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1.
Clin Exp Rheumatol ; 42(5): 1029-1034, 2024 May.
Article in English | MEDLINE | ID: mdl-38179724

ABSTRACT

OBJECTIVES: We aimed to evaluate the prevalence of non-criteria clinical features in patients with primary antiphospholipid syndrome (APS), and to assess their relationship to thrombosis and damage. METHODS: We retrospectively included 177 primary APS patients, and/or patients who only achieved the serological Sydney criteria but had thrombocytopenia and/or haemolytic anaemia. We registered demographics, serology, treatment, thrombotic/obstetric manifestations and non-criteria clinical manifestations (cutaneous, haematologic, renal, heart valve disease, and neurological). We scored the DIAPS and a modified SLICC index. We used logistic regression and reported OR with 95% CI. RESULTS: 78% were women with a median follow-up of 6.7 years. Thrombosis was found in 74% of patients, 29.3% had obstetric features, and 64% had non-criteria clinical manifestations. The frequency of the non-criteria clinical manifestation was: haematologic 40.1%, cutaneous 20.9%, neurologic 18%, cardiac 5% and renal 4.5%. Non-criteria features were associated with LA (OR 2.3, 95% 1.03-5.1) and prednisone use (OR 8.2, 95% CI 1.7-39.3). A DIAPS score ≥1 was associated with thrombosis (OR 53.1, 95% CI 17.8-15.2), prednisone use (OR 0.27, CI 95% 0.09-0.83) and neurological involvement (OR 6.4, 95% CI 1.05-39.8); whereas a modified SLICC ≥ 1 with thrombosis (OR 10.2; IC 95% 4.43-26.1), neurological involvement (OR 6.4, 95%CI 1.05-39.8), obstetric features (OR 0.32 CI 95% 0.12-0,81) and cutaneous features (OR 5.3, CI 95% 1.4-19), especially livedo reticularis (OR 5.45; IC 95% 1.49-19.8). CONCLUSIONS: Non-criteria clinical manifestations are common and associated with LA. Among them, neurologic involvement and the presence of livedo were associated with damage accrual.


Subject(s)
Antiphospholipid Syndrome , Thrombosis , Humans , Antiphospholipid Syndrome/complications , Antiphospholipid Syndrome/diagnosis , Antiphospholipid Syndrome/epidemiology , Female , Retrospective Studies , Adult , Male , Middle Aged , Thrombosis/etiology , Thrombosis/epidemiology , Risk Factors , Prevalence , Odds Ratio , Logistic Models , Anemia, Hemolytic/etiology , Anemia, Hemolytic/epidemiology , Thrombocytopenia/epidemiology , Thrombocytopenia/etiology , Nervous System Diseases/epidemiology , Nervous System Diseases/etiology , Kidney Diseases/epidemiology , Kidney Diseases/etiology , Kidney Diseases/diagnosis , Prednisone/therapeutic use , Prognosis , Time Factors , Antibodies, Antiphospholipid/blood
3.
Thromb Res ; 174: 141-147, 2019 02.
Article in English | MEDLINE | ID: mdl-30612005

ABSTRACT

OBJECTIVE: The clinical significance of anti-phosphatidylserine/prothrombin (aPS/PT) in antiphospholipid syndrome (APS) is still controversial. We assessed the prevalence of aPS/PT antibodies, their association with other anti-phospholipid antibodies (aPL) and with different APS clinical phenotypes. METHODS: We included 95 primary APS patients according to the Sydney classification criteria, and patients with thrombocytopenia and/or hemolytic anemia who also fulfilled the serological APS criteria. We tested aCL, anti-ß2GP-I and aPS/PT antibodies (both IgG and IgM isotypes) and lupus anticoagulant (LA). We used χ2 test, Spearman's correlation coefficient, Mann-Whitney U test and logistic regression. RESULTS: Seventy-seven percent of patients had thrombosis, 50% hematologic involvement and 25% obstetric events (non-exclusive groups). Twenty patients had only hematologic features. The prevalence of IgG and IgM aPS/PT antibodies was 61% and 60%, respectively. Patients with LA+ had a higher prevalence and higher titers of IgG and IgM aPS/PT antibodies. aPS/PT antibodies correlated with aPL antibodies including LA. IgG aPS/PT antibodies were associated with thrombosis (OR 8.6 [95% CI 2.13-33.8, p = 0.002]) and pure hematologic features (OR 0.2, CI 95% 0.05-0.97, p = 0.004). IgM anti-ß2GP-I antibodies conferred high risk for both hematologic (OR 7.9, 95% CI 1.88-34.61, p = 0.006) and thrombotic involvement (OR 7.4, 95% CI 1.76-31.12, p = 0.006). CONCLUSIONS: aPS/PT antibodies were highly prevalent and correlated with other aPL antibodies. IgG aPTS/PT conferred a high risk for thrombosis, but not for pure hematologic involvement. aPS/PT antibodies may be a useful serological tool in the diagnosis and phenotypic characterization of APS patients.


Subject(s)
Antibodies, Antiphospholipid/immunology , Antiphospholipid Syndrome/immunology , Adult , Female , Humans , Male , Phenotype , Prevalence
4.
Rheumatol Int ; 37(7): 1159-1164, 2017 Jul.
Article in English | MEDLINE | ID: mdl-28474094

ABSTRACT

The objective was to describe the management and risk factors for complications of antiphospholipid syndrome (APS) patients who underwent a surgical procedure in a single center. We reviewed medical records of all patients with primary or secondary APS who underwent an elective surgery during a 6-year period. Demographical data, management of anticoagulation and complications were recorded. We identified 43 patients, mean age 37.9 ± 8.9 years, who underwent a total of 48 elective surgeries. All patients had history of at least one thrombotic event and were under vitamin K antagonists. Before surgery, all patients received bridging therapy with intravenous infusion of heparin or low molecular weight heparin (LMWH). Among the LMWH group, 36 had a full anticoagulation regimen and nine prophylactic doses. In 62% of the surgeries, we identified an optimal management of periprocedural anticoagulation according to guidelines. Overall six patients had severe bleeding and three thrombotic complications (full anticoagulation regimen n = 2 and prophylactic dose group n = 1). Patients with optimal management of anticoagulation experienced less thrombotic and hemorrhagic complications (7 vs. 33%; OR 0.14, 95% CI 0.02-0.81; p = 0.040) and patients with INR ≤1.5 at surgery had fewer episodes of major bleeding (6 vs. 29%; OR 0.19, 95% CI 0.02-0.98; p = 0.050). All three thrombotic events occurred in patients with INR ≤1.5. Proper management of anticoagulation based on guidelines is associated with less complications in patients with APS. Notwithstanding the proper use of bridging therapy, some patients may develop thrombotic complications.


Subject(s)
Anticoagulants/administration & dosage , Antiphospholipid Syndrome/drug therapy , Blood Coagulation/drug effects , Heparin, Low-Molecular-Weight/administration & dosage , Perioperative Care/methods , Surgical Procedures, Operative , Adult , Anticoagulants/adverse effects , Antiphospholipid Syndrome/blood , Antiphospholipid Syndrome/complications , Antiphospholipid Syndrome/diagnosis , Chi-Square Distribution , Drug Administration Schedule , Elective Surgical Procedures , Female , Hemorrhage/chemically induced , Heparin, Low-Molecular-Weight/adverse effects , Humans , Infusions, Intravenous , Male , Medical Records , Mexico , Middle Aged , Odds Ratio , Perioperative Care/adverse effects , Retrospective Studies , Risk Factors , Surgical Procedures, Operative/adverse effects , Tertiary Care Centers , Thrombosis/etiology , Thrombosis/prevention & control , Time Factors , Treatment Outcome , Vitamin K/antagonists & inhibitors
5.
Int J Rheum Dis ; 20(6): 755-759, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28261992

ABSTRACT

AIM: To evaluate health-related quality of life (HRQoL) in primary antiphospholipid syndrome (PAPS) and correlate it with a crude estimate of accrual organ damage, comorbidity (diabetes mellitus, hypertension and dyslipidemia) and treatment (oral anticoagulation, immunosuppressors and prednisone). METHODS: We assessed HRQoL with the Short-Form 36 (SF-36) and the Lupus Quality of Life instrument (LupusQoL) and the disease burden with a modified Systemic Lupus International Collaborating Clinics/American College of Rheumatology Damage Index (SLICC/ACR SDI). As controls we used SF-36 data from a Mexican general population within the same age range. RESULTS: We included 50 PAPS patients (86% women), mean age 47.6 ± 14.5 years, median disease duration 9.4 years, median SLICC/ACR score of 1 point and 80% had thrombotic events. PAPS patients had lower HRQoL than controls. We found a positive correlation between SF-36 and LupusQoL (r = 0.85, P < 0.0001). The SLICC/ACR SDI correlated negatively with both LupusQoL and SF-36, specifically the peripheral vascular domain (r = -0.29, P = 0.03, for both). Patients on oral anticoagulants (n = 37) had lower LupusQoL, physical functioning, intimate relationships, burden to others and pain scores as well as a lower SF-36 physical functioning score. We did not find differences in HRQoL regarding comorbidities and other treatments. CONCLUSIONS: HRQoL in PAPS was related to burden of the disease specifically at the vascular peripheral area and use of anticoagulants.


Subject(s)
Anticoagulants/administration & dosage , Antiphospholipid Syndrome/drug therapy , Blood Coagulation/drug effects , Quality of Life , Administration, Oral , Adult , Antiphospholipid Syndrome/blood , Antiphospholipid Syndrome/diagnosis , Antiphospholipid Syndrome/psychology , Case-Control Studies , Comorbidity , Cost of Illness , Cross-Sectional Studies , Female , Glucocorticoids/administration & dosage , Humans , Immunosuppressive Agents/administration & dosage , Male , Mexico , Middle Aged , Prednisone/administration & dosage , Risk Factors , Severity of Illness Index , Surveys and Questionnaires , Treatment Outcome
6.
Hum Immunol ; 78(2): 146-152, 2017 Feb.
Article in English | MEDLINE | ID: mdl-28038927

ABSTRACT

Homozygote genotype V247 of the ß2-glycoprotein-I (ß2GP-I) gene has been associated with anti-ß2GP-I and thrombosis in patients with primary anti-phospholipid syndrome APS (PAPS). However, the cellular immune response to ß2GP-I247 has been little studied. OBJECTIVE: To evaluate the immune cellular proliferation in response to native and non-native ß2GP-I247 valine/leucine phenotype from Mexican patients with PAPS. METHODS: We studied 10 patients with PAPS and 10 healthy control subjects (HC). The polymorphism at position 247 of the ß2GP-I gene was determined by PCR-RFLP and the corresponding ß2GP-I protein was subsequently purified from normal human plasma by affinity chromatography. PBMC purified from patients and controls were stimulated with ß2GP-I under native and in non native (reduced) conditions. We also determined the anti-ß2GP-I production in vitro by B cell clones (EBV) generated in cocultures experiments. Differential Scanning Calorimetry (DSC) was studied to determine the structural differences between the ß2GP-I247 valine/leucine isoforms. Cytokine profile (IL-2, IL-4, IL-6, TNFα, INFγ) was evaluated in culture supernatants. RESULTS: PAPS and healthy control PBMCs had a higher proliferative response when stimulated with ß2GP-I under reduced cultures conditions compared to non-denatured conditions. PBMCs response from PAPS patients was higher. We observed more cell proliferation in response to ß2GP-I247 valine/leucine or valine isoforms in non-native conditions. In contrast, this response was not significant against ß2GP-I247 leucine. These findings were T CD4+-dependent. Similar results were obtained with B cell clones derived from PAPS patients, which showed more pronounced proliferation in non native conditions and higher against ß2GP-I247 valine. No differences were found in anti-ß2GP-I production, but high levels of IL-6 in vitro were identified. The structural analysis of both ß2GP-I247 isoforms by DSC showed a major conformational change due to a single mutation in the ß2GP-I variants. CONCLUSIONS: PAPS PBMCs had a higher cellular response against ß2GP-I247 in non-native culture conditions preferentially to the ß2GP-I247 valine phenotype. This effect is T CD4+ dependent and appears to be driven by tertiary structural changes adopted by ß2GP-I247 polymorphism.


Subject(s)
Antiphospholipid Syndrome/genetics , CD4-Positive T-Lymphocytes/immunology , beta 2-Glycoprotein I/genetics , Adult , Antiphospholipid Syndrome/immunology , Clone Cells , Cytokines/metabolism , Female , Genotype , Humans , Immunity, Cellular , Leucine , Male , Mexico , Middle Aged , Mutation/genetics , Phenotype , Polymorphism, Genetic , Valine
7.
Ann Rheum Dis ; 76(2): 450-457, 2017 Feb.
Article in English | MEDLINE | ID: mdl-27432357

ABSTRACT

OBJECTIVES: Patients with antiphospholipid syndrome (APS) are at risk for subclinical endothelial injury, as well as accelerated atherosclerosis. In the related disease systemic lupus erythematosus, there is a well-established defect in circulating endothelial progenitors, which leads to an accrual of endothelial damage over time. This defect has been at least partially attributed to exaggerated expression of type I interferons (IFNs). We sought to determine whether these pathways are important in APS. METHODS: We studied 68 patients with primary APS. Endothelial progenitors were assessed by flow cytometry and functional assay. Type I IFN activity was determined by a well-accepted bioassay, while peripheral blood mononuclear cells were scored for expression of IFN-responsive genes. RESULTS: Endothelial progenitors from patients with APS demonstrated a marked defect in the ability to differentiate into endothelial cells, a phenotype which could be mimicked by treating control progenitors with APS sera. Elevated type I IFN activity was detected in the circulation of patients with APS (a finding that was then replicated in an independent cohort). While IgG depletion from APS sera did not rescue endothelial progenitor function, the dysfunction was successfully reversed by a type I IFN receptor-neutralising antibody. CONCLUSIONS: We describe, for the first time to our knowledge, an IFN signature in primary APS and show that this promotes impaired endothelial progenitor function. This work opens the door to novel approaches that may mitigate vascular damage in APS, such as anti-IFN drugs.


Subject(s)
Antiphospholipid Syndrome/immunology , Cell Differentiation/physiology , Endothelial Progenitor Cells/physiology , Interferon-alpha/immunology , Leukocytes, Mononuclear/immunology , Adult , Aged , Antibodies, Neutralizing/pharmacology , Antiphospholipid Syndrome/physiopathology , Case-Control Studies , Cell Differentiation/drug effects , Endothelial Progenitor Cells/drug effects , Female , Flow Cytometry , Humans , Interferon Type I/immunology , Male , Middle Aged , Receptor, Interferon alpha-beta/antagonists & inhibitors , Young Adult
8.
Clin Rheumatol ; 35(1): 55-61, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26255191

ABSTRACT

The objective of this study was to determine the relationship between citrullinated proteins in synovial tissue with peripheral anti-citrullinated peptides autoantibodies (ACPA) and peptidylarginine deiminase (PADI) PADI2, PADI3, and PADI4 messenger RNA (mRNA) expressions in synovial tissue and fibroblast-like synoviocytes in rheumatoid arthritis (RA) patients. Eleven RA and 12 osteoarthritis (OA) patients who underwent knee replacement surgery were studied. We detected citrullinated proteins in synovial tissue homogenates by western blot and serum ACPA by ELISA to anti-cyclic citrullinated peptide (anti-CCP) antibodies, and PADI2, PADI3, and PADI4 mRNA expressions in synovial tissue and in fibroblast-like synoviocytes. Patients with high amount of citrullinated proteins in synovial tissue (3 out of 7) have high levels of anti-CCP in serum. However, in the remaining 4 patients, the amount of synovial citrullinated proteins was minimal and their sera showed low levels of anti-CCP antibodies. Furthermore, we observed an increase in PADI2 mRNA expression in RA synovial tissue compared with OA patients (p = 0.02). We detected PADI3 mRNA in the synovial tissue of RA patients, but not in the tissue of OA patients. Even though fibroblast-type synoviocytes in RA are not the main source of PADs in the synovial tissue, they express PADI2 mRNA moderately, PADI4 mRNA weakly, while there is no detectable expression of PADI3 mRNA. In conclusion, we found a variety of citrullinated proteins in the synovial tissue of RA patients and the amount of such proteins is related to serum concentration of anti-CCP antibodies. We identified the presence of PADI3 mRNA expression in synovial tissue and PADI2 and PADI4 mRNA expressions in fibroblast-like synoviocytes from patients with RA.


Subject(s)
Arthritis, Rheumatoid/blood , Autoantibodies/blood , Hydrolases/genetics , Osteoarthritis/blood , Peptides, Cyclic/immunology , Synovial Membrane/metabolism , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Protein-Arginine Deiminase Type 2 , Protein-Arginine Deiminase Type 3 , Protein-Arginine Deiminase Type 4 , Protein-Arginine Deiminases , RNA, Messenger/genetics
9.
Clin Exp Rheumatol ; 33(4): 503-8, 2015.
Article in English | MEDLINE | ID: mdl-26203932

ABSTRACT

OBJECTIVES: To assess the presence of acute thrombotic microangiopathy (aTMA) and chronic vascular lesions (cTMA) in lupus nephropathy, and to evaluate their association with extrarrenal lupus features, aPL positivity, antiphospholipid syndrome (APS) and renal survival. METHODS: We studied lupus patients with renal biopsy, ≥1 year of post-biopsy follow-up and at least two aCL (IgG-IgM), anti-ß2GP-I (IgG-IgM) and/or lupus anticoagulant (LAC) determinations. A blinded nephropathologist evaluated all biopsies. We retrospectively collected clinical, serological, treatment and renal survival data. We plotted survival curves and used Cox regression analysis. RESULTS: A total of 90 biopsies were included with a median disease duration 5.9 years and median follow-up 2.4 years. Eleven patients (12.2%) had cTMA and 3 (3%) aTMA. There was no difference in age, lupus duration, hypertension, drugs, APS, non-renal lupus features, low C3 or C4 aCL IgG, anti-ß2GP1-IgG or IgM and LAC between cTMA and non-cTMA groups. The cTMA group had aCL-IgM less frequently (27% vs. 66%, p=0.02), more class IV nephropathy (100% vs. 40%, p=0.01), higher activity index scores (7.5 vs. 2, p=0.03) and a tendency to need chronic dialysis (54.5% vs. 24% p=0.06). At four years of follow-up, 28% of the cTMA group and 62% of the non-cTMA group were free of dialysis (log rank p=0.03). cTMA was associated with chronic dialysis (RR 2.9, CI 95% 1.1-8.1, p=0.03). CONCLUSIONS: cTMA conferred a poor renal outcome. We found a low frequency of TMA that was not associated with with APL positivity or APS, suggesting that other factors hitherto not studied are involved in its pathogenesis.


Subject(s)
Kidney , Lupus Erythematosus, Systemic/complications , Lupus Nephritis , Renal Dialysis/methods , Thrombotic Microangiopathies , Adult , Antiphospholipid Syndrome/diagnosis , Biopsy , Disease Progression , Female , Humans , Kidney/pathology , Kidney/physiopathology , Kidney Function Tests/methods , Lupus Coagulation Inhibitor/blood , Lupus Nephritis/diagnosis , Lupus Nephritis/etiology , Lupus Nephritis/immunology , Lupus Nephritis/physiopathology , Lupus Nephritis/therapy , Male , Severity of Illness Index , Thrombotic Microangiopathies/diagnosis , Thrombotic Microangiopathies/etiology , Thrombotic Microangiopathies/metabolism , Thrombotic Microangiopathies/physiopathology , Tissue Survival , beta 2-Glycoprotein I/blood
10.
Arthritis Rheumatol ; 67(11): 2990-3003, 2015 Nov.
Article in English | MEDLINE | ID: mdl-26097119

ABSTRACT

OBJECTIVE: Antiphospholipid antibodies (aPL), especially those targeting ß2 -glycoprotein I (ß2 GPI), are well known to activate endothelial cells, monocytes, and platelets, with prothrombotic implications. In contrast, the interaction of aPL with neutrophils has not been extensively studied. Neutrophil extracellular traps (NETs) have recently been recognized as an important activator of the coagulation cascade, as well as an integral component of arterial and venous thrombi. This study was undertaken to determine whether aPL activate neutrophils to release NETs, thereby predisposing to the arterial and venous thrombosis inherent in the antiphospholipid syndrome (APS). METHODS: Neutrophils, sera, and plasma were prepared from patients with primary APS (n = 52) or from healthy volunteers and characterized. No patient had concomitant systemic lupus erythematosus. RESULTS: Sera and plasma from patients with primary APS had elevated levels of both cell-free DNA and NETs, as compared to healthy volunteers. Freshly isolated neutrophils from patients with APS were predisposed to high levels of spontaneous NET release. Further, APS patient sera, as well as IgG purified from APS patients, stimulated NET release from control neutrophils. Human aPL monoclonal antibodies, especially those targeting ß2 GPI, also enhanced NET release. The induction of APS NETs was abrogated with inhibitors of reactive oxygen species formation and Toll-like receptor 4 signaling. Highlighting the potential clinical relevance of these findings, APS NETs promoted thrombin generation. CONCLUSION: Our findings indicate that NET release warrants further investigation as a novel therapeutic target in APS.


Subject(s)
Antibodies, Antiphospholipid/immunology , Antiphospholipid Syndrome/immunology , Extracellular Traps , Neutrophils/immunology , Thrombosis/immunology , Antiphospholipid Syndrome/metabolism , Humans , Immunoglobulin G/immunology , Neutrophils/metabolism , Reactive Oxygen Species/metabolism , Signal Transduction/physiology , Thrombosis/metabolism , Toll-Like Receptor 4/metabolism
11.
Biochem Biophys Res Commun ; 453(1): 94-100, 2014 Oct 10.
Article in English | MEDLINE | ID: mdl-25256745

ABSTRACT

Several studies have shown that conformational changes of ß(2)-glycoprotein I (ß(2)GPI) when bound to negatively charged components expose cryptic epitopes and subsequent binding of anti-ß(2)GPI from patients with antiphospholipid syndrome (APS). However, the role of the carbohydrate chains of ß(2)GPI in this anti-ß(2)GPI reactivity is poorly understood. We therefore studied the reactivity and inhibition of anti-ß(2)GPI antibodies from APS patients with native, partially glycosylated ß(2)GPI (pdß(2)GPI; without sialic acid) and completely deglycosylated ß(2)GPI (cdß(2)GPI). To determine the potential biologic importance of these glycoforms and their interaction with anti-ß(2)GPI in vitro, stimulation assays were performed with the U937 cell line. Circular dichroism (CD) and fluorescence analysis of the three ß(2)GPI forms were also studied. We found an increased reactivity of anti-ß(2)GPI against pdß(2)GPI and cdß(2)GPI compared to native ß(2)GPI. Both deglycosylated ß(2)GPI isoforms showed higher inhibition of the anti-ß(2)GPI reactivity than the native protein in soluble-phase. Likewise, the antibody/ß(2)GPI/glycoform complexes increased the synthesis of IL-6, IFNγ and TNFα and the expression of HLA-DR, CD14 and CD11c in U937 cells. CD and fluorescence studies of the glycoforms yielded considerable changes in the fluorescence signals. Our work suggests that the partial or complete removal of the carbohydrate chains uncover cryptic epitopes present in ß(2)GPI. The differentiation and increased synthesis of pro-inflammatory cytokines by U937 cells in vitro may have pathogenetic implications.


Subject(s)
Antiphospholipid Syndrome/blood , Antiphospholipid Syndrome/immunology , beta 2-Glycoprotein I/chemistry , beta 2-Glycoprotein I/immunology , Adult , Antibodies, Anticardiolipin/blood , Antiphospholipid Syndrome/etiology , Case-Control Studies , Cell Differentiation , Cytokines/biosynthesis , Epitopes/chemistry , Epitopes/immunology , Female , Glycosylation , Humans , Immunoglobulin G/blood , Male , Monocytes/immunology , Monocytes/pathology , Sialic Acids/chemistry , Sialic Acids/immunology , U937 Cells , Young Adult
12.
Clin Exp Rheumatol ; 31(3): 382-8, 2013.
Article in English | MEDLINE | ID: mdl-23305633

ABSTRACT

OBJECTIVES: To ascertain rethrombotic risk factors in patients with primary antiphospholipid syndrome (PAPS). METHODS: We retrospectively evaluated 95 patients according to their rethrombotic status. We registered anticoagulation (OA) status, comorbidities, traditional thrombotic factors, prevalence of aCL (IgG-IgM), anti-ß2GP-I (IgG-IgM), LA and triple marker positivity (LA, aCL and anti-ß2GP-I). RESULTS: Forty-two patients had rethrombosis and 53 were rethrombosis-free. The median follow-up was 4.5 (0.3-26) years. There were no differences in comorbidities and traditional thrombotic factors. Patients with rethrombosis had more frequently LA (62% vs. 40%, p=0.04), were younger (41 vs. 47 years, p=0.01) and received less frequently OA (23% vs. 54%, p=0.002). A logistic regression analysis showed that the OA status (OR 0.17, 95% CI 0.05-0.57, p=0.004) and age (OR 0.94, 95% CI 0.90-0.98, p=0.01) remained significant. Patients who discontinued OA and developed rethrombosis (Group 1, n=32) vs. patients who discontinued OA, but remained rethrombosis-free (Group 2, n=24) were also analysed. We found a higher prevalence of LA and triple marker positivity in Group 1 (67% vs. 31%; OR= 4.5, 95% CI 1.3-14.9, p= 0.01 and 57% vs. 27%; OR 3.6, 95% CI 1.7-12; p=0.03), respectively. Both variables remained associated with rethrombosis when compared with the overall rethrombosis group vs. Group 2 (LA 62% vs. 31%, OR= 3.6 95% CI 1.1-11.2, p=0.03; triple marker 54% vs. 27%; OR 32 95% CI 1.01-10.2, p=0.05). CONCLUSIONS: LA positivity and triple aPL positivity confer a more severe risk of rethrombosis in PAPS patients, irrespective of their anticoagulation status and known conventional risk factors.


Subject(s)
Antibodies, Anticardiolipin/immunology , Anticoagulants/therapeutic use , Antiphospholipid Syndrome/immunology , Lupus Coagulation Inhibitor/immunology , Thrombosis/prevention & control , beta 2-Glycoprotein I/immunology , Adult , Antiphospholipid Syndrome/complications , Disease-Free Survival , Female , Follow-Up Studies , Humans , Immunoglobulin G/immunology , Immunoglobulin M/immunology , Kaplan-Meier Estimate , Male , Middle Aged , Recurrence , Retrospective Studies , Risk Factors , Thrombosis/etiology , Thrombosis/immunology
13.
Blood ; 116(16): 3058-63, 2010 Oct 21.
Article in English | MEDLINE | ID: mdl-20625007

ABSTRACT

The updated Sapporo classification criteria for antiphospholipid syndrome (APS) only include thrombosis or pregnancy morbidity as clinical criteria. To test this notion, we studied 55 patients (80% women) with hematologic manifestations. All fulfilled the laboratory criteria for primary APS. Thirty-five patients (64%) had thrombocytopenia, 14 (25%) had autoimmune hemolytic anemia, and 6 (11%) had both. Twenty-five patients (22 women, 88%) also fulfilled one clinical criterion for APS after a median follow-up of 13.2 years (range, 1.45-37 years), whereas the remaining 30 patients (22 women, 73%) have not had any thrombotic event nor pregnancy morbidity after a median follow-up of 5.4 years (range, 0.12-24 years). No patient developed systemic lupus erythematosus during follow-up. The hematologic manifestation was asynchronous with the APS onset in 84% of patients. The response to treatment was similar regardless of the APS status. Patients with definite APS were more frequently positive for the lupus anticoagulant (63%) than lupus anticoagulant-positive patients without APS (30%; odds ratio, 3.5; 95% confidence interval, 1.07-11.4; P < .02). Anticardiolipin or anti-ß(2)-glycoprotein-I antibodies were highly prevalent among the study groups. Our study suggests that, depending upon their antiphospholipid profile, patients with hemocytopenias appear to comprise a peculiar subset of patients with APS; some develop thrombotic and/or obstetric APS whereas others continue with hematologic APS.


Subject(s)
Anemia, Hemolytic, Autoimmune/complications , Anemia, Hemolytic, Autoimmune/immunology , Antibodies, Antiphospholipid/immunology , Antiphospholipid Syndrome/complications , Antiphospholipid Syndrome/immunology , Thrombocytopenia/complications , Thrombocytopenia/immunology , Adult , Antiphospholipid Syndrome/classification , Female , Follow-Up Studies , Humans , Male , Middle Aged , Pregnancy
14.
Methods Mol Biol ; 562: 61-70, 2009.
Article in English | MEDLINE | ID: mdl-19554287

ABSTRACT

This chapter describes the construction and screening of a library of single-chain variable fragments (svFv) derived from patients with autoimmune disease. The methods cover the isolation of mononuclear cells from peripheral blood, preparation of RNA, and recovery of immunoglobulin-coding sequences by polymerase chain reaction (PCR). Cloning into a phage display vector and screening of the scFv display library by a simple panning procedure are described. These methods are applicable to library construction from any patient group or (with alternative primer sets) any mammalian species.


Subject(s)
Antibodies, Anti-Idiotypic/immunology , Immunoglobulin Variable Region/immunology , Peptide Library , Prothrombin/immunology , beta 2-Glycoprotein I/immunology , Antibodies, Monoclonal/genetics , Antibodies, Monoclonal/immunology , Autoimmune Diseases/genetics , Autoimmune Diseases/immunology , DNA Primers/chemistry , Genetic Vectors , Humans , Immunoglobulin Heavy Chains/genetics , Immunoglobulin Heavy Chains/immunology , Immunoglobulin Light Chains/genetics , Immunoglobulin Light Chains/immunology , Immunoglobulin Variable Region/genetics , Polymerase Chain Reaction/methods
15.
J Neuroimaging ; 18(1): 62-5, 2008 Jan.
Article in English | MEDLINE | ID: mdl-18190498

ABSTRACT

BACKGROUND: To determine and characterize the prevalence of cerebral changes on MRI in patients with antiphospholipid syndrome (APLS) within systemic lupus erythematosus (SLE). METHODS: Seventy-one patients with SLE were prospectively studied with brain MRI: 32 with definite APLS and 39 without. Atrophy, ventricular enlargement, leukoaraiosis, interuncal distance, Evans' index, infarcts, and white matter hyperintensities (WMH) were analyzed. Demographic data, treatment, and SLE activity were analyzed. RESULTS: Groups were similar in age (32.4 vs. 32.8 years old; P= non-significant [NS]), and gender. Duration of disease was longer in patients with APLS (87.3 vs. 55.4 months; P= .064). Cortical atrophy was common in both groups (68.7% vs. 89.7%; P= NS). Leukoaraiosis was present in only 3 patients (9.4%; P= .08), all in the APLS group. WMH were found in more than 40% of the patients from both groups. Infarcts (21.9% vs. 2.6%; P= .019) and infarcts plus WHM (12.5% vs. 0; P= .037) were more common in patients with APLS. CONCLUSIONS: Although a higher prevalence of neurological involvement in SLE has been reported in APLS patients, we found gross brain changes to be similar between groups. Strokes and leukoaraiosis were more common in the APLS group, consistent with the idea of an APLS-induced prothrombotic state.


Subject(s)
Antiphospholipid Syndrome/pathology , Brain/pathology , Magnetic Resonance Imaging/methods , Adolescent , Adult , Antiphospholipid Syndrome/etiology , Case-Control Studies , Chi-Square Distribution , Female , Humans , Lupus Erythematosus, Systemic/complications , Male , Middle Aged , Predictive Value of Tests , Prevalence , Prospective Studies , Statistics, Nonparametric
16.
Autoimmun Rev ; 6(3): 169-75, 2007 Jan.
Article in English | MEDLINE | ID: mdl-17289553

ABSTRACT

Recent studies demonstrated an IL-17-producer CD4+ T cell subpopulation, termed Th17, distinct from Th1 and Th2. It represents a different pro-inflammatory Th-cell lineage. This notion is supported by gene-targeted mice studies. Mice lacking IL-23 (p19-/-) do not develop experimental autoimmune encephalomyelitis (EAE) or collagen-induced arthritis (CIA), while knockout mice for the Th1 cytokine IL-12 (p35-/-) strongly develop both autoimmune diseases. Disease resistance by IL-23 knockout mice correlates well with the absence of IL-17-producing CD4(+) T lymphocytes in target organs despite normal presence of antigen-specific-IFN-gamma-producing Th1 cells. This finding may thus explain previous contradictory reports showing that anti-IFN-gamma-treated mice, IFN-gamma- or IFNR-deficient mice develop CIA or EAE. TGF-beta, IL-6 and IL-1 are the differentiation factors of Th17 cells. IL-23 is dispensable for this function, but necessary for Th17 expansion and survival. The master regulator that directs the differentiation program of Th17 cells is the orphan nuclear receptor RORgammat. IL-27, a member of the IL-12/IL-23 family, potently inhibits Th17 development. Evidence suggesting rheumatoid arthritis and multiple sclerosis as primarily IL-17 autoimmune inflammatory-mediated diseases is rapidly accumulating. The IL-17/23 axis of inflammation and related molecules may rise as therapeutic targets for treating these and perhaps other autoimmune diseases.


Subject(s)
Arthritis, Rheumatoid/immunology , Autoimmune Diseases/immunology , T-Lymphocytes, Helper-Inducer/immunology , Animals , Arthritis, Experimental/immunology , Autoimmune Diseases/genetics , Cell Differentiation , Cell Lineage , Gene Targeting , Humans , Interleukin-17/deficiency , Interleukin-17/genetics , Interleukin-17/immunology , Interleukin-23/deficiency , Interleukin-23/genetics , Interleukin-23/immunology , Mice , Mice, Knockout , Models, Immunological , Receptors, Retinoic Acid/metabolism , Receptors, Thyroid Hormone/metabolism
17.
J Autoimmun ; 26(1): 57-65, 2006 Feb.
Article in English | MEDLINE | ID: mdl-16330187

ABSTRACT

The molecular structure of antibodies associated with autoimmune thrombosis is beginning to be understood. We describe the binding specificities and sequence analysis of anti-beta2-glycoprotein-I (anti-beta2GP-I) or anti-prothrombin (anti-PT) antibody fragments generated by phage display from a patient with primary antiphospholipid syndrome (APS). We obtained 39 positive clones, two that had the correct size reacted with beta2GP-I (Beta 1 and Beta 2). Ten clones with the same restrictive pattern recognized PT (Prot 1) and cross-reacted with beta2GP-I. All three clones recognized anionic and zwitterionic phospholipids. The V(H) regions of both anti-beta2GP-I clones are members of the VH4 family. Prot 1 has a V(H) segment of the VH3 family. The Beta 1 J(H) segments are J(H)5b and J(H)4b for Beta 2 and Prot 1. V(L) genes are V(lambda)1, 3 and 1, respectively. No J(L) was identified for Beta 1, while Beta 2 and Prot 1 carry J(lambda)3b genes. Beta 1 and Beta 2 carry highly conserved germ-line V(H) and V(L) genes. Mutations of the Prot 1 gene appear to be antigen-dependent, most are hotspot mutations located in the CDR 1 and 2 regions. Our work suggests that some anti-beta2GP-I from patients with primary APS are natural autoantibodies. Our work may also help to explain the frequent coexistence of anti-beta2GP-I and anti-PT in the same patient.


Subject(s)
Antibodies, Monoclonal/chemistry , Antiphospholipid Syndrome/immunology , Glycoproteins/immunology , Peptide Library , Prothrombin/immunology , Amino Acid Sequence , Amino Acid Substitution , Antibodies, Monoclonal/genetics , Antibody Specificity , Conserved Sequence , Humans , Immunoglobulin Variable Region/chemistry , Immunoglobulin Variable Region/genetics , Molecular Sequence Data , Mutation , Phospholipids/immunology , beta 2-Glycoprotein I
18.
Rev Invest Clin ; 57(4): 563-71, 2005.
Article in English | MEDLINE | ID: mdl-16315641

ABSTRACT

INTRODUCTION: We investigated the activated protein C resistance (APCR) phenotype and the lupus anticoagulant (LA), activity induced by anti-beta2-glycoprotein-I (anti-beta2GP-I) antibodies. PATIENTS AND METHODS: We studied plasma and sera samples from 29 patients with persistently positive anti-beta2GP-I: 22 with thrombosis (12 with primary APS, 10 with APS secondary to SLE) and seven without thrombosis (all with SLE); 25 healthy subjects were studied as controls. We detected anticardiolipin antibodies (ACA); IgG (and its subclasses) and IgM anti-beta2GP-I, on irradiated and non-irradiated plates by ELISA. APCR was assessed by the activated partial thromboplastin time (APTT)-based assay and by the modified test. The FV Leiden mutation was studied by PCR. LA determination included screening and confirmatory dRVVT. Serum anti-2 GP-I were affinity purified on sepharose columns and their isotype, subclass, and reactivity against various antigens were studied by ELISA. RESULTS: We found that titers of IgG anti-beta2GP-I on irradiated plates were higher than on non-irradiated plates (p = 0.002), IgG2 was the predominant subclass. Fifteen patients (13 with thrombosis) had LA and 15 (also 13 with thrombosis) induced the APCR phenotype. Eleven (all with thrombosis) had both. Two patients were heterozygous for the Leiden mutation. Two purified antibodies, monospecific for beta2GP-I, induced an in vitro APCR phenotype and LA activity. CONCLUSIONS: Our results seem to indicate that the inhibition of the APC anticoagulant function by IgG2 anti-beta2GP-I with LA activity may be one of the responsible mechanisms of thrombophilia in patients with APS.


Subject(s)
Activated Protein C Resistance/immunology , Autoantibodies/immunology , Glycoproteins/immunology , Immunoglobulin G/pharmacology , Lupus Coagulation Inhibitor/blood , Thrombophilia/immunology , Thrombosis/etiology , Activated Protein C Resistance/etiology , Adult , Aged , Antibodies, Anticardiolipin/blood , Antibody Specificity , Antiphospholipid Syndrome/blood , Antiphospholipid Syndrome/immunology , Autoantibodies/isolation & purification , Autoantigens/immunology , Autoimmune Diseases/blood , Autoimmune Diseases/immunology , Enzyme-Linked Immunosorbent Assay , Factor V/analysis , Factor V/genetics , Female , Humans , Immunoglobulin G/immunology , Immunoglobulin G/isolation & purification , Immunoglobulin M/pharmacology , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/complications , Lupus Erythematosus, Systemic/immunology , Male , Middle Aged , Partial Thromboplastin Time , Phenotype , Plasma , Plastics/radiation effects , Prothrombin Time , Thrombophilia/blood , Thrombophilia/etiology , Thrombophilia/genetics , Thrombosis/blood , Thrombosis/genetics , Thrombosis/immunology , beta 2-Glycoprotein I
20.
Rev. invest. clín ; 57(4): 563-571, jul.-ago. 2005. ilus, tab
Article in English | LILACS | ID: lil-632418

ABSTRACT

ABSTRACT Introduction. We investigated the activated protein C resistance (APCR) phenotype and the lupus anticoagulant (LA), activity induced by anti-β2-glycoprotein-I (anti-β2GP-I) antibodies. Patients and methods. We studied plasma and sera samples from 29 patients with persistently positive anti-β2GP-I: 22 with thrombosis (12 with primary APS, 10 with APS secondary to SLE) and seven without thrombosis (all with SLE); 25 healthy subjects were studied as controls. We detected anticardiolipin antibodies (ACA); IgG (and its subclasses) and IgM anti-β2GP-I, on irradiated and non-irradiated plates by ELISA. APCR was assessed by the activated partial thromboplastin time (APTT)-based assay and by the modified test. The FV Leiden mutation was studied by PCR. LA determination included screening and confirmatory dRVVT. Serum anti-β2GP-I were affinity purified on sepharose columns and their isotype, subclass, and reactivity against various antigens were studied by ELISA. Results. We found that titers of IgG anti-β2GP-I on irradiated plates were higher than on non-irradiated plates (p = 0.002), IgG2 was the predominant subclass. Fifteen patients (13 with thrombosis) had LA and 15 (also 13 with thrombosis) induced the APCR phenotype. Eleven (all with thrombosis) had both. Two patients were heterozygous for the Leiden mutation. Two purified antibodies, monospecific for β2GP-I, induced an in vitro APCR phenotype and LA activity. Conclusions. Our results seem to indicate that the inhibition of the APC anticoagulant function by IgG2 anti-β2GP-I with LA activity may be one of the responsible mechanisms of thrombophilia in patients with APS.


Introducción. Investigamos la resistencia a la proteína C activada (RPCA) y la actividad de anticoagulante lápico (AL), inducidas por anticuerpos anti-β2-glicoproteína-I (anti-β2GP-I). Pacientes y métodos. Estudiamos los plasmas y sueros persistentemente positivos para anti-β2GP-I de 29 pacientes: 22 tuvieron trombosis (12 con síndrome de antifosfolípidos (SAF) primario y 10 con SAF secundario a lupus erítematoso generalizado (LEG)) y siete sin trombosis (todos con LEG). Como controles estudiamos 25 sueros de personas clínicamente sanas. Detectamos anticuerpos anticardiolipina, anti-β2GP-I IgG (y sus subclases) e IgM por ELISA en placas irradiadas y no irradiadas. Evaluamos la RPCA por medio del tiempo parcial de tromboplastina activada y por la prueba modificada. Estudiamos la mutación FV de Leiden por PCR y el anticoagulante lápico con el método de dRVVT screening y confirmatorio. Después de purificar los anti-β2GP-I séricos con una columna de antígeno unido a sefarosa, analizamos por ELISA sus isotipos, subclases y reactividad contra β2GP-I y algunos fosfolípidos. Resultados. Los títulos de anti-β2GP-I IgG fueron más altos en placas irradiadas que en no irradiadas (p = 0.002), predominó la subclase IgG2. Quince plasmas (13 de pacientes con trombosis) tuvieron AL y 15 (13 también de pacientes con trombosis) indujeron el fenotipo de RPCA. Once plasmas (todos de pacientes con trombosis) indujeron ambas actividades. Dos pacientes fueron heterocigotos para la mutación de Leiden. Dos anticuerpos purificados monoespecíficos para β2GP-I indujeron el fenotipo de la RPCA y la actividad de AL in vitro. Conclusiones. Nuestros resultados sugieren que la RPCA, inducida por los anti-β2GP-I que concomitantemente tienen actividad de AL, puede tener implicaciones patogénicas en la trombofílía del SAF.


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Activated Protein C Resistance/immunology , Autoantibodies/immunology , Glycoproteins/immunology , Immunoglobulin G/pharmacology , Lupus Coagulation Inhibitor/blood , Thrombophilia/immunology , Thrombosis/etiology , Antibody Specificity , Activated Protein C Resistance/etiology , Antibodies, Anticardiolipin/blood , Antiphospholipid Syndrome/blood , Antiphospholipid Syndrome/immunology , Autoantibodies/isolation & purification , Autoantigens/immunology , Autoimmune Diseases/blood , Autoimmune Diseases/immunology , Enzyme-Linked Immunosorbent Assay , Factor V/analysis , Factor V/genetics , Immunoglobulin G/immunology , Immunoglobulin G/isolation & purification , Immunoglobulin M/pharmacology , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/complications , Lupus Erythematosus, Systemic/immunology , Partial Thromboplastin Time , Phenotype , Plasma , Prothrombin Time , Plastics/radiation effects , Thrombophilia/blood , Thrombophilia/etiology , Thrombophilia/genetics , Thrombosis/blood , Thrombosis/genetics , Thrombosis/immunology
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