Cytomegalic Inclusions Research Articles

Congenital cytomegalovirus infection.

, Sarah RossiA female infant born at 37 weeks and 2 days of gestation by vaginal delivery was admitted to the neonatal intensive care unit on July 11, 2009 with birth weight of 2,030 g, height of 41,0 cm and head circumference of 31.0 cm. The infant Apgar were 8, 9 and 9 at 1, 5 and 10 min, respectively. A diagnosis of congenital toxoplasmosis was entertained because of the mother clinical history (she IgG positive and IgM positive for toxoplasmosis with avidity of 85% during first trimester of pregnancy). At the first trimester morphology scan intracranial calcifications were seen. The newborn had intrauterine growth restriction, splenomegaly, petechias and thrombocytopenia. Treatment of toxoplasmosis was done with sulphadiazine, pyrimethamine and folinic acid. The tomography of the skull showed multiple foci of bilateral periventricular calcifications and on the base nuclei. Due to these findings the serology for cytomegalovirus of mother and child was ordered. The mother was IgM negative, but IgG positive. The newborn was IgM and IgG positive. Cytomegalic inclusions in newborn’s urine and polymerase chain reaction (PCR) for CMV had positive results. The newborn was discharged and was referred to outpatient clinic to be treated with ganciclovir. The congenital infections, syphilis, toxoplamosis, rubella, cytomegalovirus and herpes correspond to

Combined In Situ Hybridization and Immunohistochemistry for Automated Detection of Cytomegalovirus and p53.

Background: Cytomegalovirus (CMV) infection has been shown to be associated with p53 overexpression in coronary artery restenosis. We investigated the occurance of this association in other forms of CMV infection using an automated in situ hybridization (ISH) technique. Methods and Results: We performed ISH for CMV using digoxigenin-labeled or biotinylated probes followed by avidin-alkaline phosphatase and nitroblue tetrazolium color substrate. Immunohistochemistry (IHC) was then performed using an anti-p53 antibody utilizing streptavidin-immunoperoxidase and 3,3;-diaminobenzidine tetrahydrochloride as a chromogen. Sixteen cases with characteristic cytomegalic inclusions from a variety of body sites were examined. All 16 cases were positive for CMV by ISH. Nine of sixteen expressed nuclear p53. Six of these nine cases showed viral cytopathic effect in the cells with p53 expression. In an illustrative case, double colocalized staining for CMV and p53 protein was demonstrated in individual cytopathic cells. When microwave antigen retrieval was necessary, ISH was performed before IHC, and our standard microwaving time was reduced by two-thirds. Conclusions: The colocalization of p53 protein overexpression with CMV within single cells adds further evidence that this overexpression is a viral-induced phenomenon. The combined ISH and IHC assay can be carried out in a rapid automated mode, increasing the ease of investigating relationships between message and protein expression within single cells in a wide variety of settings.

CMV infection of the renal allograft is much more common than the pathology indicates: a retrospective analysis of qualitative and quantitative buffy coat CMV-PCR, renal biopsy pathology and tissue CMV-PCR.

Quantitative blood polymerase chain reaction (PCR) for cytomegalovirus (CMV) is used to direct therapy in kidney transplant patients, but cytomegalic inclusions are rarely found in allograft renal biopsies even with an elevated serum creatinine and apparent CMV disease. The relationship between quantitative blood CMV and renal allograft pathology is unknown. Thirteen biopsy samples were available for analysis from patients suspected of CMV disease, who had a buffy coat CMV-PCR drawn within 2-5 days of a renal allograft biopsy for an elevated creatinine. All were evaluated for CMV pathologically, by light microscopy, immunohistochemistry, in situ hybridization and tissue PCR. Qualitative and quantitative buffy coat CMV-PCR were positive in 10/13 (77%) patients. Tissue CMV-PCR was positive in five (50%) biopsies, including two with CMV inclusions and three with no inclusions. Quantitative buffy coat CMV-PCR levels did not correlate with detection of CMV inclusions in renal tissue. Paradoxically, quantitative buffy coat CMV-PCR was low (239 and 538 copies/microg of DNA) when CMV inclusions were detected. All five biopsies with acute rejection were associated with CMV viraemia and two of the five with allograft CMV inclusions. A quantitative buffy coat CMV-PCR of <100 copies/microg of DNA ruled out disease with CMV inclusions. CMV nephropathy is much more common than previously reported when sensitive techniques are used for detection in tissue. Acute rejection and CMV viraemia occur commonly together in patients at risk for CMV. Quantitative buffy coat CMV-PCR does not correlate with the presence of CMV inclusions. These findings have implications for management of patients who have elevated serum creatinine and are at risk for CMV disease.

Longitudinal Analysis of Plasma Cytomegalovirus DNA in a Child with Crohn's Disease and Cytomegalovirus Gastroenteritis

Longitudinal Analysis of Plasma Cytomegalovirus DNA in a Child with Crohn's Disease and Cytomegalovirus Gastroenteritis

Anémie sidéroblastique congénitale chez un nourrisson

Sideroblastic anemia is rare in infants. When it occurs, it is generally hereditary and X-linked. A microcytic anemia without iron deficiency was diagnosed in a 2-month old infant with probable acquired cytomegalic infection. Bone marrow examination disclosed ringed sideroblasts without cytomegalic inclusions. Treatment with pyridoxine was tried without effect and no other therapeutic trial was attempted. The child, now aged 5 years, remains well without blood transfusion. Hereditary and acquired sideroblastic anemias may occur in infants and children. Whatever their cause might be, the evolution is unpredictable. The only successfull treatment for the hereditary variant remains pyridoxine supplementation which brings relief in about one patient out of three. Efficiency and safety of other therapeutic regimens remain to be proven.

Combined in situ hybridization and immunohistochemistry for automated detection of cytomegalovirus and p53

Background: Cytomegalovirus (CMV) infection has been shown to be associated with p53 overexpression in coronary artery restenosis. We investigated the occurance of this association in other forms of CMV infection using an automated in situ hybridization (ISH) technique. Methods and Results: We performed ISH for CMV using digoxigenin-labeled or biotinylated probes followed by avidin-alkaline phosphatase and nitroblue tetrazolium color substrate. Immunohistochemistry (IHC) was then performed using an anti-p53 antibody utilizing streptavidin-immunoperoxidase and 3,3-diaminobenzidine tetrahydrochloride as a chromogen. Sixteen cases with characteristic cytomegalic inclusions from a variety of body sites were examined. All 16 cases were positive for CMV by ISH. Nine of sixteen expressed nuclear p53. Six of these nine cases showed viral cytopathic effect in the cells with p53 expression. In an illustrative case, double colocalized staining for CMV and p53 protein was demonstrated in individual cytopathic cells. When microwave antigen retrieval was necessary, ISH was performed before IHC, and our standard microwaving time was reduced by two-thirds. Conclusions: The colocalization of p53 protein overexpression with CMV within single cells adds further evidence that this overexpression is a viral-induced phenomenon. The combined ISH and IHC assay can be carried out in a rapid automated mode, increasing the ease of investigating relationships between message and protein expression within single cells in a wide variety of settings. (Mol Diagn 1996 Dec;1(4):291-296)

Triple Retinal Infection with Human Immunodeficiency Virus Type 1, Cytomegalovirus, and Herpes Simplex Virus Type 1: Light and Electron Microscopy, Immunohistochemistry, and In Situ Hybridization

This report describes the histopathologic and virologic findings of the retina from a 55-year-old bisexual patient with the acquired immune deficiency syndrome (AIDS), who had concurrent human immunodeficiency virus type 1 (HIV-1), cytomegalovirus (CMV), and herpes simplex virus type 1 (HSV-1) retinitis, and was treated with ganciclovir. The eyes were obtained at autopsy and processed for light microscopy and transmission electron microscopy. Immunohistochemical stains for HSV-1, CMV, HIV-1, varicella zoster virus, and glial fibrillary acidic protein were carried out using the peroxidase-antiperoxidase and streptavidin-biotin-alkaline phosphatase techniques. For in situ hybridization, a radiolabeled CMV DNA probe (Eco-RI-Y fragment of strain AD 169) was used. Results of histopathologic examination showed a full-thickness necrotizing retinitis with cytomegalic and herpes viral intranuclear inclusions in cells of the neurosensory retina, retinal vascular endothelium, and the retinal pigment epithelium. Some areas of the retina were replaced by glial tissue. The choroid contained only a few chronic inflammatory cells. Immunoperoxidase studies disclosed CMV antigens diffusely distributed throughout all layers of the retina and the retinal pigment epithelium. Herpes simplex virus type 1 antigens were present in retinal cells and the retinal vascular endothelium. Human immunodeficiency virus type 1 antigens were found in mononuclear cells in all layers of the sensory retina. Dual infections with HIV-1 and CMV of individual multinucleated giant cells of glial origin were demonstrated immunohistochemically. Transmission electron microscopy showed herpes viral particles in the vascular endothelium of the retinal vessels and the choriocapillaris. Human immunodeficiency virus particles were identified in the endothelium of the choriocapillaris. The possibility of multiple viral infections of the retina, mimicking classic CMV retinitis, should be considered in the clinical and histologic differential diagnosis of necrotizing retinitis in patients with AIDS.

Cytomegalovirus induction of tumor necrosis factor-alpha by human monocytes and mucosal macrophages.

Cytomegalovirus (CMV) is a major cause of inflammatory organ disease in immunosuppressed persons. To elucidate the mechanisms of CMV-induced inflammation, we investigated whether tumor necrosis factor-alpha (TNF-alpha) was involved in the pathogenesis of CMV colitis in patients with AIDS. In in situ hybridization experiments, TNF-alpha mRNA was shown to be abundantly present in colonic mucosa from AIDS patients with CMV colitis but not in colonic mucosa from control (AIDS and normal) subjects. The TNF-alpha transcripts, identified in macrophage-like cells containing cytomegalic inclusions, were positively associated with CMV, but not HIV-1, within the mucosa. In in vitro experiments, a patient-derived isolate of CMV, but not HIV-1Ba-L, induced human monocytes to express TNF-alpha mRNA and to release increased levels of TNF-alpha peptide following stimulation. CMV induction of TNF-alpha may play a critical role in CMV-induced inflammation and, since TNF-alpha upregulates expression of HIV-1, offers a mechanism by which CMV could serve as a co-factor for HIV-1 expression without both viruses infecting the same cell.

Intra-abdominal calcification in a fetus with cytomegalovirus infection, mimicking meconium peritonitis on ultrasound

An autopsy case is presented of a fetus with Ultra-sound (US) diagnosis of intra-abdominal calcification. Histology revealed typical Cytomegalic inclusions in nuclei and cytoplasm. A routine US detected abnormalities at 16 weeks. There was growth retardation, echogenic gut, ascites without hydrops and (paradoxically) oligohydramnios. Chromosomal analysis was normal. Meconium peritonitis was suspected. Still birth occurred at 32 weeks. At post-mortem there was no evidence of meconium peritonitis. Histology revealed cytomegalic inclusions in kidney, lung and intestinal mucosa. Numerous calco-spherites were present in luminal mucin. The features of cystic fibrosis were lacking. It appears from this case that Cytomegalovirus infection can cause changes mimicking meconium peritonitis on US.

Ganciclovir in the treatment of progressive AIDS‐related polyradiculopathy

We present 7 HIV-infected patients with a unique, subacute, progressive polyradiculopathy. All had AIDS, sacral sensory loss, acute urinary retention, and progression to flaccid paraparesis in days to weeks. Cytomegalovirus was cultured from spinal fluid of 4 patients, and postmortem examination of the 1st 5 patients disclosed an inflammatory polyradiculopathy with cytomegalic inclusions. The inclusion-bearing cells were immunocytochemically positive for cytomegalovirus. Two patients who received early anti-cytomegalovirus treatment with ganciclovir improved. Thus, early recognition and treatment with ganciclovir may be effective in this otherwise fatal condition.

SOLUBLE INTERLEUKIN-2 RECEPTOR LEVEL AS AN INDICATOR OF LIVER ALLOGRAFT REJECTION

We studied the serum levels of soluble interleukin-2 receptors (SIL-2R) in liver allograft recipients: a control group without rejection or CMV disease, a group with only rejection episodes, and a group with only cytomegalovirus disease. Rejection was diagnosed by the presence of compatible laboratory and histologic abnormalities and absence of other causes of graft dysfunction. CMV disease was diagnosed by isolation of CMV in blood or liver specimen cultures or identification of cytomegalic inclusions in the liver biopsy specimen. Of 82 consecutive recipients treated with cyclosporine and prednisone, 12 were in the control group, 20 in the rejection group, and 5 in the CMV disease group. The remaining 45 had other or multiple complications. In the control group the SIL-2R levels (determined by an ELISA) decreased by a mean of 4% per day after transplantation; in the rejection group the levels increased by a mean of 17% per day in the 10 days prior to the diagnosis of rejection; in the CMV disease group the levels tended to increase prior to the diagnosis of CMV disease. The rejection group had significantly higher SIL-2R levels than the control group at comparable times. Thus, SIL-2R levels were significantly increased at the time of allograft rejection compared with levels in a control group, and recipients with CMV disease had increased levels of SIL-2R but they were not as high as in recipients with rejection episodes.

Temporal sequence of viral antigen expression in the cochlea induced by cytomegalovirus.

Injection of guinea-pig cytomegalovirus into seronegative guinea pig cochleas results in viral labyrinthitis. The location of infected cells 1-8 days following inoculation was examined using an immunohistochemical assay for viral antigens. Mesothelial cells below the basilar membrane and those lining the scala tympani, especially below the osseous spiral lamina, were the first cells to express antigen. This occurred 2 days following inoculation. Infection then spread rapidly toward the brain and through the perilymphatic duct to involve inflammatory cells, mesothelial cells of Reissner's membrane, spiral ganglion cells, and the vestibular ganglion cells. In any one cochlea, more mesothelial cells and inflammatory cells expressed viral antigen than any other cell type. It seems then, that there is a differential susceptibility to viral infection and antigen expression in the various cell types of the cochlea. Typical cytomegalic inclusions were common in mesothelial cells. However, many cells labelled for viral antigens did not contain inclusions. The identification of viral antigens then, is a more sensitive technique for demonstrating infection than is the location of cytomegalic inclusion cells.

Gastrointestinal Cytomegalovirus Infection in a Homosexual Man With Severe Acquired Immunodeficiency Syndrome

A 44-yr-old white homosexual man with a history of Pneumocystis carinii pneumonia developed watery diarrhea and fever. Flexible sigmoidoscopy revealed yellow plaques in the sigmoid colon suggestive of pseudomembranous colitis. Stool examination for ova and parasites, Clostridium difficile toxin, and cultures for pathogens, including Clostridium difficile, were negative. Infection with multiple organisms, including Mycobacterium avium intracellulare, Toxoplasma gondii, herpes simplex, and cytomegalovirus, was demonstrated. Postmortem examination showed esophageal and colonic ulcerations with cytomegalic inclusions, and cytomegalovirus was cultured from the colonic lesions. This report describes a case of disseminated cytomegalovirus infection with esophageal and colonic involvement in a homosexual man with the acquired immunodeficiency syndrome.

An autopsy case of pure red cell aplasia with thymoma

A 76 year-old woman was hospitalized on Oct. 22, 1973, because of fatigue, anorexia, pallor of the face for the past 2 months.Laboratory data: Red cell count was extremely reduced, being 77×104 per cu mm, reticulocytes 1.0‰, Hb. 3.0g/dl, Ht. 7%. Bone marrow aspiration disclosed erythroblast 1.0%.Hematologic examination disclosed normocytichyperchromic anemia without granulocytopenia and thrombocytopenia.Serum examination revealed hypo-γ-globulinemia (9.8%), increased serum iron value (231-282γ/dl), negative for Coombs' and LE test.There was no evidence of pathologic shadow on chest X-ray examination.Clinical diagnosis was pure red cell aplasia without thymoma.Treatment was instituted with Methycobal injection, blood transfusion (total 4, 600ml) and steroids. The therapeutic effects appeared to enhance remission with further steroid therapy.The patient died on June 22, 1974, 8 months after admission due to bronchopneumonia.At autopsy, there was encapsulated solid tumor on anterior upper mediastinum -70g weight of thymoma. Histologically, tumor consisted predominantly made of spindle-shaped cells and small number of lymphocytes scattered among them separated by fibrous septa.Bone marrow was in normal hematopoeisis including erythropoeisis.The lungs showed bilateral bronchopneumonia associated with many cytomegalic inclusions and fungus infection. The similar cytomegalic infected cells were found in thymus, salivary glands, liver, tongue, esophagus, stomach, pancreas and genital organ.Electron microscopically, herpes type virus showed in squamous cells of tongue.The association of pure red cell aplasia, hypo-γ-globulinemia and thymoma in our case suggests the presence of a basic inmunologic disorders, but not any definitive evidence as autoinmune disease was obtained.Eighteen cases out of 88 cases of pure red cell aplasia have been autopsied in Japan.

Ménétrier disease and cytomegalovirus infection in childhood.

Cytomegalic inclusions, characteristic of infection with cytomegalovirus, were found in the hypertrophic gastric mucosa of a 3½-year-old child with giant hypertrophic gastritis and protein loss in the gastrointestinal tract (Menetrier disease). There was a tardy 16-fold rise of complement fixation titers against cytomegalovirus soon after the acute phase of the illness. To our knowledge, the association of Menetrier disease and cytomegalovirus has been reported only once before.

Identification of a cytomegalovirus recovered from field mice.

A virus recovered in murine tissue cultures from the salivary glands of &lt;i&gt;Microtus arvalis &lt;/i&gt;specimens, with cytomegalic inclusions in the duct cells, has been identified as a murine cytomegalovirus, distinct from the previously isolated Smith and Raynaud strains.

Benign lymphoepithelial lesion of the parotid gland associated with cytomegaly.

A 16-year-old girl had a benign lymphoepithelial lesion in the left parotid gland, associated with cytomegalic inclusions which were confined solely to the area of the lesion. To our knowledge, no analogical case has been described. The possible relationship between these two conditions is discussed, and pertinent literature is reviewed.

Two unusual cases of mural endocarditis, with a review of the literature.

Two unusual cases of mural endocarditis are described. The first was that of a 13-year-old boy with infected burns on the legs. Autopsy revealed mural vegetations in both ventricles and cytomegalic viral inclusions were demonstrated in the vegetations, heart muscle, and endothelial cells of various organs. This appears to be the first reported case of cytomegalic inclusion viral endocarditis. A review of the literature revealed only six previous reports of cytomegalic viral myocarditis, which was also present in this case. The second case was that of a 4-year-old girl with bronchiectasis. Autopsy revealed mural vegetations in the right ventricle. Group C beta-hemolytic streptococcus was isolated from the mural vegetations, as well as from the diseased lungs. Only two previous reports of endocarditis caused by this organism are found in the literature.

Hemorrhagic cystitis and cytomegalic inclusions in the bladder associated with cyclophosphamide therapy

A case of severe hemorrhagic cystitis resulting from cyclophosphamide administration is presented. This complication is of import because it may lead to fatal hemorrhage if the etiology is unrecognized. The lesion appears to result from direct contact with urinary metabolites of the drug, although its lack of universal occurrence may indicate associated genetic or constitutional factors. Prompt discontinuation of the drug and conservative measures usually alleviate symptoms, but cystostomy with tamponade or urinary diversion may be required in cases of persistent bleeding. The resemblance of cyclophosphamide cystitis to changes produced by irradiation is noted. Although the major alterations occur within the lamina propria, atypical epithelial changes may be confused with carcinoma. A unique finding included the presence of cytomegalovirus inclusions in the bladder. It is possible that bladder damage was potentiated by this infection or that the injured bladder offered a site of preferential localization for the virus.

The incidence of cytomegaloviruria and postmortem cytomegalic inclusions in children with acute leukemia

Summary Attempts were made to detect cytomegaloviruria in 97 children with acute leukemia. Two of these patients (2 per cent) yielded cytomegalovirus, whereas none of 50 control patients yielded cytomegalovirus. Review of autopsy material from 82 leukemic children revealed 2 cases (3 per cent) with disseminated cytomegalic inclusions. These studies demonstrate the infrequent ocurrence of cytomegalovirus infection in children with leukemia in the population studied. The incidence was too low to have a meaningful correlation with their clinical state; however, both patients with cytomegaloviruria were receiving immuno suppressive therapy and had neutropenia.