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Toll-Like Receptor-Targeting Immunotherapy for Oral Cancer

Oral Science International.2004;1(1):1-15.

It is important to augment the anti-cancer host response in cancer treatment. Recent studies have suggested that the signaling that occurs via the Toll-like receptors (TLRs), which are newly identified receptor molecules recognizing many pathogens, are involved in the induction of anti-cancer immunity. OK-432, a penicillin-killed and lyophilized preparation of Streptococcus pyogenes, is being successfully used as an immunotherapeutic agent in many types of malignancies. However, the molecular mechanisms of OK-432, a whole bacterial preparation, and the active components which make it effective against cancer, remain uncertain. We have succeeded in isolating the active component of OK-432 (lipoteichoic acid-related molecule, OK-PSA) by affinity chromatography of a butanol extract of OK-432 on the CNBr-activated sepharose 4B bound TS-2 monoclonal antibody that neutralizes the interferon (IFN)-γ-inducing activity of OK-432. OK-PSA induced Th1-type cytokines both in human and in mice, and elicited an anti-cancer effect in tumor-bearing mice via TLR4. Furthermore, our clinical study revealed that TLR4 signaling is intimately involved in the anti-cancer effect achieved by OK-432 in patients with oral cancer. We elucidated that OK-432 is first captured and digested by phagocytes, such as dendritic cells and macrophages, and then its active component, OK-PSA, which is released from the phagocytes, stimulates TLR4 signaling. It is strongly suggested that OK-PSA is the molecule most responsible for the anti-cancer effect of OK-432, and that TLR4 may be a definite molecular target for cancer immunotherapy with OK-432/OK-PSA.

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Molecular Analysis of Salivary Gland Branching Morphogenesis

Takayoshi Sakai ; Melinda Larsen ; Mikihiko Kogo ; Kenneth M. Yamada

Oral Science International.2004;1(1):16-21.

Recently, clinicians and scientists have focused on tissue engineering for regenerative medical therapy. This approach promises to provide remarkable clinical breakthroughs for the future. In oral and craniofacial medicine, most scientific approaches to tissue engineering currently involve tooth and bone, while little progress has been made toward regenerating organs such as salivary gland. To develop strategies for salivary gland regeneration, it will be important to understand the molecular mechanisms of normal salivary development. This mini-review describes a recently developed and tested set of approaches for identifying and characterizing molecules essential for branching morphogenesis and other developmental processes. It shows the value of using laser microdissection and the new process of T7-SAGE for gene discovery of putative candidate molecules that may be crucial regulators or mediators. We describe a stepwise series of associated strategies for reliable identification and functional testing of a candidate molecule, as well as its successful application to a specific candidate molecule originally identified by T7-SAGE.

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The Relationship between Subcellular Localization of Actinin-4 and Cell Motility in Oral Squamous Cell Carcinoma

Yutaka Iida ; Hiroshige Chiba

Oral Science International.2004;1(1):30-37.

To elucidate the mechanism of invasion of oral squamous cell carcinoma, we newly established two different cell lines with a high-motility phenotype (designated HI type) and low-motility phenotype (LI type) from CA-9-22, a human oral squamous cell carcinoma cell line, through cell invasion assay (Boyden chamber assay). When we examined the subcellular localization and protein expression of actinin-4 using these cell lines, although the growth curves were not significantly different between the HI type and LI type, more invasion was seen in the HI-type than in the LI-type on Boyden chamber assay (p<0.0001). Morphologically, a larger number of sharply extended cell processes and spindle formation were observed in the HI-type than in the LI-type, and actinin-4 was mainly distributed in these processes. Western analysis showed that the expression level of actinin-4 was almost equivalent between the HI and LI types. These findings suggest that subcellular localization of actinin-4 might be involved in cell motility and cancer invasion by regulating the actin cytoskeleton at the cell processes in oral squamous cell carcinoma.

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Computed Tomographic Analysis of Inflammatory Spread in Adult Parotitis

Masahiro Izumi ; Yoshiko Ariji ; Akitoshi Katsumata ; Masakazu Gotoh ; Munetaka Naitoh ; Kenichi Kurita ; Kazuo Shimozato ; Masami Fujishita ; Eiichiro Ariji

Oral Science International.2004;1(1):38-44.

Adult parotitis exhibits various appearances on images. The purpose of this study was to investigate the imaging features of adult parotitis, focusing on the inflammatory spread in and around the parotid gland on CT images. We investigated 21 patients who were clinically and radiologically diagnosed with parotitis, including patients with concomitant occurrences of sialolith or Sjören's syndrome. Intra-glandular spread was divided into 13 diffuse types and 8 focal types. There was a significant difference in duration from the first onset of symptoms between the diffuse and focal types. Six of 8 patients showing focal type spread were associated with the concomitant occurrence of sialolith or Sjören's syndrome. Extra-glandular spread was observed in 13 (62%) patients. Of them, 12 (92%) showed subcutaneous fat tissue and/or masticator space involvement. Involvement of the parapharyngeal space and the superficial area just below the gland was observed in 4 and 7 patients, respectively. The appearance of extra-glandular spread was influenced by intra-glandular appearance and concomitant disease. The spread of adult parotitis possessed characteristic features in relation to clinical findings and concomitant disease. CT examination appeared to be effective for these diagnoses.

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Application of the Digital Radiographic Bone Trabecular Structure Analysis to the Mandible using Morphological Filter

Sumio Sakoda ; Ryota Kawamata ; Takashi Kaneda ; Isamu Kashima

Oral Science International.2004;1(1):45-53.

[Purpose] To develop a digital radiographic bone trabecular structure analyzing system and to applied it to mandible. [Methods] Structural change was simulated by removing spongy bone in human dried mandible. Using a morphological filter processing, the skeletal features were extracted from digital radiographic image data obtained by computed radiography before and after the removal of spongy bone. The skeletal volume, surface, thickness, number, separation, complexity, spacing continuity and connectivity on the skeletal binary images were determined by morphometric indices calculation, star volume analysis and node-strut analysis. Using these quantified skeletal feature parameters, a mapping sheet for the structural evaluation was produced. The correlation between the fluctuation rate of each parameter after the removal of spongy bone with respect to the value before the removal of spongy bone and simulated structural change was examined. [Results] The skeletal binary image data quantified sixteen skeletal feature parameters using morphometric indices calculation, star volume analysis and node-strut analysis. These parameters agreed with the theoretical fluctuation pattern in the simulated skeletal structure deterioration of the mandibular bone trabeculae. [Conclusion] Using a morphological filter and bone histomorphometry, the radiological bone-morphometric analyzing system is useful for the evaluation of the mandibular trabecular structure.

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Crucial Role of Tissue-specific Apoptosis on the Development of Primary Sjögren's Syndrome

Yoshio Hayashi ; Rieko Arakaki ; Naozumi Ishimaru

Oral Science International.2004;1(2):55-64.

Primary Sjögren's syndrome is an autoimmune disorder characterized by lymphocytic infiltrates and destruction of the salivary and lacrimal glands, and systemic production of autoantibodies to the ribonucleoprotein (RNP) particles SS-A/Ro and SS-B/La, leading to clinical symptoms of dryness of the mouth and eyes (sicca syndrome). Autoreactive T cells bearing the CD4 molecule may recognize an unknown self antigen, triggering autoimmunity in the salivary and lacrimal glands. Although several candidate autoantigens including α-fodrin have been reported in Sjögren's syndrome, the pathogenic roles of the autoantigens in initiation and progression of SS are still unclear. It is possible that individual T cells activated by an appropriate self antigen can proliferate and form a restricted clone. Recent evidence suggests that the apoptotic pathway plays a central role in making T cells tolerant to tissue-specific self antigen, and may drive the autoimmune phenomenon. We recently reported that tissue-specific apoptosis in estrogen-deficient mice may contribute to autoantigen cleavage, leading to the development of autoimmune exocrinopathy. The studies reviewed imply that tissue-specific apoptosis and caspase-mediated α-fodrin proteolysis are involved in the progression of autoimmune lesions in Sjögren's syndrome. Moreover, Fas ligand (FasL) and its receptor Fas are essential in the homeostasis of the peripheral immune system. It is considered that a defect in activation-induced cell death (AICD) of effector T cells may result in the development of autoimmune exocrinopathy in Sjögren's syndrome.

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Relationship between Degenerative Bone Changes of Condylar Surface and Articular Disc Disorders in Symptomatic Osteoarthrosis of Temporomandibular Joints

Kousuke Honda ; Yoshiko Natsumi ; Masahiro Urade

Oral Science International.2004;1(2):65-70.

Purpose: We investigate the relation of bone changes of the condylar surface to disc displacement and discuss the development of joint symptoms in osteoarthrosis of temporomandibular (TM) joints.Subjects and Methods: Seventy-seven patients with an image diagnosis of degenerative bone changes of the unilateral condylar surface accompanied with joint symptoms were studied. The bone changes were assessed by panoramic radiographs and classified into two groups: pathologic bone changes (PBC) including erosion, osteophyte and deformity, and adaptive bone changes (ABC) including flattening and concavity. Magnetic resonance imaging was performed on the subjective TM joints to examine the configuration and position of articular discs. A visual analogue scale was used for evaluation of joint pain.Results: Erosion and deformity showed significantly higher prevalence than the other three kinds of bone changes in the joints with anterior disc displacement without reduction (ADWoR) as compared to those with anterior disc displacement with reduction (ADWR). The cases with the vertical disc position to the condyle ranging from 60° to less than 150° were more frequent than those ranging from 0° to less than 60° in the PBC group, whereas the cases with the vertical disc position to the condyle ranging from 0° to less than 60° were more frequent than those ranging from 60° to less than 150° in the ABC group. The average degree of joint pain when chewing but not jaw opening was higher in the joints with ADWoR than in those with ADWR, and in the PBC group than in the ABC group.Conclusion: The bone changes of the condylar surface diagnosed as PBC tended to induce more advanced disc displacement and chewing pain than those diagnosed as ABC.

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Alterations in G1 Phase Cell Cycle Regulation during the Development of Benzo[a]pyrene-induced Epithelial Dysplasia in the Murine Tongue

Misaki Ota-Sanada ; Daisuke Ito ; Ming-Heng Li ; Takeshi Odani ; Ai Kawamata ; Masayasu Iwase ; Masao Nagumo

Oral Science International.2004;1(2):71-79.

The environmental contaminant benzo[a]pyrene (B[a]P) has been regarded as one of the pathogens of oral premalignant and malignant lesions. To elucidate the pathogenesis of oral premalignancies, B[a]P-induced dysplasia of the murine tongue was investigated for G1-associated cell cycle regulation. B[a]P solution was applied orally up to six weeks to induce epithelial dysplasia of the tongue. BrdU incorporation and the expression of p21, cyclin D1, and CDK4 were examined by immunohistochemistry and Western blotting. Rb phosphorylation and E2F-Rb binding were examined by immunoprecipitation and Western blotting. B[a]P treatment resulted in dysplastic changes and active DNA synthesis in the tongue epithelia. Immunohistochemical analyses showed p21 up-regulation and cyclin D1/CDK4 overexpression in B[a]P-induced dysplasia. Rb hyperphosphorylation and E2F release were caused by B[a]P treatment. Thus, dysregulation of G1-phase regulation is likely to be an important event in the development of oral epithelial dysplasia in mice.

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Detection of DNA and Immunoreactivity Specific for Mycoplasma fermentans in Synovial Fluid and Sera from Patients with Temporomandibular Joint Disorders

Masako Suga ; Eiro Kubota ; Takanori Shibata

Oral Science International.2004;1(2):80-88.

This study investigated whether the presence of Mycoplasma fermentans (M. fermentans) in the temporomandibular joint (TMJ) could be associated with the pathology of temporomandibular joint disorders (TMD). One hundred fifteen synovial fluid (SF) samples from patients with TMD were evaluated for the presence of DNA of M. fermentans by polymerase chain reaction (PCR) assay. Specific antibody against M. fermentans was also detected in the SF as well as sera by Western blot analysis. M. fermentans DNA was identified in 37.4% of the SF samples from the TMD patients. There was no difference between PCR-positive and -negative rate regarding sex and disease categories, e.g., internal derangement (ID) and osteoarthritis (OA). However, the prevalence of M. fermentans DNA in ID patients was higher in elderly patients (73.3%) than in younger patients (31.8%). Anti-M. fermentans immunoreactivities (IgG) specific for lipoproteins with various molecular sizes, 56 kilo-Dalton (kDa), 48 kDa, 38 kDa, and 29 kDa, were also identified in the SF. The immunoreactivity was also detected in the patients'sera. The reactivity patterns of the anti-M. fermentans antibodies were, however, different between the SF and the sera; reactivities to 48 kDa and 29 kDa lipoproteins were prominent in the former, while the reactivities to those of 56 kDa, 48 kDa, and 29 kDa were evidently increased in the latter. The presence of specific DNA and antibody for M. fermentans in the TMJ implies that M. fermentans could possibly induce joint specific immunoreaction, thus perpetuating the inflammatory reaction in the diseased TMJ.

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Nodular Fasciitis in the Buccal Mucosa

Tetsuro Ikebe ; Yuichi Ogata ; Yasuo Takamune ; Kazutoshi Ota ; Takehisa Obayashi ; Masanori Shinohara

Oral Science International.2004;1(2):89-92.

A case of nodular fasciitis which arose from the buccal submucosal region is reported. One week after an incisional biopsy, the lesion enlarged alarmingly and protruded from the submucosa. Although a sarcoma was suspected because of rapid growth, the diagnosis of the biopsy was nodular fasciitis showing a haphazard arrangement of plump fibroblasts without atypical mitoses. After complete resection, no signs of recurrence were seen.

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