ABSTRACT
Immunological and angiogenetic factors enhance the implantation of endometrial cells in the peritoneal cavity. The aim of this work was to determine the role of the CXCL12-CXCR4 axis in the attraction and the peritoneal implantation of endometriotic stromal cells in deep infiltrating endometriosis (DIE). Biopsies of DIE nodules were obtained from 14 patients undergoing surgical treatment for DIE with low rectal involvement and from 12 patients without macroscopic endometriosis undergoing laparoscopy. CXCR4 expression was evaluated by Western blot analysis and flow cytometry in eutopic endometrial cells and DIE stromal cells in primary cultures derived from the biopsies. CXCL12-induced migration of DIE eutopic endometrial stromal cells was evaluated by transwell migration. CXCL12 was assayed in peritoneal fluids by ELISA. CXCR4 expression was higher in eutopic endometrial stromal cells than in control endometrial cells (p<0.05) and in DIE stromal cells (p<0.05). Eutopic endometrial stromal cells were more attracted by CXCL12 than control cells (p<0.01). CXCL12 was higher in DIE peritoneal fluids than in controls (p<0.05). CXCR4 was down-regulated in deep infiltrating endometriotic stromal cells. The CXCL12-CXCR4 axis plays a role in the attraction of eutopic endometrial cells into the peritoneal cavity, and the down-regulation of CXCR4 in resident endometriotic cells could cause their arrest in situ.
Subject(s)
Chemokine CXCL12/immunology , Endometriosis/pathology , Endometrium/cytology , Receptors, CXCR4/immunology , Ascitic Fluid/cytology , Cell Movement , Cells, Cultured , Chemokine CXCL12/biosynthesis , Endometriosis/immunology , Endometrium/physiology , Female , Humans , Inflammation/immunology , RNA, Messenger/biosynthesis , Receptors, CXCR4/biosynthesis , Receptors, CXCR4/genetics , Signal Transduction/geneticsABSTRACT
Organotellurides are newly described redox-catalyst molecules with original pro-oxidative properties. We have investigated the in vitro and in vivo antitumoral effects of the organotelluride catalyst LAB027 in a mouse model of colon cancer and determined its profile of toxicity in vivo. LAB027 induced an overproduction of H(2)O(2) by both human HT29 and murine CT26 colon cancer cell lines in vitro. This oxidative stress was associated with a decrease in proliferation and survival rates of the two cell lines. LAB027 triggered a caspase-independent, ROS-mediated cell death by necrosis associated with mitochondrial damages and autophagy. LAB027 also synergized with the cytotoxic drug oxaliplatin to augment its cytostatic and cytotoxic effects on colon cancer cell lines but not on normal fibroblasts. The opposite effects of LAB027 on tumor and on non-transformed cells were linked to differences in the modulation of reduced glutathione metabolism between the two types of cells. In mice grafted with CT26 tumor cells, LAB027 alone decreased tumor growth compared with untreated mice, and synergized with oxaliplatin to further decrease tumor development compared with mice treated with oxaliplatin alone. LAB027 an organotelluride catalyst compound synergized with oxaliplatin to prevent both in vitro and in vivo colon cancer cell proliferation while decreasing the in vivo toxicity of oxaliplatin. No in vivo adverse effect of LAB027 was observed in this model.
Subject(s)
Antineoplastic Agents/therapeutic use , Colonic Neoplasms/drug therapy , Naphthoquinones/therapeutic use , Organometallic Compounds/therapeutic use , Animals , Antineoplastic Agents/toxicity , Apoptosis , Caspases/metabolism , Catalysis , Cell Line, Tumor , Cell Proliferation/drug effects , Colonic Neoplasms/metabolism , Disease Models, Animal , Glutathione/metabolism , Humans , Hydrogen Peroxide/metabolism , Mice , Naphthoquinones/toxicity , Organometallic Compounds/toxicity , Organoplatinum Compounds/toxicity , Oxaliplatin , Oxidation-Reduction , Oxidative Stress , Tellurium/chemistry , Transplantation, HeterologousSubject(s)
Immunosuppressive Agents/administration & dosage , Lupus Erythematosus, Systemic/drug therapy , Mycophenolic Acid/analogs & derivatives , Area Under Curve , Humans , Immunosuppressive Agents/blood , Immunosuppressive Agents/pharmacokinetics , Mycophenolic Acid/administration & dosage , Mycophenolic Acid/blood , Mycophenolic Acid/pharmacokineticsABSTRACT
Ro52 antigen has recently been identified as TRIM21 protein, but the clinical significance of anti-Ro52/TRIM21 antibodies remains controversial. The aim of this multicentric study was to investigate the significance of anti-Ro52 antibodies without anti-SSA/Ro60 antibodies in various connective diseases. Sera were selected by each laboratory using its own method (ELISA, immunodot or Luminex technology), and then performed with ANA Screen BioPlex™ reagent (BIO-RAD). Among the 247 screened sera, 155/247 (63%) were confirmed as anti-Ro52 positive and anti-SSA/Ro60 negative. These sera were analyzed for the detection of other antibodies in relation with clinical settings. Isolated anti-Ro52 antibodies were detected in 89/155 (57%) sera. For the remaining sera (66/155), the main antibodies associations were Sm/SmRNP or Chromatin (n=38; 57%), Jo1 (n=17; 26%) and CenpB (n=9; 14%). Clinical data from the 155 patients showed high prevalence in autoimmune diseases (73%) including myositis or dermatomyositis (n=30), lupus (n=23); Sjögren and/or sicca syndrome (n=27); CREST or Systemic sclerosis (n=11) and autoimmune hepatitis (n=11). We found that pulmonary manifestations were often associated with the presence of anti-Ro52 antibodies (n=34, 22%), in addition with anti-tRNA synthetases, anti-SRP or anti-Ku antibodies (18/34) or isolated in half of cases (16/34). Separate detection of anti-Ro52 antibodies might be useful in related antisynthetase syndrome diagnosis. The presence of anti-Ro52 antibodies should probably precede development of autoimmune disease and must induce sequential follow-up of positive patients, particularly in interstitial lung disease progression.
Subject(s)
Antibodies/blood , Autoimmune Diseases/blood , Lung Diseases/blood , Adolescent , Adult , Aged , Aged, 80 and over , Antibodies/immunology , Autoantibodies/blood , Autoantibodies/immunology , Autoimmune Diseases/immunology , Female , Humans , Lung Diseases/immunology , Male , Middle Aged , Ribonucleoproteins/immunology , Young AdultABSTRACT
Oxidative stress plays a role in the regulation of cancer cell metastasis which involves cell invasion and adhesion that could be supported by ADAM proteins through the activities of their metalloprotease and disintegrin domains. We hypothesized that oxidative stress could act through the induction of ADAM9 protein in some cancer cells. Indeed, Western blot analysis for ADAM9 performed on A549 cells exposed to H(2) O(2) reveals a dose-dependent induction of two proteins (80 and 68 kDa) correlated with a sharp increase of the ADAM protease activity measured in supernatant while the activity measured on the cell layer was slightly affected. The 80kDa protein corresponds to the mature form of ADAM9. Immunoprecipitation analysis performed on concentrated supernatants revealed that the 68 kDa protein is a secreted form of ADAM9. When exposed to H(2) O(2) , A549 cells cocultured with confluent endothelial vascular cells resulted in a 5.5 fold (p < 0.001) increase in the number of adherent cells. Similarly, matrigel assay revealed a 3.25 fold (p < 0.01) increase in the number of invasive cells. The suppression of ADAM9 expression by specific small interfering RNA reduced oxidative stress-induced invasiveness and adhesiveness. These functions could be mediated by an interaction between ADAM9 and ß1 integrin because each of them were inhibited when the experiment is performed in presence of mAbs targeting ADAM9 ectodomain or ß1-integrin. These results emphasize the importance of oxidative stress in the regulation of cancer cell metastasis and suggest that ADAM9 and its secreted isoform can be important determinants in the ability of cancer cells to disseminate.
Subject(s)
ADAM Proteins/metabolism , Cell Membrane/metabolism , Lung Neoplasms/pathology , Membrane Proteins/metabolism , Oxidants/pharmacology , Oxidative Stress , ADAM Proteins/antagonists & inhibitors , ADAM Proteins/genetics , ADAM10 Protein , ADAM17 Protein , Amyloid Precursor Protein Secretases/genetics , Amyloid Precursor Protein Secretases/metabolism , Biocompatible Materials , Blotting, Western , Cells, Cultured , Collagen/metabolism , Drug Combinations , Endothelium, Vascular/cytology , Endothelium, Vascular/metabolism , Humans , Hydrogen Peroxide/pharmacology , Integrin beta1/genetics , Integrin beta1/metabolism , Laminin/metabolism , Lung Neoplasms/metabolism , Membrane Proteins/antagonists & inhibitors , Membrane Proteins/genetics , Metalloproteases/metabolism , Protein Isoforms , Proteoglycans/metabolism , RNA, Small Interfering/geneticsABSTRACT
Lung fibrosis is considered a severe manifestation of microscopic polyangiitis (MPA). Antimyeloperoxidase (anti-MPO) antibodies in MPA patients' sera can activate MPO and lead to the production of reactive oxygen species (ROS). While high levels of ROS are cytotoxic, low levels can induce fibroblast proliferation. Therefore, we hypothesised that the oxidative stress induced by anti-MPO antibodies could contribute to lung fibrosis. 24 MPA patients (45 sera) were enrolled in the study, including nine patients (22 sera) with lung fibrosis. Serum advanced oxidation protein products (AOPP), MPO-induced hypochlorous acid (HOCl) and serum-induced fibroblast proliferation were assayed. AOPP levels, MPO-induced HOCl production and serum-induced fibroblast proliferation were higher in patients than in healthy controls (p<0.0001, p=0.0001 and p=0.0005, respectively). Increased HOCl production was associated with active disease (p=0.002). Serum AOPP levels and serum-induced fibroblast proliferation were higher in patients with active MPA and lung fibrosis (p<0.0001). A significant linear relationship between fibroblast proliferation, AOPP levels and HOCl production was observed only in patients with lung fibrosis. Oxidative stress, in particular the production of HOCl through the interaction of MPO with anti-MPO antibodies, could trigger the fibrotic process observed in MPA.
Subject(s)
Antibodies/immunology , Microscopic Polyangiitis/immunology , Oxidative Stress , Peroxidase/immunology , Peroxidase/metabolism , Pulmonary Fibrosis/immunology , Adult , Aged , Blood Proteins/metabolism , Cell Proliferation , Female , Fibroblasts/metabolism , Humans , Hypochlorous Acid/blood , Male , Microscopic Polyangiitis/enzymology , Middle Aged , Oxidation-Reduction , Pulmonary Fibrosis/enzymology , Severity of Illness IndexABSTRACT
The IgM antibodies from rheumatoid arthritis (RA) patients' sera were screened for peptide hydrolyzing activity. Recovery of structurally intact IgM antibodies (Abs), in a single step, was achieved using a weak anion-exchange methacrylate monolith disk. The IgM Abs from patients' sera hydrolyzed the Pro-Phe-Arg-4-methyl-coumaryl-7-amide (PFR-MCA) substrate appreciably compared to the healthy donors. The apparent K(m) values of IgM Abs from patients' sera were between 0.4 and 0.7 mM. Furthermore, IgM Abs displayed 5 to 10-folds greater proteolysis activity than IgG Abs, recovered from the same pathological serum. The proteolysis activity, as a function, was found to be independent of IgM-RF titer value. Affinity labeling approach targeted at the catalytic site histidine was studied, using a specific irreversible inhibitor, N-α-tosyl-L-lysine chloromethyl ketone (TLCK). Despite modification of catalytic His, observation of serine protease like activity suggest presence of an atypical catalytic framework in a few pathological IgM Abs.
Subject(s)
Antibodies/chemistry , Antibodies/metabolism , Arthritis, Rheumatoid/blood , Immunoglobulin M/chemistry , Immunoglobulin M/metabolism , Protein Processing, Post-Translational , Affinity Labels/pharmacology , Antibodies/blood , Antibodies/isolation & purification , Arthritis, Rheumatoid/immunology , Catalysis , Catalytic Domain/drug effects , Chromatography, Affinity/methods , Enzyme-Linked Immunosorbent Assay/methods , Histidine/metabolism , Humans , Immunoglobulin G/metabolism , Immunoglobulin M/blood , Immunoglobulin M/isolation & purificationABSTRACT
The efficacy of drugs acting within lymphocytes depends on their intracellular concentrations, which could be modulated by membrane efflux transporters including P-glycoprotein (P-gp), encoded by the MDR1 gene. In particular, P-gp induction may compromise the efficacy of its substrates. Rifampicin and phenobarbital have been shown to induce P-gp in hepatic and intestinal cells through the activation of the nuclear receptors PXR and CAR. However, controversial data exist in human lymphocytes. We investigated the effect of these drugs on P-gp activity and expression in lymphocytes in vitro and ex vivo. CCRF-CEM cells and peripheral blood mononuclear cells (PBMCs) from healthy volunteers were incubated in the presence of rifampicin, phenobarbital, or without any drug. P-gp activity was measured by flow cytometry using DiOC(6) efflux. MDR1, PXR and CAR mRNA expression were measured by quantitative RT-PCR. Neither P-gp activity nor MDR1 mRNA expression were modified by rifampicin or phenobarbital both in CCRF-CEM cells and PBMCs. Moreover, P-gp protein expression at the membrane was neither detectable nor induced. The very weak PXR and CAR mRNA expression levels in these cells could partly explain these results. Therefore, P-gp induction by rifampicin and phenobarbital may play a negligible role in drug interactions occurring within lymphocytes.
Subject(s)
ATP Binding Cassette Transporter, Subfamily B, Member 1/metabolism , Lymphocytes/drug effects , Phenobarbital/pharmacology , Rifampin/pharmacology , ATP Binding Cassette Transporter, Subfamily B , ATP Binding Cassette Transporter, Subfamily B, Member 1/genetics , Constitutive Androstane Receptor , Drug Interactions , Flow Cytometry , HL-60 Cells , Humans , Lymphocytes/metabolism , Phenobarbital/metabolism , Pregnane X Receptor , RNA, Messenger/metabolism , Receptors, Cytoplasmic and Nuclear/metabolism , Receptors, Steroid/metabolism , Rifampin/metabolism , Time Factors , Transfection , Up-RegulationABSTRACT
Anti-cyclic citrullinated peptides (anti-CCP) are highly characteristics of rheumatoid arthritis (RA). Since 2006, anti-CCP assays have been included in both French and European recommendations. We have evaluated the analytical and clinical performances of the anti-CCP assay on the Elecsys analyzer (Roche Diagnostics). Two plasma pools (target values: 17.2 and 363.0 U/mL) and two quality controls (target values: 24.5 and 157.0 U/mL) were tested; we also analyzed three hundred plasma samples from healthy subjects (n = 86) and diseased patients (presenting with RA, non rheumatoid disorders, or undifferentiated arthritis: n = 214). Analytical performances (intra- and inter-assay precisions) and clinical performances (ROC analysis and method comparison) were evaluated. Elecsys assay was compared to Immunoscan RA(R) assay using contingency tables. Intra- and inter-assay precisions showed coefficients of variation less than 5%. ROC analysis showed an area under the curve at 0.886. Considering the value of 17 U/mL as the optimal cut-off, we found sensibility and specificity at 75% and 95%, respectively. Comparison of the Elecsys anti-CCP assay with the Immunoscan RA(R) assay showed an overall agreement of 98,3%. We conclude that the the Elecsys anti-CCP assay displayed a high precision and clinical performances comparable to that of the efficient anti-CCP assay Immunoscan RA(R).
Subject(s)
Arthritis, Rheumatoid/diagnosis , Autoanalysis/methods , Peptides, Cyclic/immunology , Arthritis, Rheumatoid/blood , Autoantibodies/blood , Equipment Design , Humans , Peptides, Cyclic/blood , ROC Curve , Reference Values , Reproducibility of ResultsABSTRACT
OBJECTIVE: Contradictory results have been reported regarding vasculogenesis in systemic sclerosis (SSc). Our aim was to investigate bone marrow-derived circulating endothelial precursors (EPCs) and activated circulating endothelial cells (CECs) in SSc patients. METHODS: Peripheral blood from consecutive patients with SSc hospitalised for systemic follow-up was analysed and compared with blood from patients with active refractory rheumatoid arthritis (RA) and osteoarthritis (OA). EPCs were quantified by cell sorting and flow cytometry and were identified as circulating CD34+CD133+ cells. Activated CECs were defined as CD105+CD62+CD105+CD102+CD105+CD106+ cells. RESULTS: Patients with SSc had higher putative EPC levels than OA patients, but lower levels than RA patients. In SSc patients, EPC levels increased with European disease activity score. Activated CEC levels were high in SSc patients and RA patients, but not correlated with EPC levels. CONCLUSION: These results together and previous data suggest that EPCs may be recruited during active vascular disease but that the sustained ischaemic conditions of SSc may eventually lead to EPCs depletion.
Subject(s)
Blood Cell Count , Endothelial Cells/cytology , Scleroderma, Systemic/blood , Stem Cells/cytology , AC133 Antigen , Adult , Aged , Antigens, CD , Antigens, CD34 , Arthritis, Rheumatoid/blood , Cell Adhesion Molecules , Cell Differentiation , E-Selectin , Endoglin , Female , Flow Cytometry , Glycoproteins , Humans , Male , Middle Aged , Osteoarthritis/blood , Peptides , Receptors, Cell Surface , Vascular Cell Adhesion Molecule-1ABSTRACT
OBJECTIVES: To investigate the role of reactive oxygen species (ROS) in the development of the various patterns of systemic sclerosis (SSc) and the mechanisms of ROS production by endothelial cells and fibroblasts. METHODS: Production of hydrogen peroxide (H(2)O(2)), nitric oxide (NO) and cellular proliferation were determined following incubation of endothelial cells and fibroblasts with 56 SSc and 30 healthy sera. Correlations were established between those markers, the type and the severity of the clinical involvements, and the response to treatment. The factors leading to ROS production were determined. RESULTS: H(2)O(2) production by endothelial cells and fibroblasts was higher after incubation with SSc sera than with normal sera (p<0.001) and with sera from SSc patients with severe complications than sera from other patients (p<0.05). Sera from patients with lung fibrosis triggered the proliferation of fibroblasts more than other SSc sera (p<0.001), whereas sera from patients with vascular complications exerted no proliferative effect on fibroblasts, but inhibited endothelial cell growth (p<0.05) and induced NO overproduction (p<0.05). Bosentan reduced NO release by 32%, whereas N-acetylcystein potentiated 5-fluorouracil (5FU) to inhibit fibroblast proliferation by 78%. Those serum-mediated effects did not involve antibodies but advanced oxidation protein products that selectively triggered cells to produce H(2)O(2) or NO. CONCLUSIONS: SSc sera induce the production of different types of ROS that selectively activate endothelial cells or fibroblasts, leading to vascular or fibrotic complications. Assaying serum-induced ROS production allows clinical activity of the disease to be followed and appropriate treatments to be selected.
Subject(s)
Reactive Oxygen Species/metabolism , Scleroderma, Systemic/metabolism , Skin/metabolism , Adult , Aged , Autoantibodies/blood , Biomarkers/blood , Cell Line , Cell Proliferation , Cells, Cultured , Endothelial Cells/immunology , Endothelial Cells/metabolism , Endothelial Cells/pathology , Female , Fibroblasts/immunology , Fibroblasts/metabolism , Fibroblasts/pathology , Humans , Hydrogen Peroxide/blood , Immunosuppressive Agents/therapeutic use , Longitudinal Studies , Male , Middle Aged , Nitric Oxide/blood , Prognosis , Scleroderma, Systemic/drug therapy , Scleroderma, Systemic/pathology , Skin/pathologyABSTRACT
The aim of the present study was to investigate the presence of anti-fibroblast antibodies in patients with idiopathic or scleroderma-associated pulmonary arterial hypertension (PAH) and healthy controls. PAH was documented by right-heart catheterisation (mean pulmonary artery pressure at rest >25 mmHg). Serum immunoglobulin (Ig)G and IgM reactivities of patients with idiopathic PAH (n = 35), scleroderma-associated PAH (n = 10), diffuse (n = 10) or limited cutaneous (n = 10) scleroderma without PAH and age- and sex-matched healthy individuals (n = 65) were analysed by cell-based ELISA and immunoblotting on normal human fibroblasts. As assessed by ELISA, 14 out of 35 (40%) patients with idiopathic PAH and three out of 10 (30%) patients with scleroderma-associated PAH expressed anti-fibroblast IgG antibodies. IgG from all individuals bound to one major 40-kDa protein band. IgG from patients with idiopathic PAH bound to two 25- and 60-kDa bands with a higher intensity than IgG from other individuals. In conclusion, immunoglobulin G anti-fibroblast antibodies are present in the serum of patients with pulmonary arterial hypertension. Immunoglobulin G from patients with idiopathic pulmonary arterial hypertension or scleroderma-associated pulmonary arterial hypertension express distinct reactivity profiles with fibroblasts antigens, suggesting distinct target antigens.
Subject(s)
Fibroblasts/immunology , Hypertension, Pulmonary/immunology , Immunoglobulin G/blood , Immunoglobulin M/blood , Scleroderma, Systemic/immunology , Adult , Cell Line , Enzyme-Linked Immunosorbent Assay , Female , Humans , Hypertension, Pulmonary/etiology , Immunoblotting , Male , Middle Aged , Scleroderma, Systemic/complicationsABSTRACT
BACKGROUND: It has previously been shown that IgG antibodies from patients with limited cutaneous systemic sclerosis (SSc) bind to specific microvascular endothelial cell antigens. Since patients with limited cutaneous SSc are prone to develop pulmonary arterial hypertension (PAH), and since endothelial cell activation is involved in the pathogenesis of idiopathic PAH (IPAH), a study was undertaken to examine the presence of anti-endothelial cell antibodies in patients with idiopathic or SSc associated PAH. METHODS: PAH was confirmed by right heart catheterisation (mean pulmonary artery pressure at rest >25 mm Hg). Serum IgG and IgM reactivities were analysed by immunoblotting on human macrovascular and microvascular lung and dermal endothelial cells from patients with IPAH (n = 35), patients with PAH associated with SSc (n = 10), patients with diffuse (n = 10) or limited cutaneous (n = 10) SSc without PAH, and 65 age and sex matched healthy individuals. RESULTS: IgG antibodies from patients with IPAH bound to a 36 kDa band in macrovascular endothelial cell extracts with a higher intensity than IgG from other patient groups and controls. IgG antibodies from patients with IPAH bound more strongly to a 58 kDa band in microvascular dermal endothelial cells and to a 53 kDa band in microvascular lung endothelial cells than IgG antibodies from other patients and controls. IgG antibodies from patients with limited cutaneous SSc with or without PAH, but not from other groups or from healthy controls, bound to two major bands (75 kDa and 85 kDa) in microvascular endothelial cells. CONCLUSION: IgG antibodies from patients with idiopathic or SSc associated PAH express distinct reactivity profiles with macrovascular and microvascular endothelial cell antigens.
Subject(s)
Autoantibodies/immunology , Hypertension, Pulmonary/immunology , Scleroderma, Systemic/immunology , Adult , Antigens/immunology , Cells, Cultured , Endothelial Cells/immunology , Female , Humans , Immunoglobulin G/blood , Immunoglobulin M/blood , Male , Umbilical Veins/immunologyABSTRACT
OBJECTIVE: To investigate the prevalence and clinical significance of antiphospholipid antibodies in patients with systemic sclerosis (SSc). METHODS: Autoantibodies against cardiolipin (aCL) and beta2-glycoprotein 1 (beta2-GPI) were detected by enzyme-linked immunoabsorbent assays (ELISAs) in successively hospitalised SSc patients admitted during a 24-month period. These patients were compared to patients with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA). RESULTS: 108 SSc patients were included: 61 had limited cutaneous SSc, 47 had the diffuse sub-type, 16 had primitive pulmonary arterial hypertension (PAH) and 34 had digital ulcerations. The control groups consisted of 37 RA and 38 SLE patients. The prevalence of aCL positivity was lower in SSc patients vs SLE patients (14 vs 47%; p < 0.001), lower in RA patients vs SLE patients (19 vs 47%; p < 0.001), and not different in SSc vs RA patients (14 vs 19%; NS). The mean aCL titer was also lower in SSc vs SLE patients (8+/-10 vs 15+/-20; p < 0.001). In SSc patients, positivity for aCL was associated with PAH (p = 0.009) and the aCL titer correlated with that of the von Willebrand antigen factor (r= 0.23; p = 0.045). The prevalence of anti beta2-GPI positive patients (IgG and/or IgM) was 5% in the SSc group, 18% in the SLE group and 5% in the RA group (SLE vs SSc and SLE vs RA; p = 0.005). CONCLUSION: We found that the prevalence of antiphospholipid antibodies in SSc patients was low. However, aCL antibodies were associated with PAH and endothelial injury.
Subject(s)
Antibodies, Anticardiolipin/analysis , Endothelium, Vascular , Hypertension, Pulmonary/immunology , Scleroderma, Diffuse/immunology , Scleroderma, Limited/immunology , Adult , Aged , Aged, 80 and over , Arthritis, Rheumatoid/complications , Arthritis, Rheumatoid/diagnosis , Arthritis, Rheumatoid/immunology , Endothelium, Vascular/pathology , Enzyme-Linked Immunosorbent Assay , Female , Glycoproteins/immunology , Humans , Hypertension, Pulmonary/complications , Hypertension, Pulmonary/physiopathology , Lupus Erythematosus, Systemic/complications , Lupus Erythematosus, Systemic/diagnosis , Lupus Erythematosus, Systemic/immunology , Male , Middle Aged , Pulmonary Artery/pathology , Pulmonary Artery/physiopathology , Scleroderma, Diffuse/complications , Scleroderma, Diffuse/diagnosis , Scleroderma, Limited/complications , Scleroderma, Limited/diagnosis , beta 2-Glycoprotein IABSTRACT
OBJECTIVE: To study whether interleukin-8 (IL-8) mRNA in vaginal secretions is associated with congenital infection and preterm delivery in the case of preterm labor with intact membranes. METHODS: This prospective clinical study in a tertiary referral center included 280 patients who gave birth to 360 infants from 1997 through 1999. IL-8 mRNA in vaginal secretions was determined with reverse transcriptase polymerase chain reaction. Logistic regression was used to examine the association between vaginal IL-8 mRNA and congenital infection independently of the time of birth. Main outcome measures were congenital infection and delivery before 37 and 33 weeks' gestation. RESULTS: A total of 100 women (100/280 (35.7%)) gave birth before 37 weeks. A total of 54 children (54/360 (15%)) had congenital infection. IL-8 mRNA in vaginal secretions was associated with delivery within 14 days of the sampling (24 (15.6%) vs. 7 (5.6%) p < 0.01), but not with delivery within 48 h, 7 days (p = 0.07) or before 37 or 33 weeks. There were more congenital infections in the group with detectable IL-8 mRNA (37 (19.3%) than in the negative group (17 (10.1%); p < 0.05). IL-8 mRNA was associated with congenital infection independently of the time of birth (OR: 2.6 (1.3-5.1)). This test had a sensitivity for predicting neonatal infection of 69%. Its specificity was 49%, its positive predictive value 19%, and its negative predictive value 90%. CONCLUSION: IL-8 mRNA could be a prenatal noninvasive vaginal marker of congenital infection.
Subject(s)
Cervix Uteri/metabolism , Interleukin-8/genetics , Obstetric Labor, Premature , Pregnancy Complications, Infectious/diagnosis , Prenatal Diagnosis , Adult , Biomarkers , DNA Primers , Female , Humans , Infant, Newborn , Predictive Value of Tests , Pregnancy , Pregnancy Outcome , Pregnancy Trimester, Third , Prospective Studies , RNA, Messenger/analysis , Reverse Transcriptase Polymerase Chain Reaction , Sensitivity and SpecificityABSTRACT
OBJECTIVES: To investigate autoantibody induction in rheumatoid arthritis (RA) patients treated with infliximab. METHODS: We included 59 refractory RA patients treated with infliximab in combination with low-dose prednisone and methotrexate or leflunomide. We tested the sera of the patients for antinuclear antibodies (ANA), rheumatoid factor (RF), anti double-stranded DNA antibodies (anti dsDNA), anti-histone and anti-extractable nuclear antigen antibodies (aENA) at baseline and before infusion at weeks 6 and 30. Infliximab, initiated at a dose of 3 mg/kg, was increased to 5 mg/kg if insufficient improvement was observed after three infusions. RESULTS: At week 6, only the frequency of anti-histone IgM antibody-positive patients had significantly increased (19 vs 42%, p = 0.009). At week 30, the frequency of patients with ANA had increased from 29% to 69% (p < 0.001), that of patients with anti-dsDNA antibodies had increased from 0% to 3% for IgG (NS) and from 0% to 32% for IgM (p < 0.001); the frequency of antihistone IgG detection had increased from 22% to 32% (p = 0.04) and that of IgM detection, from 18% to 79% (p < 0.001). No lupus-like syndrome was observed. RF decreased significantly (87 IU to 52.5 IU, from baseline to week 30; p < 0.001). No significant difference was observed between the 16 non-responders and the responders, in terms of autoantibody status at baseline and changes with infliximab therapy. CONCLUSION: Infliximab therapy lead to the selective and delayed induction of autoantibodies. This induction was not associated with clinical symptoms until week 30 and did not differ between responders and non-responders.
Subject(s)
Antibodies, Monoclonal/administration & dosage , Antirheumatic Agents/administration & dosage , Arthritis, Rheumatoid/drug therapy , Arthritis, Rheumatoid/immunology , Autoantibodies/blood , Adult , Aged , Antibodies, Monoclonal/adverse effects , Antibodies, Monoclonal/immunology , Antirheumatic Agents/adverse effects , Antirheumatic Agents/immunology , Drug Therapy, Combination , Female , Humans , Immunosuppressive Agents/administration & dosage , Infliximab , Isoxazoles/administration & dosage , Leflunomide , Male , Methotrexate/administration & dosage , Middle Aged , Prednisolone/administration & dosage , Prospective StudiesABSTRACT
BACKGROUND: Hyperacute rejection (HAR) of discordant xenografts in the pig-to-human combination can be prevented using tranplants expressing transgenic molecules that inhibit human complement. Hypodermin A (HA), a serine esterase that degrades C3, was tested in the guinea-pig-to-rat and in the pig-to-human combinations. METHODS: Hypodermin A was tested in vitro, ex vivo, and in vivo models of HAR in the guinea-pig-to-rat combination. Hamster ovary cells (CHO) and a line of porcine aortic endothelial cells (PAEC11) were transfected with HA complementary DNA (cDNA). RESULTS: The pattern of degradation of rat and human C3 by HA was different (multiple bands lower than 40 kDa) from the physiologic pattern observed after spontaneous degradation of rat C3 or physiologic activation of human C3. The CH50 activity in serum was significantly lower in rats treated with 3.2 mg HA/kg than in untreated rats (45 +/- 16 U/ml vs. 700 +/- 63 U/ml, P < 0.05). Sera from rats injected with 3.2 mg/kg of HA were less effective in lysing guinea-pig endothelial cells (12 +/- 7%) than normal rat sera (79 +/- 3%; P < 0.001). Ex vivo, guinea-pig hearts perfused by rat serum supplemented with HA survived longer than those perfused by non-treated serum (210 +/- 34 and 154 +/- 71 min, respectively; P < 0.05). In vivo, guinea-pig hearts transplanted into HA treated rats survived longer than in non-treated rats (27 +/- 5 min vs. 13 +/- 4 min; P < 0.001). In the presence of human serum, smaller amounts of C6 and C5b-9 were deposited onto HA-transfected CHO cells than onto control cells. The mHA-PAEC11 cells were significantly more resistant to lysis by human C than control PAEC11 cells. CONCLUSIONS: These data suggest that transgenic HA could be used to prevent hyperacute xenogeneic rejection.
Subject(s)
Complement Inactivator Proteins/pharmacology , Graft Rejection/prevention & control , Heart Transplantation/immunology , Serine Endopeptidases/pharmacology , Transplantation, Heterologous/immunology , Acute Disease , Amino Acid Sequence , Animals , CHO Cells , Cell Survival , Complement C3/metabolism , Complement C6/metabolism , Complement Membrane Attack Complex/metabolism , Cricetinae , Endothelium, Vascular/cytology , Graft Survival/immunology , Humans , Molecular Sequence Data , Rats , Serine Endopeptidases/chemistry , Serine Endopeptidases/isolation & purification , Swine , TransfectionABSTRACT
OBJECTIVE: To determine the usefulness of reagent test strips for screening inflammatory synovial fluid (SF). METHODS: Consecutive patients undergoing diagnostic arthrocentesis, attending the Department of Rheumatology of a large tertiary care hospital were evaluated. All SF specimens obtained were tested using two techniques: (i) white blood cell (WBC) count with the differential according to standard practice (which is considered the gold standard) (an inflammatory SF was defined as a WBC count > or =2000 cells/mm3); and (ii) reagent strips used to test urine (Multistix 8 SG, Bayer Diagnostics) for the presence of leucocytes (a positive test was defined as a strip showing more than a trace for leucocytes). Sensitivity, specificity, predictive values and likelihood ratio (LR) of the reagent strip in diagnosing inflammatory SF were determined. RESULTS: Two hundred and eight samples of SF were tested. The results of using the reagent strip were: sensitivity 76.9% (95% CI, 66.0-85.7%), specificity 86.9% (95% CI, 79.9-92.2%); positive LR, 5.88 (95% CI, 3.71-9.31) and negative LR, 0.27 (95% CI, 0.18-0.40). In 13 of the 19 false negative results, the differential cell count showed a predominance (> or =50%) of lymphocytes. CONCLUSION: This study suggests that, in daily practice, the evaluation of SF by reagent strips could be of use to discriminate between inflammatory and non-inflammatory SF.
Subject(s)
Reagent Strips , Synovial Fluid/cytology , Synovitis/diagnosis , Humans , Leukocyte Count , Leukocytes/pathology , Predictive Value of Tests , Reproducibility of ResultsABSTRACT
BACKGROUND: In multiple sclerosis (MS), case control studies have shown that anticardiolipin antibodies (aCL Ab) are more frequent than in the general population and that aCL Ab positivity may be associated with specific clinical characteristics. OBJECTIVES: To determine whether patients with MS who are positive for aCL Ab have specific characteristics. METHODS: 285 consecutive patients with MS were tested for aCL Ab positivity. Patients also underwent complete autoimmune screening and were systematically evaluated for clinical characteristics and individual or family history of autoimmune disease. RESULTS: aCL Ab positivity was found in 42 patients (15%). The main isotype was aCL IgM (32 patients, 11%). Demographics and clinical characteristics including sex, age at onset, course of the disease, expanded disability status scale score, and progression index were not different between aCL Ab positive and aCL Ab negative patients. Clinical systems involved at onset or during the course of the disease were not different from what is usually observed in MS. aCL Ab positivity was not associated with an increased frequency of autoimmune disease and was not predictive of a family history of autoimmune disease. Patients positive for aCL IgM were more frequently positive for the presence of non-organ specific antibodies (53% v 39%, respectively, p = 0.02). CONCLUSIONS: These results do not support the hypothesis that patients with MS with aCL Ab constitute a subgroup of MS according to demographic clinical and familial characteristics. The greater frequency of other antibodies in aCL Ab positive patients suggests that they only reflect a more general autoimmune activation in MS.
Subject(s)
Antibodies, Anticardiolipin/blood , Multiple Sclerosis/classification , Multiple Sclerosis/immunology , Adult , Age of Onset , Autoimmune Diseases/genetics , Autoimmune Diseases/immunology , Biomarkers/analysis , Demography , Disease Progression , Female , Humans , Immunoglobulin M/blood , Male , Medical History Taking , Middle Aged , Multiple Sclerosis/pathologyABSTRACT
BACKGROUND & AIMS: Acute liver failure (ALF) of viral origin results from massive hepatocyte apoptosis induced by the interaction between Fas expressed on hepatocytes and Fas ligand on activated T lymphocytes. Because Fas-induced apoptosis of hepatocytes involves mitochondrial damages and potential reactive oxygen species (ROS) overproduction, we investigated whether manganese III tetrakis (5,10,15,20 benzoic acid) (MnTBAP), a nonpeptidyl mimic of superoxide dismutase (SOD), can inhibit Fas-induced ALF. METHODS: An agonist anti-Fas monoclonal antibody was used to induce hepatocyte apoptosis in vitro and ALF in vivo. RESULTS: Preventive and curative treatments by MnTBAP significantly increased survival rates and significantly reduced aspartate aminotransferase levels and parenchymal lesions. ROS generation was suggested by those beneficial effects and significant increases in SOD and Gpx activities after anti-Fas injection. Flow cytometry of isolated hepatocytes incubated with anti-Fas monoclonal antibody showed that ROS production was associated with the collapse of transmembrane potential and loss of cardiolipin content. After injection of anti-Fas monoclonal antibody, mitochondrial Bcl-2 was decreased, cytochrome c released, and caspase-3 activated. Mitochondrial alterations and their consequences were abrogated by MnTBAP. CONCLUSIONS: ROS are key executioners in Fas-induced hepatocyte apoptosis. This finding explains why a nonpeptidyl mimic of SOD can cure ALF in a model of viral hepatitis, pointing out the potential interest of this molecule in humans.