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1.
Nutr Metab Cardiovasc Dis ; 24(1): 34-41, 2014 Jan.
Article in English | MEDLINE | ID: mdl-24418377

ABSTRACT

BACKGROUND/AIM: Obesity is associated with changes in adiponectin and pro-inflammatory adipokines. Sodium intake can affect adipokine secretion suggesting a role in cardiovascular dysfunction. We tested if long-term dietary sodium restriction modifies the expression of adiponectin and ameliorates the pro-inflammatory profile of obese, diabetic mice. METHODS/RESULTS: Db/db mice were randomized to high sodium (HS 1.6% Na+, n = 6) or low sodium (LS 0.03% Na+, n = 8) diet for 16 weeks and compared with lean, db/+ mice on HS diet (n = 8). Insulin levels were 50% lower in the db/db mice on LS diet when compared with HS db/db (p < 0.05). LS diet increased cardiac adiponectin mRNA levels in db/db mice by 5-fold when compared with db/db mice on HS diet and by 2-fold when compared with HS lean mice (both p < 0.01). LS diet increased adiponectin in adipose tissue compared with db/db mice on HS diet, achieving levels similar to those of lean mice. MCP-1, IL-6 and TNF-α expression were reduced more than 50% in adipose tissue of db/db mice on LS diet when compared with HS db/db mice (all p < 0.05), to levels observed in the HS lean mice. Further, LS db/db mice had significantly reduced circulating MCP-1 and IL-6 levels when compared with HS db/db mice (both p < 0.01). CONCLUSION: In obese-diabetic mice, long-term LS diet increases adiponectin in heart and adipose tissue and reduces pro-inflammatory factors in adipose tissue and plasma. These additive mechanisms may contribute to the potential cardioprotective benefits of LS diet in obesity-related metabolic disorders.


Subject(s)
Adiponectin/blood , Diabetes Mellitus/diet therapy , Diet, Sodium-Restricted , Sodium, Dietary/administration & dosage , Adipokines/blood , Adipokines/metabolism , Adipose Tissue/metabolism , Animals , Blood Glucose/metabolism , Body Weight , Chemokine CCL2/blood , Diet , Heart/physiology , Insulin/blood , Insulin Resistance/physiology , Interferon-gamma/blood , Interleukin-6/blood , Male , Mice , Mice, Obese , Obesity/blood , Triglycerides/blood , Tumor Necrosis Factor-alpha/blood
2.
Cancer Genomics Proteomics ; 7(4): 217-29, 2010.
Article in English | MEDLINE | ID: mdl-20656987

ABSTRACT

The ubiquitous cytokine transforming growth factor-beta1 (TGF-beta1) is one of the most potent metastatic inducers. Functional interactomic mapping using high-throughput proteomic and genomic data provides valuable insights into the regulation of tumor suppressive and metastatic attributes of TGF-beta1. Polarity changes of the TGF-beta1 interactome at a given time contributes to these contrasting effects. Differential expression profiles of pivotal interactomic nodes contribute to these polarity changes. These insights are of immense value in the development of effective cancer therapeutics. Moreover, TGF-beta1 interactomic nodes are useful in discovering novel cancer biomarkers. This review describes an initial version of the TGF-beta1 interactome in relation to tumor progression and metastasis. Thus, this review embodies an important step towards the mapping of comprehensive and individualized TGF-beta1 interactomes that will assist in the development of personalized cancer therapeutics.


Subject(s)
Neoplasms/therapy , Proteome/analysis , Transforming Growth Factor beta1/metabolism , Animals , Humans , Neoplasm Metastasis/prevention & control , Neoplasms/blood supply , Neoplasms/chemistry , Neoplasms/metabolism , Neovascularization, Pathologic , Signal Transduction
3.
Kidney Int ; 70(10): 1759-68, 2006 Nov.
Article in English | MEDLINE | ID: mdl-17021606

ABSTRACT

Prospective, placebo-controlled clinical trials suggest that estrogen may have adverse effects on the vascular system in women. The goal of this study was to determine if 17beta-estradiol (E2) would have adverse effects on the renovasculature in a rat model of renal injury characterized by low nitric oxide (NO) and high angiotensin II (AngII). We studied female Wistar rats that were sham-operated (sham), ovariectomized (OVX), or ovariectomized and replaced with E2 (OVX/E2). All rats were maintained on a high salt diet and renovascular injury was caused by treating rats with an inhibitor of NO synthase, N(omega)-nitro-L-arginine-methyl-ester (L-NAME), for 14 days, plus AngII on days 11 through 14. L-NAME/AngII treatment, as compared to placebo, caused proteinuria, glomerular injury, and fibrinoid necrosis of renal arterioles in sham-operated rats. Ovariectomy reduced L-NAME/AngII-induced renal damage, whereas E2 treatment increased L-NAME/AngII-induced damage in OVX rats. In rats treated with L-NAME/AngII, levels of AngII type 1 receptor (AT(1)R) protein were higher in the renal cortex of sham and OVX/E2 rats than in OVX rats. AT(1)R protein correlated with renal injury. E2 treatment also increased expression of AT(1)R mRNA. Thus, under conditions of low NO and high AngII, E2 exacerbated renal injury. E2-mediated increases in renal cortical AT(1)R expression may represent a novel mechanism for the adverse renovascular effects of estrogen.


Subject(s)
Angiotensin II/pharmacology , Estradiol/adverse effects , Kidney/metabolism , NG-Nitroarginine Methyl Ester/pharmacology , Proteinuria/metabolism , Receptor, Angiotensin, Type 1/metabolism , Animals , Enzyme Inhibitors/pharmacology , Estrogen Receptor alpha/genetics , Estrogen Receptor alpha/metabolism , Estrogen Receptor beta/genetics , Estrogen Receptor beta/metabolism , Female , Gene Expression Regulation/drug effects , Kidney/blood supply , Kidney/drug effects , Kidney/pathology , Nitric Oxide/metabolism , Ovariectomy , Proteinuria/pathology , RNA, Messenger/genetics , RNA, Messenger/metabolism , Rats , Rats, Wistar , Receptor, Angiotensin, Type 1/genetics , Vasoconstrictor Agents/pharmacology
4.
BJU Int ; 90(6): 513-7, 2002 Oct.
Article in English | MEDLINE | ID: mdl-12230607

ABSTRACT

OBJECTIVES: To determine if levels of inter-alpha-trypsin inhibitor (I alpha TI)-trimer differ in normal individuals based on age, gender or hormonal status, as the regulation of calcium oxalate (CaOx) crystallization inhibitors, e.g. by sex steroids, could be a mechanism contributing to the differences in CaOx urolithiasis between the sexes. SUBJECTS AND METHODS: Voided urine samples were collected from normal males and females. In Experiment 1 samples were grouped by gender and age, i.e. paediatric (PED) < or = 10 years, male (M) 21, female (F) 14; young adult (YGAD) 20-30 years, M 23, F 18; adults (AD), 35-50 year, M 25, F 13; adults aged > or = 60 years (> 60), M 24, F 16 (totals, M 93, F 61). In Experiment 2 samples were grouped by gender, age and hormonal status, i.e. PED, M 24, F 17; AD, M 24, F 22; > 60 and not on hormonal therapy, M 23, F 30; M > 60 and on androgen deprivation therapy (ANDEP) 18; and F > 60 on oestrogen supplementation, F+EST, 18 (total M 89, F 85). Levels of urinary I alpha TI-trimer were determined by immunoblotting and enhanced chemiluminescence, and relative densities of the bands determined. RESULTS: In both experiments the relative levels of I alpha TI-trimer were 2-7 times higher in M-PED than in all other groups of males (P < or = 0.007). Among adult males, I alpha TI-trimer levels were similar in all groups, including ANDEP (P > or = 0.9). There were no differences in the relative levels of I alpha TI-trimer among any of the groups of females, regardless of age or hormonal status (P > or = 0.7). CONCLUSIONS: In males a decrease in I alpha TI-trimer was associated with the onset of adulthood and entry into the 'stone-forming years'. Females did not show this decrease, and neither sex showed an increase in I alpha TI-trimer in the > 60 group, when the incidence of CaOx urolithiasis is supposedly declining. While changes in urinary I alpha TI-trimer levels in males may reflect maturational changes in the kidney, overall these data do not support the hypothesis that the age-related changes in the incidence of urolithiasis are paralleled by changes in the expression I alpha TI-trimer. Additionally, the sex steroids do not appear to acutely regulate the expression of I alpha TI-trimer in adults, making differences in I alpha TI-trimer levels unlikely to be the reason for the disparity in the incidence of CaOx urolithiasis between the sexes.


Subject(s)
Alpha-Globulins/urine , Calcium Oxalate/metabolism , Gonadal Steroid Hormones/physiology , Kidney Calculi/etiology , Urinary Calculi/etiology , Adolescent , Adult , Age Factors , Aged , Child , Female , Humans , Immunoblotting , Male , Middle Aged , Sex Characteristics
6.
Clin Chim Acta ; 302(1-2): 161-70, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11074073

ABSTRACT

The objective of this study was to detect myocardial injury defined by an increase of plasma cardiac troponin I (cTnI) following percutaneous transluminal coronary angioplasty (PTCA) and compare plasma cTnI with the risk of cardiac complications at 30 days. Plasma cTnI, creatine kinase (CK) MB, and total CK were determined in 83 patients before (baseline) and 6, 12 and 24 h after PTCA. Thirty-eight patients underwent conventional PTCA, 39 PTCA-stent and six rotational atherectomy. Patients with acute myocardial infarction (AMI) and increased pre-procedural cTnI >0.8 microg/l were categorized into group 1 (n=23). The remaining 60 patients (pre-procedural cTnI=0.8 microg/l) were categorized as follows: group 2 (n=15) AMI; group 3 (n=20) unstable angina (UA); group 4 (n=25) coronary artery disease (CAD). Twelve hours post-procedure, all three cardiac markers were more frequently increased over baseline in group 2 patients (40-60%) compared to patients in group 3 (5-29%, P<0.03) or group 4 (0.5-5%, P<0.01). This was also true for patients undergoing PTCA-stent compared to conventional PTCA or rotational atherectomy (27-40 vs. 4-14%, P<0.02). cTnI was more sensitive (60%) to detect release of myocardial protein after PTCA compared to total CK (47%) or CKMB (43%). A moderate increase of cTnI (0.8-1.5 microg/l) in groups 2, 3 and 4 was associated with higher risk of complications 30 days post-procedure.


Subject(s)
Angioplasty, Balloon, Coronary/adverse effects , Heart Diseases/etiology , Troponin I/blood , Aged , Angina, Unstable/etiology , Creatine Kinase/blood , Female , Heart Arrest/etiology , Humans , Isoenzymes/blood , Male , Middle Aged , Risk Factors , Time Factors , Treatment Outcome
8.
Ann Clin Lab Sci ; 30(2): 185-90, 2000 Apr.
Article in English | MEDLINE | ID: mdl-10807163

ABSTRACT

A fully automated immunoassay for total plasma homocysteine assay was evaluated at four centers. To measure total homocysteine, oxidized forms of homocysteine in serum and plasma were reduced by dithiothreitol and assayed by a competitive fluorescence polarization technique. The assay had within-run precision from 0.9 to 3.0% and total precision from 2.8 to 4.1% for control materials with homocysteine concentrations of approximately 7, 12.5, and 25 micromol/L, a sensitivity of 0.35 micromol/L, good parallelism upon dilution, and analytical recovery ranging from 97.4 to 103.8%. The immunoassay correlated with four different HPLC assays for homocysteine, yielding a slope of 0.98, an intercept of -0.19 micromol/L, and a correlation coefficient of 0.966 for 440 paired samples. The reference range, determined with plasma samples from 609 males and 600 females, yielded a mean of 9.17+/-2.86 micromol/L, with a central 95% range of 4.78-15.43 micromol/L. The immunoassay is a suitable alternative to HPLC and may be useful in screening persons with high risk of coronary artery disease.


Subject(s)
Coronary Disease/diagnosis , Homocysteine/analysis , Homocysteine/blood , Immunoassay/methods , Chemistry, Clinical/methods , Chemistry, Clinical/standards , Chromatography, High Pressure Liquid , Coronary Disease/blood , Evaluation Studies as Topic , Humans , Immunoassay/standards , Reference Values , Regression Analysis , Sensitivity and Specificity
9.
Rev Urol ; 2(4): 232-5, 2000.
Article in English | MEDLINE | ID: mdl-16985759

ABSTRACT

Fibrous pseudotumor of the bladder, a rare, benign, and proliferative lesion of the submucosal stroma, can be mistaken on gross examination for a malignant lesion and must be differentiated on histologic examination from several bladder malignancies. Radiographic examination alone cannot establish a definitive diagnosis. Complete transurethral resection of such lesions appears to be curative.

10.
Clin Chem ; 45(12): 2129-35, 1999 Dec.
Article in English | MEDLINE | ID: mdl-10585344

ABSTRACT

BACKGROUND: The expression of multiple cardiac troponin T (cTnT) isoforms has been demonstrated in diseased human skeletal muscle. However, cardiac troponin I (cTnI) expression has been described only in heart muscle. The goal of this study was to determine whether mRNA for cTnT, slow skeletal troponin T (sTnT), or cTnI was expressed in skeletal muscle biopsies obtained from patients with end-stage renal disease (ESRD) and Duchenne muscular dystrophy (DMD). METHODS: Total mRNA was extracted from healthy human heart (n = 4), healthy human skeletal muscle (n = 5), and skeletal muscle from patients with ESRD (n = 7) and DMD (n = 5). Total RNA (1 microg) was reverse-transcribed using Moloney murine leukemia virus reverse transcriptase. The reverse-transcribed cDNAs were amplified by PCR using oligonucleotide primers specific for cTnT, sTnT, and cTnI sequences (GenBank accession numbers X74819, m19308, and X54163, respectively). RESULTS: In all heart specimens, a 150-bp cTnT amplicon was detected. Skeletal muscle from four of seven patients with ESRD and two of five patients with DMD showed expression of a 150-bp amplicon. Using DNA sequencing and a comparison program, the 150-bp amplicons found in heart and diseased skeletal muscle specimens were 100% identical and specific to the cTnT mRNA sequence. No cTnT mRNA expression was found in healthy skeletal muscle. No evidence of sTnT mRNA was found in heart muscle. A 200-bp sTnT amplicon specific to a human sTnT sequence was detected in all skeletal muscle specimens. A 250-bp cTnI amplicon specific to the cTnI sequence was detected in all heart specimens. However, no cTnI mRNA expression was found in healthy or diseased skeletal muscle specimens. cTnT mRNA expression in both heart and diseased skeletal muscles corresponded with cTnT isoform expression, respectively, as determined by Western blot analysis. CONCLUSION: Our findings demonstrate cTnT mRNA expression, but no cTnI mRNA expression, by reverse transcription-PCR in diseased human skeletal muscle that expresses cTnT isoforms.


Subject(s)
Muscle, Skeletal/metabolism , Muscular Dystrophy, Duchenne/metabolism , RNA, Messenger/analysis , Troponin T/genetics , Base Sequence , Gene Expression , Humans , Kidney Failure, Chronic/metabolism , Molecular Sequence Data , Muscular Dystrophy, Duchenne/genetics , Myocardium/metabolism , Protein Isoforms/genetics , RNA, Messenger/biosynthesis , Reverse Transcriptase Polymerase Chain Reaction , Troponin I/genetics
11.
12.
Clin Chem ; 45(2): 206-12, 1999 Feb.
Article in English | MEDLINE | ID: mdl-9931042

ABSTRACT

We evaluated the AxSYM troponin I (cTnI) immunoassay for assisting in the detection of acute myocardial infarction (AMI). At four sites, the total imprecision (CV) over 20 days was 6.3-10.2%. The minimum detectable concentration was 0.14 +/- 0.05 microgram/L. Comparison of cTnI measurements between the AxSYM and Stratus (n = 406) over the dynamic range of the AxSYM assay demonstrated good correlation, r = 0.881, with a proportional bias: AxSYM cTnI = 3.50(Stratus cTnI) - 1. 10. The confidence intervals (95%) for the slope and intercept were 3.39-3.64 and -1.32 to -0.95, respectively. The expected cTnI concentration in healthy individuals was /=96%, in skeletal muscle injury, chronic renal disease, and same-day noncardiac surgery patients.


Subject(s)
Immunoenzyme Techniques/standards , Myocardial Infarction/diagnosis , Troponin I/blood , Evaluation Studies as Topic , Humans , Immunoenzyme Techniques/methods , Myocardial Infarction/blood , ROC Curve , Reproducibility of Results , Sensitivity and Specificity
13.
Eur Heart J ; 19 Suppl N: N30-3, 1998 Nov.
Article in English | MEDLINE | ID: mdl-9857936

ABSTRACT

The purpose of this study was to determine whether the two monoclonal anti-cardiac troponin T (cTnT) antibodies used in the second generation cTnT assay (capture Ab, M11.7; detection Ab, M7) would detect expression of cTnT isoforms in skeletal muscle from chronic renal disease patients. Skeletal muscle biopsies obtained from 45 chronic renal disease patients (as well as human heart muscles and normal human skeletal muscles) were prepared for Western blot analysis and blotted with the following anti-cTnT antibodies: M 11.7; M7; JS-2, Lakeland Biomedical; 13-11, Duke University) and anti-cTnI antibody JS-1. Using the M11.7 Ab, 20 of 45 renal skeletal muscles demonstrated one to three cTnT isoforms, MW34 39 kDa. These findings were confirmed by both the Lakeland and Duke antibodies. However the BM M7 antibody detected, in two of 45 muscles, only a protein with a MW of approximately 39 kDa. All four antibodies demonstrated equivalence in detection the 39 kDa cTnT isoform expressed in heart muscle. None detected any isoforms in normal skeletal muscle. A single cTnI isoform, MW 25 kDa, was detected by JS-1 only in normal adult myocardium. Based on the antibody configuration of the second generation cTnT assay, we conclude that while cTnT isoforms are expressed in human skeletal muscle obtained from chronic renal disease patients, if released into the circulation, they would not be detected.


Subject(s)
Kidney Failure, Chronic/metabolism , Muscle, Skeletal/chemistry , Troponin T/analysis , Adult , Aged , Aged, 80 and over , Antibodies, Monoclonal , Blotting, Western , False Positive Reactions , Female , Humans , Immunoassay , Male , Middle Aged , Protein Isoforms
14.
Clin Chem ; 44(9): 1919-24, 1998 Sep.
Article in English | MEDLINE | ID: mdl-9732977

ABSTRACT

The purpose of this study was to determine whether the two monoclonal anti-cardiac troponin T (cTnT) antibodies (MAbs) used in the second generation cTnT assay by Boehringer Mannheim (BM, capture Ab, M11.7; detection Ab, M7) would detect cTnT isoforms expressed in human skeletal muscle in response to chronic renal disease (CRD). cTnT expression was examined in skeletal muscle biopsies obtained from 45 CRD patients, as well as nondiseased human heart (n = 3) and skeletal muscle (n = 3). cTnT proteins were resolved by modified 7.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to nitrocellulose, and probed with the following anti-cTnT MAbs: M11.7; M7; JS-2, Lakeland Biomedical; and 13-11, Duke University. All four antibodies detected the cTnT isoforms (Ta, Te) expressed in human myocardium. In 20 of 45 skeletal muscle biopsies, MAb M11.7 recognized its epitope in one to three proteins, molecular mass 34-36 kDa, designated Te, Td, and Tc; the strongest signal was that of Te. The same proteins were recognized by MAbs JS-2 and 13-11. The BM M7 antibody did not detect the cTnT isoforms in the molecular mass range of 34-36 kDa. However, MAb M7 did detect a cTnT isoform, molecular mass 39 kDa, in 2 of 45 biopsies. This isoform had an electrophoretic mobility similar to the predominant heart cTnT isoform, Ta. We conclude that cTnT isoforms are expressed in the skeletal muscle of CRD patients. However, given the epitopes recognized by the BM MAbs M7 and M11.7 and the variable presence of these cTnT isoforms in skeletal muscle, the second generation BM cTnT assay will not detect these isoforms if they are released from skeletal muscle into the circulation.


Subject(s)
Kidney Failure, Chronic/metabolism , Muscle, Skeletal/metabolism , Myocardium/metabolism , Troponin/analysis , Adult , Aged , Aged, 80 and over , Antibodies, Monoclonal/immunology , Blotting, Western , Epitopes/immunology , False Positive Reactions , Female , Humans , Male , Middle Aged , Reagent Kits, Diagnostic , Troponin/biosynthesis , Troponin/immunology , Troponin T
15.
Am J Clin Pathol ; 110(2): 241-7, 1998 Aug.
Article in English | MEDLINE | ID: mdl-9704624

ABSTRACT

We studied the distribution of cardiac troponins I (cTnI) and T (cTnT) in ischemic left ventricular (LV) tissue in 7 infarct zones, 7 remote nonischemic LV areas, and 7 nonischemic areas each from the right ventricle and circumflex in an acute coronary artery occlusion dog model to correlate myocardial loss of troponins with infarct size 3 weeks after the infarction and to determine whether the decrease of troponins in ischemic myocardium can be used to assess the infarct size in dogs after coronary occlusion. The serum profiles for time vs mean cTnI and cTnT concentrations in 6 dogs after occlusion showed peak concentrations at 1 day and 5 days, respectively. The concentrations of troponins were similar in all nonischemic zones. However, cTnI and cTnT decreased significantly in the LV ischemic tissues. Loss of cTnT, but not cTnI, in ischemic LV tissues correlated significantly with infarct size 3 weeks after the infarction. Biochemical alterations suggest that the increases in serum troponins after the infarction parallel the decreases in tissue concentrations of troponins.


Subject(s)
Myocardial Infarction/metabolism , Myocardial Infarction/pathology , Myocardium/metabolism , Troponin I/metabolism , Troponin/metabolism , Animals , Dogs , Female , Male , Models, Cardiovascular , Myocardium/pathology , Osmolar Concentration , Troponin/blood , Troponin I/blood , Troponin T
16.
Clin Chem ; 43(6 Pt 1): 990-5, 1997 Jun.
Article in English | MEDLINE | ID: mdl-9191551

ABSTRACT

Cardiac troponin T (cTnT) and troponin I (cTnI) have been suggested as new, more specific markers of myocardial cellular damage. The objective of this study was to examine how the distributions of cTnI and cTnT were affected in postinfarction left ventricular remodeled (LVR) myocardium. At 2 months postinfarct in a porcine heart failure model, both Western blot and biochemical assay analyses were performed on left ventricular myocardium remote from the infarct zone in ligation animals (n = 8). Results were compared with data from the left ventricular myocardium from similar sized healthy (control) pigs (n = 7). Autoradiograms from Western blot analysis showed that the protein mass for cTnI and cTnT in LVR hearts decreased 80% (P < 0.001) and 40% (P < 0.02), respectively, when compared with nondiseased tissue. Similarly, the concentrations for cTnI and cTnT in LVR hearts decreased 42% (P < 0.05) and 70% (P < 0.001), respectively, compared with nondiseased normal tissue. The clinical assumption is that the appearance of cTnI and cTnT in the blood is proportional to chronic loss of cTnI and cTnT from injured myocardium associated with left ventricular remodeling.


Subject(s)
Myocardial Infarction/metabolism , Myocardial Infarction/physiopathology , Myocardium/metabolism , Troponin I/metabolism , Troponin/metabolism , Ventricular Function, Left , Animals , Blotting, Western , Cardiac Volume , Disease Models, Animal , Heart Ventricles/physiopathology , Immunoassay , Swine , Troponin T
17.
J Autoimmun ; 10(2): 181-91, 1997 Apr.
Article in English | MEDLINE | ID: mdl-9185880

ABSTRACT

The Ro ribonucleoprotein particle (RoRNP) is the target of a variety of specific anti-protein autoantibodies produced by patients with systemic autoimmune diseases. RoRNPs, the function of which remains elusive, appear to be rather heterogeneous in terms of their molecular composition. Among several Ro proteins, a protein of 52 kD (Ro52) has been identified, but its association with RoRNPs remains questionable. In this study, we first mapped the Ro52 regions that are accessible at the surface of the isolated protein, by means of antibodies raised in rabbits, against 39 overlapping synthetic peptides covering the whole Ro52 molecule and using several different methods, including various ELISA formats and Western blotting (based on the use of recombinant Ro52) and immunoprecipitation of in vitro translated Ro52. Then, the whole set of anti-peptide antibodies was used to attempt to immunoprecipitate RoRNPs from a cell extract of K562 cells. RoRNPs, especially Ro(hY3)RNPs, were effectively immunoprecipitated by certain anti-peptide antibodies, indicating that Ro52 protein is associated with at least a subset of RoRNP particles. As expected, a number of epitopes available on the isolated Ro52 molecule are no longer accessible when Ro52 is associated with the RoRNP. Conversely, neotopes, among them at least one corresponding to a previously characterized epitope recognized by patients' antibodies, are only present at the surface of the particle and not detectable on the isolated Ro52 protein.


Subject(s)
Antibodies, Antinuclear/immunology , Autoantigens/immunology , Epitopes/immunology , Peptides/immunology , RNA, Small Cytoplasmic , Ribonucleoproteins/immunology , Animals , Antibodies, Antinuclear/chemistry , Antigen-Antibody Reactions , Autoantigens/chemistry , Enzyme-Linked Immunosorbent Assay , Epitope Mapping , Epitopes/chemistry , Humans , Immune Sera/biosynthesis , Immune Sera/chemistry , Molecular Weight , Peptides/chemical synthesis , Precipitin Tests , Rabbits , Recombinant Proteins/chemistry , Recombinant Proteins/immunology , Ribonucleoproteins/chemistry , Tumor Cells, Cultured
18.
Clin Immunol Immunopathol ; 77(3): 282-90, 1995 Dec.
Article in English | MEDLINE | ID: mdl-7586738

ABSTRACT

Human T-cell leukemia virus type I (HTLV-I) is associated with a large spectrum of clinical manifestation including adult T-cell leukemia (ATL) and tropical spastic paraparesis or HTLV-I-associated myelopathy (TSP/HAM). In most cases, however, infected patients remain asymptomatic. The participation of the immune system in the pathogenesis of TSP/HAM has been suggested. In this study the IgG antibody response of HTLV-I-infected individuals has been investigated using both ELISA with a panel of nuclear and cytoplasmic proteins and peptides known to be recognized by antibodies from patients with various systemic autoimmune diseases, and immunoprecipitation of ribonucleoproteins from HeLa cell extracts. The results were compared with the reactivity of sera from individuals with non-HTLV-I-related neurological diseases and healthy blood donors. Raised levels of autoantibodies reacting with several nuclear and cytoplasmic antigens were found in TSP/HAM and ATL patients. In asymptomatic HTLV-I-seropositive individuals, both the prevalence and level of IgG antibodies were lower and directed only against a restricted set of antigens. The mechanism of induction of these antibodies still remains obscure. However, the results show that a significant autoimmune response exists in these patients and it may contribute to the pathogenesis of the disease.


Subject(s)
Autoantibodies/blood , Autoantibodies/immunology , Autoantigens/immunology , HTLV-I Infections/immunology , Immunoglobulin G/blood , Immunoglobulin G/immunology , Adult , Antibodies, Antinuclear/blood , Antibodies, Antinuclear/immunology , Enzyme-Linked Immunosorbent Assay , Female , HeLa Cells , Humans , Male , Peptides/immunology , Precipitin Tests , RNA/immunology , Ribonucleoproteins/immunology
19.
J Autoimmun ; 7(5): 611-21, 1994 Oct.
Article in English | MEDLINE | ID: mdl-7530962

ABSTRACT

The possible pathological role of antibody subsets specific for different regions of 60-KD and 52-KD Ro/SSA proteins (Ro60 and Ro52) and La protein is unclear. Previously, we have shown that in patients with Sjögren's syndrome, the fine specificity of Ro60 and Ro52 antibodies, as characterized with synthetic peptides, varied considerably according to the origin of sera. We therefore looked for possible associations of HLA class I and class II alleles with Ro and La IgG antibodies in patients with primary Sjögren's syndrome (n = 24) and secondary Sjögren's syndrome associated with systemic lupus erythematosus (n = 25). Ro60 and Ro52 antibodies were tested by ELISA with the complete parent proteins and with 10 to 24 residue-long peptides of these proteins. Anti-Ro60 antibodies were more frequent in DR3-positive patients and in DQ1-negative patients whereas the presence of Ro52 and La IgG antibodies was significantly increased in patients with A1/B8/DR3 haplotype. Certain HLA associations observed with antibodies reacting with the whole Ro52 protein were not found with antibodies reacting with certain Ro60 and Ro52 peptides and, reciprocally, certain anti-peptide antibodies were linked to particular haplotypes whereas antibodies to the respective parent proteins were not linked to these haplotypes. Thus the production of antibody subsets reacting with different parts or presentations of Ro proteins is, at least in part, influenced immunologically by different HLA haplotypes, and this predisposition may play a role in the pathological development of the disease.


Subject(s)
Antibodies, Antinuclear/immunology , Autoimmune Diseases/immunology , HLA Antigens/immunology , RNA, Small Cytoplasmic , Sjogren's Syndrome/immunology , Adult , Aged , Aged, 80 and over , Autoantigens/immunology , Disease Susceptibility/immunology , Epitopes/immunology , Ethnicity/genetics , Female , Genetic Predisposition to Disease , HLA Antigens/genetics , HLA-D Antigens/genetics , HLA-D Antigens/immunology , Haplotypes/genetics , Humans , Immunoglobulin G/immunology , Lupus Erythematosus, Systemic/complications , Lupus Erythematosus, Systemic/immunology , Male , Middle Aged , Peptide Fragments/immunology , Ribonucleoproteins/immunology , Sjogren's Syndrome/etiology , SS-B Antigen
20.
Clin Exp Immunol ; 95(3): 397-407, 1994 Mar.
Article in English | MEDLINE | ID: mdl-7511075

ABSTRACT

The reactivity of autoantibodies present in the sera of 489 patients with Sjögren's syndrome (SS), systemic lupus erythematosus (SLE) and other autoimmune diseases was investigated by ELISA using recombinant 52-kD SSA/Ro protein (rRo52) and 39 overlapping synthetic peptides representing the entire sequence of Ro52. We report that IgG antibodies reacting with rRo52 were present in the sera of a large number of patients with SS (67% of patients with primary SS and 46% of patients with SS associated with SLE), whereas they were less frequent (10-25%) in SLE, rheumatoid arthritis (RA), juvenile chronic arthritis (JCA) and mixed connective tissue disease (MCTD), and absent in scleroderma. Among the 39 peptides tested, five were recognized by sera from 30-65% of patients with SS, namely peptides representing residues 2-11, 107-122, 107-126, 277-292 and 365-382. Patients with JCA had raised levels of IgG antibodies reacting with peptides 2-11 and 365-382, and 51% of patients with MCTD had raised levels of IgG antibodies reacting with peptide 365-382. None of the five peptides was recognized by more than 20% of sera from patients with SLE and RA. Interestingly, and of importance in the field of diagnostic tests based on peptides, the reactivity of antibodies to the Ro52 synthetic peptides varied greatly according to the origin of sera. Inhibition experiments using either patients' sera or antibodies induced in rabbits against Ro52 peptides showed that the four domains 2-11, 107-122, 277-292 and 365-382 are accessible on the surface of the Ro52 protein. These regions may thus be involved in the induction of specific antibodies in autoimmune patients.


Subject(s)
Antibody Specificity , Autoantibodies/blood , Autoantigens/immunology , Autoimmune Diseases/immunology , B-Lymphocytes/immunology , Epitopes , RNA, Small Cytoplasmic , Ribonucleoproteins/immunology , Amino Acid Sequence , Enzyme-Linked Immunosorbent Assay/methods , Humans , Immunoblotting , Immunoelectrophoresis , Lupus Erythematosus, Systemic/immunology , Molecular Sequence Data , Peptide Fragments/immunology , Sjogren's Syndrome/immunology
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