Rapid generation of functional hepatocyte-like cells from human minor salivary gland-derived stem cells.

Liver failure is one of the major risk factors for death worldwide, and the only effective liver transplantation is currently very limited. Adult stem cells can be induced into hepatocytes in vitro and implanted into the body to repair damaged liver. However, most of the induction time in vitro is relatively long, which is not suitable for practical application. Therefore, search for new seed cells that can rapidly differentiate into functional hepatocytes is crucial for the clinical application of cell transplantation therapy. In this study, we explored a three-step protocol to rapidly induce human minor salivary gland mesenchymal stem cells (hMSG-MSCs) into hepatocytes in vitro, and finally obtained hepatocyte-like cells within 6 days. After a series of relevant detection from gene, protein and functional levels, we confirmed that the finally induced cells were mature hepatocyte-like cells with certain hepatocyte functions to some extent. Besides, we injected the preliminary induced cells into mice with acute liver injury, showing a good repair effect on the damaged liver. All these results indicate that the hMSG-MSCs have potential to be a kind of seed cells for rapid hepatic differentiation.

Sjögren's Syndrome: Concerted Triggering of Sicca Conditions.

AIM: The aim of this study was to evaluate the expression of persistence of mumps virus and some cells that interact with viral infection in the focus of the autoimmune epithelitis and peripheral blood of Sjögren's syndrome patients in comparison to patients with rheumatoid arthritis (RA) and nonautoimmune sicca syndrome (nSS). MATERIALS AND METHODS: 126 patients (119 women and 7 men) were grouped into four groups: (1) patients with primary Sjögren's syndrome (pSS), (2) patients with secondary Sjögren's syndrome due to rheumatoid arthritis (sSS), (3) patients with rheumatoid arthritis (RA), and (4) patients with nonautoimmune sicca syndrome (nSS). Immunohistochemical analysis of immune response to the suggested silent persistence of mumps virus in the minor labial salivary gland biopsies and flow cytometric analysis of blood cells was done. RESULTS: Immunohistochemical signs of mumps virus persistence were found in the minor salivary glands of all study groups. Also, a significantly different immune response to virus infection (protein IFI16, interferons gamma and beta, dendritic cells, and receptor for natural killers) was revealed in the minor salivary glands of the study groups. Cytometric analysis of the blood cells revealed a dropping amount of circulating natural killers and dendritic cells in patients with SS. Significant correlations between immunohistochemical staining and serological findings were revealed. CONCLUSIONS: Abundant immunohistochemical signs of mumps virus protein in the salivary glands and depletion of circulating immune cells make a background for thought of presumable mumps or/and other virus participation in epithelial damage causing sicca syndrome in predisposed patients.

Comparison of p63/p40 Expression With Myoepithelial Markers in Minor Salivary Gland Tumors.

The present study aimed to compare the expression of p63/p40 with smooth muscle actin (SMA) and vimentin (VIM) by myoepithelial cells in minor salivary gland tumors. Fifty-two formalin-fixed paraffin-embedded samples of minor salivary gland tumors derived from intercalated duct (pleomorphic adenoma [PA], adenoid cystic carcinoma [ACC], epithelial-myoepithelial carcinoma [EMC], polymorphous adenocarcinoma [PAC], and secretory carcinoma [SC]) and 3 samples of minor salivary gland tumors derived from excretory duct (mucoepidermoid carcinoma [MEC]) were evaluated by means of immunohistochemistry. The data were analyzed qualitatively. The results indicated that p63 and p40 expression were detected in myoepithelial cells present in PA, ACC, and EMC. However, both proteins were also observed in squamous areas of PA and all cases of MEC. SMA were noticed in some myoepithelial cells of PA, ACC, and EMC. Expression of SMA was negative in the other salivary gland tumors evaluated. VIM was constantly expressed by myoepithelial cells in PA, ACC, and EMC. VIM was also observed in cells of PAC and SC, but not in squamous areas of PA and MEC. In conclusion, p63 expression is almost comparable with VIM in detecting myoepithelial cells, an immunolabeling pattern not followed by p40, and consequently, caution has to be taken during the interpretation of salivary gland tumor exhibiting an p63/p40 phenotype in order to avoid a misdiagnosis.

Association of high 5-hydroxymethylcytosine levels with Ten Eleven Translocation 2 overexpression and inflammation in Sjögren's syndrome patients.

Here, we determined the 5-hydroxymethylcytosine (5hmC), 5-methylcytosine (5mC), Ten Eleven Translocation (TETs), and DNA methyltransferases (DNMTs) levels in epithelial and inflammatory cells of labial salivary glands (LSG) from Sjögren's syndrome (SS)-patients and the effect of cytokines on HSG cells. LSG from SS-patients, controls and HSG cells incubated with cytokines were analysed. Levels of 5mC, 5hmC, DNMTs, TET2 and MeCP2 were assessed by immunofluorescence. In epithelial cells from SS-patients, an increase in TET2, 5hmC and a decrease in 5mC and MeCP2 were observed, additionally, high levels of 5mC and DNMTs and low levels of 5hmC were detected in inflammatory cells. Cytokines increased TET2 and 5hmC and decreased 5mC levels. Considering that the TET2 gene.promoter contains response elements for transcription factors activated by cytokines, together to in vitro results suggest that changes in DNA hydroxymethylation, resulting from altered levels of TET2 are likely to be relevant in the Sjögren's syndrome etiopathogenesis.

Spatial organization of the excretory ducts and sections of microcirculatory blood flow of the labial salivary glands in older adults.

OBJECTIVE: Introduction: Salivary glands have a significant impact on the state of the homeostasis of the human body, oral cavity in particular, sensitively responding to pathological processes. The reactivity of the salivary glands in response to pathological processes that are organically linked to morphology and functions of the organ's structures, and particularly the excretory ducts of the glands and their microcirculatory blood flow, is one of the problems which have not been solved to date. The aim of the paper was to elucidate the features of the stereomicroscopic structure of the excretory ducts and sectors of the microcirculatory blood flow in labial glands of older adults. PATIENTS AND METHODS: Materials and methods: The object of the study was the labial mucosa of the older adults, which was cut into 3х3 mm pieces and fixed in the buffered 4% glutaraldehyde solution with subsequent 2-hour fixation in osmium. Once the pieces were washed and dehydrated they were embedded into the Epon-812. The series of the semi-thin sections, made from the obtained epoxy blocks, were stained in phosphate buffered 0,1% toluidine blue solution. The serial semi-thin sections were subjected to histological and cytological studies and multilayered plastic reconstruction. RESULTS: Results and conclusions: The series of histological epoxy semi-thin sections, as well as graphic and plastic reconstruction of the sectors of microcirculatory blood flow and excretory ducts of the human labial glands have demonstrated a range of morphological facts that can be used to clarify the intertissue stereological relationships. They also determined the syntopic proximity of the capacitive sectors of microcirculatory blood flow to the excretory ducts of the gland. Such pattern is especially notable in the collecting venules and intralobular ducts. It has been shown that the biggest venule is the collecting venous vessel. Anastomoses between the intralobular arterioles and collecting venules have been found in the microcirculatory blood flow of the labial glands.

The use of morphological study technique for investigation of labial and palatine glands.

INTRODUCTION: Due to the deterioration of environmental conditions that promotes the onset of inflammatory and autoimmune diseases, and the progress in the diagnosis, the frequency of registration of intercurrent pathology of salivary glands has markedly risen in recent years, demonstrating the increased scientific interest in the research of the common and distinctive features of their structure. THE AIM: The paper was aimed at the development of the method of morphological study of human minor salivary (labial and palatine) glands by the use of plastic wax reconstruction to obtain the plastic model of the acini and ducts of human minor salivary glands. MATERIALS AND METHODS: Specimens of the glandular area of the hard palate mucosa and labial mucosa in its middle third have been studied. To gain the objective of the investigation the technique for morphological study of the human minor salivary (labial and palatine) glands is to be developed, encompassing the analysis of the spatial organization of the glandular epithelium of the labial and palatine glands together with blood microcirculatory flow by fixing the obtained specimens of the minor salivary glands in 4% glutaraldehyde solution and osmium tetroxide with subsequent embedding into the Epon-812, staining the serial semi-thin sections with phosphate buffered 0,1% toluidine blue solution, photomacrography of the distinguished boundaries of the investigated structures and obtaining of photoreconstructions. RESULTS AND CONCLUSIONS: Thus, the use of suggested technique enables to obtain the megascopic reconstruction of the acini and ducts of the labial and palatine glands, which can be studied from different sides, getting the full visualization of the shape and size, as well as to explore the glands' inner configuration, the geometry of the lumen of the epithelial excretory ducts, to determine changes in the thickness of the wall, to get a visual representation of the microtopographic interactions between the different parts of blood microcirculatory flow and excretory ducts of the minor salivary glands.

Therapeutic potential of human minor salivary gland epithelial progenitor cells in liver regeneration.

Liver disease is a serious problem affecting millions of people with continually increasing prevalence. Stem cell therapy has become a promising treatment for liver dysfunction. We previously reported on human minor salivary gland mesenchymal stem cells (hMSGMSCs), which are highly self-renewable with multi-potent differentiation capability. In this study, keratinocyte-like cells with self-regeneration and hepatic differentiation potential were isolated and characterized, and named human minor salivary gland epithelial progenitor cells (hMSG-EpiPCs). hMSG-EpiPCs were easily obtained via minor intraoral incision; they expressed epithelial progenitor/stem cell and other tissue stem cell markers such as CD29, CD49f, cytokeratins, ABCG2, PLET-1, salivary epithelial cell markers CD44 and CD166, and the Wnt target related gene LGR5 and LGR6. The cells were induced into functional hepatocytes in vitro which expressed liver-associated markers ALB, CYP3A4, AAT, and CK18. Upon transplantation in vivo, they ameliorated severe acute liver damage in SCID mice caused by carbon tetrachloride (CCl4) injection. In a two-thirds partial hepatectomy mouse model, the transplanted cells survived at least 4 weeks and exhibited hepatic potential. These findings demonstrate that hMSG-EpiPCs have potential as a cellular therapy basis for hepatic diseases, physiological and toxicology studies and regenerative medicine.

Insight into pathogenesis of Sjögren's syndrome: Dissection on autoimmune infiltrates and epithelial cells.

Sjögren's syndrome (SS) is a chronic autoimmune disease with broad clinical spectrum, extending from benign exocrinopathy to severe systemic disease and lymphoma development. The glandular and extraglandular dysfunction of SS is associated with lymphocytic infiltrates that invade the epithelial structures of affected organs. The in-depth study of autoimmune lesions in the minor salivary glands (MSG), which are the major target-organ of SS responses, revealed that the lymphocytic infiltrates vary in severity and composition among SS-patients, are full-blown at diagnosis and remain unchanged thereafter. Although the pathogenetic pathways underlying SS have not yet elucidated, it is well-established that glandular epithelial cells are central regulators of local autoimmune responses. Moreover, chronic inflammation affects epithelial function and phenotype, which strengthens or weakens their immunoregulatory/secretory function, leading to deterioration of autoimmune phenomena. Herein, the current findings regarding the autoimmune lesions, the role of epithelial cells and their interaction with infiltrating lymphocytic cells are discussed.

Characteristics of primary Sjögren's syndrome patients with IgG4 positive plasma cells infiltration in the labial salivary glands.

The purpose of this study was to investigate the characteristics of primary Sjögren's syndrome (pSS) patients with IgG4 positive (IgG4+) plasma cell infiltration in labial salivary glands (LSGs). Paraffin sections of LSGs from 336 pSS patients were stained with IgG4 and IgG monoclonal antibodies. According to the infiltration of IgG4+ plasma cells, patients were divided and clinical and serological characteristics were analyzed and compared. Based on the infiltration of IgG4+ plasma cells in the LSGs, patients were divided into three subgroups, low IgG4, moderate IgG4, and high IgG4 groups. A negative association between the number of infiltrated IgG4+ plasma cells and the disease characteristics was observed. We found that the higher the IgG4+ expression in plasma cells, the lower the positive rates of serum anti-SSA antibodies, anti-SSB antibodies, antinuclear antibodies (ANA), and rheumatoid factor (RF). Besides, patients from the high IgG4 group had the highest frequency of interstitial lung disease (ILD, 30.6%) and tubulointerstitial nephritis (TIN, 13.9%), but the lowest frequency of leucopenia (13.9%), thrombocytopenia (11.1%), and abnormal thyroidal function (0%). PSS patients with different IgG4+ plasma cells infiltration in the LSGs had distinctive clinical and laboratory characteristics. It may help us to further understand the role of IgG4+ plasma cells in pSS.

Analysis of the cell populations composing the mononuclear cell infiltrates in the labial minor salivary glands from patients with rheumatoid arthritis and sicca syndrome.

OBJECTIVES: Sicca symptoms occur in around 30% of rheumatoid arthritis (RA) patients. Herein, we examined the characteristics of RA patients bearing sicca symptomatology (RA-sicca) with a special focus on the immunohistopathological features of their labial minor salivary gland (LMSG) biopsies. METHODS: Our cohort included 100 consecutive RA patients which were interrogated using a sicca symptoms questionnaire. Positive responders were evaluated for ocular and oral dryness and underwent an LMSG biopsy. All samples were immunohistochemically evaluated for the presence and distribution of specific leukocyte subsets using appropriate markers and for the expression of certain immunoregulatory molecules by salivary gland epithelial cells. Positively stained and total mononuclear cells (MNC) were counted in the entire section. Counts were expressed as cell frequency (percentage of cell type number/total infiltrating MNC number). RESULTS: In the majority (86.1%) of the 44 RA-sicca cases, periductal infiltrates were observed in LMSG biopsies. The frequencies of infiltrating cell subtypes and their correlation with lesion severity were different from that previously described in primary Sjögren's syndrome (pSS). Moreover, DCs and ΜΦs frequencies were increased in RA-sicca patients who had a biopsy focus score <1 and absence of anti-Ro/anti-La autoantibodies, in contrast to what was observed for B cells. In about half of the biopsies, salivary gland epithelial cells expressed CD80/B7.1 molecules, most commonly in patients with a positive biopsy or anti-Ro/anti-La autoantibodies. CONCLUSION: LMSG infiltrates composition in RA-sicca patients is distinct from that described in pSS. These differences, further attest to diverse pathophysiologic processes operating in these two entities.

Characterization of a Self-renewing and Multi-potent Cell Population Isolated from Human Minor Salivary Glands.

Adult stem cells play an important role in maintaining tissue homeostasis. Although these cells are found in many tissues, the presence of stem cells in the human minor salivary glands is not well explored. Using the explant culture method, we isolated a population of cells with self-renewal and differentiation capacities harboring that reside in the human minor salivary glands, called human minor salivary gland mesenchymal stem cells (hMSGMSCs). These cells show embryonic stem cell and mesenchymal stem cell phenotypes. Our results demonstrate that hMSGMSCs have the potential to undergo mesodermal, ectodermal and endodermal differentiation in conditioned culture systems in vitro. Furthermore, in vivo transplantation of hMSGMSCs into SCID mice after partial hepatectomy shows that hMSGMSCs are able to survive and engraft, characterized by the survival of labeled cells and the expression of the hepatocyte markers AFP and KRT18. These data demonstrate the existence of hMSGMSCs and suggest their potential in cell therapy and regenerative medicine.

Presence and distribution of leptin and its receptor in the minor salivary glands of the donkey.

Leptin is a hormone widely diffused in the mammalian body in which it plays functions that go far beyond control of appetite and energy metabolism. The finding that it is present in the major salivary glands of various animal species is of interest for the functional implications that it may imply. Since there are no data on the presence of leptin and its receptor in the minor salivary glands, the aim of this study was to demonstrate their presence and distribution in such glands of donkeys. This latter was chosen as species of reference because the monogastric herbivore shows intense salivation during their masticatory activity. Tissue samples were collected from four adult donkeys, of both sexes, following slaughter. Samples were fixed, embedded in paraffin, and processed for immunohistochemical analysis using primary antibodies directed against leptin and its receptor. Controls for non-specific staining were always included. Leptin and its receptor were found in the minor salivary glands. Their distribution was similar to that described in the major salivary glands of animal species that have been investigated to date. We hypothesized that leptin can play a role in regulating gland function, via an autocrine/paracrine mechanism.

Establishment of functional acinar-like cultures from human salivary glands.

Disorders of human salivary glands resulting from therapeutic radiation treatment for head and neck cancers or from the autoimmune disease Sjögren syndrome (SS) frequently result in the reduction or complete loss of saliva secretion. Such irreversible dysfunction of the salivary glands is due to the impairment of acinar cells, the major glandular cells of protein, salt secretion, and fluid movement. Availability of primary epithelial cells from human salivary gland tissue is critical for studying the underlying mechanisms of these irreversible disorders. We applied 2 culture system techniques on human minor salivary gland epithelial cells (phmSG) and optimized the growth conditions to achieve the maintenance of phmSG in an acinar-like phenotype. These phmSG cells exhibited progenitor cell markers (keratin 5 and nanog) as well as acinar-specific markers-namely, α-amylase, cystatin C, TMEM16A, and NKCC1. Importantly, with an increase of the calcium concentration in the growth medium, these phmSG cells were further promoted to acinar-like cells in vitro, as indicated by an increase in AQP5 expression. In addition, these phmSG cells also demonstrated functional calcium mobilization, formation of epithelial monolayer with high transepithelial electrical resistance (TER), and polarized secretion of α-amylase secretion after ß-adrenergic receptor stimulation. Taken together, suitable growth conditions have been established to isolate and support culture of acinar-like cells from the human salivary gland. These primary epithelial cells can be useful for study of molecular mechanisms involved in regulating the function of acinar cells and in the loss of salivary gland function in patients.

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 TGFß1 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 (TGFß) pathway in their maintenance.

Cholera toxin B subunit-binding and ganglioside GM1 immuno-expression are not necessarily correlated in human salivary glands.

OBJECTIVE: To determine and compare the presence and in situ localization of the glycosphingolipid ganglioside GM1 in human salivary glands using the biomarkers for GM1: cholera toxin and antibodies against GM1. MATERIALS AND METHODS: Immunohistochemical analyses were performed on sections of adult human submandibular, parotid and palatinal glands using cholera toxin sub-unit B and two polyclonal antibodies against ganglioside GM1 as biomarkers. RESULTS: Immunofluorescence microscopy showed that the toxin and antibodies were co-localized in some acini but not in others. The cholera toxin mainly reacted with the cell membranes of the mucous acini in the submandibular gland, while incubation with the antibody against GM1 gave rise to a staining of the cytoplasm. The cytoplasm in some secretory acinar cells in the parotid gland was stained by the cholera toxin, whereas only small spots on the plasma membranes reacted with anti-GM1. The plasma membranes in the parotid excretory ducts appeared to react to anti-GM1, but not to cholera toxin. CONCLUSIONS: Cholera toxin induces the expression of ion channels and carriers in the small intestine and increases the production of secretory mucins. Although their mutual immunohistochemical localization may differ, both cholera toxin and ganglioside GM1 are present in the mucin-producing acini from salivary glands. This could point to a relationship between ganglioside expression and production of salivary mucins.

Frecuencia de complicaciones y rédito de la biopsia de glándula salival menor / Frequency of complications and usefulness of the minor salivary gland biopsy

El hallazgo de anticuerpos específicos y datos histopatológicos son indispensables para llegar al diagnóstico de síndrome de Sjögren (SS). La biopsia de glándulas salivales menores (BGSM), si bien es un procedimiento sencillo, debe ser realizada en una institución a fin de evitar complicaciones. Objetivo: Estimar la frecuencia de complicaciones mediatas e inmediatas y el rédito de la técnica. Materiales y métodos: Se incluyeron los pacientes derivados al Hospital Rivadavia para realización de biopsia, entre octubre del 2007 y mayo del 2010. Los pacientes fueron citados a la semana y al mes del procedimiento para control de la lesión. Resultados: Frecuencia de complicaciones inmediatas (n = 186): 15 pacientes: 8,1%, IC del 95%, 4,7-13,2 (sangrado 7,5%, lipotimia 3,2%, hematomas 2,7%; no hubo accidentes). Complicaciones mediatas (n = 164): 16 pacientes: 9,75%, IC del 95%, 5,9-15,6 (dolor 7,32%, signos inflamatorios 3,66%, trastornos de sensibilidad 3,05%, granuloma 1,22%). No hubo casos de infecciones, ni dehiscencia del punto de sutura. Rédito microscópico: total 154 biopsias: se obtuvo tejido glandular en el 90,9%, IC del 95%, 85-95 (típica, sialoadenitis, infiltrado grado III y IV). Conclusiones: La BGSM presenta una baja frecuencia de complicaciones mediatas e inmediatas y un alto rédito en el estudio anatomo-patológico (AU)

Findings of specific antibodies and histopathology data are essential for the diagnosis of Sjögren syndrome (SS). Although the minor salivary gland biopsy (MSGB) is technically simple, it needs to be performed in a medical institution to avoid complications. Objective: To determine the frequency of complications and the usefulness of this technique. Materials and methods: Patients who underwent a minor salivary gland biopsy for a possible diagnosis of SS at Rivadavia Hospital between October 2007 and May 2010 where included. The patients were seen a week and a month after the procedure for follow up. Results: Frequency of acute complications (n = 186): 15 patients; 8.1%, 95%CI: 4.7-13.2 (Bleeding 7.5%, syncope 3.2%, hematoma 2.7%. No accidents occurred). Medium term complications (n = 164): 16 patients: 9.75%, 95%CI: 5.9-15.6 (pain 7.32%, inflammation 3.66%, sensitivity disorders 3.05%, granuloma 1.22%,). No infections or suture dehiscence occurred. Microscopic results: 154 biopsy reports were received: glandular 90.9%, 95%CI: 85-95 (typical, sialadenitis, grade III and IV infiltration). Conclusions: MSGB has very low frequency of medium term and acute complications and it has high usefulness (AU)

Anatomical and histological structure of the tongue and histochemical characteristics of the lingual salivary glands in the Chukar partridge (Alectoris chukar, Gray 1830).

1. The aim of the study was to examine the morphology of the tongue and the histochemical features of the lingual salivary glands in this species. 2. The tongue was elongated, terminating in a rather sharp, dagger-like apex. On the surface of the tongue and situated between the body and root of the tongue, two rows of conical papillae, the sharp apices of which pointed towards the posterior part of the tongue, were observed. The keratinised epithelium lining the dorsal surface lacked typical gustatory papillae. However, it was observed that taste buds were present in the epithelium of the lingual body and root. The tongue was supported by a structure composed of hyaline cartilage, the paraglossum, which extended from the lingual root to the apex. Simple branched tubular glands, which were encapsulated by connective tissue, were embedded within the submucosa in the body (anterior salivary glands) and root (posterior salivary glands) of the tongue. It was observed that the secretion of the lingual glands contained neutral mucins, proteoglycans containing carboxylic acid, weak and strong sulphated groups, N-acetylated sialomucins, but lacked glycogen. 3. It was demonstrated that, the general morphological features, papillary distribution of the tongue and the histological structure of the mucosa epithelium and the supportive elements displayed similarity to those of other domestic avian species. It was also determined that, in view of the particular feeding types, in the partridge, the presence of the papillary crest was not correlated with diet.

The salivary gland epithelial cells of patients with primary Sjögren's syndrome manifest significantly reduced responsiveness to 17ß-estradiol.

Several lines of evidence indicate that salivary gland epithelial cells (SGEC) play an important role in the pathogenesis of primary Sjogren's syndrome (SS). Normal SGEC have been shown to possess functional estrogen receptors, however, the estrogenic response of SGEC in patients with SS has not been previously assessed. To address this issue, we comparatively tested cultured non-neoplastic SGEC lines from SS patients (SS-SGEC, n = 8) and from disease controls (control-SGEC, n = 12) in a standard estrogenic inhibition assay of cytokine-induced adhesion molecule expression, where the modulation of the expression of constitutive and interferon-gamma (IFNγ)-induced CD54/ICAM.1 molecules following treatment with 17ß-estradiol (E2) was evaluated by flow cytometry. Similarly high ICAM.1 expression was induced by IFNγ in control-SGEC and SS-SGEC lines. E2-treatment did not modify the constitutive ICAM.1 expression in either control-SGEC or SS-SGEC lines. In line with previous results, E2-pretreatment of control-SGEC was found to impede significantly the IFNγ-induced upregulation of ICAM.1 (p = 0.003). However, such inhibition was not observed in the SS-SGEC lines (p = 0.55). Such aberrant response of SS-SGEC to estrogens did not appear to associate with altered expression of estrogen receptor (ER) proteins, as no discernible differences could be revealed by immunoblotting and immunohistochemistry in the patterns or the intensity of ERα and ERß (ERß1- and ERß2-isoforms) protein expression in SGEC lines or minor salivary gland tissues between SS patients and disease controls. The deficient estrogenic responsiveness of SS-SGEC likely represents a manifestation of the intrinsic epithelial activation that characterizes SS and possibly indicates the perturbation of the immunoregulatory potential of estrogens in SS-epithelia.

Histochemical analyses of glycoconjugates and antimicrobial substances in goat labial glands.

Saliva is known to protect the oral cavity and contains glycoproteins and antimicrobial substances. The distribution of these salivary secretions was studied in the labial glands of the Japanese miniature (Shiba) goat using lectin histochemical and immunohistochemical methods. The mucous acinar cells of the labial glands exhibited glycoconjugates with different saccharide residues, such as GalNAcα1-3GalNAc, Galß1-4GalNAc, ß-D-GlcNAc and sialic acid linked to α2-6Gal/GalNAc. Furthermore, α-D-Man, α-L-Fuc, α-D-GalNAc, ß-D-Gal and sialic acid residues were present, in particular, in the serous demilunar cells. Antimicrobial substances (lysozyme, IgA, lactoferrin and ß-defensin) were shown to be mainly immunolocalized in the serous demilunes and duct cells. The results obtained are discussed with regard to the functional role of labial glands. The secretory compounds demonstrated may play an important role in the maintenance of oral health with regard to saliva.

Electron microscopic immunogold localization of statherin in human minor salivary glands.

In this study, which supplements a recent article on the localization of statherin in human major salivary glands, we investigated the intracellular distribution of this peptide in minor salivary glands by immunogold cytochemistry at the electron microscopy level. In the lingual serous glands of von Ebner, gold particles were found in serous granules of all secreting cells, indicating that statherin is released through granule exocytosis. In buccal and labial glands, mostly composed of mucous tubuli, statherin reactivity was detected in the serous element, which represents only a small population of the glandular parenchyma. In these serous cells, however, statherin labeling was absent in secretory granules and restricted to small cytoplasmic vesicles near or partially fused with granules. Vesicle labeling could be related to the occurrence of an alternative secretory pathway for statherin in buccal and labial glands.