Detection of proviruses and viral RNA in the early stages of feline immunodeficiency virus infection in cats: a possible model of the early stage of HIV infection.

Feline immunodeficiency virus (FIV) infection in cats has been reported to be a useful animal model for human AIDS studies, especially in the early stages of infection. We examined the temporal changes in provirus detection in peripheral blood mononuclear cells (PBMC) and the distribution of FIV-DNA and RNA in feline tissues by the polymerase chain reaction at 10, 35, 70 days after intravenous inoculation of FIV. Viral DNA in the PBMC was detected three to four weeks after infection and its fluctuation was demonstrated for the first time. Ten days after infection, before seroconversion, proviruses were detected only in the mesenteric lymph nodes and intestines. At 35 and 70 days after infection, after seroconversion, proviruses were detected in most lymphoid organs and the salivary glands, but the expression of FIV-RNA was limited to the thymus at 70 days after infection. These results show that FIV-RNA is transcribed from proviral DNA exclusively in the thymus at this stage. We suggest that the quantitative changes in detectable proviruses in the PBMC depend on the relation between the decrease in infected cells caused by cytolytic T lymphocytes and/or apoptosis and their increase caused by the release of a new supply of lymphocytes from the thymus.

Serosal balls detected immunocytochemically in peritoneal lavage obtained during surgery.

Using the immunoperoxidase technique, we studied "serosal balls," which have features resembling those of cells from primary and metastatic tumors, and may thus complicate cytodiagnosis. Serosal balls were detected in 32 (18%) of 174 peritoneal washings. The balls consisted of oval clusters of cells in solid masses surrounded by flattened cells. The interior of the serosal balls was stained green with Papanicolaou method, showing the presence of homogeneous amorphous material, sometimes stained in a filamentous pattern. Almost all serosal balls were stained immunocytochemically for both keratin and vimentin. The interior was stained with antibodies against collagen types I and III. Therefore, these balls were fragments of serous membrane, and contained fibrous tissue and mesothelial cells.

Cytodiagnosis of parvovirus B19 infection from ascites fluid of hydrops fetalis: report of a case.

The cytologic changes in the smears of fetus ascites fluid with parvovirus B19 infections are described. Cytology revealed the ground-glass appearance of the nuclei having a perinuclear halo, and the nuclear chromatin was spread as a thin rim around the inclusion. Subsequently, monoclonal antibodies for parvovirus B19 were applied to identify the specific antigen in the Papanicolaou-destained specimens. Positive staining reaction products were found throughout the cytoplasm of cells. Similar phenomena were confirmed in paraffin-embedded tissue sections of many organs in autopsy materials. It is hoped that recognition of those virus-infected cells in the body cavity fluid of the fetus will be helpful in making a diagnosis of hydrops fetalis associated with parvovirus B19 infections of the pregnant woman.

Cytology of cardiac myxomas: presence of Ulex europaeus agglutinin-I (UEA-I) lectin by immunoperoxidase staining.

Cytological findings are presented of seven cases of cardiac myxomas. Avidin-biotin-peroxidase complex (ABC) method was employed to demonstrate Ulex europaeus agglutinin-I (UEA-I) lectin in imprint smears as well as in paraffin-embedded tissue sections in cardiac myxomas. The cytology was characterized by tumor cells with polyhedral or stellate and mucinous background with lymphocytes, neutrophils, and hemosiderin-laden macrophages. In smears as well as tissue sections, UEA-I lectin was detected throughout the cytoplasm of myxoma cells. This study established the applicability of the immunoperoxidase staining for cardiac myxoma as an aid in cytopathological diagnosis.

Detection of cytomegalovirus DNA in pulmonary specimens: confirmation by in situ hybridization in two cases.

Papanicolaou stained bronchial brush and imprint pulmonary smears containing intranuclear and cytoplasmic inclusion bearing alveolar pneumocytes suggestive of cytomegalovirus infection were destained and reprocessed for in situ hybridization using a biotinylated probe for cytomegalovirus DNA. Two cases were processed in this way. A hybridization signal for viral DNA was noted in each case. However, no reddish brown staining reaction products were noted in any of the control samples. This simple and rapid nonradioactive detection system is a valuable supplement to routine pulmonary cytology for the definitive diagnosis of this virus infection, and this technique is also appropriate for retrospective study.

Cytological detection of atypical cells by routine urinalysis in a cardiovascular center.

During the past 9 yr, 187,529 Sternheimer-Malbin-stained urinary sediments were examined as routine urinalysis specimens from patients attending the National Cardiovascular Center in Osaka, Japan. Abnormal cells were found in 20 patients who did not have clinical diagnoses of malignancy. Malignant cytological changes in 18 patients resulted in a rate of 1 case in 6,751 patients; the two remaining specimens with abnormal cells showed polyomavirus infection. This article describes our experience in the diagnosis of malignant cells of the urinary tract through the cooperation of the clinical and cytological laboratories. Since in Japan, the rate of death for bladder cancer is similar to 1 in 6,751, this method seems to be of great use in the diagnosis of urinary tract malignancies.

Cytological diagnosis of herpesvirus keratitis by means of in situ hybridization: report of a case.

Typical herpesvirus keratitis that developed in a 12-year-old boy was initially diagnosed cytologically by the Papanicolaou method demonstrating the 'ground-glass' appearance of the nuclei with multinucleated syncytial cells. Subsequently, the in situ hybridization technique was applied to identify the herpes simplex virus (HSV) DNA in the Papanicolaoudestained cellular samples. Positive hybridization was found with intense staining for the HSV DNA in the nuclei of cells having a 'ground-glass' appearance. In situ hybridization has been shown to be a useful technique for the identification of HSV in corneal scrapes, and similar studies may be carried out in cellular samples from the other body sites.

Detection of human polyomavirus DNA in Papanicolaou stained smears of urinary sediment by in situ hybridization.

Papanicolaou stained smears of urinary sediment containing inclusion bearing urothelial cells suggestive of human polyomavirus infection were destained and reprocessed for in situ hybridization using a biotinylated probe for human polyomavirus DNA. Seven slides were processed in this way. A hybridization signal for viral DNA was noted in each case, even in smears that had previously been stored for 11 years. This simple and rapid non-radioactive detection system is a valuable supplement to routine urinary cytology for the definitive diagnosis of this virus infection.

Immunocytochemical demonstration of glial fibrillary acidic protein in imprint smears of human brain tumors.

Papanicolaou-destained imprint smears from 24 brain tumors were investigated by means of avidin-biotin-peroxidase complex method (ABC) with the use of monoclonal antibodies against glial fibrillary acidic protein (GFAP). Positive staining reaction to GFAP antibody has been demonstrated in cells from the following tumors: astrocytoma, anaplastic astrocytoma, glioblastoma multiforme, mixed glioma, and ependymoma. The reaction for GFAP was negative for the following tumors: medulloblastoma, neurilemmoma, melanoma, hemangioblastoma, and metastatic tumors. In astrocytoma, the cell bodies and processes were positive with delicate fibrillary patterns; in anaplastic astrocytoma, cytoplasm and the processes were intensively stained. In glioblastoma multiforme, the staining patterns were also mixed, and the short, thickened processes were characteristic. Use of both a smear preparation and the immunoperoxidase staining technique is of great value in diagnosis of tumors of the central nervous system.

Immunocytochemical localization of carcinoembryonic antigen in cerebrospinal fluid with metastatic carcinoma of the stomach: report of four cases.

A study was undertaken to evaluate the cellular findings of gastric carcinoma in cerebrospinal fluid (CSF). Immunocytochemical localization of carcinoembryonic antigen (CEA) was performed on four cases of metastatic gastric carcinoma cells in CSF samples. A positive peroxidase-antiperoxidase (PAP) reaction was obtained in all cases, with intense staining for CEA in the CSF samples as well as in the paraffin-embedded tissue sections from the primary gastric tumors. Cellular morphology and the results of immunoperoxidase staining can be studied simultaneously. We believe that the PAP method for CEA increases diagnostic accuracy of cytology in the CSF.

Demonstration of a Marek's disease virus-specific antigen in tumour lesions of chickens with Marek's disease using monoclonal antibody against a virus phosphorylated protein.

By use of monoclonal antibody against a Marek's disease virus (MDV) serotype 1-specific phosphorylated protein, MDV antigen-positive cells were demonstrated in tumour lesions of various visceral organs of chickens with Marek's disease. However, these tumour lesions did not appear to have the MDV glycoproteins gA and gB, which are considered to be late gene products of the virus genome gA and gB as well as the phosphorylated protein were detected in the feather follicle epithelium, which is a permissive site for MDV replication.

Immunoperoxidase staining for the detection of herpes simplex virus antigen in cervicovaginal smears.

Destained cervicovaginal smears from eight patients with herpes simplex virus (HSV) infections were stained by means of the peroxidase-antiperoxidase (PAP) technique to demonstrate the presence of the HSV type 2 (HSV-2) antigen. Positive results were obtained in six of the eight cases, with intense staining for the HSV-2-specific antigen throughout the cytoplasm and nuclei of cells having a ground-glass nuclear appearance as well as in multinucleated giant cells. Virus isolation was successfully performed for the HSV-2-positive case that also had a histologically confirmed squamous-cell carcinoma of the cervix. The combined use of cytology and the PAP staining technique was of great value in the demonstration of cervical HSV infections.

Induction of acute myoclonic encephalopathy in hamsters by subacute sclerosing panencephalitis virus.

Myoclonus is a characteristic neurological sign of subacute sclerosing panencephalitis (SSPE). Attempts were made to induce myoclonus in a large proportion of hamsters with a cell-associated strain of SSPE virus (the Biken strain) and thereby to establish an experimental model for study of the mechanism of development of this condition. When injected intracerebrally, Biken virus induced myoclonus within two to 14 days in 84% of the three- to nine-week-old hamsters tested. Electroencephalographic traces showed a periodic and synchronous discharge consisting of high-voltage slow waves and spikes that appeared coincidentally with myoclonus. Neurons in the cortex and thalamus of the affected animals had severely degenerated cytoplasm. Inflammatory changes, such as perivascular cuffing or infiltration of mononuclear cells, were not detected. Staining with immunoperoxidase revealed measles viral antigens in the cytoplasm and dendrites of the affected neurons. SSPE virus with the same properties as the parent virus was recovered from brain cells of sick animals by cocultivation with Vero cells.

Co-existence of malignant squamous cells and herpes simplex virus type 2-infected cells.

A case is presented in which malignant squamous cells and herpes virus infected cells were recognized concomitantly at routine cytologic examination for detection of cervical cancer. Further examinations on admission revealed Stage Ib carcinoma of the cervix and characteristic pathologic changes of herpetic infection in the tumor cell nests. Virus was isolated from the cancerous tissues in the ectocervix following radical hysterectomy, and was identified as herpes simplex virus type 2 (HSV-2). The typical HSV-2 particles were detected by electron microscopic observation on the infected FL monolayer cells with isolation. Neutralizing antibody levels against HSV-2 declined and remained low after hysterectomy. The HSV-2, isolated and identified, might be considered as "passenger" virus. The role of this isolated virus as an etiologic agent of the cervical cancer was not clear.