Cells In Minor Salivary Glands Research Articles

Mycoplasma bovis May Travel Along the Eustachian Tube to Cause Meningitis in Japanese Black Cattle

Mycoplasma bovis (M. bovis) is a common inhabitant of the upper and lower respiratory tracts of cattle and is considered to be the main aetiological agent of otitis media in calves. The eustachian tube appears to be the most common portal for pathogens to enter the middle ear. We investigated the transmission route of M. bovis causing otitis media that progressed to meningitis or meningoencephalitis in Japanese Black cattle. M. bovis was detected in 10 cases by a loop-mediated isothermal amplification method or by immunohistochemistry. One case of caseonecrotic granulomatous meningoencephalitis, one case of caseonecrotic granulomatous meningitis, one case of suppurative meningoencephalitis, eight cases of eustachitis, nine cases of tonsillitis and six cases of suppurative bronchopneumonia were identified by histopathological examination. M. bovis antigen was detected in the eustachian tubes of eight cases. In nine cases, M. bovis was also detected in tonsillar epithelial crypts and lumina, in intraluminal inflammatory cells and in the epithelial cells of minor salivary glands located around the eustachian tubes and tonsils. The results suggest that M. bovis can infect and colonize the tonsils and enter the eustachian tubes, causing otitis media, which, in cases of chronic infection, can progress to meningitis.

Diminished CXCR5 expression in peripheral blood of patients with Sjögren's syndrome may relate to both genotype and salivary gland homing.

Genetic investigations of Sjögren's syndrome (SS) have identified a susceptibility locus at p23.3 of chromosome 11, which contains the CXCR5 gene. C-X-C motif chemokine receptor 5 (CXCR5) is a chemokine receptor expressed on B and T cell subsets, and binds the chemotactic ligand C-X-C motif chemokine ligand 13 (CXCL13). In this study we aimed to link the genetic association with functional effects and explore the CXCR5/CXCL13 axis in SS. Expression quantitative trait loci analysis of the 11q23.3 locus was performed using B cell mRNA expression data from genotyped individuals. Lymphocyte surface markers were assessed by flow cytometry, and CXCL13 levels by a proximity extension assay. CXCR5+ and CXCL13+ cells in minor salivary glands were detected using immunohistochemistry. Our results demonstrated that SS-associated genetic polymorphisms affected the expression of CXCR5 (P<0·01). Notably, a decreased percentage of CXCR5+ cells, with lower CXCR5 expression, was observed for most circulating B and T cell subsets in SS patients, reaching statistical significance in CD19+ CD27+ immunoglobulin (Ig)D+ marginal zone (P<0·001), CD19+ CD27+ IgD- memory (P<0·05) and CD27-IgD double-negative (P<0·01) B cells and CD4+ CXCR3- CCR6+ Th17 cells (P<0·05). CXCL13 levels were increased in patient plasma (P<0·001), and immunohistochemical staining revealed expression of CXCL13 and higher numbers of CXCR5+ cells (P<0·0001) within focal infiltrates and interstitially in salivary glands of SS patients. In conclusion, we link a genetic susceptibility allele for SS to a functional phenotype in terms of decreased CXCR5 expression. The decrease of CXCR5+ cells in circulation was also related to homing of B and T cells to the autoimmune target organ. Therapeutic drugs targeting the CXCR5/CXCL13 axis may be useful in SS.

Ductal cells of minor salivary glands in Sjögren's syndrome express LINE-1 ORF2p and APOBEC3B.

Type I interferon activation is a hallmark event in Sjögren's syndrome. L1 retroelements stimulate plasmacytoid dendritic cells, activating the type I interferons, and are regulated by various mechanisms, including the APOBEC3 deaminases. As L1s are potential trigger factors in autoimmunity, we aimed to investigate the immunohistochemical localization of L1 ORF2p and its inhibitor APOBEC3B protein in minor salivary glands of Sjögren's syndrome patients. Twenty minor salivary gland-tissue samples from 20 Sjögren's syndrome patients, classified according to Tarpley's histological criteria, and 10 controls were evaluated for L1 ORF2p and APOBEC3B expression via immunohistochemistry. L1 ORF2p was expressed in 17/20 SS patients and all controls. APOBEC3B expression was observed in 15/20 Sjögren's syndrome patients, 5/5 chronic sialadenitis, and 3/5 normal minor salivary glands. Both antibodies stained the cytoplasm of the ductal epithelial cells. Negative staining was observed in the acinar cells. L1 ORF2p-positive immunostaining was significantly lower in Tarpley IV Sjögren's syndrome patients than controls (P = .039), and APOBEC3B-positive staining was significantly lower in Tarpley I compared to Tarpley II Sjögren's syndrome patients (P = .008) and controls (P = .035). L1 ORF2p and APOBEC3B are expressed in the ductal epithelial cells of minor salivary glands that are among the key targets in Sjögren's syndrome. L1 ORF2p expression may promote the L1 ability to act as an intrinsic antigen in Sjögren's syndrome. The potential future use of L1 ORF2-reverse transcriptase inhibitors in autoimmunity supports further investigation of L1 epigenetic regulation by APOBEC3 enzymes.

Defining the Localization and Molecular Characteristic of Minor Salivary Gland Label-Retaining Cells

Adult stem cells (SCs) are important to maintain homeostasis of tissues including several mini-organs like hair follicles and sweat glands. However, the existence of stem cells in minor salivary glands (SGs) is largely unexplored. In vivo histone2B green fluorescent protein pulse chase strategy has allowed us to identify slow-cycling, label-retaining cells (LRCs) of minor SGs that preferentially localize in the basal layer of the lower excretory duct with a few in the acini. Engraftment of isolated SG LRC in vivo demonstrated their potential to differentiate into keratin 5 (basal layer marker) and keratin 8 (luminal layer marker)-positive structures. Transcriptional analysis revealed activation of TGFβ1 target genes in SG LRC and BMP signaling in SG progenitors. We also provide evidence that minor SGSCs are sensitive to tobacco-derived tumor-inducing agent and give rise to tumors resembling low grade adenoma. Our data highlight for the first time the existence of minor SG LRCs with stem cells characteristic and emphasize the role of transforming growth factor beta (TGFβ) pathway in their maintenance.

Inflammatory cells in minor salivary glands of patients with chronic hepatitis C: Immunophenotype, pattern of distribution, and comparison with liver samples

ObjectivesTo characterize the immunophenotype and the distribution of the inflammatory infiltrate (INF) in salivary glands (SG) of patients with chronic hepatitis C, comparing with laboratorial data (genotype, viral load, METAVIR, and HCV RNA in SG), and liver. MethodsINF was classified as diffuse or focal. Immunohistochemistry for CD3, CD20, CD8, CD4, CD57, CD68, and S100 was performed in 61 SG and 59 livers. ResultsDiffuse INF was more common in SG than in liver. CD3+, CD20+, and CD8+ were the most frequent cells in both tissues, with few CD57+, CD68+, and S100+ cells. CD4+ cells were common in liver, but rare in SG. Liver presented higher indexes for all markers, except S100+ (p<0.05). Higher CD3+, CD20+, and CD8+ (p<0.05) were observed in SG with focal infiltrate than with diffuse infiltrate. In liver, CD20+ and CD3+ were higher in focal infiltrate, and CD68+ in diffuse infiltrate (p<0.05). Comparisons with laboratorial data did not show statistical significance. ConclusionsThe INF in SG was mainly composed by T and B lymphocytes, mostly cytotoxic T cells. The glandular INF can present differences in composition according to its distribution. A more intense inflammation was observed in liver, but similar cell types were identified in SG, except for CD4+.

Modes of epithelial cell death and repair in Sjögren's syndrome (SS).

We evaluated possible modes of epithelial cell destruction and restoration in minor salivary gland biopsies from patients with SS. Minor salivary gland biopsies from 10 primary Sjögren's syndrome (pSS) patients and eight control individuals were evaluated by immunohistochemical staining for the expression of apoptosis-related molecules, substances released by activated cytotoxic T cells, as well as proteins involved in epithelial cell repair. The results were analysed by computer screen analysis and they were expressed as average percentages. Apoptosis-promoting molecules, Fas antigen and Fas ligand were observed in ductal and acinar epithelial cells as well as in infiltrating mononuclear cells of minor salivary glands from SS patients in comparison with control biopsies. Bax protein, which acts as a death-promoter message, was expressed in the ductal and acinar epithelial cells and in mononuclear infiltrating cells of SS patients compared with control individuals, while Bcl-2, an inhibitor of apoptosis, was primarily found in the lymphocytic infiltrates. In situ DNA fragmentation assay (TUNEL) revealed that epithelial cells were apoptotic in patients with SS compared with control subjects. Immunohistochemical staining for perforin and granzyme B, released from granules of activated cytotoxic lymphocytes, revealed their presence in lymphocytic infiltrates of patients with SS compared with control biopsies. pS2, a member of the trefoil protein family which functions as promoter of epithelial cell repair and cell proliferation, was expressed in epithelial cells in biopsies from SS patients. These studies suggest that the functional epithelium of minor salivary glands in patients with SS appears to be influenced by both intrinsic and extrinsic mechanisms of destruction, while a defensive mechanism of epithelial restoration seems to be active.

Characteristics of Salivary Diffuse Infiltrative Lymphocytosis Syndrome in West Africa Human Immunodeficiency Virus-Associated Salivary Ductal Epithelial Atypia?

c Objective.—To determine the prevalence of diffuse infil- trative lymphocytosis syndrome (DILS) in the minor sali- vary glands of 30 African Cameroonian adults with the ac- quired immunodeficiency syndrome (AIDS). Design.—Salivary gland tissue was analyzed using a modified classification system that was developed to aid the diagnosis of Sjogren syndrome. The advantages and dis- advantages of this approach are discussed. Materials and Methods.—Formalin-fixed, paraffin-em- bedded, hematoxylin-eosin-stained biopsy sections were prepared for 30 patients with AIDS, 26 healthy individuals who declined human immunodeficiency virus (HIV) test- ing, and 4 seronegative healthy controls. Tissues were im- munostained for CD4/CD8 1 lymphocytes and cytomega- lovirus (CMV), and transmission electron microscopy was performed to locate viral particles. Patients were tested for HIV-1 and HIV-2 by the HIV/Chek System 3 or CAMSTIX- HIV-1 and HIV-2 assay. Results.—Severe salivary ductal atypia (96%) was the feature most strongly associated with AIDS, and the lym- phocytic focus score was the second histologic feature most strongly correlated with AIDS. Forty-eight percent of patients with HIV-1 infection had more than 1 lymphocytic focus in a minor salivary gland. These lymphocytes were primarily CD8 1 . We report, to the best of our knowledge, the first case of multinucleated salivary duct epithelial cells in minor salivary glands also containing enveloped virus particles. All cases were negative for CMV. Conclusions.—The prevalence of DILS in West Africans with AIDS appears higher than the prevalence reported in whites from the United States and Europe and in blacks from the United States, a group that has been reported to have a greater incidence of DILS than whites. This discrep- ancy may be related to differences in patient selection cri- teria. The determination of lymphocytic focus score, as used in the diagnosis of Sjogren syndrome, with the ad- junct of ductal atypia is useful for assessing DILS. The im- pact of patient selection, drug therapy, and parasites on salivary gland pathology is discussed. (Arch Pathol Lab Med. 2000;124:1773-1779)

Characteristics of Salivary Diffuse Infiltrative Lymphocytosis Syndrome in West Africa

To determine the prevalence of diffuse infiltrative lymphocytosis syndrome (DILS) in the minor salivary glands of 30 African Cameroonian adults with the acquired immunodeficiency syndrome (AIDS). Salivary gland tissue was analyzed using a modified classification system that was developed to aid the diagnosis of Sjögren syndrome. The advantages and disadvantages of this approach are discussed. Formalin-fixed, paraffin-embedded, hematoxylin-eosin-stained biopsy sections were prepared for 30 patients with AIDS, 26 healthy individuals who declined human immunodeficiency virus (HIV) testing, and 4 seronegative healthy controls. Tissues were immunostained for CD4/CD8+ lymphocytes and cytomegalovirus (CMV), and transmission electron microscopy was performed to locate viral particles. Patients were tested for HIV-1 and HIV-2 by the HIV/Chek System 3 or CAMSTIX-HIV-1 and HIV-2 assay. Severe salivary ductal atypia (96%) was the feature most strongly associated with AIDS, and the lymphocytic focus score was the second histologic feature most strongly correlated with AIDS. Forty-eight percent of patients with HIV-1 infection had more than 1 lymphocytic focus in a minor salivary gland. These lymphocytes were primarily CD8+. We report, to the best of our knowledge, the first case of multinucleated salivary duct epithelial cells in minor salivary glands also containing enveloped virus particles. All cases were negative for CMV. The prevalence of DILS in West Africans with AIDS appears higher than the prevalence reported in whites from the United States and Europe and in blacks from the United States, a group that has been reported to have a greater incidence of DILS than whites. This discrepancy may be related to differences in patient selection criteria. The determination of lymphocytic focus score, as used in the diagnosis of Sjögren syndrome, with the adjunct of ductal atypia is useful for assessing DILS. The impact of patient selection, drug therapy, and parasites on salivary gland pathology is discussed.

Professional Antigen Presenting Cells in Minor Salivary Glands in Sjögren's Syndrome: Potential Contribution to the Histopathological Diagnosis?

Sjögren's syndrome is an autoimmune disease in which lymphocytic infiltrates develop in the salivary and lacrimal glands. We have shown that dendritic cells (DC) infiltrate the submandibular gland of the nonobese diabetic (NOD) mouse, a mouse model for Sjögren's syndrome, before lymphocytic infiltration, suggesting that these antigen-presenting cells (APC) may play a role in the initiation of Sjögren's syndrome. In later stages, DC and macrophages also form an important part of the infiltrate of the NOD sialoadenitis. To find out if DC and macrophages form part of the infiltrate in Sjögren's syndrome as well, and to determine whether they may be useful in the histopathological diagnosis of Sjögren's syndrome, we studied their presence in minor salivary glands (MSG) of patients with Sjögren's syndrome and patients with focal lymphocytic sialoadenitis (FLS), but without clinical or serological criteria of Sjögren's syndrome. Immunohistochemistry was applied, followed by semiquantitative analysis. DC and macrophages were present in all MSG; however, there were clear differences in marker expression between Sjögren's syndrome and FLS, on the one hand, and control tissue, on the other hand. CD1a+ DC and RFD9+ macrophages were mainly observed in MSG in which a focal lymphocytic infiltrate was present. In fact, the diffuse presence of single CD1a+ DC and RFD9+ macrophages correlated closely with the presence of a focal lymphocytic infiltrate in the MSG. This indicates that these cells could be of help during the evaluation of a MSG. Because the detection of APC is technically less cumbersome than a focal score, this parameter may perhaps replace the focal score in the histopathological diagnosis of Sjögren's syndrome. This study therefore prompts further investigation focusing on the presence of CD1a+ and RFD9+ cells in the MSG of a large cohort of patients.

Expression of GLUT1 in stratified squamous epithelia and oral carcinoma from humans and rats.

Most cells express facilitative glucose transporters. Four isoforms (GLUT1-4) transporting D-glucose across the plasma membrane show a specific tissue distribution, which is the basis for tissue-specific patterns in glucose metabolism. GLUT1 is expressed at high levels in tissue barriers such as the blood-brain barrier, and this isoform has been suggested as an indicator of such barriers. GLUT1 has been found in basal layers of human epidermis where no such tissue barrier is present. To further clarify these issues, we examined the distribution of GLUT1 and GLUT4 in skin, different types of oral mucosa from rat and man, and a human oral carcinoma by indirect immunofluorescence microscopy. The results showed that GLUT1 was expressed in the basal and parabasal layers of the different stratified squamous epithelia, with some variations between keratinized and non-keratinized subtypes. GLUT1 was also expressed in ductal- and myoepithelial cells of minor salivary glands and perineural sheath located in the lamina propra, and furthermore in the cells of an oral carcinoma. GLUT4 was not expressed in any of the tissues examined. This distribution of GLUT1 does not fit with the idea of GLUT1 as a general indicator of tissue barriers. In contrast, our results support the prevailing, but limited knowledge of glucose metabolism in squamous stratified epithelia, a metabolism believed to depend mostly on glycolysis, especially in the basal layers. High-level expression seemed to be confined to keratinocytes without glycogen stores.

Sjögren's Syndrome: An Autoimmune Epithelitis

Sjögren's syndrome is a chronic autoimmune disorder characterized by mononuclear cell infiltration proximally to epithelial cells of exocrine glands. In recent years, several studies have tried to address the function of the components of the immunopathologic lesion in Sjögren's syndrome. The majority of the mononuclear infiltrating cells are CD4 positive T lymphocytes (60-70%) whereas B cells constitute one fourth of the infiltrating cells. Macrophages and natural killer cells are poorly represented in the lesion. Epithelial cells of minor salivary glands of patients with Sjögren's syndrome express proinflammatory cytokines (IL-1 beta, IL-6), protooncogenes (c-myc) and costimulatory molecules (B71, B72). The destruction of epithelial cells of Sjögren's syndrome patients is probably due to activation of several apoptotic pathways since epithelial cells express different apoptosis related molecules such as Fas, FasL, Bax, while mononuclear cells express Bcl-2, Perforin and Granzymes. Finally epithelial cells seem to exert a regenerative effort since they express trefoil proteins (pS2). The above properties give epithelial cells a significant role in the pathophysiology of the syndrome but the exact events which drive the immune system towards an autoimmune reaction remain obscure.

Lewis Blood Group Antigens in Salivary Glands and Stratified Epithelium: Lack of Regulation of Lewis Antigen Expression in Ductal and Buccal Mucosal Lining Epithelia1

The expression of Lewis antigens is thought to be controlled by the Secretor and Lewis genes. While secretor status is known to regulate the expression of ABH antigens in many tissues, few studies have attempted to correlate Lewis antigen expression on erythrocytes and saliva with that of epithelial tissues. We examined the expression of Lewis a and b and related antigens in human epithelium of minor salivary glands and labial nonkeratinized oral mucosae from 16 individuals by immunohistology using monoclonal antibodies. The expression of these antigens, as detected by monoclonal antibodies (MAbs) used, was correlated with erythrocyte phenotype and saliva secretor status. In acinar cells of glands, Leb antigen was expressed only in secretors, and Lea only in nonsecretors. However, in gland ducts and oral mucosae, Leb was found in both secretors and nonsecretors, as well as in 2 cases of Lea-b-, secretors. Thus, antigen expression in acinar cells of minor salivary glands was correlated with the predicted genotypes, whereas inappropriate expression of Leb antigen was found in epithelial cells of gland ducts and oral mucosae. The present data indicate that the Lewis blood group phenotype is regulated differently in duct cells and stratified epithelium than in saliva and erythrocytes.

Lewis Blood Group Antigens in Salivary Glands and Stratified Epithelium: Lack of Regulation of Lewis Antigen Expression in Ductal and Buccal Mucosal Lining Epithelia

The expression of Lewis antigens is thought to be controlled by the Secretor and Lewis genes. While secretor status is known to regulate the expression of ABH antigens in many tissues, few studies have attempted to correlate Lewis antigen expression on erythrocytes and saliva with that of epithelial tissues. We examined the expression of Lewis a and b and related antigens in human epithelium of minor salivary glands and labial nonkeratinized oral mucosae from 16 individuals by immunohistology using monoclonal antibodies. The expression of these antigens, as detected by monoclonal antibodies (MAbs) used, was correlated with erythrocyte phenotype and saliva secretor status. In acinar cells of glands, Le^b antigen was expressed only in secretors, and Le^a only in nonsecretors. However, in gland ducts and oral mucosae, Le^b was found in both secretors and nonsecretors, as well as in 2 cases of Le^a-b-, secretors. Thus, antigen expression in acinar cells of minor salivary glands was correlated with the predicted genotypes, whereas inappropriate expression of Le^b antigen was found in epithelial cells of gland ducts and oral mucosae. The present data indicate that the Lewis blood group phenotype is regulated differently in duct cells and stratified epithelium than in saliva and erythrocytes.

Keratin proteins identified in epithelial fragments of salivary mucoceles

Immunohistochemically detected keratin proteins in mucoceles of oral mucosa were used to served as markers to identify epithelial-derived cells of minor salivary glands. Normal ducts of minor salivary glands showed an intense keratin staining. Epithelial lining and or epithelial fragments in cystic walls of mucoceles displayed a strong reaction to keratin proteins too, whereas granulation tissue or connective tissue fibers of the walls were not seen. Foamy macrophages present in cystic cavities signify higher staining with the use of keratin proteins. Mucoceles in oral minor salivary glands are probably caused by ductal obstructions leading to continuous mucous secretion. Keratin proteins were used as an epithelial marker of ductal segments.