ABSTRACT
BACKGROUND: Trials typically group cancers of the gastro-oesophageal junction (GOJ) with oesophageal or gastric cancer when studying neoadjuvant chemoradiation and perioperative chemotherapy, so the results may not be fully applicable to GOJ cancer. Because optimal neoadjuvant treatment for GOJ cancer remains controversial, outcomes with neoadjuvant chemoradiation versus chemotherapy for locally advanced GOJ adenocarcinoma were compared retrospectively. METHODS: Data were collected from all patients who underwent neoadjuvant treatment followed by surgery for adenocarcinoma located at the GOJ at a single high-volume institution between 2002 and 2017. Postoperative major complications and mortality were compared between groups using Fisher's exact test. Overall survival (OS) and disease-free survival (DFS) were assessed by log rank test and multivariable Cox regression analyses. Cumulative incidence functions were used to estimate recurrence, and groups were compared using Gray's test. RESULTS: Of 775 patients, 650 had neoadjuvant chemoradiation and 125 had chemotherapy. These groups were comparable in terms of clinical tumour and lymph node categories, although the chemoradiation group had greater proportions of white men, complete pathological response to chemotherapy, and smaller proportions of diffuse cancer, poor differentiation, and neurovascular invasion. Postoperative major complications (20.0 versus 17.6 per cent) and 30-day mortality (1.7 versus 1.6 per cent) were not significantly different between the chemoradiation and chemotherapy groups. After adjustment, type of therapy (chemoradiation versus chemotherapy) was not significantly associated with OS (hazard ratio (HR) 1.26, 95 per cent c.i. 0.96 to 1.67) or DFS (HR 1.27, 0.98 to 1.64). Type of recurrence (local, regional, or distant) did not differ after neoadjuvant chemoradiation versus chemotherapy. CONCLUSION: In patients undergoing surgical resection for locally advanced adenocarcinoma of the GOJ, OS and DFS did not differ significantly between patients who had neoadjuvant chemoradiation compared with chemotherapy.
Treating advanced cancer of the gastro-oesophageal junction (GOJ) poses a challenge given its location in the distal oesophagus and proximal stomach, and whether it should be treated as oesophageal or gastric cancer. Given the indistinct location, it is unclear whether GOJ cancer should be treated with neoadjuvant chemoradiation, which is the treatment of choice for advanced oesophageal cancers, or perioperative chemotherapy, which is the treatment of choice for advanced gastric cancers. Few studies have addressed treatment options specifically for GOJ cancers. This study investigated whether there was a difference in survival between patients with GOJ cancer who were treated with chemoradiation versus chemotherapy.
Subject(s)
Adenocarcinoma/therapy , Antineoplastic Agents/therapeutic use , Esophageal Neoplasms/therapy , Esophagectomy/adverse effects , Esophagogastric Junction , Neoplasm Staging , Adenocarcinoma/diagnosis , Adenocarcinoma/mortality , Aged , Chemoradiotherapy, Adjuvant , Esophageal Neoplasms/diagnosis , Esophageal Neoplasms/mortality , Female , Follow-Up Studies , Humans , Incidence , Male , Middle Aged , New York/epidemiology , Postoperative Complications/epidemiology , Prognosis , Retrospective Studies , Survival Rate/trendsABSTRACT
BACKGROUND: Immune checkpoint inhibitors (ICIs) and thoracic radiotherapy are increasingly used to treat advanced cancers. Despite data indicating exaggerated radiation toxicities in patients with autoimmune disease, the safety of thoracic radiotherapy in patients with prior ICI-associated immune-related adverse events (irAEs) is undefined. PATIENTS AND METHODS: Patients treated from 2014 to 2020 with ICIs were queried for receipt of corticosteroids and radiotherapy. Patients who received thoracic radiation after symptomatic irAEs were assessed for ≥grade 2 radiation pneumonitis (RP). Characteristics predictive of RP were assessed using logistic regression and response relationships were modeled. RESULTS: Among 496 assessed patients, 41 with irAE history subsequently treated with thoracic radiotherapy were analyzed. Most irAEs were grade 2 (n = 21) and 3 (n = 19). Median time from irAE onset to radiotherapy was 8.1 months. Most patients received stereotactic body radiation therapy (n = 20) or hypofractionated radiotherapy (n = 18). In total, 25 patients (61%) developed ≥grade 2 RP at a median of 4 months from radiotherapy and 11 months from onset of irAEs. Three months from RP onset, 16 of 24 (67%) assessable patients had persistent symptoms. Among patients with prior ICI pneumonitis (n = 6), five patients (83%) developed ≥grade 2 RP (grade 2, n = 3; grade ≥3, n = 2). The mean lung radiation dose (MLD) predicted for RP (odds ratio: 1.60, P = 0.00002). The relationship between MLD and RP was strong (area under the receiver-operating characteristic curve: 0.85) and showed an exaggerated dose-response. Among patients with an MLD >5 Gy (n = 26), 21 patients (81%) developed ≥grade 2 RP. CONCLUSION: This is the first study assessing the toxicity of radiotherapy among patients with prior irAEs from ICIs. Patients with prior irAEs were found to be at very high risk for clinically significant and persistent RP from thoracic radiotherapy. Careful consideration should be given to the possibility of an increased risk of RP, and close monitoring is recommended in these patients.
Subject(s)
Carcinoma, Non-Small-Cell Lung , Lung Neoplasms , Radiation Pneumonitis , Humans , Immune Checkpoint Inhibitors , Radiation Pneumonitis/epidemiology , Radiation Pneumonitis/etiology , Retrospective StudiesABSTRACT
AIM: Significant recent changes in management of locally advanced rectal cancer (LARC) include preoperative staging, use of extended neoadjuvant therapies and minimally invasive surgery (MIS). This study was aimed at characterizing these changes and associated short-term outcomes. METHOD: We retrospectively analysed treatment and outcome data from patients with T3/4 or N+ LARC ≤ 15 cm from the anal verge who were evaluated at a comprehensive cancer centre in 2009-2015. RESULTS: In total, 798 patients were identified and grouped into five cohorts based on treatment year: 2009-2010, 2011, 2012, 2013 and 2014-2015. Temporal changes included increased reliance on MRI staging, from 57% in 2009-2010 to 98% in 2014-2015 (P < 0.001); increased use of total neoadjuvant therapy, from 17% to 76% (P < 0.001); and increased use of MIS, from 33% to 70% (P < 0.001). Concurrently, median hospital stay decreased (from 7 to 5 days; P < 0.001), as did the rates of Grade III-V complications (from 13% to 7%; P < 0.05), surgical site infections (from 24% to 8%; P < 0.001), anastomotic leak (from 11% to 3%; P < 0.05) and positive circumferential resection margin (from 9% to 4%; P < 0.05). TNM downstaging increased from 62% to 74% (P = 0.002). CONCLUSION: Shifts toward MRI-based staging, total neoadjuvant therapy and MIS occurred between 2009 and 2015. Over the same period, treatment responses improved, and lengths of stay and the incidence of complications decreased.
Subject(s)
Disease Management , Neoadjuvant Therapy/trends , Patient Care Team/trends , Proctectomy/trends , Rectal Neoplasms/therapy , Aged , Female , Humans , Length of Stay/trends , Male , Margins of Excision , Middle Aged , Neoplasm Grading , Neoplasm Staging , Rectal Neoplasms/pathology , Retrospective Studies , Time Factors , Treatment OutcomeABSTRACT
Food and feed safety assessment is not enhanced by performing protein expression analysis on stacked trait products. The expression levels of six proteins in cotton matrices from four single cotton events and three conventionally stacked trait cotton products are reported. Three proteins were for insect control; two proteins confer herbicide tolerance; and one protein was a transformation-selectable marker. The cotton matrices were produced at three U.S., five Brazil, and two Argentina field trials. Similar protein expression was observed for all six proteins in the stacked trait products and the single events. However, when two copies of the bar gene were present in the stacked trait products, the expression level of phosphinothricin acetyl transferase herbicide tolerance was additive. Conventional breeding of genetically engineered traits does not alter the level or pattern of expression of the newly introduced proteins, except when multiple copies of the same transgene are present.
Subject(s)
Gossypium/genetics , Plant Proteins/genetics , Acetyltransferases/genetics , Acetyltransferases/metabolism , Gossypium/drug effects , Gossypium/metabolism , Herbicides/pharmacology , Hybridization, Genetic , Plant Proteins/metabolismABSTRACT
OBJECTIVES: To evaluate outcomes of our breast frozen section (FS) practice in its first 5 years, including our specialized FS of margins (FSM) procedure for breast conserving therapy (BCT) patients. METHODS: One thousand two hundred and forty eight patients undergoing 1303 breast FSM and/or sentinel lymph node (SLN) FS were included. Clinicopathologic features were assessed by chart review. RESULTS: Use of SLN FS declined, from 43.5% of FS cases before to 19.2% of FS cases after 2012. FSM patients had a decline in overall reexcision to 12.3% in 2013-2014 (p = 0.063). There was also decline in reexcision for focally close margins (p < 0.0001) but no change in reexcision for extensively close margins. Reexcision was significantly associated with lobular subtype, multifocality and larger (≥T2) size. False negative FSM cases were most often influenced by extensively close or positive final (reexcised) margins sent for permanent section only (96/148; 64.9%). CONCLUSIONS: Despite changing surgical practices, FSM remains a valuable service that reduces reexcision in BCT patients.
Subject(s)
Breast Neoplasms/pathology , Carcinoma, Ductal, Breast/pathology , Carcinoma, Intraductal, Noninfiltrating/pathology , Carcinoma, Lobular/pathology , Frozen Sections/statistics & numerical data , Margins of Excision , Adolescent , Adult , Aged , Aged, 80 and over , Breast Neoplasms/surgery , Breast Neoplasms, Male/pathology , Breast Neoplasms, Male/surgery , Carcinoma, Ductal, Breast/surgery , Carcinoma, Intraductal, Noninfiltrating/surgery , Carcinoma, Lobular/surgery , Female , Frozen Sections/trends , Humans , Intraoperative Period , Male , Mastectomy, Segmental/methods , Middle Aged , Reoperation , Sentinel Lymph Node/pathology , Sentinel Lymph Node Biopsy/methods , Surgicenters , Tumor Burden , Young AdultABSTRACT
This study looks at toxicity and survival data when chemoradiation (CRT) is delivered using intensity-modulated radiation therapy (IMRT) after induction chemotherapy. Forty-one patients with esophageal adenocarcinoma treated with IMRT from March 2007 to May 2009 at Memorial Sloan-Kettering Cancer Center were analyzed. All patients received induction chemotherapy prior to CRT. Thirty-nine percent (n = 16) of patients underwent surgical resection less than 4 months after completing CRT. Patients were predominantly male (78%), with a median age of 68 years (range 32-85 years). The majority of acute treatment-related toxicity was hematologic or gastrointestinal, with 17% of patients having grade 3+ hematologic toxicity and 12% of patients having grade 3+ gastrointestinal toxicity. Only two patients developed grade 2-3 pneumonitis (5%) and 5 patients experienced post-operative pulmonary complications (29%). Eight patients (20%) required a treatment break. With a median follow up of 41 months for surviving patients, 2-year overall survival was 61%, and the cumulative incidences of local failure (LF) and distant metastases were 40% and 51%, respectively. This rate of LF was reduced to 13% in patients who underwent surgical resection. Surgery and younger age were significant predictors of decreased time to LF on univariate analysis. Induction chemotherapy followed by CRT using IMRT in the treatment of esophageal cancer is well tolerated and is not associated with an elevated risk of postoperative pulmonary complications. The use of IMRT may allow for integration of more intensified systemic therapy or radiation dose escalation for esophageal adenocarcinoma, ultimately improving outcomes for patients with this aggressive disease.
Subject(s)
Adenocarcinoma/secondary , Adenocarcinoma/therapy , Chemoradiotherapy, Adjuvant/adverse effects , Esophageal Neoplasms/pathology , Esophageal Neoplasms/therapy , Neoplasm Recurrence, Local , Radiotherapy, Intensity-Modulated , Adult , Age Factors , Aged , Aged, 80 and over , Antineoplastic Combined Chemotherapy Protocols/adverse effects , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Camptothecin/administration & dosage , Camptothecin/analogs & derivatives , Chemoradiotherapy, Adjuvant/methods , Esophagectomy/adverse effects , Female , Fluorouracil/administration & dosage , Humans , Induction Chemotherapy/adverse effects , Irinotecan , Lymphatic Metastasis , Male , Middle Aged , Neoadjuvant Therapy , Paclitaxel/administration & dosage , Platinum Compounds/administration & dosage , Radiotherapy, Intensity-Modulated/adverse effects , Survival Rate , Tumor BurdenABSTRACT
BACKGROUND: Retroviral infection has been implicated in the pathogenesis of primary Sjögren's syndrome. OBJECTIVE: To examine the efficacy of the reverse transcriptase inhibitor lamivudine in patients with this syndrome. METHODS: 16 patients with primary Sjögren's syndrome were randomised to receive either lamivudine 150 mg twice daily or placebo for three months. Measures of lacrimal and salivary function, including minor salivary gland biopsies, were obtained before and after treatment. RESULTS: Treatment with lamivudine did not result in significant improvement in the primary outcome measure of unstimulated whole salivary flow or other secondary measures, including minor salivary gland biopsy focus scores. CONCLUSION: Lamivudine is not effective in patients with primary Sjögren's syndrome, suggesting either that a retroviral aetiology is not present or that it may be important only in early disease.
Subject(s)
Lamivudine/therapeutic use , Reverse Transcriptase Inhibitors/therapeutic use , Sjogren's Syndrome/drug therapy , Adult , Biopsy , Double-Blind Method , Female , Humans , Longitudinal Studies , Male , Middle Aged , Salivary Glands, Minor/pathology , Salivation/drug effects , Severity of Illness Index , Sjogren's Syndrome/physiopathology , Sjogren's Syndrome/virology , Tears/metabolism , Treatment FailureABSTRACT
Xerostomia is a common symptom with various causes that, if ignored, can lead to serious oral consequences. Clinical evaluation of patients complaining of dry mouth must include some additional history and specific examination of the salivary glands, oral mucosa, and teeth. Additional evaluation may include consultation with the patient's physician, request for microbial culture, or labial salivary gland biopsy. No one form of treatment for patients with chronic xerostomia is sufficient, but comprehensive treatment is effective in improving patient oral comfort and function and preventing unnecessary loss of teeth. This treatment must include ongoing dental caries prevention and treatment, salivary flow stimulation, recognition and treatment of oral candidiasis, selective use of saliva substitutes, and possible changes in the patients' prescription and nonprescription drug use.
Subject(s)
Xerostomia/diagnosis , Candidiasis, Oral/diagnosis , Candidiasis, Oral/prevention & control , Chronic Disease , Dental Caries/prevention & control , Humans , Medical History Taking , Pharmaceutical Preparations/administration & dosage , Physical Examination , Saliva, Artificial/therapeutic use , Salivary Glands/metabolism , Secretory Rate , Tooth Loss/prevention & control , Xerostomia/microbiology , Xerostomia/physiopathology , Xerostomia/prevention & control , Xerostomia/therapyABSTRACT
Polyreactive antibodies bind to a variety of different self and non-self antigens. The B cells that make these antibodies express the polyreactive lg receptor on their surface. To determine the frequency of polyreactive antigen-binding B cells in peripheral blood, we incubated two different antigens, one (insulin) labeled with fluorescein isothiocyanate and the other (beta-galactosidase) with phycoerythrin, with peripheral B cells. The percentage of cells that bound these antigens was determined with the fluorescence-activated cells sorter. Approximately 21% of adult B cells bound insulin, 28% bound beta-galactosidase, and 11% bound both antigens. In contrast to B cells in the adult repertoire, 49% of B cells in cord blood bound insulin, 54% bound beta-galactosidase, and 33% bound both antigens. The properties of polyreactive antigen-binding B cells in adult and cord blood were similar, except for the fact that almost all the polyreactive antigen-binding B cells in cord blood were CD5 positive (93%), whereas only 40% of the polyreactive antigen-binding B cells in adult peripheral blood were CD5 positive, indicating that the CD5 marker is not directly linked to polyreactivity. The percentage of polyreactive antigen-binding B cells in patients with Sjögren's syndrome, systemic lupus erythematosus and rheumatoid arthritis was equal to or slightly below that found in the normal adult B cell repertoire. It is concluded that polyreactive antigen-binding B cells are a major constituent of the normal adult B cell repertoire and are the predominant cell type in the newborn B cell repertoire.
Subject(s)
B-Lymphocytes/immunology , Infant, Newborn/immunology , Adolescent , Adult , Aged , Arthritis, Rheumatoid/immunology , Child , Child, Preschool , Female , Fetal Blood/cytology , Humans , Infant , Lupus Erythematosus, Systemic/immunology , Male , Sjogren's Syndrome/immunologyABSTRACT
We have used a human salivary gland cell line (HSG) as a possible in vitro model to evaluate the effects of IFN-gamma on human salivary gland epithelium (Wu et al., 1994, 1996, 1997). In the present study, we examined the JAK-STAT signal-transduction pathway in IFN-gamma-treated HSG cells. We demonstrate that JAK2 and Stat1 are phosphorylated at tyrosine residues in a time- and concentration-dependent manner following exposure to IFN-gamma. In addition, we show that activation of this signalling pathway is decreased by the addition of a blocking antibody to the IFN-gamma receptor. The same maneuver is also able to reduce by approximately 50-70% the surface expression of two IFN-gamma-induced immunoregulatory molecules: HLA-DR and ICAM-1. These results demonstrate that the JAK2 and Stat1 signalling pathway is active in salivary-derived epithelial cells and may contribute to their immunopathologic destruction.
Subject(s)
DNA-Binding Proteins/metabolism , Interferon-gamma/pharmacology , Protein-Tyrosine Kinases/metabolism , Proto-Oncogene Proteins , Salivary Glands/metabolism , Signal Transduction , Trans-Activators/metabolism , Antibodies/immunology , Blotting, Western , Cell Line , Fluorescent Antibody Technique , HLA-DR Antigens/immunology , HLA-DR Antigens/metabolism , Humans , Intercellular Adhesion Molecule-1/immunology , Intercellular Adhesion Molecule-1/metabolism , Janus Kinase 2 , Phosphorylation , Phosphotyrosine/metabolism , Precipitin Tests , Receptors, Interferon/antagonists & inhibitors , Receptors, Interferon/immunology , STAT1 Transcription Factor , Salivary Glands/cytology , Interferon gamma ReceptorABSTRACT
Gelatinases have been shown to be regulated by many cytokines and growth factors, and have been implicated in the pathogenesis of certain autoimmune diseases via tissue destruction. High levels of several cytokines, including IFN-gamma and TNF-alpha, have been demonstrated in the salivary gland microenvironment of patients with Sjogren's syndrome (SS). How these cytokines may be contributing to the pathogenesis of this disease is not well understood. We hypothesized that IFN-gamma with or without (+/-) TNF-alpha could be playing a role in the pathogenesis of SS via the regulation of matrix metalloproteinase (MMP) levels. This study examined the role of IFN-gamma and (+) TNF-alpha in the regulation of the matrix metalloproteinases, MMP-2 (72 kD gelatinase A) and MMP-9 (92 kD gelatinase B). A human salivary gland cell line (HSG) has been used as a possible in vitro model to study the role of IFN-gamma + TNF-alpha in the pathogenesis of SS. The HSG cell line, in the presence of IFN +/- TNF-alpha, displays increased MMP-2 and MMP-9 gelatinolytic activity, protein and RNA levels. The increase in MMP activity was partially blocked with an antibody against the IFN-gamma receptor, and this was associated with a complete inhibition of the previously described IFN-gamma +/- TNF-alpha antiproliferative effect. However, incubation of IFN-gamma treated HSG cells with the synthetic MMP inhibitor BB94 did not alleviate this antiproliferative effect. In addition, we demonstrate that there are very high levels of MMP-9 in the saliva of patients with SS when compared to healthy control subjects. These data suggest that cytokines could be regulating MMP production by salivary epithelial cells and thus indicate a potential role for these cells in the pathogenesis of SS.
Subject(s)
Collagenases/metabolism , Gelatinases/metabolism , Interferon-gamma/pharmacology , Metalloendopeptidases/metabolism , Parotid Gland/cytology , Animals , Antibodies, Monoclonal , Binding, Competitive/immunology , Blotting, Northern , Cell Division/drug effects , Cell Line/cytology , Cell Line/drug effects , Cell Line/enzymology , Collagenases/genetics , Epithelial Cells , Epithelium/drug effects , Epithelium/enzymology , Gelatinases/genetics , Humans , Matrix Metalloproteinase 2 , Matrix Metalloproteinase 9 , Metalloendopeptidases/genetics , Mice , Parotid Gland/enzymology , Phenylalanine/analogs & derivatives , Phenylalanine/pharmacology , Protease Inhibitors/pharmacology , RNA, Messenger/analysis , Receptors, Interferon/immunology , Saliva/enzymology , Sjogren's Syndrome/metabolism , Thiophenes/pharmacology , Tumor Necrosis Factor-alpha/pharmacologyABSTRACT
Interferon-gamma (IFN-gamma) +/- tumor necrosis factor-alpha (TNF-alpha) induces antiproliferation and intracellular Ca2+ store depletion in a human submandibular ductal cell line (HSG), which can be reversed on cytokine removal [A. J. Wu, G. C. Chen, B. J. Baum, and I. S. Ambudkar. Am. J. Physiol. 270 (Cell Physiol. 39): C514-C521, 1996]. Here we have examined a possible mechanism for the IFN-gamma-induced intracellular Ca2+ store depletion. There was a time-dependent decrease in thapsigargin-dependent internal Ca2+ release after exposure of the cells to the cytokines. The intracellular Ca2+ pump [sarco(endo)plasmic reticulum Ca(2+)-ATPase (SERCA)] protein in lysates and membranes of cells treated with IFN-gamma +/- TNF-alpha, but not with TNF-alpha alone, showed a similar time-dependent decrease (examined using a SERCA2 antibody). Removal of the cytokines, which resulted in recovery of cell growth and refill of internal Ca2+ stores, also increased the level of SERCA protein. The decrease in SERCA is not a result of decreased cell proliferation, since thapsigargin, 2,5-di-(t-butyl)-1,4-hydroquinone, or serum-free growth conditions induced antiproliferative effects on HSG cells without any corresponding decrease in SERCA. We suggest that the IFN-gamma-induced decrease in the level of SERCA accounts for the depleted state of internal Ca2+ stores in cytokine-treated HSG cells. These data suggest a novel mechanism for the inhibition of HSG cell growth by IFN-gamma.
Subject(s)
Calcium-Transporting ATPases/biosynthesis , Calcium/metabolism , Interferon-gamma/pharmacology , Submandibular Gland/enzymology , Cell Division/drug effects , Cell Line , Cell Membrane/enzymology , Culture Media, Serum-Free , Cytosol/metabolism , Enzyme Inhibitors/pharmacology , Gene Expression Regulation, Enzymologic/drug effects , Humans , Hydroquinones/pharmacology , Kinetics , Recombinant Proteins , Submandibular Gland/cytology , Thapsigargin/pharmacology , Time Factors , Tumor Necrosis Factor-alpha/pharmacologyABSTRACT
Increased levels of several cytokines, including interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha), have been demonstrated in the salivary gland microenvironment of patients with Sjögren's syndrome (SS). How these cytokines may be contributing to the pathogenesis of the disease is not well understood. This study examined the role of IFN-gamma +/- TNF-alpha on cellular death in a cultured human salivary gland cell line (HSG). Cells treated long-term with IFN-gamma +/- TNF-alpha demonstrate a profound antiproliferative effect with a decrease in cell number to below that initially plated. Treatment of HSG cells with TNF-alpha alone did not have any significant effects on growth but did increase the expression of the IFN-gamma receptor. Cells labelled with propidium iodide and anti-digoxigenin dUTP/dATP were examined by flow cytometry to determine the percentage of cells exhibiting low DNA content and DNA strand breaks. The percentage of cells exhibiting subdiploid DNA and DNA strand breaks increased with increased time of exposure to the cytokines. The maximum percentage of cells exhibiting DNA degradation at 12 days was 58% for cells treated with IFN-gamma + TNF-alpha, 31% for IFN-gamma treated cells, and < 5% for TNF-alpha-treated and untreated cells. The cells with subdiploid ( < 2n) DNA were subsequently demonstrated to represent two populations, both with evidence of increased DNA strand breaks but with differing light scatter characteristics. One population had features of cells undergoing necrosis, whereas the second population exhibited features of apoptosis. These findings were confirmed by transmission electron microscopy. Cells not exposed to cytokines did not exhibit significant evidence of either death process. We conclude that long-term exposure of a human salivary gland epithelial cell line to IFN-gamma +/- TNF-alpha leads to increased DNA degradation and subsequent cell death. This suggests a potential SS disease mechanism and implicates the role of the epithelial cell in this disease as an important area for future study.
Subject(s)
Interferon-gamma/pharmacology , Salivary Glands/cytology , Cell Count , Cell Death/drug effects , Cell Death/immunology , Cell Line/cytology , Cell Line/drug effects , Cell Line/ultrastructure , DNA/analysis , DNA Damage/drug effects , DNA Damage/immunology , Electrophoresis , Epithelial Cells , Epithelium/drug effects , Epithelium/ultrastructure , Flow Cytometry , Humans , Microscopy, Electron , Receptors, Interferon/analysis , Receptors, Interferon/physiology , Tumor Necrosis Factor-alpha/pharmacology , Up-Regulation/physiologyABSTRACT
Interferon-gamma (IFN-gamma), in the presence of tumor necrosis factor-alpha (TNF-alpha), decreases proliferation of a human salivary gland ductal cell line, HSG (Wu, A., R. Kurrasch, J. Katz, P. Fox, B. Baum, and J. Atkinson. J. Cell. Physiol. 161:217-226, 1994). We examined the possible effects of these cytokines (1,000 U/ml IFN-gamma +/- 20 U/ml TNF-alpha for 7 days) on Ca2+ mobilization in HSG cells. In HSG cells, fetal bovine serum (10%) or carbachol (100 microM) stimulated rapid increases in cytosolic Ca2+ concentration ([Ca2+]i), apparently mobilized from different thapsigargin-sensitive intracellular Ca2+ stores. Serum induced a proliferative effect on HSG cells, which was suppressed (> 90%) by treatment with IFN-gamma +/- TNF-alpha, but not with TNF-alpha alone. Serum-, carbachol-, and thapsigargin-stimulated [Ca2+]i elevations were reduced by 90, 60, and > 65%, respectively, in cells treated with IFN-gamma +/- TNF-alpha and 30, 45, and 45%, respectively, in cells treated with TNF-alpha. Removal of the cytokines from the growth medium induced recovery of both cell proliferation and Ca2+ mobilization responses within 7 days. Treatment of HSG cells with thapsigargin (0.02-2 nM) induced a dose-dependent decrease in cell proliferation. Additionally, acute treatment (< 10 min) of cells with IFN-gamma did not affect [Ca2+]i or alter carbachol-, thapsigargin-, or serum-induced changes in [Ca2+]i. These data demonstrate that prolonged treatment of HSG cells with IFN-gamma +/- TNF-alpha leads to a persistent depletion of intracellular Ca2+ stores. We suggest that this may have a role in cell growth.
Subject(s)
Calcium/antagonists & inhibitors , Interferon-gamma/pharmacology , Salivary Glands/drug effects , Salivary Glands/metabolism , Blood Physiological Phenomena , Calcium-Transporting ATPases/antagonists & inhibitors , Carbachol/pharmacology , Cell Division/drug effects , Cell Line , Humans , Salivary Glands/cytology , Thapsigargin/pharmacology , Time Factors , Tumor Necrosis Factor-alpha/pharmacologyABSTRACT
BACKGROUND: Previous morphological studies have shown that both the human parotid and submandibular glands display age-related reductions in the proportion of fluid secreting acinar cells. In contrast, short-term functional studies of fluid secretion do not show such a consistent disparity among different-aged persons. This study compared the ability of a population of healthy young and old individuals to secret saliva from the parotid and submandibular glands for an extended period of time under conditions of intense gustatory stimulation. METHODS: Saliva was collected over 30 minutes from 30 healthy, unmedicated individuals using 10% citric acid as a gustatory stimulus. Of the 30 subjects, 15 were young (age range 27-40 years) and 15 were old (age range 60-97 years). Parotid salivary flow rates were determined every minute, and submandibular flow rates were determined at 2, 10, 20, and 30 minutes. A comparison was made between the slopes of the two groups. The slopes were derived from the average salivary flow rate at each time point. RESULTS: There was no difference in the ability of the parotid glands of young and old individuals to secrete saliva. In contrast, the submandibular glands of the elderly individuals did not show the same pattern of secretion when compared to their younger counterparts, who demonstrated increasing flow rates over the test period. CONCLUSIONS: The parotid glands of young and old persons are able to maintain high secretory ability under the stress of intense stimulation for an extended period of time. Conversely, the submandibular gland demonstrates an age-related, statistically significant difference in the pattern of fluid secretion with an intense gustatory stimulus.
Subject(s)
Aging/physiology , Parotid Gland/metabolism , Saliva/metabolism , Submandibular Gland/metabolism , Adult , Aged , Aged, 80 and over , Humans , Middle Aged , Secretory RateABSTRACT
Interferon-gamma (IFN-gamma) is a product of activated T-lymphocytes, and tumor necrosis factor-alpha (TNF-alpha) is a product of both lymphocytes and macrophages. These cell types are often present at sites of tissue damage secondary to chronic infection or autoimmune disease. The purpose of this study was to characterize the effects of TNF-alpha and IFN-gamma on a human submandibular gland epithelial cell line (HSG). IFN-gamma caused a concentration-dependent decrease in HSG cell growth (approximately 70% in 6 days). Conversely, TNF-alpha alone had little effect on the growth of these cells. When these cytokines were added in combination (20 units/ml TNF-alpha and 1,000 units/ml of IFN-gamma), there was a synergistic antiproliferative effect; no apparent cell growth was observed. The cytokine-induced antiproliferative effect was reversible. After the apparent cessation of cell growth for 3-6 days, removal of the cytokines permitted complete growth recovery. Further, cells that recovered and exhibited growth patterns that were similar to control cells remained susceptible to the antiproliferative effects of the cytokines. Flow cytometry revealed that the percentage of cells in G0/G1 with the combination of cytokines was significantly increased by 24 h. The antiproliferative effect of IFN-gamma alone and that of IFN-gamma and TNF-alpha in combination were blocked completely using an antibody to the IFN-gamma receptor. A hypothesized mechanism of tissue damage in autoimmune inflammatory disorders is via up-regulation of cell surface markers such as intercellular adhesion molecule type I (ICAM-1) and histocompatibility antigen HLA-DR which can exacerbate the inflammatory process. Treatment of HSG cells with IFN-gamma, with or without TNF-alpha, resulted in increased levels of ICAM-1 and the acquisition of HLA-DR expression. These aggregate data suggest that IFN-gamma alone can regulate the expression of cell surface markers involved in the inflammatory process as well as cause a potent yet reversible inhibition of HSG cell growth that is modulated by the presence of TNF-alpha.
Subject(s)
Interferon-gamma/pharmacology , Submandibular Gland/cytology , Tumor Necrosis Factor-alpha/pharmacology , Antibodies/immunology , Antibodies/pharmacology , Cell Cycle/drug effects , Cell Division/drug effects , Cells, Cultured , Cytokines/pharmacology , Humans , Receptors, Interferon/immunology , Receptors, Tumor Necrosis Factor/immunology , Time Factors , Interferon gamma ReceptorABSTRACT
Stimulated parotid salivary flow rates were compared in elderly normotensive, hypertensive, and controlled hypertensive African-Americans, the latter group taking hydrochlorothiazide (HCTZ). The normotensive group consisted of 15 healthy unmedicated subjects with systolic blood pressures of less than 150 mm Hg and diastolic pressures less than 90 mm Hg. The hypertensive group consisted of 10 unmedicated subjects with systolic pressures greater than 160 mm Hg and diastolic pressures greater than 100 mm Hg. The controlled hypertensive group consisted of 20 subjects taking HCTZ (50 mg, daily) with controlled blood pressures similar to the normotensive control group. Stimulated parotid salivary samples were collected from each subject. A 2% citrate solution applied to the dorsum of the tongue was used for stimulation. The results showed no significant differences in stimulated parotid flow rates between normotensive and uncontrolled hypertensive subjects. However, the medicated, controlled hypertensive subjects had a significant reduction of stimulated parotid salivary flow rates compared to both the normotensive and hypertensive groups.
Subject(s)
Hydrochlorothiazide/adverse effects , Hypertension/physiopathology , Parotid Gland/metabolism , Saliva/metabolism , Salivation/drug effects , Aged , Analysis of Variance , Baltimore , Black People , Case-Control Studies , Citrates/pharmacology , Citric Acid , Electrolytes/analysis , Female , Humans , Hypertension/drug therapy , Immunoglobulin A, Secretory , Male , Middle Aged , Parotid Gland/drug effects , Saliva/chemistry , Salivary Proteins and Peptides/analysis , Secretory Rate/drug effects , Stimulation, ChemicalABSTRACT
Although hypertension is a prevalent condition among the elderly, little is known with respect to the influence of hypertension on oral health and function. Therefore a study was conducted that compared stimulated parotid salivary flow rates in elderly persons (65 years and older) from two diverse populations who are normotensive, mild, and severe hypertensive. The normotensive group consisted of 45 healthy subjects with systolic blood pressures of less than 140 mm Hg and diastolic pressures less than 90 mm Hg. The mildly hypertensive group consisted of 14 otherwise healthy subjects with either systolic pressures greater than 140 mm Hg or diastolic pressures greater than 90 mm Hg. The severely hypertensive group consisted of 10 otherwise healthy subjects with either systolic pressures greater than 180 mm Hg and/or diastolic pressures greater than 100 mm Hg. All three groups were not taking any prescription or nonprescription medications. Samples of 2% citrate-stimulated parotid saliva were collected from each subject. The results showed no significant differences in stimulated parotid flow between normotensive, mildly hypertensive, and severely hypertensive subjects. These results suggest that hypertension per se has no influence on stimulated parotid salivary gland flow rates in otherwise healthy, elderly unmedicated white and African-American persons.
Subject(s)
Blood Pressure , Hypertension/physiopathology , Parotid Gland/physiopathology , Saliva/metabolism , Aged , Aging/physiology , Analysis of Variance , Baltimore , Black People , Citrates/pharmacology , Citric Acid , Female , Humans , Hypertension/ethnology , Male , Parotid Gland/physiology , Secretory Rate , Stimulation, Chemical , White PeopleABSTRACT
This article reviews the diagnostic criteria for Sjörgen's syndrome (SS), as well as some of the more common associated signs and symptoms. Dermatological manifestations associated with SS will be evaluated. Finally, general treatment strategies will be discussed.