Yukiko Ohyama

Oral involvement in chronic graft-versus-host disease after allogeneic bone marrow transplantation

We examined 37 patients who had undergone an allogeneic bone marrow transplantation and compared their oral findings to their systemic involvement with chronic graft-versus-host disease. Among the clinical signs and symptoms in their oral region, only the presence of oral lichenoid lesions had a statistically significant relationship to the diagnosis of chronic graft-versus-host disease. The histologic findings in the labial salivary glands and buccal mucosa closely reflected the status of chronic graft-versus-host disease. Statistically, the presence of diffuse and periductal lymphocytic infiltration in labial salivary glands, subepithelial lymphocytic infiltration and epithelial changes in buccal mucosa also showed a significant relationship to the diagnosis of chronic graft-versus-host disease. The present study suggests that a systematic oral examination, especially pathologic examination of the labial salivary glands and buccal mucosa, is useful in evaluating the status of chronic graft-versus-host disease.

Cytokine/chemokine profiles contribute to understanding the pathogenesis and diagnosis of primary Sjögren's syndrome

To investigate the pathogenesis of localized autoimmune damage in Sjögrens syndrome (SS) by examining the expression patterns of cytokines, chemokines and chemokine receptors at sites of autoimmune damage. mRNA expression of these molecules in the labial salivary glands (LSGs) and peripheral blood mononuclear cells (PBMCs) from 36 SS patients was examined using a real‐time polymerase chain reaction‐based method. Subsets of the infiltrating lymphocytes and chemokines/chemokine receptors expression in the LSG specimens were examined by immunohistochemistry. Cytokines/chemokine concentrations in the saliva were analysed using flow cytometry or enzyme‐linked immunosorbent assay. mRNA expression of T helper type 1 (Th1) cytokines, chemokines and chemokine receptors was higher in LSGs than in PBMCs. In contrast, mRNA expression of Th2 cytokines, chemokines [thymus and activation‐regulated chemokine (TARC/CCL17), macrophage‐derived chemokine (MDC/CCL22)] and chemokine receptor (CCR4) was associated closely with strong lymphocytic accumulation in LSGs. Furthermore, TARC and MDC were detected immunohistochemically in/around the ductal epithelial cells in LSGs, whereas CCR4 was detected on infiltrating lymphocytes. The concentrations of these cytokines/chemokines were significantly higher in the saliva from SS patients than those from controls, and the concentrations of Th2 cytokines/chemokines were associated closely with strong lymphocytic accumulation in LSGs. These results suggest that SS might be initiated and/or maintained by Th1 and Th17 cells and progress in association with Th2 cells via the interaction between particular chemokines/chemokine receptors. Furthermore, the measurement of cytokines/chemokines in saliva is suggested to be useful for diagnosis and also to reveal disease status.

Accumulation of Common T Cell Clonotypes in the Salivary Glands of Patients with Human T Lymphotropic Virus Type I-Associated and Idiopathic Sjögren’s Syndrome

To clarify the pathogenesis of human T lymphotropic virus type I (HTLV-I)-associated Sjögren’s syndrome (SS), the TCR Vβ gene usage by the infiltrating lymphocytes in the target organ was examined. The Vβ families predominantly used in the labial salivary gland (LSG) from the HTLV-I-seropositive (HTLV-I+) SS patients were more restricted than those from the HTLV-I-seronegative (idiopathic) SS patients, and were commonly Vβ5.2, Vβ6, and Vβ7. The single-strand conformation polymorphism analysis revealed that T cell clonotypes with Vβ5.2, Vβ6, and Vβ7 accumulate in the LSG from the HTLV-I+ and idiopathic SS patients. Among junctional sequences of the most dominant Vβ7 transcripts, the conserved amino acid motif (QDXG: X is any amino acid) was found in six of the five HTLV-I+ SS patients and was also detected in two of the five idiopathic SS patients. Using the probes specific to the motif, the Vβ7 transcripts with the motif were detected in the LSG from all of the seven HTLV-I+ and five of the six idiopathic SS patients, but not from eight healthy subjects. The Vβ7 transcripts with this motif were also detected in the HTLV-I-infected T cell lines obtained from the LSG of an HTLV-I+ SS patient. The accumulation of HTLV-I-infected T cells expressing TCR with the conserved motif was thus indicated. These T cells were commonly present in patients with idiopathic SS and are strongly suggested to most likely be involved in the pathogenesis of both HTLV-I-associated and idiopathic SS. This work was supported in part by grants from the Ministry of Education, Science, and Culture of Japan. Address correspondence and reprint requests to Dr. Seiji Nakamura, Second Department of Oral and Maxillofacial Surgery, Faculty of Dentistry, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. E-mail address: seiji@dent.kyushu-u.ac.jp Abbreviations used in this paper: HTLV-I, human T lymphotropic virus type-I; SS, Sjögren’s syndrome; LSG, labial salivary gland; PG, parotid gland; SSCP, single-strand conformation polymorphism; HAM/TSP, HTLV-I-associated myelopathy/tropical spastic paraparesis; HAAP, HTLV-I-associated arthropathy; CDR3, complementarity-determining region 3.

A comparison of glandular involvement between chronic graft-versus-host disease and Sjögren's syndrome.

Patients with chronic graft-versus-host disease (cGVHD) occasionally suffer from symptoms of xerostomia and xerophthalmia, which are also features of Sjógrens syndrome (SS). To identify differences in the glandular involvement between cGVHD and SS, we measured the proportions of infiltrating lymphocyte subsets and the expression of HLA-DR antigen and cell adhesion molecules in labial salivary glands (LSG). In cGVHD, more than 90% of the infiltrating lymphocytes were T cells with a slight predominance of CD8+ over CD4+ cells. In SS, CD4+ cells were predominant, and B cells accounted for 10-30% of the infiltrating lymphocytes. Ductal epithelial cell associated with lymphocytic infiltration expressed HLA-DR antigen in both cGVHD and SS. In SS alone, HLA-DR antigen expression also occurred without associated lymphocytic infiltration. The expression of adhesion molecules on ductal epithelial cells, especially vascular cell adhesion molecule 1, was more intense in SS than in cGVHD, while that on endothelial cell was similar in cGVHD and SS. These data suggest that the pathogenesis of glandular involvement of cGVHD is different from that of SS.

Severe Focal Sialadenitis and Dacryoadenitis in NZM2328 Mice Induced by MCMV: A Novel Model for Human Sjögren’s Syndrome

The genetic and environmental factors that control the development of Sjögren’s syndrome, an autoimmune disease mainly involving the salivary and lacrimal glands, are poorly understood. Viruses which infect the glands may act as a trigger for disease. The ability of sialotropic murine CMV (MCMV) to induce acute and chronic glandular disease was characterized in an autoimmune-prone mouse strain, NZM2328. MCMV levels were detectable in the salivary and lacrimal glands 14–28 days after i.p. infection and correlated with acute inflammation in the submandibular gland. After latency, virus was undetectable in the glands by PCR. At this stage, NZM2328 female mice developed severe chronic periductal inflammation in both submandibular and lacrimal glands in contrast to the much milder infiltrates found in female B6-lpr and male NZM2328. The focal infiltrates consisted of CD4+ and B220+ cells as opposed to diffuse CD4+, CD8+, and B220+ cells during acute infection. Salivary gland functional studies revealed a gender-specific progressive loss of secretory function between days 90 and 125 postinfection. Latent MCMV infection did not significantly affect the low incidence of autoantibodies to Ro/SSA and La/SSB Ags in NZM2328 mice. However, reactivities to other salivary and lacrimal gland proteins were readily detected. MCMV infection did not significantly alter the spontaneous onset of kidney disease in NZM2328. Thus, chronic inflammation induced by MCMV with decreased secretory function in NZM2328 mice resembles the disease manifestations of human Sjögren’s syndrome.

Accumulation of human T lymphotropic virus type I-infected T cells in the salivary glands of patients with human T lymphotropic virus type I- associated Sjogren's syndrome

OBJECTIVE To clarify the involvement of human T lymphotropic virus type I (HTLV-I) in the pathogenesis of Sjogrens syndrome (SS). METHODS In HTLV-I-seropositive patients with SS, HTLV-I proviral DNA in the labial salivary glands (SG) was detected by polymerase chain reaction (PCR) amplification of the extracted cellular DNA, and the localization in the SG was examined by in situ PCR hybridization. RESULTS The cellular DNA extracted from the SG contained full HTLV-I proviral DNA, which was present in the nucleus of the infiltrating T cells, but not in either the SG epithelial cells or the acinar cells. Furthermore, the viral loads in the SG were approximately 8 times to 9 x 10(3) times higher than those in the peripheral blood mononuclear cells. CONCLUSION Accumulation of HTLV-I-infected T cells in the SG suggests that HTLV-I likely causes the self-reactive T cells to proliferate, which, as a result, induces SS.

Clinical characteristics of Mikulicz's disease as an IgG4-related disease

ObjectivesMikulicz’s disease (MD) was considered to be a subtype of Sjögren’s syndrome (SS), based on histopathological similarities. However, recent studies have indicated that patients with MD show high serum IgG4 concentration, and suggested that MD is one of “IgG4-related disease” and distinguishable from SS. Therefore, we clinically and histopathologically examined the disease states of MD and SS in detail.Materials and methodsTwenty patients with Mikulicz’s disease and 18 with SS were comparatively studied to determine clinical characteristics in MD patients.ResultsSialography in MD patients did not show the “apple-tree sign” typically seen in SS. Serologically, high serum IgG4 levels but not anti-SS-A or anti-SS-B antibodies were observed in MD. SS showed lymphocytic infiltration of various subsets with atrophy or severe destruction of the acini, while MD showed selective infiltration of IgG4+ plasma cells with hyperplastic germinal centers and mild acini destruction. Corticosteroid treatment of MD reduced IgG and IgG4 levels and improved salivary function. A negative correlation between disease duration and increasing rate of salivary flow was observed in MD.ConclusionsThese results suggested that the pathogenesis of MD might be different from those of SS. Clinical Relevance: early diagnosis and treatment of MD is important for the improvement of salivary function.

Sonographic diagnosis for Mikulicz disease

OBJECTIVE The aim was to investigate the diagnostic imaging characteristics of Mikulicz disease (MD), especially sonographic ones, and to clarify the differences between them and those in Sjögren syndrome (SS), based on new criteria of MD. STUDY DESIGN The sonographic and sialographic images, as well as clinical, histopathologic, and serologic findings of 9 patients satisfying the new criteria of MD were analyzed and compared with those in SS. RESULTS All swollen submandibular glands showed bilateral nodal hypoechoic areas with high vascularization on sonograms and a parenchymal defect on sialograms, whereas parotid glands showed normal or slight change on both images. Nodal areas in submandibular gland sonograms were unclear on computerized tomography and on magnetic resonance imaging, but showed accumulation on gallium scintigraphy. CONCLUSION Mikulicz disease showed a high rate of bilateral nodal change in submandibular glands, which was completely different from SS. For detection and follow-up of these changes, sonography may be the best imaging modality.

T-cell receptor Vβ gene usage by T cells reactive with the tumor-rejection antigen SART-1 in oral squamous cell carcinoma

We recently described that the SART‐1690–698 peptide could induce HLA‐A24‐restricted cytotoxic T lymphocytes (CTLs), which recognize the SART‐1  259+ tumor cells from peripheral blood mononuclear cells (PBMCs) of HLA‐A24+ cancer patients. In our study, in 5 of 14 HLA‐A24+ patients with oral squamous cell carcinomas (SCCs), CTLs could be induced with the SART‐1690–698 peptide from the PBMCs. In 2 of the patients from whom the highest CTL activities were induced, the T‐cell receptor (TCR) Vβ repertoire expressed by the SART‐1690–698‐specific CTLs was found to be restricted and multiple Vβ families were predominantly expressed in each patient. Although the predominant Vβ families were different between the 2 patients, Vβ7 was highly and commonly predominant. The same predominant Vβ families were also detected in the tumor‐infiltrating lymphocytes (TILs) from each patient, and each Vβ family contained one or more unique T‐cell clonotypes. The unique T‐cell clonotypes were found to be common between the TILs and SART‐1690–698‐specific CTLs from each patient, and especially 2 T‐cell clonotypes with Vβ7 were identical even in the 2 patients. One of the 2 T‐cell clonotypes with Vβ7 was detected in the TILs from 11 of 14 HLA‐A24+ patients and another was found in those from 8 of HLA‐A24+ patients, while none of 10 HLA‐A24− patients demonstrated both T‐cell clonotypes. These results strongly suggest that the T‐cell clonotypes with Vβ7 are major TCR Vβ genes expressed by SART‐1690–698‐specific CTLs. Furthermore, autologous tumor cells from one of the HLA‐A24+ patients stimulated the PBMCs and regional lymph node cells (LNCs) to expand the same T‐cell clonotypes as those in the SART‐1690–698‐specific CTLs. These results strongly suggest that the SART‐1690–698‐specific CTLs clearly accumulate in vivo, especially in the TILs, as a consequence of in situ antigenic stimulation by autologous tumor cells. The identification of the unique TCR Vβ genes used by SART‐1259‐specific CTLs should help to improve the diagnosis of the specific immune response in patients with SART‐1  259+ cancers, especially during anticancer immunotherapy.

Sonographic diagnosis of Sjögren syndrome: evaluation of parotid gland vascularity as a diagnostic tool

OBJECTIVE To evaluate the usefulness of the vascularity in parotid glands in sonographic diagnosis for Sjögren syndrome. STUDY DESIGN Sonographic images of 72 cases of previously suspected Sjögren syndrome (including 43 actual cases) were analyzed retrospectively for the abnormal vascularity in the parotid gland parenchyma. The relationships between the vascularity and the results of sialographic, serologic, and histopathologic examinations were analyzed. We also compared the diagnostic accuracy of B-mode only with that of B-mode plus Doppler-mode. RESULTS Sjögren-positive cases showed significantly higher vascularity. As the grade of vascularity became higher, the rate of the Sjögren-negative cases became lower. The highest mean vascular score could be observed both in the initial stage and in the cavitary-destructive stage in the sialographic grades. Sensitivity and accuracy were markedly improved with vascular information. CONCLUSION By using vascular information, sonographic diagnosis for Sjögren syndrome can be improved.