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Cytomegalovirus infection in dialysis patients and personnel.

In a 12-month prospective study of cytomegalovirus infection on an acute hemodialysis unit, 10 of 80 patients (13%) and none of 26 staff developed active cytomegalovirus infection. Seven infections were coincidental with renal allograft rejection; three occurred 3 to 6 weeks after the transfusion of multiple units of conventional blood into seronegative patients. No person-to-person transmission was documented. In contrast to the effects of transfusing conventional blood, all 21 patients who entered dialysis without detectable cytomegalovirus antibody and received 2 to 10 U of frozen deglycerolyzed erythrocytes (total of 157 U) remained seronegative. Transmission of cytomegalovirus infection with transfusion with conventional blood is probably secondary to passage of leukocyte-borne virus that is lost during the freezing and deglycerolization procedure. Frozen erythrocytes prepared by cytoagglomeration procedures appear to be free of viable leukocytes and appear to carry a minimal risk of transmitting cytomegalovirus infection.

Acute Disease

Experimental cytomegalovirus infection and the developing mouse inner ear: in vivo and in vitro studies.

Mouse cytomegalovirus causes perilabyrinthitis in the cochlea following intracranial inoculation of the newborn mouse. In spite of infection of cranial nerve ganglion cells and Schwann cells the organ of Corti achieves normal adult morphology including efferent nerve endings. No involvement of neuroaxons is seen. In comparison human cytomegalovirus infection causes endolabyrinthitis without involving periotic connective tissues or nerves even in the presence of meningoencephalitis. The lack of epithelial infection and susceptibility of periotic connective tissues to mouse cytomegalovirus was confirmed by infection of fetal mouse otocysts in organ culture. The mouse model of cytomegalovirus ear infection shows greater similarities to other herpesvirus infections of the ear such as varicella-zoster than to human cytomegalovirus and thus remains extremely useful for the study of herpetic neuronal and ear infection.

Animals

Controlled clinical trial of prophylactic human-leukocyte interferon in renal transplantation. Effects on cytomegalovirus and herpes simplex virus infections.

A double-blind, placebo-controlled trial of interferon prophylaxis against viral infections was conducted in renal-transplant recipients receiving standard immunosuprressive therapy with or without antithymocyte globulin. Interferon was administered for six weeks, beginning on the day of transplantation. Cytomegalovirus excretion began earlier and viremia was more frequent in placebo-treated than in interferon-treated patients. Cytomegalovirus viremia correlated with clinical syndromes was more frequent in recipients of antithymocyte globulin. In contrast, neither interferon nor antithymocyte globulin altered excretion of herpes simplex virus. Reversible leukopenia and thrombocytopenia occurred in seven interferon recipients. Patient and graft survival were comparable in interferon and placebo groups. There preliminary results suggest that a six-week course of prophylactic interferon delays shedding of cytomegalovirus and decreases the incidence of viremia after transplantation. In contrast, antithymocyte globulin appears to increase the severity of infection from cytomegalovirus among these patients.

Adult

Reactivation of herpes simplex virus type 2 from a quiescent state by human cytomegalovirus.

The ability of human cytomegalovirus to stimulate replication of herpes simplex virus type 2 (HSV-2) was examined. The system used involved HSV-2-infected human embryonic lung cells under conditions (39.5-40 degrees C) in which HSV-2 remains undetectable. Reactivation of HSV-2 was maximal and persisted for the longest duration when cultures were superinfected with 0.02 plaque-forming unit of human cytomegalovirus per cell. Infectious HSV-2 appeared 2 days after superinfection with human cytomegalovirus and ranged from 10(2) to 10(6) plaque-forming units per culture. Virus reactivated from these cultures was neutralized by rabbit immune serum produced against HSV-2. The specificity of this interaction was demonstrated by various criteria: production of HSV-2 was not observed in cultures treated with mock infecting fluid, and inactivation of human cytomegalovirus by heat, ultraviolet irradiation, or immune serum prior to superinfection eliminated its ability to induce HSV-2 replication. These results sugges that interaction between these two human herpesviruses may be of importance in herpesvirus latency in vivo.

Cells, Cultured

Murine model for immunoprophylaxis of cytomegalovirus infection. I. Efficacy of immunization.

Murine cytomegalovirus was utilized as a model for human cytomegalovirus, which had no experimental animal, to study immunoprophylaxis of the cytomegalovirus infections. (1) Murine cytomegalovirus (MCMV) serially propagated in mouse embryonic fibroblasts had lost pathogenicity for weanling mice including neonatally thymectomized mice. (2) The cell culture-adapted MCMV was effective as a "live, attenuated virus vaccine" against challenge by virulent, mouse-passaged MCMV. (3) The immunization via intraperitoneal route protected mice from every parameter of MCMV infection. These included clinical signs, virus replication, histopathology and mortality. (4) The protective immunity was active against the virulent MCMV which was not neutralized by the rabbit anti-attenuated MCMV serum.

Animals

Enhancement of mouse cytomegalovirus infection during host-versus-graft reaction.

C3H/He mice chronically infected with murine cytomegalovirus were given skin allografts from histoincompatible BALB/c donors. A significant increase in cytomegalovirus titers occurred within 3 days after placement of the graft in the spleens and kidneys of the allograft recipients as compared with control animals. No significant changes in virus titers were detected in the salivary gland, lung, liver, or blood of allograft recipients. These results indicate that the host-versus-graft reaction alone can enhance murine cytomegalovirus in a chronically infected host and may help explain the high incidence of cytomegalovirus infection seen after renal and other allograft transplantation in man.

Animals

Acute encephalopathy with liver dysfunction, chylous ascites and cytomegalovirus infection.

A child with acute encephalopathy and liver dysfunction subsequently developed acute chylous ascites. Titers for cytomegalovirus increased from less than 1:2 to 1:32 during the illness, and cytomegalovirus was isolated from the urine. The case is the first one possibly linking cytomegalovirus and acute encephalopathy and liver dysfunction in a child. In our patient, enlargement of the abdominal lymph nodes, as seen on a lymphangiogram, resulted in a severe obstruction of abdominal lymphatic flow, producing a transudation of lymph into the peritoneal cavity. The acute chylous ascites associated with mesenteric lymphadenitis was likely caused by the cytomegalovirus infection.

Brain Diseases

Evidence for the association of cytomegalovirus with carcinoma of the prostate.

A human genital isolate of cytomegalovirus is shown to have transformed human embryonic lung cells in vitro. These cells produce tumors when injected into athymic nude mice. Two cell lines derived from tissue from human prostatic carcinoma have survived more than 20 passages in vitro and demonstrate cytomegalovirus-specific membrane antigen. Significant humoral antibody titers against cytomegalovirus have been demonstrated. Cell-mediated lymphocytotoxicity against these transformed cells has been demonstrated in patients with urinary tract tumors. This evidence indicates that an association between cytomegalovirus and human prostatic cancer may be more than coincidental.

Adenocarcinoma

Clinical efficacy of adenine arabinoside in therapy of cytomegalovirus infections in renal allograft recipients.

Three renal allograft recipients, two with cytomegalovirus pneumonia and one with cytomegalovirus retinitis, were treated with adenine arabinoside. The dose was 5-10 mg/kg per day administered for four to six days. There was no clinical improvement in any of the patients. One patient died from overwhelming interstitial viral pneumonia. Cytomegalovirus was readily isolated from two patients after therapy (autopsied lung of one and the saliva of the other). Suppression of viruria was observed and quantitated in one patient (10(4.0)-10(2.5) 50% tissue culture infective doses/0.2 ml). However, the suppression was transient, and viral titers returned to the levels before therapy within two months after completion of therapy. Treatment was stopped in two cases when hematologic toxicity (drop in hematocrit and platelet and leukocyte counts) was noted in the patients five days after initiation of therapy with adenine arabinoside. The drug appears to be ineffective and possibly toxic in immunosuppressed renal allograft recipients infected with cytomegalovirus.

Adult

An analysis of cytomegalovirus infection and HLA antigen matching on the outcome of renal transplantation.

Eighty-five recipients and donors of renal allografts were examined for evidence of cytomegalovirus infection before and repeatedly after transplantation. The recipients were also divided into two group on the basis of HLA antigen matching. Better allograft survival was noted in patients well matched for HLA antigens (0-2) mismatched antigens) compared to those poorly matched (three or more antigens mismatched), and in patients free of cytomegalovirus compared to those infected. Cytomegalovirus infection had a more marked influence on allograft survival than did HLA antigen matching. The differing rates of success of transplantation, apparently dependent on blood relationship between donor and recipient, have been assumed largely to be due to inherited factors. This study, however, revealed an important factor to be the disparate incidence of cytomegalovirus infection in sibling, parental, and cadaveric categories of transplantation. The mechanism of this disparity can be explained on the basis of the incidence of latent CMV infection in the recipients and various categories of kidney donors.

Adult

Cellular basis for susceptibility to mouse cytomegalovirus: evidence from tracheal organ culture.

Tracheal organ cultures were prepared from strains of mice with known susceptibility to the lethal effects of mouse cytomegalovirus. When organ cultures from susceptible mice were infected with this virus, characteristic c.p.e. developed and virus titres greater than 10(5) p.f.u./ml were produced 2 to 3 weeks after infection. Identically infected tracheal organ cultures from resistant mice consistently failed to develop significant virus c.p.e. and produced 10- to 100-fold lower titres of virus. When fibroblast cultures were infected with mouse cytomegalovirus, however, no consistent differences between susceptible and resistant mouse strains were observed. Since cytomegalovirus replicates in epithelial cells both in vivo and in tracheal organ culture, these results suggest that resistance may be based in part upon innate genetic susceptibility of epithelial cells to cytomegalovirus infection.

Animals

Effect of ribavirin on murine cytomegalovirus infection.

Ribavirin (1-beta-d-ribofuranosyl-1,2,4-triazole-3-carboxamide), a new synthetic nucleoside, inhibited murine cytomegalovirus in cell culture. This was shown by the inhibition of viral cytopathic effect and plaque formation, as well as reduction in the yield of virus. Despite this in vitro antiviral effect, ribavirin did not protect mice from mortality produced by a high inoculum of cytomegalovirus. When a lower inoculum was used to initiate a chronic infection, the administration of ribavirin for 9 days had no effect on the titer of cytomegalovirus in the salivary gland, kidney, liver, and spleen. Thus, ribavirin was ineffective in the treatment of both acute and chronic murine cytomegalovirus infections.

Acute Disease

Cell-mediated and humoral immune responses to herpes simplex virus and cytomegalovirus in renal transplant patients.

Cell-mediated immunity to herpes simplex virus and cytomegalovirus, using the lymphocyte transformation test and interferon induction in lymphocytes, was studied in 59 patients from 1 day to 7 years after allotransplantation and compared with the results in normal subjects. Both parameters were permanently depressed with regard to cytomegalovirus. With herpes simplex virus, interferon production was also permanently depressed, whereas the transformation reaction was normal during the first year after transplantation and only slightly depressed in patients more than 1 year after transplantation. In 6 patients the above-mentioned assays and the complement fixation reaction were performed serially and related to the clinical signs of herpes simplex virus and cytomegalovirus infection. The relationship between depression of the transformation reaction and interferon production in lymphocytes and the occurrence of clinically evident herpes simplex virus and cytomegalovirus infections was, however, equivocal. The humoral immune response to herpes simplex virus was measured by the complement fixation test and the more sensitive antibody-dependent, cell-mediated cytotoxicity reaction, and a good correlation was found between these two tests, although only a few persons were found to be negative in the antibody-dependent, cell-mediated cytotoxicity reaction. The suggestion is made that only a few adults are "true" herpes simplex virus seronegative.

Adolescent

Human cytomegalovirus: glycoproteins associated with virions and dense bodies.

The glycoproteins associated with the membranes of cytomegalovirions and dense bodies were characterized by their relative mobility, percentage of glucosamine incorporation, and molecular weight. Eight glycopolypeptides were repeatedly detectable. Three glycopolypeptides of higher molecular weight with low levels of glucosamine incorporation were occasionally detectable. These latter glycopolypeptides may be precursors or aggregates of the glycopolypeptides with lower molecular weights. The glycoproteins associated with the membranes were on the surface, as determined by iodination with 125I of virions and dense bodies partially purified in gradients of D-sorbitol. Velocity centrifugation in linear gradients of D-sorbitol was used to obtain concentrated and partially purified preparations of infectious cytomegalovirus. Viral infectivity and the membranes of cytomegalovirions and dense bodies were stable in gradients of sorbitol, but cellular contaminants were not completely removed. Additional centrifugation in CsCl separated both cellular contaminants and viral nucleocapsids from virions and dense bodies. Many dense bodies, which are considered to be aberrant forms of cytomegalovirus, had the same size, sedimentation properties, and density as virions. Consequently, they were not separable from virions by various centrifugation techniques. Electron microscopy demonstrated that purified virions and dense bodies were qualitatively free of extraneous material and that each dense body was bounded by a membrane, as evidenced by its double-tract appearance. Antisera to a preparation of purified virions and dense bodies, or to their glycoproteins, contained antibodies that neutralized viral infectivity and reacted with antigens in cells infected with cytomegalovirus. However, these same antisera did not contain antibodies that reacted with uninfected cells. The glycoproteins associated with the membranes of cytomegalovirions and dense bodies are considered to be specified by the cytomegalovirus genome.

Antibodies, Viral

Sequence of protein synthesis in cells infected by human cytomegalovirus: early and late virus-induced polypeptides.

At least 10 distinct early virus-induced polypeptides were synthesized within 0 to 6 h after infection of permissive cells with cytomegalovirus. These virus-induced polypeptides were synthesized before and independently of viral DNA replication. A majority of these early virus-induced polypeptides were also synthesized in nonpermissive cells, which do not permit viral DNA replication. The virus-induced polypeptides synthesized before viral DNA replication were hypothesized to be nonstructural proteins coded for by the cytomegalovirus genome. Their synthesis was found to be a sequential process, since three proteins preceded the synthesis of the others. Synthesis of all early cytomegalovirus-induced proteins was a transient process; the proteins reached their highest molar ratios before the onset of viral DNA replication. Late viral proteins were synthesized at the time of the onset of viral DNA replication, which was approximately 15 h after infection. Their synthesis was continuous and increased in molar ratios with the accumulation of newly synthesized viral DNA in the cells. The presence of the amino acid analog canavanine or azetadine during the early stage of infection suppressed viral DNA replication. The amount of viral DNA synthesis was directly correlated to the relative amount of late viral protein synthesis. Because synthesis of late viral proteins depended upon viral DNA replication, the proteins were not detected in permissive cells treated with an inhibitor of viral DNA synthesis or in nonpermissive cells that are restrictive for cytomegalovirus DNA replication.

Canavanine

Cytopathogenicity of cytomegalovirus to human ecto- and endocervical epithelial cells in vitro.

The effect of cytomegalovirus on cell cultures initiated from human uterine endo- and ectocervix was studied. Pure epithelial cultures were obtained which differed in morphology depending on the source. The ectocervical cells grew in mosaic-like regular epithelial patterns, whereas endocervical cultures had a poorer intercellular cohesion, irregular polygonal cell form and curved cytoplasmic processes. Occasional fibroblastic colonies grew in three out of 32 ectocervical cultures and in none of the endocervical cultures. In the latter no cells with fibroblast-specific surface antigens could be seen. Ectocervical cells were resistant to cytomegalovirus infection; only in three out of 20 cultures could individual altered cells be detected. In contrast, endocervical cultures supported the growth of cytomegalovirus and at best 30 per cent of the explanted colonies showed cytologic alterations as well as virus-specific immunofluorescence. The various types of altered cells are described and it si concluded that endocervical epithelial cells may be the site of replication of human cytomegalovirus so commonly found in cervical secretions.

Antigens, Viral

[Hepatic disease in childhood and cytomegalovirus infection (author's transl)].

135 children with hepatitis and 179 patients with hepatomegaly of undetermined etiology were investigated for complement fixing antibodies to cytomegalovirus. 69 patients with hepatitis (51%) and 74 patients with hepatomegaly (41%) had a titer of 1:4 or above as compared with a control group of children without hepatic pathology in whom positive response occurred in only 20 resp. 18%. The mean titer found in the sero-positive patients was also significantly higher than that in the control group. In contrast, the incidence of positive sera in newborns with hyperbilirubinemia of unknown etiology did not differ significantly from that in a healthy control collective. Of 57 patients with liver disease, 20 excreted cytomegalovirus which was isolated 55 times from 103 urine specimens. These patients included 9 with hepatitis and 11 with hepatomegaly. Of 129 children without any indication of liver disease, cytomegalovirus could be demonstrated in only 3. Our results point toward a causal connection between the presence of cytomegalovirus and the development of hepatic disease in childhood.

Adolescent

Cytomegalovirus as a frequent cause of Guillain-Barré syndrome.

In 10 patients with Guillain-Barré syndrome a preceeding cytomegalovirus infection could be demonstrated by virus-specific IgM antibodies that were present in high titers in 9 of the 10 patients in the first serum specimen. The IgM antibodies to cytomegalovirus were detected by a sensitive "double" indirect immunofluorescence technique. In most of our cases (8 of 10) the complement-fixing antibody titers had already reached high levels on admission into a hospital, and significant titer changes were not observed. Cytomegalovirus was isolated from the urine of five patients.

Adult