Mitsuru Urata

Effects of 5-fluorouracil and the combination of 5-fluorouracil and human leukocyte interferon on human salivary gland adenocarcinoma cell line in culture

The growth inhibitory effects of 5-fluorouracil (5-Fu) and the combination of 5-Fu and human leukocyte interferon (HuIFN-alpha) on the human salivary gland adenocarcinoma cell line HSG in culture were examined by measuring colony formation in the semi-solid agar medium and cell proliferation in the monolayer culture. As a consequence, the colony-forming ability of HSG cells in the agar medium containing 5-Fu was found to be markedly inhibited as compared with the untreated control. The concentration of 5-Fu yielding 50% inhibition of colony-forming ability of HSG cells under the presence of 5-Fu was 0.08 micrograms/ml. When the growth inhibitory effects of the combination of 5-Fu and HuIFN-alpha on HSG cells were examined, the colony forming ability of HSG cells was synergistically inhibited, whereas effects of the combined treatment on HSG cells in the monolayer culture were less sensitive than those on the colony formation. These findings strongly suggest that the combination of 5-Fu and HuIFN-alpha or 5-Fu alone is selectively effective on the neoplastic cells of HSG cell population but not on the non-neoplastic cells.

Therapeutic effect of human fibroblast interferon on premalignant lesions arising in oral mucosa. A pilot study.

Human fibroblast interferon (HuIFN-beta) was topically administered to 20 premalignant lesions histopathologically showing epithelial dysplasia such as leukoplakia and lichen planus which arose in the oral mucosa. HuIFN-beta was prepared in the water-soluble gel form containing 2% carboxymethylcellulose, 45% glycelin, 0.1 M citrate buffer (pH 4.5) and 0.2% SDS as stabilizing agents. This preparation was found to be effective for herpetic gingivostomatitis and zostal lesions arising along the intercostal nerve. Thus, the HuIFN-beta preparation (10(4) to 5 X 10(3) IU) was applied to the oral mucosal lesion for 1 h twice a week. The lesion with topical administration of HuIFN-beta was covered tightly with the mucosal bandage which was coated with carboxymethylcellulose, glycelin and CaCl2 on vinyl acetate matrix. The 14 oral lesions with erosion or ulcer formation accompanied by severe pain by touch, had complete remission after approximately 10 successive applications of this preparation. Although subjective symptoms such as irritation pain in the other 6 patients with severe hyperkeratotic lesion subsided, white coatings and streaks could not be completely removed by this therapy. No other side-effects excluding slight pain and reddish swelling which occurred intermittently during HuIFN-beta administration were observed.

Interferon activity and its characterization in the sera of patients with head and neck cancer

The interferon (IFN) assay of the sera from the 40 patients with head and neck cancer was performed by the plaque‐reduction assay with vesicular stomatitis virus in FL cells derived from human amniotic membrane. The patients mainly had Stage III or IV lesion without distant metastasis, and previously had not received any cancer therapy. All of the patients were histologically diagnosed as squamous cell carcinoma. When the serum IFN activity was characterized by acid treatment, significant increases of IFN‐α/β/γ (n = 24, P < 0.05) and acid‐labile IFN (n = 24, P < 0.001), and significant decrease of acid‐stable IFN (n = 24, P < 0.001) in the cancer patients of 50‐to‐79‐year age group were found, as compared with those in the normal controls of the same age group (n = 20). When IFN titers including various immunologic parameters of the patients and normal controls were simultaneously assayed prior to the beginning of the cancer therapy, the titers of IFN‐α/β/γ, acid‐stable IFN, and acid‐labile IFN were significantly correlated with some immunologic parameters such as natural killer (NK) activity, the absolute number of Tγ lymphocytes, the percentages of β‐ and γ‐globulin, and the amounts of IgA, IgG, IgM, and β2 microglobulin. To define further the nature of this IFN, both sera of the patients and normal donors of 50‐to‐79‐year age group were characterized by a neutralization assay with an antiserum to HuIFN‐α and HUIFN‐β. The IFN activity left when the testing sera were neutralized with these antisera was expressed as γ‐like IFN. The titers of γ‐like IFN in the sera of patients (n = 24, P < 0.0001) showed a highly significant increase as compared with the normal controls (n = 20). When the correlation between prognosis of the disease and titers of serum IFN were investigated by measuring γ‐like IFN and acid‐stable IFN in the sera of patients, all of nine patients with good prognosis after the cancer treatment showed significant decreased levels of γ‐like IFN (P < 0.01) and acid‐stable IFN (P < 0.05) as compared with those on the time before cancer therapy. On the other hand, titers of γ‐like IFN in the sera of six patients with recurrent disease showed a significant increase as compared with those on the IFN measurement before cancer therapy (P < 0.05). These findings led us to suggest that the serum IFN levels, especially γ‐like IFN activity, may be associated with a function of some loops of the immune system in the patients with head and neck cancer.

Interferon activity and its characterization in the sera of patients with premalignant lesions arising in oral mucosa

The interferon (IFN) assay of the sera from the 26 patients with premalignant lesions such as lichen planus and leukoplakia arising in oral mucosa was performed by the plaque-reduction assay with vesicular stomatitis virus in FL cells derived from human amniotic membrane. When the serum IFN activity was characterized by acid treatment, significant increase of acid-stable IFN in the patients was found as compared with those in the normal controls. The titers of gamma-like IFN defined by anti-HuIFN-alpha and anti-HuIFN-beta in the sera of patients of 50-79 years age group (n = 17, P less than 0.002) showed a highly significant increase as compared with the relevant normal controls (n = 20). All of the 26 patients were treated with topical administration of HuIFN-beta. When the correlation between prognosis of the disease and titers of serum IFN was investigated by measuring gamma-like IFN and acid-stable IFN in the sera of patients, all of 13 patients with good prognosis after the HuIFN-beta therapy showed significantly decreased levels of gamma-like IFN (P less than 0.01), whereas the serum level of acid-stable IFN after HuIFN-beta therapy showed a significant increase compared to that before the therapy (P less than 0.05). These findings indicate that the endogenous IFN system may be associated with the pathophysiology in patients with the oral mucosal lesions.

Sensitivity of a neoplastic epithelial duct cell line from a human submandibular salivary gland to human leukocyte interferon as assessed by an in vitro semi-solid agar technique

A neoplastic epithelial duct cell line from a human submandibular salivary gland, HSG, was assayed by an in vitro semi-solid agar technique for sensitivity to human leukocyte interferon (HuIFN-alpha). The clone which showed the most stable growth and the greatest ability of colony formation in the agar medium, was isolated from the colonies of HSG cells and used in the present study. This clone was designated as HSG-1. As a consequence, cultivation of HSG-1 cells in the agar medium containing HuIFN-alpha of 1000 units/ml resulted in the inhibition of 47.6% of the colony-forming ability of HSG-1 cells, as compared with the untreated control. Addition of increasing concentrations of HuIFN-alpha (between 100 units/ml and 1000 units/ml) in the agar medium caused a linear reduction in the number of the colonies, ranging from 5.2% to 47.6%. Moreover, addition of HuIFN-alpha showed an inhibitory effect on the multiplication of HSG-1 cells in the monolayer culture. These observations strongly suggest that HuIFN-alpha has antineoplastic and antiproliferative effects on HSG-1 cells.

An odontogenic myxofibroma related to an embedded third molar of the mandible: Report of a case

A case of odontogenic myxofibroma involving the angle of the mandible is reported. This case arose in the area closely associated with the pericoronal portion of the embedded third molar. When the mandible resected was histopathologically examined, the tumor mass containing various shapes of epithelial cell nests and amorphous calcified bodies in the myxomatous fibrous tissue was found to be formed in the localized area well-demarcated by the enamel of the embedded tooth. These findings strongly suggest that this lesion is of odontogenic origin.