Minor Salivary Gland Biopsy Specimens Research Articles

Distinctive Patterns of Transthyretin Amyloid in Salivary Tissue

Accurate typing of amyloidosis is still a major issue for pathologists and clinicians. Besides clinical data and immunohistochemistry, the histologic distribution of amyloid could represent a useful tool to prevent typing errors, such as the misdiagnosis of hereditary and senile amyloidosis as light chain-related amyloidosis (AL). Minor salivary gland biopsy (MSGB) is a widely performed procedure for amyloidosis diagnosis and typing. In the largest clinicopathologic series of amyloid-containing MSGB specimens to date, we investigated for the first time whether amyloidosis subtypes can be distinguished according to their pattern of salivary amyloid deposition. The histologic distribution and semiquantification of amyloid within salivary tissue were thoroughly reassessed for each case using Congo red-fluorescence. Clinical data were retrospectively collected. The cohort included 92 patients with amyloid-containing minor salivary gland biopsies. The type of amyloidosis was AL in 51 patients (55.4%), non-V30M mutant ATTR in 10 (10.9%), V30M mutant ATTR in 8 (8.7%), serum amyloid A-derived amyloidosis (AA) in 6 (6.5%), wild-type ATTR in 4 (4.3%), gelsolin in 3 (3.3%), and unclassified in 10 (10.9%). Amyloid was more abundant in AL and AA compared with ATTR amyloidosis, because of more extensive basement membranes and vascular deposits. Conversely, non-V30M mutant ATTR and wt-ATTR were strongly associated with peculiar amyloid nodules located in close contact with salivary excretory ducts, with a specificity of 91.7%. In conclusion, our study suggests for the first time that MSGB, in addition to its high sensitivity for amyloidosis diagnosis, is a simple and effective tool for the recognition of ATTR amyloidosis.

Detection of HTLV-1 in the Labial Salivary Glands of Patients with Sjögren’s Syndrome: A Distinct Clinical Subgroup?

To examine whether patients with Sjögren's syndrome (SS) can be distinguished based on the expression of human T cell lymphotrophic virus type I (HTLV-1) and, if so, whether the subgroups differ in their clinical features and serological measures. Polymerase chain reaction (PCR) and nested PCR were used to amplify viral DNA from peripheral blood mononuclear cells (PBMC) in 53 patients with SS, using primers from the HTLV-1 pX, p19, pol, and tax regions. Minor salivary gland biopsy specimens from 33 patients with SS were examined for the presence of HTLV-1 p19 or tax proteins immunohistochemically. The sociodemographic, glandular, and extraglandular manifestations, and laboratory findings including autoantibodies, complement, and immunoglobulin levels, were analyzed. The HTLV-1 tax gene was detected in PBMC samples from 2 of 53 patients (3.8%), whereas the HTLV-1 pX, p19, and pol genes were not expressed. As well, 100% of PBMC samples from 4 family members of patients in whom the tax gene was detected also expressed the tax gene. Immunohistochemical staining for HTLV-1 p19 and tax was seen in 10 out of 33 (30.3%) patients with SS each. Overall, 14 (42.4%) patients expressed HTLV-1 p19 or tax proteins, and they had lower rheumatoid factor and C3 levels (p = 0.015 and p = 0.005, respectively) and higher lymphocyte counts (p = 0.016). The prevalence of glandular and extraglandular manifestations did not differ between the HTLV-1-positive and negative patients. Our findings suggest that HTLV-1 in the salivary glands is involved in the pathogenesis of a subpopulation of SS, and HTLV-1-associated SS might have different immunological patterns than idiopathic SS.

Whole stimulated salivary flow: Correlation with the pathology of inflammation and damage in minor salivary gland biopsy specimens from patients with primary Sjögren's syndrome but not patients with sicca

To determine which measure of the salivary flow rate, stimulated or unstimulated, is most strongly associated with pathologic changes in minor salivary gland (MSG) biopsy specimens, and to explore the correlation of salivary flow with oral surface damage, disease duration, and symptom severity in patients with primary Sjögren's syndrome (SS). In all patients (n = 32), a biopsy of the MSG was performed, and stimulated salivary flow was assessed. Beginning in 2002, unstimulated salivary flow was also assessed. Scores for the severity of symptoms, according to the decayed/missing/filled teeth (DMF) index, were recorded. Associations between measures of salivary flow and covariates characterizing pathology were examined. A definite association between stimulated salivary flow and the MSG focus score, the grade of MSG fibrosis, the duration of dry mouth symptoms, and the DMF score was observed. In contrast, unstimulated salivary flow was not associated with fibrosis, atrophy, the DMF score, or the duration of dry mouth symptoms. In patients with primary SS, the DMF score was associated with pathologic changes in the MSG. Among patients with sicca, 57.9% had an abnormal unstimulated salivary flow rate (versus 82.4% of patients with primary SS), and 15.2% had an abnormal stimulated salivary flow rate (versus 61.8% of patients with primary SS). Among patients with sicca, neither stimulated salivary flow nor unstimulated salivary flow was associated with the degree of fibrosis or atrophy or with the DMF score. Compared with unstimulated salivary flow, stimulated salivary flow appeared to be a better measure of inflammation (according to the focus score) and fibrosis. In patients with sicca, the unstimulated salivary flow rate appeared to be abnormal more commonly compared with the stimulated salivary flow rate. In the future, stimulated salivary flow may serve as a noninvasive surrogate biomarker of inflammation and fibrosis as well as a measure of response to treatment in patients with primary SS.

What Ophthalmologists Should Know About Sjögren’s Syndrome

About one-quarter of patients with dry-eye syndrome (DES) have an underlying rheumatic condition, most commonly Sjogren’s syndrome (SS). SS is a common, frequently underdiagnosed cause of dry eye with multisystemic involvement. The immunological background of SS involves activation of both innate and acquired immune systems. According to the 2002 American and European classification systems, presence of any four of the six criteria (i.e. symptoms of dry eye, symptoms of dry mouth, ocular signs of dry eye, objective salivary gland involvement, typical histopathology findings in minor salivary gland biopsy specimens and presence of serum autoantibodies) as long as either the biopsy or serology is positive or presence of any three of the four objective criteria is considered diagnostic of SS. The main goal of treatment of ocular involvement is the alleviation of dry-eye symptoms with replacement therapy, stimulation of tear secretion and supportive surgical procedures to conserve tears. Preservative-free tear substitutes, topical steroids, cyclosporine A, autologous serum eye-drops and punctal plugs are commonly used for this purpose.

Salivary gland epithelial cells: A new source of the immunoregulatory hormone adiponectin

Adiponectin is an adipocytokine that displays insulin-sensitizing and immunoregulatory properties. Adipocyte development in association with fibrosis is frequently detected in primary Sjögren's syndrome lesions, connoting a healing process. The aim of this study was to examine the expression of adiponectin in minor salivary gland biopsy specimens obtained from patients with primary SS and controls. The expression of adiponectin in minor salivary gland biopsy specimens and in long-term-cultured non-neoplastic salivary gland epithelial cell (SGEC) lines obtained from patients with primary SS and control subjects was examined, using immunohistochemistry and immunoblotting, respectively. The expression of adiponectin, adiponectin receptor 1 (AdipoR1), and AdipoR2 messenger RNA (mRNA) by SGECs was investigated by reverse transcription-polymerase chain reaction. Immunohistochemical analysis for adiponectin revealed positive staining of adipocytes from primary SS lesions as well as ductal epithelial cells from both patients with primary SS and controls. All of the SGEC lines tested were shown to express adiponectin, AdipoR1, and AdipoR2 mRNA, whereas adiponectin protein expression was detected by immunoblotting in SGECs from patients with primary SS but not in those from controls. The analysis of concentrated culture supernatants also revealed increased adiponectin expression by SGECs from patients with SS compared with controls. Our findings provide novel evidence that adiponectin is produced by SGECs. The high constitutive expression of adiponectin by SGECs from patients with primary SS is likely attributable to aberrant activation of these cells. Although the significance of adiponectin expression remains unknown, it is possible that adiponectin functions in an autocrine manner, as suggested by concurrent expression of the relevant receptors.

Epithelial CXCR3-B Regulates Chemokines Bioavailability in Normal, but Not in Sjögren’s Syndrome, Salivary Glands

Expression of CXCR3-targeting chemokines have been demonstrated in several diseases, suggesting a critical role for CXCR3 in recruiting activated T cells to sites of immune-mediated inflammation. Sjögren's syndrome (SS) is an autoimmune disease characterized by a mononuclear cell infiltrate of activated T cells around the duct in the salivary gland. Analysis of minor salivary gland biopsy specimens from 20 healthy subjects and 18 patients with primary SS demonstrated that CXCR3, in particular, the B form of this receptor, is constitutively expressed by human salivary gland epithelial cells. Salivary gland epithelial cell cultures demonstrated that CXCR3 participate in removing relevant amount of agonists from the supernatant of exposed cells without mediating calcium flux or chemotaxis while retaining the ability to undergo internalization. Although in normal salivary gland epithelial cells, CXCR3 behaves as a chemokine-scavenging receptor, its role in SS cells is functionally impaired. The impairment of this scavenging function might favor chemotaxis, leading to heightened immigration of CXCR3-positive T lymphocytes. These findings suggest that epithelial CXCR3 may be involved in postsecretion regulation of chemokine bioavailability. They also support a critical role for CXCR3 in the pathogenesis of SS and identify its agonists as potential therapeutic targets.

Relapse of respiratory insufficiency one year after organising pneumonia

A 66-yr-old nonsmoker female presented in June 1998 with a 3-week history of progressive dyspnoea. Her past medical history was noteworthy for idiopathic bilateral uveitis without sequelae between 1978 and 1993, hypercholesterolaemia and angina pectoris. Her medication included fluvastatin, which started in June 1997 (60 mg·day−1), trinitrin patch and acetylsalicylic acid. She had no particular work-related or environmental dust exposure. Examination of the chest revealed bilateral basal crackles. There was no digital clubbing or extrapulmonary signs. At admission, chest radiograph and thoracic computed tomography (CT) scan (fig. 1a⇓) revealed patchy consolidations, predominantly involving the bilateral lower lobes. Fig. 1.— Computed tomography (CT) scans of 1-mm thickness at lung window. a) CT scan revealed irregularly marginated patchy alveolar consolidations with an air bronchogram. The lesions were bilateral and predominantly involved the lower lobes. b) After 1 yr, the consolidations disappeared almost completely. CT scan revealed some linear opacities of irregular thickness. c) CT scan from September 1999. d) CT scan revealed the same lesions with some improvement in May 2003. Arterial blood gas analysis under supplemental oxygen (6 L·min−1) revealed a partial pressure of oxygen of 8.3 kPa (62 mmHg), a partial pressure of carbon dioxide of 4.3 kPa (32 mmHg) and a pH of 7.46. As a result of rapidly progressive respiratory failure, she was admitted to the intensive care unit. Bronchoalveolar lavage (BAL) analysis showed a 140×106 cells·L−1 increased lymphocyte rate (38%), low CD4/CD8 ratio (0.18), increased neutrophil rate (17%), 44% of macrophages and 1% of eosinophils. Transbronchial lung biopsy, including 20 alveoli, revealed typical lesions of organising pneumonia (fig. 2a⇓). Minor salivary gland biopsy specimens found no sign of Sjogren's syndrome. Antinuclear antibodies were positive at a titre of 1:50 of speckled pattern; anti-JO1 antibodies were positive. Neither clinical nor biological signs of muscular involvement were found. Fluvastatin was stopped. Fig. 2.— a) Transbronchial biopsy …

Lymphoma development in Sjögren's syndrome: novel p53 mutations.

Sjögren's syndrome (SS) is a chronic autoimmune disease characterized by lymphocytic infiltrations of the exocrine glands. Disease progression may lead to uncontrolled clonal proliferation of B lymphocytes and development of lymphoma. This study was undertaken to examine the possible involvement of the cell cycle checkpoint genes p53 and p21 in the pathophysiology of the syndrome. Protein expression of p53 and p21 was studied, by immunohistochemistry and Western blot analysis, in minor salivary gland (MSG) biopsy specimens from 7 patients with SS and 5 control subjects. In addition, sequence analysis of the p53 gene was performed on DNA samples obtained from MSG biopsy samples of the same 7 patients with SS and from 4 patients with SS and in situ non-Hodgkin's lymphoma (NHL). The study revealed increased protein expression of p53 and p21 in MSG biopsy specimens from patients as compared with controls, while sequence analysis showed that the p53 gene was of the wild type. Furthermore, sequence analysis of the p53 gene from patients with SS and in situ NHL revealed 2 novel mutations in exon 5 of the p53 gene. These mutations are single-base substitutions and appear to be functional since exon 5 is included in the coding region of the p53 gene. This is the first report on wild-type p53 gene activation in SS. Our findings indicate a probable role for the DNA damage response genes in the pathogenesis of this syndrome. The novel mutations of the p53 gene implicate dysregulation of this tumor suppressor gene as a possible mechanism for lymphoma development in SS.

Detection of Epstein-Barr virus in salivary gland specimens from Sjögren's syndrome patients.

The cause of Sjögren's syndrome remains unclear, but several environmental and genetic factors have been implicated. The Epstein-Barr virus (EBV), among others (e.g., cytomegalovirus, human herpesvirus 6, and retroviruses), has been widely studied in connection with Sjögren's syndrome without conclusive results. To determine the role of EBV infection in patients with Sjögren's syndrome, the presence of EBV deoxyribonucleic acid (DNA) in major and minor salivary gland biopsy specimens was investigated by means of sulfur 35 in situ hybridization and polymerase chain reaction. Additionally, the presence of latent virus proteins EBV latent membrane protein and Epstein-Barr nuclear antigen 2 was analyzed by immunohistochemical methods. Viral DNA, detected by in situ hybridization, was found in 19% of patients with a diagnosis of Sjögren's syndrome and in 3% of controls. All tissues studied were found to be negative for EBV DNA by polymerase chain reaction. EBV latent membrane protein-positive staining was seen in 17% of patients and 22% of control subjects, while Epstein-Barr-positive staining was found in 25% of patients and 39% of controls. The low frequency of EBV DNA detected in the biopsy specimens does not indicate that the virus itself is the cause of Sjögren's syndrome. However, the possibility that the virus acts as a cofactor cannot be ruled out.

The Significance of Oral Mucosal and Salivary Gland Pathology After Allogeneic Bone Marrow Transplantation

Lichen planus—like oral mucosal findings as well as xerostomia are common after allogeneic bone marrow transplantation (BMT).¹Mucosa and salivary glands are sampled to establish or confirm the diagnosis of chronic graft-vs-host disease (GVHD). We studied mucosal epithelial and minor salivary gland biopsy specimens obtained for this purpose, using a four-level grading scheme (<b>Table</b>), similar to that for acute cutaneous disease, to investigate the clinical relevance of histologic findings for BMT outcome and to find correlation with other aspects of mucocutaneous disease after allogeneic BMT. <h3>Materials and Methods.</h3> Sixty-six specimens from 59 patients, taken an average of 136 days after allogeneic BMT (range, 65 to 201 days), were reviewed. Type of malignant neoplasm, preparative regimen, major histocompatibility complex disparity, and medications during and after hospitalization varied among the patients. Specimens were simultaneously evaluated semiquantitatively and graded by two observers (E.B.R., R.L.C.). Factors associated with survival were selected based on

Association of serum IgM kappa monoclonicity in patients with Sjogren's syndrome with an increased proportion of kappa positive plasma cells infiltrating the labial minor salivary glands.

Minor salivary gland biopsy specimens from 11 patients with primary Sjögren's syndrome with circulating monoclonal IgM kappa cryoglobulins, seven without cryoglobulins, and four patients with rheumatoid arthritis and Sjögren's syndrome (one with monoclonal and three with polyclonal cryoglobulins) were examined by the peroxidase antiperoxidase bridge technique, using antihuman kappa and lambda antibodies. In 6/11 patients with primary Sjögren's syndrome and in one patient with Sjögren's syndrome and rheumatoid arthritis with monoclonal cryoglobulins a predominance of plasma cells containing intracytoplasmic kappa light chains was found (kappa:lambda greater than 3:1). Two of those seven patients had immunohistological features of immunocytomas. In the other five patients with circulating monoclonal cryoglobulins the kappa:lambda ratio of positive cells did not exceed 3:1, while six out of seven patients without cryoglobulins and the patients with rheumatoid arthritis and Sjögren's syndrome with polyclonal cryoglobulins had almost equal numbers of kappa and lambda stained cells. One of seven patients with primary Sjögren's syndrome without cryoglobulins had an increased number of lambda light chain positive cells, indicating a non-secretory lambda monoclonal population. These findings suggest that the main area of B cell monoclonal expansion in primary Sjögren's syndrome may be the affected exocrine glands.

HLA-DR expression by labial minor salivary gland tissues in Sjogren's syndrome.

Minor salivary gland biopsy specimens from patients with Sjögren's syndrome (primary and secondary) and from normal controls were examined with the four step biotin-avidin-immunoperoxidase assay. The composition of the infiltrating cells was similar in patients with both primary and secondary Sjögren's syndrome, consisting primarily of T lymphocytes with predominance of T helper/inducer cells. B lymphocytes (Leu-14) were approximately 20-35% of the infiltrating lymphocytes, while only a few OKM1 (monocytes/macrophages) cells and Leu-7+ (natural killer; NK) cells were observed. The majority of infiltrating lymphocytes expressed HLA-DR antigens. In the biopsy specimens of the controls there were no infiltrates; the scattered lymphocytes, however, were also predominantly T lymphocytes. Finally, the glandular epithelial cells (ducts and acini) were inappropriately expressing HLA-DR antigens, in contrast with controls where minimal HLA-DR expression was found.

Anti-Ro(SSA) positive rheumatoid arthritis (RA): a clinicoserological group of patients with high incidence of D-penicillamine side effects.

The clinical, laboratory, histological, and radiological manifestations of 90 Greek patients with anti-Ro(SSA) negative rheumatoid arthritis (RA) were compared with those of 15 Greek patients with anti-Ro(SSA) positive RA. Anti-Ro(SSA) positive RA patients had the same articular and extra-articular manifestations as anti-Ro(SSA) negative patients. However, they were predominantly females with lower rheumatoid factor titres and a high incidence of positive minor salivary gland biopsy specimens for Sjögren's syndrome. Finally, anti-Ro(SSA) positive RA patients frequently experienced penicillamine side effects.