cytomegalovirus IgG positive individuals were analysed. No correlation was found between islet cell antibodies and cytomegalovirus antibodies in newly-diagnosed diabetic patients. The results do not support the hypothesis that primary cytomegalovirus infections could initiate the cascade leading to autoimmune destruction of the beta cells.Abbreviations IDDM Insulin-dependent diabetes mellitus - ICA islet cell autoantibodies - CMV cytomegalovirus - EIA enzyme immunoassay - EIU enzyme immunoassay unit     

Risk factors for Pneumocystis carinii pneumonia in kidney transplant recipients: a case–control study

Objective. To analyze risk factors for Pneumocystis carinii pneumonia (PCP) in kidney transplant recipients. Study design. In a case–control study, 17 PCP cases diagnosed between July 1994 and July 2000 were matched with two controls each (previous and subsequent kidney transplant recipients who did not develop PCP during the same follow‐up period). Demographics, organ origin, human leukocyte antigen (HLA) mismatches, use of poly‐ or monoclonal anti‐CD3 antibodies (Po/MoAb) for induction or rejection treatment, rejection episodes, cumulative steroid dose for rejection treatment, immunosuppressive regimens, and other infections were analyzed. Results. No significant differences were seen in gender (male 10 vs. 15), mean age (39.7 vs. 35.4 years), organ origin (cadaver donor 13 vs. 19), HLA mismatches, or Po/MoAb use in induction treatment. Significant differences were observed in PCP cases for rejection history (P=0.02), and median and total number of rejection episodes (P=0.0018). The relative risks for PCP for 1, 2, and ≥3 rejection treatments vs. no such treatment were 1, 1.05, and 6.30, respectively (P=0.021). The relative risk for PCP for steroid‐resistant rejection was 4.34 (95% confidence interval [CI], 1.04–18.89) (P=0.019), and that for the use of Po/MoAb for rejection treatment was 7.23 (95% CI, 1.28–49.34) (P=0.006). The relative risk for PCP for 0, 1, and ≥2 previous or concomitant cytomegalovirus (CMV) infection vs. no such infections were 1.0, 2.32, and 13.0, respectively (P=0.012). The relative risks for PCP for tuberculosis (TB) was 18 (95% CI, 1.76–852.03), that for bacterial pneumonia was 14.22 (95% CI, 2.16–150.23), and that for hepatitis C virus infection was 5.25 (95% CI, 1.03–28.91). Immunosuppressive regimens with tacrolimus, mycophenolate mofetil (MMF), steroids (P=0.06), and MMF as a single variable (P=0.05) were more frequently used in cases. Primary trimethoprim‐sulfamethoxazole prophylaxis failure was observed in 12 patients in association with heavy immunosuppression and concomitant infections. Conclusions. The risk of PCP in kidney transplant recipients is related to the number and type of rejection treatments. It is also related to the occurrence of CMV infection, and to other immunomodulating infections such as TB and hepatitis C, and might also be increased with the use of newer and more potent immunosuppressive agents. Primary prophylaxis failure may occur in association with some of these risk factors.     

Prevention dof recurrent cytomegalovirus disease in renal and liver transplant recipients: effect of oral ganciclovir

Background: Although the primary treatment of symptomatic cytomegalovirus (CMV) disease in organ transplant recipients is successful in >90% of individuals, relapsing disease, particularly in those with primary infection, remains an important problem. Previously, we had observed that the rate of symptomatic recurrence was >60% in those with primary disease (seronegative for CMV prior to transplant), and approximately 20% in those who were seropositive prior to transplant. The present study was undertaken to determine whether a maintenance regimen of oral ganciclovir for 2–3 months added to the routine 14–21 days of intravenous ganciclovir would further prevent symptomatic CMV recurrence. Methods: From May 1995 until June 1998, all kidney and liver transplant recipients with confirmed tissue‐invasive CMV disease or CMV syndrome were treated with 14–21 days of intravenous ganciclovir (5 mg/kg b.i.d. with dose adjusted for renal dysfunction) followed by 2–3 months of oral ganciclovir (2 g daily). The incidence of recurrence of CMV disease and/or viremia during and after oral therapy was then determined over a mean follow‐up of 530.6 days. Results: Thirty‐seven patients, 19 kidney and 18 liver transplant recipients, were studied; 5 had biopsy‐proven tissue‐invasive disease (13.5) and 32 suffered a CMV syndrome (86.5). Twenty‐one of these patients (58.6) were seronegative for CMV prior to transplant and received an allograft from a seropositive donor (D+/R?). Overall, 10 patients (27.0) developed CMV recurrence. Eight of 21 patients who were D+/R? for CMV (38.1) developed recurrence as opposed to 2 of 16 patients with other serologic status (12.5) (P=0.14). Patients with recurrent CMV disease and/or viremia had a peak antigenemia assay titer during their initial CMV event of 319.2 positive cells/2 slides compared with 109.8 positive cells/2 slides for patients without recurrent CMV infection (P=0.14); the trend of having a higher peak antigenemia assay titer among patients who recurred occurred both in patients who were at risk of primary CMV infection (D+/R? for CMV) and in those who were not. Two patients developed recurrent infection with strains of CMV that were resistant to ganciclovir. Conclusions: This new therapeutic regimen of oral ganciclovir following intravenous ganciclovir slightly reduced the overall rate of recurrent CMV disease and/or viremia, but it still did not adequately prevent CMV recurrence in patients who are at risk of primary infection prior to transplant. Of particular concern, 2 patients with primary infection treated with this regimen developed ganciclovir‐resistant recurrent disease ( Note Presented in part at the American Society of Transplant Physicians Meeting, May 1999, Chicago, Illinois.
).     

Defensins: Natural component of human innate immunity

The widespread use of antibiotics has contributed to a huge increase in the number of resistant bacteria. New classes of drugs are therefore being developed of which defensins are a potential source. Defensins are a group of antimicrobial peptides found in different living organisms, involved in the first line of defense in their innate immune response against pathogens. This review summarizes the results of studies of this family of human antimicrobial peptides (AMPs). There is a special emphasis on describing the entire group and individual peptides, history of their discovery, their functions and expression sites. The results of the recent studies on the use of the biologically active peptides in human medicine are also presented. The pharmaceutical potential of human defensins cannot be ignored, especially considering their strong antimicrobial activity and properties such as low molecular weight, reduced immunogenicity, broad activity spectrum and resistance to proteolysis, but there are still many challenges and questions regarding the possibilities of their practical application.     

Allium sativum‐derived allitridin inhibits treg amplification in cytomegalovirus infection

This study investigated the effects of allitridin compound on murine cytomegalovirus (MCMV)‐induced regulatory T cell (Treg; CD4⁺CD25⁺Foxp3⁺) amplification in vivo and in vitro. One hundred twenty MCMV‐infected mice were allocated at random into two groups for treatment with allitridin or placebo. Another 120 mock‐infected mice were randomly allocated as controls for the allitridin treatment and placebo treatment groups. The mice were euthanized at various time points after infection (out to 120 days) to evaluate the effects of treatment on Treg presence and function, as well as MCMV infective load. Co‐culture with mouse embryo fibroblasts (MEF) and MCMV was performed to evaluate allitridin‐mediated Treg and anti‐CMV effects. The maximum tolerance concentration (MTC) of allitridin was used to treat cells for 3 days. Changes in Foxp3 mRNA and protein levels, percentages of T cell subsets, and Treg‐related cytokines (IL‐10 and TGF‐β) were measured. Allitridin treatment did not influence Foxp3 expression and Treg proportion in uninfected mice, but did down‐regulate each in infected mice during the chronic infection period. Additionally, allitridin treatment reduced the MCMV load in salivary glands. MTC allitridin treatment of co‐cultures partially blocked MCMV induction of Foxp3 mRNA and protein expression. In vitro treatment with allitridin also increased significantly the percentages of Tc1, Tc2, and Th1, reduced the secreted levels of IL‐10 and TGF‐β1, and significantly suppressed viral loads. In conclusion, allitridin can promote MCMV‐induced Treg expansion and Treg‐mediated anti‐MCMV immunosuppression. Therefore, allitridin may be useful as a therapeutic agent to enhance the specific cellular immune responses against CMV. J. Med. Virol. 85:493–500, 2013. © 2013 Wiley Periodicals, Inc.     

Comparison of two nucleic acid extraction and testing systems for HCMV-DNA detection and quantitation on whole blood specimens from transplant patients

Quantitative detection of human cytomegalovirus (HCMV) DNA on whole blood is currently the primary choice for virological monitoring in transplant patients and for determining the appropriate antiviral strategy, however specific issues of variability remain in terms of extraction methods, amplification efficiency, and variability. This study compared the performance characteristics of two nucleic acid extraction and testing systems for HCMV-DNA quantitation, the artus^® CMV QS-RGQ kit, associated with a fully automated DNA extraction and assay set up by Qiagen (system 1) and the Q-CMV Real Time Complete kit by Nanogen, associated with a semiautomated nucleic acid extraction system by Biomérieux (system 2) in 189 specimens from transplant patients and 10 from 2012 HCMV Quality Control for Molecular Diagnostics (QCMD). The two systems exhibited a 80.4% concordance. Differences between the two systems were within ±1 log₁₀ copies/ml of the averaged log₁₀ results for 88.9% of the tested specimens. For all qualitatively discordant specimens, mean viral load was ≤3 log₁₀ copies/ml. Considering viral load measurement, system 1 gave earlier positives that system 2, with a 14.8% of specimens resulted positive at low viral loads with system 1 and negative with system 2. In QCMD specimens, difference was below 0.7 log₁₀ copies/ml for both the systems.