Immunohistochemical characterization of epithelial cells in human lacrimal glands

The distribution of cytokeratins (CK), actin, lactoferrin (Lf), lysozyme (Ly), vimentin and S-100 protein was immunohistochemically investigated in paraffin-embedded specimens of five inflammatory and five neoplastic lesions of lacrimal glands (LGs). Atrophic acini in dacryoadenitis reacted with antibodies (ABs) KL1 and Pkk1 (CK 7, 8, 17, 18) in a manner similar to ducts. Apart from myoepithelial cells and some luminal-duct cells, the remaining epithelia in dacryoadenitis were negative with AB 34 beta E12 (CK 5). The number of AB HHF35 (actin)-positive myoepithelial cells was not altered in dacryoadenitis. Epithelia in dacryoadenitis reacted weakly but consistently with Lf while revealing weak and inconsistent staining for Ly. Vimentin was negative in epithelial cells in dacryoadenitis except in one case. S-100 protein was detected only in epithelia of inflammatory major LGs. Epimyoepithelial islands in lymphoepithelial proliferation reacted variably for CKs, Lf, Ly and vimentin and remained negative for actin and S-100. In pleomorphic adenomas, neoplastic cells showing duct-like differentiation (luminal) reacted consistently with CK 7, 8, 17, 18 and S-100 protein and inconsistently with CK 5, Lf and Ly but remained negative for actin and vimentin. Other neoplastic cells (ovoid/peripheral cells) stained consistently for CK 5, vimentin and S-100 protein and focally for CK 7, 8, 17, 18, actin, Lf and Ly. Spindle-form neoplastic cells found in the stroma exhibited vimentin and S-100 protein and, less frequently, actin. Determination of these antigens in pleomorphic LG adenomas may help to evaluate their prognosis.     

Demonstration of nerve fibers in human accessory lacrimal glands

To provide morphologic evidence for the innervation of accessory lacrimal glands, glands were biopsied and examined using standard transmission electron microscopic techniques. Non-myelinated nerve fibers were found in the connective tissue between the glandular epithelia where they made contact with glandular epithelial cells, myoepithelial cells, vascular endothelial cells, plasma cells and fibroblasts. The distances measured between axons and target cells ranged from 30 to 130 nm. Where nerve fibers approached cells sustaining a basement membrane, their basement membranes fused to form a discrete unit resembling so-called synapses à distance. Cells with no basement membrane were situated in direct contact with the basement membrane of a nerve fiber. Single axons were identified between glandular epithelial cells and cells of intralobular ducts. Most of these axons contained many small clear vesicles and a few large, dense core vesicles, a finding considered typical of cholinergic parasympathetic nerve fibers. In addition, one of the axons identified contained small dense core vesicles typical of sympathetic nerve fibers. Human accessory lacrimal glands are therefore definitely innervated, with parasympathetic structures morphologically prevailing over sympathetic structures.Supported by Aktion Kampf der Erblindung     

Secretoglobins: sexually dimorphic expression of androgen-binding protein mRNA in mouse lacrimal glands

PURPOSE: This study was conducted to seek sex differences in mRNA expression in normal mouse lacrimal glands. Gene expression differences in the lacrimal gland may contribute to susceptibility to lacrimal gland or ocular surface disease. METHODS: A differential display analysis was performed on poly(A)+ RNA isolated from male and female Swiss Webster mouse exorbital lacrimal glands. Four potential gender-specific products were subcloned and sequenced. Full-length cDNAs of each product were obtained using RACE-PCR. 32P-labeled fragments of each clone were hybridized to a blot of male and female mouse poly(A)+ RNA isolated from harderian, lacrimal, submandibular, sublingual, and parotid glands and the liver. RESULTS: GenBank database alignments indicated that the four clones were members of the secretoglobin family. The most closely related sequences were the mouse salivary androgen-binding protein (ABP) subunits alpha, beta, and gamma. We named the four lacrimal clones the delta, epsilon, zeta, and eta subunits of ABP. Northern blot analysis showed that mRNAs for each of these four ABP subunits were lacrimal-gland-specific. The delta and zeta subunits of ABP were expressed primarily in male mouse lacrimal gland. CONCLUSIONS: Sequence attributes predict that the ABP subunits expressed in lacrimal glands comprise proteins that are secreted in tears. These data imply compositional differences in ABPs secreted by mouse lacrimal and submandibular glands, and in ABPs secreted by male and female mouse lacrimal glands.     

Estrogen up-regulation of metalloproteinase-2 and -9 expression in rabbit lacrimal glands

Increased levels of the matrix metalloproteinases (MMPs)-2 and -9 have been found in tear fluids of patients with dry eye disease, suggesting that these MMPs may be implicated in the pathogenesis of this disease. One of the main causes of dry eye disease is lacrimal gland insufficiency. However, the contribution of the lacrimal gland (LG) to the expression and production of MMP-2 and MMP-9 in tears is not known. Since dry eye disease occurs more frequently in women, sex hormones, especially estrogens, have also been implicated in the pathogenesis of this disease. Estrogens have been shown to regulate the synthesis levels of MMP-2 and MMP-9 in several tissues, Thus, the purpose of these studies was to determine if: (1) rabbit lacrimal glands secrete MMP-2 and MMP-9; (2) MMP-2 and MMP-9 are produced by lacrimal epithelial cells and/or lacrimal lymphocytes; and (3) the expression, activity and level of these enzymes are regulated by sex hormones. Lacrimal epithelial cells (LEC) and lacrimal lymphocytes (LL) from sexually mature New Zealand White female rabbits were isolated, purified and cultured with and without 10(-6)M dihydrotestosterone (DHT) or 10(-6), 10(-8), 10(-9) and 10(-10)M 17beta-estradiol (E2). The culture supernatants were analyzed by zymography and western blotting (WB) using polyclonal anti-human MMP-2 and MMP-9 antibodies. LGs were also collected from rabbits 7 days after being sham-operated, ovariectomized (OVX), OVX treated with 4 mg/kg DHT, and OVX treated with 0.5 mg/kg of E2. LGs were collected and processed for RNA extraction as well as protein determination using WB and immunocytochemistry. The pro-forms of MMP-2 and MMP-9 were detected in primary LEC and LL culture medium by zymography and WB. Pro-MMP-2 and pro-MMP-9 were also detected at the gene and protein levels in the lacrimal glands of all four treatment groups, with the highest levels and gene expression found in the estrogen-treated group. These results suggest that both pro-MMP-2 and pro-MMP-9 are secreted by the lacrimal gland and appear to be up-regulated by estrogen. The role of the lacrimal MMPs in the pathogenesis of dry eye disease needs to be further investigated.     

Angiolymphoid hyperplasia with eosinophilia with bilateral involvement of the lacrimal glands

BACKGROUND: Angiolymphoid hyperplasia with eosinophilia (ALHE) and Kimura's disease are two rarely occurring disorders very similar to each other; however, they are individual nosological entities. For a long time they were considered as a single disease due to the frequency of similar characteristics. The majority of authors have recently considered these diseases as two individual entities with some reciprocal specifications, both clinical as well as morphological. PATIENT: We report on the case of a 76-year old male white European who had suffered for more than six years from subcutaneous tumour formations in different parts of the body. Eyelid oedema on the right side with palpable resistance under the upper temporal edge of the orbit initially occurred five years after the first symptoms of the disease. A similar finding occurred on the left side after eight months. The tumours on both sides were surgically removed and sent for histological analysis. A unilateral recurrence of the finding appeared after one year, followed by surgical intervention and histological examination. RESULTS: Tumour infiltrate in the first two orbita excisions was topically related to lacrimal gland structures; these structures were not found in the third excision. The case was histologically diagnosed as ALHE despite some similar characteristics with Kimura's disease. The diagnosis of ALHE was histologically supported, especially by the absence of lymphatic follicle formations and fibrotisation in the infiltrate, and the identification of the appearance of epitheloid up to histiocytoid of proliferating endothelium, which forms small lumens. The diagnosis of ALHE was additionally supported by the fact that the incidence of Kimura's disease in white Europeans is very rare. CONCLUSIONS: According to our findings, both entities overlap one another, even in some characteristics considered to be distinguishing. The question arises, therefore, whether the strict separation of these nosological entities can indeed be determined with one hundred percent accuracy.     

Development of T cell lineages in rat lacrimal glands

PURPOSE. The purpose of this study was to examine T cell development in rat lacrimal glands, determine whether the thymus is the source of immature T cells in this tissue and compare lacrimal gland T lymphocytes with other T cell subpopulations. METHODS. Mononuclear cells were isolated from lacrimal glands of normal or thymectomized female Fischer 344 rats and stained for flow cytometric analysis. RESULTS. The lacrimal gland T lymphocyte population included large percentages of cells with an activated phenotype and also subpopulations of immature, naive and memory T cells. The numbers of immature (Thy-1(+)) lacrimal gland T cells were unchanged following short-term adult thymectomy. In comparison, spleen had large percentages of naive T cells, only a small subpopulation of activated T cells, and similar percentages of immature (Thy-1(+)) T cells, which were nearly eliminated after thymectomy. Lacrimal gland T cells had small subpopulations of TCRgammadelta(+) and CD8alphaalpha( +) T cells, a large subpopulation of NKT cells and many integrin alphaEbeta7( +) T cells. CONCLUSIONS. Lacrimal gland T cells are composed of a variety of subpopulations whose composition is distinct from splenocytes. The marked reduction of immature splenic T cell percentages eleven days after adult thymectomy indicates that these cells were mostly derived from thymic precursors. In contrast, the unchanged percentages of immature lacrimal gland T cells following thymectomy indicate that they may have an extrathymic source. These studies provide a foundation for further investigation into the cellular basis of lacrimal gland immunobiology.     

唇唾液腺和泪腺活检诊断Sjogren综合征

目的探讨唇唾液腺和泪腺活检组织的局灶性腺炎在诊断Ｓｊｏｇｒｅｎ综合征的价值。方法对１０５例可疑Ｓｊｏｇｒｅｎ综合征患者进行活检。Ｓｊｏｇｒｅｎ综合征的诊断按照Ｆｏｘ标准，活检诊断按照Ｃｈｉｓｈｏｌｍ－Ｍａｓｏｎ分级和Ｇｒｅｅｎｓｐａｎ分灶标准。结果肌上皮岛和重度淋巴细胞浸润伴生发中心仅出现在泪腺活检组织中。采用双重组织诊断Ｓｊｏｇｒｅｎ综合征较采用单一组织诊断明显有效。结论对可疑Ｓｊｏｇｒｅｎ综合征患者同时行唇唾液腺和泪腺活检可增加诊断的正确性     

Characterization of the insulin-signaling pathway in lacrimal and salivary glands of rats

PURPOSE: Insulin has been acknowledged as a mediator of several physiological events in lacrimal and salivary glands. We investigated the presence of insulin receptors and of insulin-induced autophosphorylation of the insulin receptor and activation of elements involved in the early steps of insulin signaling in lacrimal and salivary glands of rats. METHODS: Lacrimal and salivary glands of Wistar rats were removed and processed for immunohistochemistry using anti-insulin receptor and anti-IGF-1 receptor antibodies. The activation of insulin receptors following insulin treatment, and the involvement of insulin receptor substrates-1 and -2, Shc, JAK-2 and STAT-1, were analyzed by immunoprecipitation, followed by SDS-PAGE and immunoblotting of rat lacrimal and salivary glands after exposure to insulin. RESULTS: Insulin and IGF-1 receptors were present in rat lacrimal and salivary glands and were located predominantly in the cytoplasm and plasma membrane. Functional studies demonstrated that insulin induced a dose-dependent phosphorylation of the insulin receptor, IGF-1R, insulin receptor substrates-1 and -2, Shc, and STAT-1. In rats with streptozotocin-induced diabetes mellitus there was a significant reduction in insulin-induced insulin receptor and STAT-1 phosphorylation in the lacrimal gland but not in the salivary gland; there was no influence on Shc phosphorylation in either tissue. CONCLUSIONS: The present results indicate that insulin and IGF-1 receptors are expressed in lacrimal and salivary glands, and that insulin can induce the phosphorylation of its receptor and activate elements involved in the early steps of insulin signaling in both tissues.     

Distribution of adrenergic nerves in the lacrimal glands of guinea-pig and rat

The distribution of adrenergic nerves in the ex- and intraorbital lacrimal glands of guinea-pig and rat was studied using the sucrose-potassium phosphate-glyoxylic acid (SPG) technique. Blood vessels and secretory acini of guinea-pig lacrimal glands were demonstrated as having a rich adrenergic innervation. Adrenergic fibers in the rat were, however, much more sparse, and most of them were seen in association with glandular blood vessels, with only a few being found between secretory acini. Pretreatment of animal with a monoamine oxidase inhibitor and L-dopa did not change the morphological distribution of catecholamine fluorescent fibers, although the treatment improved the fluorescence, especially in the rat. Extirpation of the ipsilateral superior cervical ganglion eliminated all the fluorescent fibers in both normal and pretreated animals. The presence of adrenergic innervation of the lacrimal glands, especially in close connection with secretory acini, supports the theory that catecholamine-containing nerves play a role in the regulation of lacrimal secretion.