Electron microscopic, immunofluorescent and virological studies on a rhabdomyosarcoma in epidermodysplasia verruciformis.

A subcutaneous tumor of a patient with epidermodysplasia verruciformis was studied by the light microscopy, the electronmicroscopy and the immunofluorescent test. The tumor cells were histologically pleomorphic and electronmicroscopically contained varying amounts of cytoplasmic filaments without Z-band formation. The antimyosin serum stained the tumor cells, showing their myogenic origin. No virus or virus-like particles were observed in the tumor. Tumor antigens stainable by the patient’s serum were not detected. Hamsters inoculated with the tumor extract at birth developed no noticeable diseases. ∗PMID: 4264056 [PubMed indexed for MEDLINE] Copyright c ©OKAYAMA UNIVERSITY MEDICAL SCHOOL Acta Med. Okayama 25, 643-648 (1971) ELECTRONMICROSCOPIC, IMMUNOFLUORESCENT AND VIROLOGICAL STUDIES ON A RHABDOMYOSARCOMA IN EPIDERMODYSPLASIA VERRUCIFORMIS* Yoshiro YABE and Hiroko KOYAMA Department oj Virology, Cancer Institute, Okayama University Medical School, Okayama 700, Japan (Director: Prof: Y. Yabe) Received jor publication, Oct. 22, 1971 Presence of C.type virus-like particles in a rhabdomyosarcoma of a patient with epidermodysplasia verruciformis was reported by OKAMOTO et al. (1). The authors, one of whom (Y. V,) was a coauthor of that report, thoroughly re-examined the tumor of the same patient by the electronmicroscope and the immunofluorescent test, and found out that the tumor was convincingly a rhabdomyosarcoma but had no virus or virus-like particles in it. With this tumor, further virological studies were done by the immunofluorescent test and by inoculating its tissue extract into newborn hamsters. The present paper concerns the results of these studies. MATERIALS AND METHODS Case: The same patient as reported by OKAMOTO et al. (1); a 40-year-old man, K. K., with the typical skin lesions of epidermodysplasia verruciformis and a 4 x 5 x 3 cm subcutaneous tumor on the right lateral chest wall. The subcutaneous tumor was removed surgically. Three months later, a metastatic tumor was observed in the right inguinal region and removed surgically. The patient died of widely disseminated tumor metastases 9 months after the first operation. Autopsy could not be done. The primary tumor and a metastatic tumor were used for the present study. Histological examination: The tissue was fixed in 10 % formalin and embedded in paraffin. The sections were stained with hematoxylin-eosin and Heidenhain's iron-hematoxylin. Elertronmicroscop),: A part of the tumor was fixed in 6 % glutaraldehyde, post-fixed in 1 % osmium tetroxide, dehydrated, embedded in Epon 812, sectioned and stained doubly with uranyl acetate and lead citrate. immunofluorescence for m)losin: The human myosin and the guinea-pig antimyosin serum were prepared by the methods similar to those described by HIRAMOTO * This wor~ was supported in part by grants from the Ministry of Education and the Ministry of Health and Welfare of Japan. 643 1 Yabe and Koyama: Electron microscopic, immunofluorescent and virological studies Produced by The Berkeley Electronic Press, 1971 644 Y. YABE and H. KOYAMA et at. (2). The fluorescein isothiocyanate-labeled rabbit antiserum against guineapig globulin was obtained from the Institute of Medical Science, Tokyo University. Using these two antisera, detection of myosin was done by the indirect method (2). Immun~fluorescence for tumor antigens: The patient's sera were obtained at the time of operation of the primary tumor and at the late stage with wide-spread metastases. The fluorescein isothiocyanate-labeled rabbit antiserum against human globulin was obtained from the Institute of Medical Science, Tokyo University. Using these sera, frozen sections fixed in acetone were stained by the indirect method. Tissue extract: Tumor tissue was ground in a chilled mortar, suspended in phosphate-buffered saline to 20 %, centrifuged at 3,500 rpm for 15 minutes and the supernatant was preserved at -60°C for 1-3 days before inoculation. Onetwentieth milliliter of thus prepared extract was inoculated into the muscular part of the thigh of hamsters within 24 hours after birth.