Immunoprofile of focal lymphocytic infiltration in minor salivary glands of healthy individuals

Aim: To characterize the immunohistochemical profile of the inflammatory cells included in the focal lymphocytic infiltration in the minor salivary glands of healthy individuals. Materials and Methods: Tissue samples of the labial and palatal salivary glands from 46 postmortem subjects, demonstrating the presence of focal lymphocytic infiltration were quantitatively evaluated for the presence of T‐ and B‐cell lymphocytes, plasma cells and macrophages by immunohistochemical and morphometric methods. Results: B‐cell lymphocytes, the predominant cell population in labial (67.5%) and palatal salivary glands (60.8%), were more frequent than T‐cell lymphocytes in both glands (P < 0.001). Among the T‐cell lymphocytes, CD₄‐positive cells were significantly more prevalent than the CD₈‐positive cells (P < 0.001). Plasma cells were almost absent, comprising only 0.01% of the focal lymphocytic infiltration cells of the labial and palatal salivary glands. Conclusions: Focal lymphocytic infiltration in the samples of the salivary glands obtained from healthy individuals is devoid of plasma cells. This can serve as an additional means to differentiate between focal lymphocytic infiltration in patients with Sjögren's syndrome, in which plasma cells are abundant, and focal lymphocytic infiltration in individuals with other causes of focal sialadenitis.     

HIV-associated parotid lymphoepithelial cysts

BACKGROUND: An outstanding feature of the diffuse infiltrative lymphocytosis syndrome, or DILS, a subset of HIV-1 disease, is asymptomatic bilateral parotid swelling. Recognition of the entity is important because people with this disease will seek routine dental care. CASE DESCRIPTION: The authors present a classic case of DILS. The patient exhibited bilateral parotid swellings caused by lymphoepithelial cysts, cervical lymphadenopathy, a CD8 circulating lymphocytosis and a CD8 lymphocytic infiltration into the labial salivary glands. A right superficial parotidectomy had been performed several years previously. However, no intervention was advised for the remaining left parotid because of its benign course. CLINICAL IMPLICATIONS: Since patients with DILS can develop lymphomas, periodic observation is mandatory. Any change in the growth pattern requires that a fine-needle aspiration biopsy be performed.     

Chronic GVHD in minor salivary glands and oral mucosa: histopathological and immunohistochemical evaluation of 25 patients

BACKGROUND: Graft-vs.-host disease (GVHD) is the major cause of morbidity and mortality in patients undergoing allogeneic Bone Marrow Transplantation (BMT). The aim of our study was to identify the most relevant histological features for diagnosis of chronic Graft-vs.-Host Disease (cGVHD) in oral mucosa and minor salivary glands of 25 patients, as well as to evaluate the immunophenotype of the inflammatory cells. METHODS: Sixteen patients that were submitted to allogeneic BMT but did not present cGVHD were selected as a control group. The sections were studied on H & E and CD68, CD45, CD4, CD8, CD20 staining. RESULTS: The most frequent histologic findings in oral mucosa at the day of diagnosis of cGVHD were: hydropic degeneration of the basal layer of the epithelium, apoptotic bodies, lymphocytic infiltration, and focal or total cleavage between the epithelial and connective tissue. In the labial salivary glands (LSG), lymphocytic infiltration, acinar loss and fibrosis were the main alterations. Cytotoxic CD8-T cells and macrophages were predominant both in the epithelium and connective tissue, as well as in minor salivary glands. CONCLUSIONS: Histological features were useful in the diagnosis of oral cGVHD. It is suggested that CD8-T cells and macrophages play important role in the pathogenesis of the disease.     

Immunohistochemical detection of insulin-like growth factor-I in the labial salivary glands of patients with Sjögren's syndrome

OBJECTIVE: The purpose of the present study was to investigate the presence of insulin‐like growth factor‐I (IGF‐I) in the labial salivary glands of patients with Sjögren's syndrome and healthy controls and to determine if there are any differences between these two groups. DESIGN: An immunohistochemical study. SUBJECTS AND METHODS: Twenty‐five patients with Sjögren's syndrome, 20 healthy controls and 20 patients with mucoceles of the lip were used in this study. All individuals underwent a systemic evaluation and a lip biopsy. Sections from the lip biopsies were stained with haema‐toxylin and eosin (H&E). Immunohistochemical staining was also performed using a three‐step indirect immunoperoxidase for IGF‐I. RESULTS: The light microscopic examination revealed the presence of a mononuclear infiltration in the labial salivary glands of patients with Sjögren's syndrome. Most of the infiltrates were lymphocytes. Immunohistochemically an intense staining result was apparent in the same group. In contrast sections of labial salivary glands of healthy individuals and of patients with mucoceles revealed very weak staining. CONCLUSIONS: The above findings and the fact that both lymphocytic infiltration and IGF‐I were predominantely seen in ductal regions, suggest that IGF‐I may be a target of autoimmunity in Sjögren's syndrome.     

An association between salivary gland disease and serological abnormalities in Sjögren's syndrome

In the labial salivary glands (LSGs) of 16 primary and 18 secondary Sjögren's syndrome (SS) patients, infiltrating lymphocytes were histologically and immunohistochemically examined: also, the serum levels of rheumatoid factor, antinuclear antibodies, anti-DNA antibodies, anti-SS-A and anti-SS-B antibodies, and immunoglobulins (including IgG, IgM and IgA) were all assayed. An immunohistochemical analysis of the lymphocyte subsets in LSGs revealed that severe lymphocytic infiltration was frequently accompanied by marked B cell accumulation both in primary and secondary SS patients. Furthermore, local B cell accumulation was also closely associated with elevated levels of anti-SS-A and anti-SS-B antibodies and IgG, and this association was statistically significant in the group with primary SS but not in the group with secondary SS. Thus, local lymphocytic infiltration, especially B cell accumulation, in the salivary glands is suggested to be involved in serological abnormalities in primary SS. while complicated autoimmune diseases other than SS may also be involved in serological abnormalities in secondary SS.     

PLUNC protein expression in major salivary glands of HIV‐infected patients

Oral Diseases (2011) 17 , 258–264 Objective: To analyse and compare the expression of Palate, Lung, and Nasal Epithelium Clone (PLUNC) proteins in salivary glands from patients with and without AIDS (control group) using autopsy material. Methods: We analysed the expression of PLUNCs using immunohistochemistry in parotid (n = 45), submandibular (n = 47) and sublingual gland (n = 37) samples of AIDS patients [30 with normal histology, 21 with mycobacteriosis, 14 with cytomegalovirus (CMV) infection, 30 with chronic non‐specific sialadenitis, and 30 HIV‐negative controls. In situ hybridization (ISH) for SPLUNC 2 in the HIV‐negative group was performed. Results: SPLUNC 1 expression was detected in the mucous acini of submandibular and sublingual glands, and SPLUNC 2 were seen in the serous cells. LPLUNC 1 expression was only positive in the salivary ducts. There was a higher expression of SPLUNC 2 in AIDS patients with CMV infection and mycobacteriosis when compared with all other groups. The intensity of staining for SPLUNC 2 was greater around the lesions than the peripheral ones. ISH for SPLUNC 2 showed perinuclear positivity in the serous cells in all HIV‐negative cases. Conclusions: SPLUNC 1 and LPLUNC 1 proteins were similarly expressed in the salivary glands of AIDS patients and non‐HIV patients. CMV infection and mycobacteriosis increase SPLUNC 2 expression in serous cells in the salivary gland of AIDS patients.     

Characterization of peripheral blood and salivary gland lymphocytes in secondary Sj?gren's syndrome.

Secondary Sj?gren's syndrome (SS) is defined as a condition of patients with sicca symptoms in association with a connective tissue disease such as rheumatoid arthritis. This study was designed to investigate the peripheral blood and affected minor salivary gland (SG) tissue lymphocytes with monoclonal antibodies in patients with secondary SS having rheumatoid arthritis. Minor SG lymphocytes of the patients and normals were determined in the fresh-frozen sections of the minor SG biopsy samples using monoclonal antibodies with immunoperoxidase technique. Peripheral blood of secondary SS patients revealed significant reduction in CD3+ and CD8+ cells. CD4/CD8 radio, HLA-DR+ cells, and B-cells were unchanged. SG biopsies showed varying degrees of lymphocytic infiltration with predominance of CD3+CD4+ cells located at the periductal areas. CD8+ cells were found to be in low numbers within the infiltrates. IgG- and IgA-producing plasma cells were both numerous in the biopsy samples. Our findings suggest that there is an alteration of lymphocyte subpopulations at the local site of inflammation in the salivary glands without, however, a corresponding alteration in the peripheral blood.     

Histologic lesion in labial salivary glands of patients with systemic lupus erythematosus

In an attempt to evaluate the presence or incidence of Sj?gren's syndrome in patients with systemic lupus erythematosus (SLE), 72 randomly selected patients with SLE, regardless of sicca manifestations, underwent labial salivary gland biopsy. Thirty-seven patients (51%) had normal labial salivary gland tissue. Seventeen patients (24%) had a mild perivascular infiltration of 15 to 20 lymphocytes per focus. In the remaining 18 patients (25%) heavy infiltration of more than 50 lymphocytes per focus was observed. From the latter group five specimens showed perivascular distribution of the infiltrates. About 40% of patients with any infiltration had keratoconjunctivitis sicca. Patients with severe lymphocytic infiltration in minor salivary gland tissue rarely had kidney involvement and often had lymphadenopathy and circulating rheumatoid factor, cryoglobulins, and antibodies to SS-A (Ro) and SS-B (La) antigens. Vasculitis was not found more frequently in any group of patients. Our results suggest that perivascular lymphocytic infiltrates in the labial salivary glands of patients with SLE may be the initial histologic lesion of Sj?gren's syndrome.     

Salivary gland changes in chronic fatigue syndrome: a case-controlled preliminary histologic study

OBJECTIVE: The purpose of this preliminary study is to compare labial salivary gland changes of 11 patients with chronic fatigue syndrome with control subjects. STUDY DESIGN: Changes in labial salivary glands were graded from 0 to 3+ for acinar dilatation, ductal dilatation, periductal fibrosis, plasmacytic infiltrate, lymphocytic infiltrate, mast cell infiltrate, and lymphocytic aggregates or foci. RESULTS: Four of the 11 subjects had 2+ to 3+ changes in at least 4 of the 7 parameters examined. Only the presence of mast cells was statistically significant between the 2 groups. Two of these 4 patients had 1 lymphocytic focus per 4 mm(2) of tissue. CONCLUSIONS: The salivary gland changes in patients with chronic fatigue syndrome show varying degrees of ductal and acinar dilatation, periductal fibrosis, lymphoplasmacytic infiltrates, and occasional lymphocytic foci, all suggestive of primary gland damage. The one parameter that showed statistical significance was the presence of mast cells (Fisher exact test, 0.0125).     

Characterization of lymphoid infiltrates in chronic obstructive sialadenitis associated with sialolithiasis

BACKGROUND: Chronic obstructive sialadenitis is characterized by acinar atrophy, lymphocytic infiltrates and progressive fibrosis. The immunological mechanisms involved in the pathogenesis of this disease are, for the most part, unknown. The aim of the present study was to characterize the lymphocytic infiltrates in chronic obstructive sialadenitis associated with sialolithiasis. METHODS: Paraffin-embedded tissue samples from 23 affected submandibular glands were immunostained for T-cells (CD3, CD4, CD8), cytotoxic T-cells (granzyme B), B-cells (CD20), plasma cells (CD38) and macrophages (Ki-M1P). RESULTS: CD4-positive subsets were the predominant cells, and they were located mainly periductally. Isolated intraepithelial CD8-positive cytotoxic T-cells associated with ductal epithelial cell destruction were observed in all cases. B lymphocytes were restricted to lymphoid follicles located periductally and around intralobular ducts. In early stages of the disease, a large number of CD38-positive plasma cells were distributed diffusely in the periacinar area. With progression of the disease, conspicuous clusters of plasma cells were located especially between atrophic acini adjacent to fibrotic tissue. An intimate relation between the lymphocytic infiltrates and the ductal epithelium, the target of the inflammatory process, was observed. CONCLUSION: The composition and distribution of inflammatory cells suggest that intraepithelial infectious agents may be the cause of the inflammatory reaction and the progressive fibrosis in this disease.     

Histological study of parotid,submaxillary and von Ebner salivary glands in chronic alcoholics

Samples from parotid, submaxillary, and von Ebner salivary glands of six chronic alcoholic individuals who had died of alcoholic hepatic cirrhosis were analyzed by topographic and histochemical routine stains and marked for cytokeratins; two normal adult individuals were used as control. Modifications in the acinar cells were found, but the most evident changes were observed in the ductal system: enlargement of major ducts, heterogeneous expression of cytokeratins and athrophy in epithelial cells, desquamated cells and stasis of content, and ductal hyperplasia in von Ebner glands. The lymphoplasmocytic infiltration does not represent the typical lymphocytic focus on Sj?gren's syndrome or other connective tissue pathologies. Our findings indicate that functional and structural variations are produced both in serous acini and ducts parotid, submaxilar and von Ebner glands affected by alcoholic sialosis.     

Salivary gland disease in HIV/AIDS and primary Sjögren''s Syndrome: analysis of collagen I distribution and histopathology in American and African patients

Background: Salivary gland disease (SGD) in HIV/AIDS is clinically and histopathologically very similar to Sjögren's Syndrome (SS), although the mechanism of tissue damage is unknown. The aim of this study is to determine the prevalence of SGD in primary SS and in HIV/AIDS in USA and in West African patients, and to seek distinguishing histopathologic features that may help to elucidate underlying mechanisms. Methods: Histologic sections of minor salivary glands from 164 HIV‐positive and ‐negative patients from Cameroon and the US, and from 17 US patients with primary SS, were evaluated following salivary gland biopsy for inflammatory changes. To confirm the presence of fibrosis, collagen I, which is the most abundant collagen type, was assessed immunohistochemically in H&E‐stained sections. Results: Forty‐eight per cent of patients with HIV from Cameroon had severe SGD, while it was only in 6% of patients from the US. Patients with HIV in the US had less fibrosis and collagen I deposits than Cameroonians. Seventy‐six per cent of US HIV‐positive patients had received anti‐retroviral therapy, while none of the African patients had. SS and AIDS patients had a tendency for lymphocytes to locate in a perivascular rather than in a periductal distribution. Conclusions: The prevalence of SGD and the presence of fibrosis and collagen I in Cameroonians with HIV is significantly higher than in HIV‐positive American patients, and is similar to US patients with primary SS. The impact of patient selection, anti‐retroviral therapy, and pathogenic mechanisms on salivary gland pathology is discussed.     

Loss of PKCδ results in characteristics of Sjögren's syndrome including salivary gland dysfunction

Oral Diseases (2011) 17 , 601–609 Objectives: Chronic infiltration of lymphocytes into the salivary and lacrimal glands of patients with Sjögren’s Syndrome (SS) leads to destruction of acinar cells and loss of exocrine function. Protein kinase C‐delta (PKCδ) is known to play a critical role in B‐cell maintenance. Mice in which the PKCδ gene has been disrupted have a loss of B‐cell tolerance, multiple organ lymphocytic infiltration, and altered apoptosis. To determine whether PKCδ contributes to the pathogenesis of SS, we quantified changes in indicators of SS in PKCδ?/? mice as a function of age. Salivary gland histology, function, the presence of autoantibodies, and cytokine expression were examined. Materials and methods: Submandibular glands were examined for the presence of lymphocytic infiltrates, and the type of infiltrating lymphocyte and cytokine deposition was evaluated by immunohistochemistry. Serum samples were tested by autoantibody screening, which was graded by its staining pattern and intensity. Salivary gland function was determined by saliva collection at various ages. Results: PKCδ?/? mice have reduced salivary gland function, B220+ B‐cell infiltration, anti‐nuclear antibody production, and elevated IFN‐γ in the salivary glands as compared to PKCδ+//+ littermates. Conclusions: PKCδ?/? mice have exocrine gland tissue damage indicative of a SS–like phenotype.     

Focal lymphocytic infiltration in the human labial salivary glands: a postmortem study

Focal lymphocytic infiltration in the human labial salivary glands was examined in a series of 190 postmortem subjects after suitable exclusion had been made. Focal lymphocytic infiltration, with or without a slight degree of parenchymal atrophic change, was found in 22.4% of the males and in 35.7% of the females. Of these, 9.0% (12 subjects) of the males and 10.7% (6 subjects) of the females with focal lymphocytic infiltration did not show any atrophic changes of the parenchyma. In the series reported here, the prevalence of focal lymphocytic infiltration apparently differs from the results of earlier investigators who had reported that none of the postmortem subjects without autoimmune diseases or connective tissue diseases showed focal lymphocytic infiltration in minor salivary glands. Although the pathological significance of focal lymphocytic infiltration in the minor salivary glands remains obscure, its diagnostic value for Sjögren's syndrome is discussed.     

Focal lymphocytic infiltration in the human labial salivary glands: a postmortem study

Focal lymphocytic infiltration in the human labial salivary glands was examined in a series of 190 postmortem subjects after suitable exclusion had been made. Focal lymphocytic infiltration, with or without a slight degree of parenchymal atrophic change, was found in 22.4% of the males and in 35.7% of the females. Of these, 9.0% (12 subjects) of the males and 10.7% (6 subjects) of the females with focal lymphocytic infiltration did not show any atrophic changes of the parenchyma. In the series reported here, the prevalence of focal lymphocytic infiltration apparently differs from the results of earlier investigators who had reported that none of the postmortem subjects without autoimmune diseases or connective tissue diseases showed focal lymphocytic infiltration in minor salivary glands. Although the pathological significance of focal lymphocytic infiltration in the minor salivary glands remains obscure, its diagnostic value for Sj?gren's syndrome is discussed.     

Clinical and laboratorial profile and histological features on minor salivary glands from patients under investigation for Sj?gren’s syndrome

Diagnosis of Sjögren’s syndrome (SS) is complex and the usefulness of labial minor salivary glands biopsy in this process remains controversial. Objectives: to evaluate the clinical and laboratorial profile and histological features on labial minor salivary glands from patients under investigation of SS. Study Design: clinical charts from 38 patients under suspicion of SS and submitted to labial minor salivary glands biopsies were reviewed. Clinical and laboratorial data were retrieved from the clinical files and the HE-stained histological slides were reviewed under light microscopy. Results: mean age of the patients was 56.5 years and 97% were females; histological analysis showed that 42% of the cases showed ductal dilatation, lymphocytic foci were found in 52.6% and, from this group, 80% of the cases presented a foci/lobules ratio above 0.8. Acinar/ductal ratio was considered diminished in 39.5% of the samples. Thirty six (95%) and 32 (84%) patients, respectively, complained about xerostomia and xerophthalmia. A study of the time interval of the symptoms that led to SS investigation showed a mean of 116 months. Moreover, sixty-six percent of the patients had already been submitted to immunosuppressive therapy prior to the labial minor salivary gland biopsy. Age of the patients, scintigraphic alterations on salivary function, antinuclear factor (ANF), anti-Ro and anti-La did not show statistical significant association with the histological features. Lobules/foci ratio above 0.8 was the only histological parameter statistically associated with Sjögren’s syndrome diagnosis (p<0.0001). Conclusions: in the studied sample, lymphocytic foci on salivary glands were the only histological parameter associated to the diagnosis of SS. Early indication of labial minor salivary gland biopsy to patients under investigation of SS could limit the effects of immunosuppressive therapy on the histological features associated with the evolution of salivary gland involvement in SS. Key words:Sjögren syndrome, minor salivary glands, biopsy, lymphocytic foci.     

Parotid gland swelling in HIV diffuse infiltrative CD8 lymphocytosis syndrome

The signs and symptoms of diffuse infiltrative CD8 lymphocytosis syndrome (DILS), a subset of HIV, include parotid swelling, cervical lymphadenopathy and a serologic CD8 elevation. A case report is used to illustrate the condition. Patients with the syndrome will be seen in the dental office. Recognition and appropriate referral are responsibilities of the dental practitioner.     

Multiple parotid lymphoepithelial cysts in patients with HIV-infection: report of two cases

OBJECTIVE: Bilateral and multiple lymphoepithelial cysts (LECs) of major salivary glands, in particular of parotid glands, are quite rare and have been reported in human immunodeficiency virus (HIV) infected patients with an incidence of about 3-6%. These lesions represent an early manifestation of HIV infection and are rarely found in patients with advanced acquired immunodeficiency syndrome. MATERIALS: Two cases of parotid LECs, the first occurring in a middle-age white woman and the second in a young white boy, both in advanced phases of HIV infection, are reported. RESULTS: Clinical, cytological, histological and immunohistochemical (cytokeratin AE1/AE3, CD20, CD45RA, CD8, kappa and lambda immunoglobulin light chains, S-100, MLA and Ki67) features are described. CONCLUSIONS: Fine needle aspiration (FNA), a relatively non-traumatic procedure, could represent both a diagnostic and a therapeutic tool in parotid LECs. No surgical therapy is usually required for these lesions and aspiration of cystic fluid with FNA is quite resolutive, although evidence of further relapses does exist. Surgical excision may become necessary when pain, because of persistent and progressive swelling of the parotid gland, occurs.     

骨髓基质细胞抗原2在原发性舍格伦综合征中的表达及意义

目的: 检测骨髓基质细胞抗原2(bone marrow stromal antigen-2,BST-2)在原发性舍格伦综合征（primary Sjögren's syndrome,pSS）患者唇腺及外周血淋巴细胞中的表达,探讨BST-2对淋巴细胞增殖的作用,以期为原发性舍格伦综合征的发病机制研究及药物治疗靶点的开发提供理论基础。方法: 应用实时定量PCR技术检测30例pSS患者及30例对照患者外周血单个核细胞（peripheral blood mononuclear cells,PBMCs）和唇腺组织中BST-2的表达水平,并与临床指标进行相关性分析。采用免疫组织化学对唇腺组织中的BST-2进行定位及半定量分析。通过分选外周血中的淋巴细胞,实时定量 PCR明确BST-2异常表达的淋巴细胞亚群,唇腺免疫组织化学连续切片染色验证。慢病毒转染人B淋巴细胞系以过表达BST-2,CCK8检测BST-2对细胞增殖活性的影响,流式细胞术分析其对细胞凋亡的影响。采用SPSS 16.0软件包对数据进行统计学分析。结果: 与对照患者相比,pSS患者唇腺组织中BST-2表达升高,阳性细胞主要是浸润腺体的B淋巴细胞。同时,pSS患者外周血CD19⁺ B细胞BST-2与对照组相比表达升高。过表达B细胞中的BST-2可以促进细胞增殖,抑制细胞凋亡。结论: BST-2在pSS患者外周血及唇腺组织中表达升高,BST-2主要定位于B淋巴细胞。推测BST-2通过促进B细胞在腺体中的增殖和浸润,从而参与pSS的发生与发展。