Michihiro Kamide

Establishment of Epstein-Barr virus (EBV)-associated nuclear antigen (EBNA)-positive nasopharyngeal carcinoma hybrid cell line (NPC-KT)

SummaryPrimary nasopharyngeal carcinoma (NPC) cells were fused to hypoxanthine-guanine phosphoribosyltransferase (HGPRT)-defective cells derived from adenoid tissues using Sendai virus. Some of the fused cells developed into epithelial-like hybrid cells in a selective HAT medium. The hybrid cells (NPC-KT) were Epstein-Barr virus (EBV)-associated nuclear antigen (EBNA)-positive cells. There have been no reports on the establishment of EBNA-positive epithelial cell lines derived from NPC. Thus, the epithelial-like hybrid cells might serve as an in vitro model for studying the biologic activity of NPC-associated EBV.

Significance of Intravenous Olfaction Test Using Thiamine Propyldisulfide (Alinamin) in Olfactometry

The significance of intravenous olfaction test in olfactometry was studied by injection of thiamine propyldisulfide (Alinamin) intravenously. An original solution of Alinamin is a thiol-type derivative of vitamin B1, and releases a mercaptan smell (garlic smell) in expired air when it arrives at the olfactory epithelium via the nasopharynx. In the intravenous olfaction test (Alinamin test), the latent time which is a period between the initiation of injection and recognition of garlic smell, and duration time which is a period between the recognition and disappearance of smell are measured. Our results indicated that latent time is influenced by olfactory acuity and duration time depends on olfactory adaptation phenomenon. Central olfactory disorders were highly suspected in hyposmia patients with duration time of less than 15 sec, and nonresponders in Alinamin test always showed poor prognosis in the recovery of olfactory acuity. It was considered that the Alinamin test is useful not only for estimating the degree of olfactory disorders, but also for differential diagnosis of impaired lesions and olfactory prognosis.

Electro-Olfactogram (EOG) in Olfactometry

The electro-olfactogram (EOG) is a potential originated in olfactory epithelium evoked by odorous stimulation, which has been demonstrated and termed by Ottoson (1959). In order to ascertain the clinical application of EOG, we attempted to record similar potentials in human subjects. In some preliminary experiments using rabbits, it was quite easy to produce Ottosons findings. In human subjects, however, there were sometimes great technical difficulties owing to the hidden anatomical position of the olfactory epithelium. Then we utilized Olympus selfoscope endoscopy for placing and retaining the electrode to the olfactory portion precisely and safely, with a tube for sending airborne odor. As results, we succeeded in recording negative spike discharge similar to Ottosons findings. We examined a total of 27 nasal cavities in 22 patients with peripheral olfactory disorders and 11 nasal cavities in 12 patients with central olfactory disorders. The positive EOG rate in patients with peripheral or central olfactory disorders was 28 and 69%, respectively. Moreover, the positive rate was decreased in proportion to the degree of olfactory disorders in chronic rhinosinusitis. No positive EOG was observed in anosmic nasal cavities, while it was 68% in normal persons and resulted in an equal value to anosmia group caused by the damage of central olfactory pathway. From these results, it is considered that the positive EOG study is the only method which provides objective finding for the differential diagnosis of anosmia caused by disorders of olfactory epithelium or disorders of central olfactory tract.

Importance of Unilateral Examination in Olfactometry

Hyposmia, the decreased sense of smell, and anosmia, the loss of sense of smell, may be unilateral or bilateral. If the olfactory acuity examined by means of bilateral test is normal, olfactory disorders are not found; unilateral examination is therefore necessary for definite evaluation of olfactory acuity. As evidence, 7 cases out of 94 patients with chronic rhinosinusitis and 6 cases out of 12 patients who received the surgery of anterior cranial fossa showed definite different olfactory threshold between nasal cavities, and there were no patients who recognized the diminished sense of smell in spite of unilateral high olfactory threshold. Additionally, we have experienced that a patient with brain tumor was diagnosed by the help of unilateral olfactory test. We thus strongly recommend the unilateral olfactometry as a method for simple and reliable test in clinical measurement of the sense of smell.

Three cases of inter-sterno-costo-clavicular ossification associated with tonsillar infection

Three cases of inter-sterno-costo-clavicular ossification (ISCCO) associated with chronic tonsillitis are reported. Two cases had additional symptoms of pustulosis palmaris et plantaris (PPP). All the patients underwent tonsillectomy. The histological changes of the palatine tonsils of case 1 and case 2 were characterized by so-called focal small lacunar ulcers and periarteriolar fibrosis (onion-skin lesion). Some multinucleated giant cells were observed in case 2. Following tonsillectomy, symptoms in the clavicular region and the PPP improved. It is possible to consider that ISCCO is caused by focal tonsillar infection.

Unusual Case of Plasma Cell Tumor with Monoclonal Gammopathy of the Sino-Nasal Cavity and Clavicular Bone

A rare case of extramedullary plasmacytoma in the sino-nasal cavity is presented. During a prolonged clinical course, the tumor has recurred several times after irradiation, chemotherapy and surgery, and presented in the clavicular bone as well. Immunohistochemical staining of each tumor showed IgA kappa phenotype, and immunoelectrophoresis of the serum showed overproduction of IgA with kappa light chains. Although an elevation of serum IgA persists, the patient has remained well without evidence of overt multiple myeloma.

Epstein-Barr Virus (EBV) Infection by Cocultivation of Fibroblast-Dominant Cell Line and EBV-Transformed Lymphocytes

A long-term cell culture epitheloid cell line (NPC-F/L) was established from the fibroblast-dominant cell line derived from nasopharyngeal carcinoma tissue by cocultivation with an Epstein-Barr virus (EBV)-transformed lymphoblastoid cell line of adenoid origin. The cells were investigated by karyotyping, by immunofluorescent testing for EBV-associated nuclear antigen (EBNA), early antigen (EA), and viral capsid antigen (VCA) and by a light microscopy. The cells were shown to be epithelial not only by morphology but also by immunocytochemical staining with monoclonal antibody to cytokeratins and they expressed EBNA and EA but not VCA. The cells developed histologically undifferentiated squamous cell carcinoma when inoculated subcutaneously into the back of athymic nude BALB/c(nu/nu) mice, which were similar to the human tumors from which the fibroblastoid cell line was established. The presence of EBNA in touch smears prepared both from the biopsy tissue of human and nude mice was demonstrated. Karyotyping analysis indicated an aneuploid chromosome with a model number of 74.

Inhibitory Effects of Kampo Medicine on Epstein-Barr Virus Antigen Induction by Tumor Promoter

We have investigated the possible interaction of Japanese Kampo medicine; Shosaiko-to with Epstein-Barr virus (EBV) induction by TPA (12-O-tetra-decanoyl-phorbol-13-acetate) in EBV-genome harboring cells. The effect of ether extract of Shosaiko-to was also studied on the induction of EBV by superinfection with EBV from P3HR-1 cells (P3H-EBV). Concomitant treatment of EBV-latently-infected cells (Raji and A2L/AH) with TPA and ether extract resulted in effective inhibition of EBV-antigen induction. However, in the experiments to test the influence of ether extract on EBV induction by superinfecting Raji cells with P3H-EBV, ether extract did not affect the percentage of EBV-antigen-positive cells. These data indicate that Shosaiko-to is effective to inhibit antigen synthesis induced by chemical promoters without affecting EBV-antigen induction after superinfection.

A case of pharyngeal syphilis and surveillance of sexually transmitted disease.

Lesions of secondary syphilis were found in the right tonsil and on the palatoglossal arch. The pharyngeal mucous membrane and tonsil exhibited severe erosion with white-gray coating. Several Treponema pallida were seen in smears of the tonsillar exudate. Serological tests revealed high VDRL and TPHA titers. Histological examination showed severe inflammation with many lymphocytes and plasma cells, and thickened vascular endothelium. We also describe the present program of surveillance of sexually transmitted diseases in Japan and emphasize that ENT-physicians should be more aware of them in patients with otorhinolaryngological symptoms and signs.

[Cytotoxic effector cells specific for Epstein-Barr nuclear antigen positive nasopharyngeal hybrid cells are present in patients with infectious mononucleosis].

Peripheral lymphoid cells, from 3 cases of acute infectious mononucleosis (IM) and 2 cases of healthy individuals, were tested in a colony inhibition assay for cytotoxic activity against two cell lines that did or did not carry the Epstein-Barr virus (EBV) genome, A2L/AH and Ad cells respectively. T-lymphocyte fraction of IM was specifically cytotoxic for EBV-genome carrying cells and the residual lymphocyte fraction had high ADCC but low NK activity compared to the normal donors. When the A2L/AH cells which were forced to enter the EBV-cycle by n-butyrate treatment used as target cells, B-cell enriched fraction showed increased cytotoxic effect that was more remarkable in NK activity than ADCC in normal donors. These results were considered to be useful for possible specific immuno-therapy in nasopharyngeal carcinoma.