ABSTRACT
Intra-operative acute hypersensitivity reactions require a decision to be made regarding whether to proceed with or abandon the planned surgical procedure once the patient has stabilised. Using retrospective case controls, we examined all cases (223) of proven acute hypersensitivity reactions from 2005 to 2014 in Western Australia, in which the syndrome was recognised by the treating clinician before or during surgery, to determine whether recovery outcomes were adversely affected by proceeding with the planned procedure. Surgery proceeded in 104 patients (47%) and was abandoned in 119 (53%). The severity of acute hypersensitivity reactions was Société Française d'Anesthésie et de Réanimation grade 1 or 2 in 56 patients (25%), grade 3 in 128 (56%) and grade 4 in 39 (17%). Abandoning surgery was more common in patients with increasing severity of hypersensitivity. The rate of major hypersensitivity-related complications for all patients was zero for grade 1 and 2 reactions, 4.7% for grade 3 and 12.8% for grade 4. There were no deaths. Patients in whom surgery was completed were not observed to have a higher frequency of major hypersensitivity-related complications when compared with cases of similar severity in whom surgery was abandoned. For patients admitted to the intensive care unit, proceeding with surgery was not associated with an increased duration of mechanical ventilation of the lungs. Our results suggest that, once initial resuscitation has been achieved and if resuscitative efforts can be re-instituted if required, continuing with planned surgery in grade 1, 2 and 3 immediate hypersensitivity was not associated with poorer outcomes. After grade 3 reactions, there was a significant incidence of complications attributable to acute hypersensitivity regardless of whether surgery proceeded or was abandoned. Surgery was frequently abandoned in grade 4 immediate hypersensitivity and was associated with a high rate of complications.
Subject(s)
Anaphylaxis/complications , Drug Hypersensitivity/complications , Intraoperative Complications/chemically induced , Resuscitation , Surgical Procedures, Operative , Acute Disease , Aged , Anaphylaxis/therapy , Case-Control Studies , Drug Hypersensitivity/therapy , Female , Humans , Intraoperative Complications/therapy , Male , Middle Aged , Retrospective Studies , Treatment Outcome , Western AustraliaABSTRACT
BACKGROUND: RA patients have a higher incidence of cardiovascular diseases compared to the general population. Serum amyloid A (SAA) is an acute-phase protein, upregulated in sera of RA patients. AIM: To determine the effects of medications on SAA-stimulated human coronary artery endothelial cells (HCAEC). METHODS: HCAEC were preincubated for 2 h with medications from sterile ampules (dexamethasone, methotrexate, certolizumab pegol, and etanercept), dissolved in medium (captopril) or DMSO (etoricoxib, rosiglitazone, meloxicam, fluvastatin, and diclofenac). Human recombinant apo-SAA was used to stimulate HCAEC at a final 1000 nM concentration for 24 hours. IL-6, IL-8, sVCAM-1, and PAI-1 were measured by ELISA. The number of viable cells was determined colorimetrically. RESULTS: SAA-stimulated levels of released IL-6, IL-8, and sVCAM-1 from HCAEC were significantly attenuated by methotrexate, fluvastatin, and etoricoxib. Both certolizumab pegol and etanercept significantly decreased PAI-1 by an average of 43%. Rosiglitazone significantly inhibited sVCAM-1 by 58%. CONCLUSION: We observed marked influence of fluvastatin on lowering cytokine production in SAA-activated HCAEC. Methotrexate showed strong beneficial effects for lowering released Il-6, IL-8, and sVCAM-1. Interesting duality was observed for NSAIDs, with meloxicam exhibiting opposite-trend effects from diclofenac and etoricoxib. This represents unique insight into specific responsiveness of inflammatory-driven HCAEC relevant to atherosclerosis.
Subject(s)
Coronary Vessels/cytology , Endothelial Cells/metabolism , Serum Amyloid A Protein/pharmacology , Endothelial Cells/cytology , Endothelial Cells/drug effects , Enzyme-Linked Immunosorbent Assay , Humans , Interleukin-6/metabolism , Interleukin-8/metabolism , Plasminogen Activator Inhibitor 1/metabolism , Vascular Cell Adhesion Molecule-1/metabolismABSTRACT
Association of vitamin D receptor (VDR) gene polymorphisms with sepsis risk and mortality was studied. VDR FokI CC genotype was associated with increased sepsis risk (OR = 13.396, p = .000009) compared to the TT genotype. Results suggest possible role of VDR FokI (rs2228570) as a molecular biomarker of increased sepsis risk.
Subject(s)
Genetic Association Studies , Genetic Predisposition to Disease , Receptors, Calcitriol/genetics , Sepsis/genetics , Adult , Aged , Alleles , Female , Genotype , Haplotypes , Humans , Male , Middle Aged , Polymorphism, Single Nucleotide , Risk Factors , Sepsis/mortality , Sepsis/physiopathologyABSTRACT
AIM: Evaluation of secondary hyperparathyroidism (SHPT) and its prognostic impact on all-cause mortality in elderly males with heart failure (HF). METHODS: Seventy three males (67 ± 7 years old) with systolic HF were included. Baseline PTH was measured. Patients were grouped according to PTH cut-off levels of 65 pg/ml (>65 pg/ml = SHPT vs. normal PTH). All-cause mortality was evaluated at 6-year follow-up. RESULTS: SHPT was diagnosed in 43 (59 %) patients. They were more severe compared to the patients with normal PTH regarding NYHA functional class (2.4 ± 0.5 vs. 2.1 ± 0.2, p = 0.001), quality of life score (34 ± 14 vs. 24 ± 12, p = 0.005), 6-min walking distance (378 ± 79 vs. 446 ± 73 m, p < 0.0001), left ventricular ejection fraction (27 ± 8 vs. 31 ± 7 %, p = 0.019), and NT-proBNP [2452 (3399) vs. 918 (1372) pg/ml, p < 0.0001]. No differences in age, vitamin D status, and renal function were noted between studied groups. A total of 41 (56 %) patients died within 6 years of follow-up. Kaplan-Meier survival analysis showed impaired long-term survival in patients with SHPT versus patients with normal PTH (p = 0.009). The rate of death was highest (75 %) in the group of patients with SHPT and NT-proBNP levels above median value (p = 0.003). Cox regression analysis demonstrated that NT-proBNP was the single independent predictor of all-cause mortality at 6-year follow-up [HR 3.698 (1.927-7.095), p < 0.0001]. CONCLUSION: SHPT was highly prevalent in elderly males with HF and was associated with impaired survival. HF patients with SHPT had more severe disease compared to the patients with normal serum PTH. Determination of serum PTH levels provided additional value to NT-proBNP for risk stratification in these patients.
Subject(s)
Heart Failure/physiopathology , Hyperparathyroidism, Secondary/epidemiology , Quality of Life , Aged , Biomarkers/metabolism , Heart Failure/complications , Humans , Hyperparathyroidism, Secondary/diagnosis , Hyperparathyroidism, Secondary/metabolism , Male , Middle Aged , Prevalence , Prognosis , ROC Curve , Serbia/epidemiology , Survival RateABSTRACT
Oxidative stress has been shown to play a role in modifying antibodies in favor of higher auto-immunoreactivity. We studied the immunoreactivity of oxidized IgG (oxIgG) to ß2-glycoprotein I (ß2GPI), six peptide sequences corresponding to amino acid clusters on its different domains, to determine their effects on human coronary artery endothelial cells (HCAEC). Human IgG was purified from seven donors, electro-oxidized and checked for immunoreactivity and avidity to ß2GPI and to peptides by ELISA. Conformational stability and antibody-antigen complex formation of oxIgG was analyzed by fluorescence spectroscopy and dynamic light scattering. Resting and activated sub-confluent HCAEC were stimulated with oxIgG or IgG. Secreted cytokines were measured by ELISA. Immunoreactivity of seven oxIgG samples increased to 7.5-fold against ß2GPI and to 3.8-fold against six peptides as compared to IgG. oxIgG showed low avidity "properties." Conformational changes and exposure of protein hydrophobic regions were confirmed by an elevation in fluorescence (2.4- to 5.0-fold) on bis-ANS dye binding to oxIgG. oxIgG significantly elevated the release of GROα and IL-8 in resting and activated states of HCAEC. Oxidation alters IgG in favor of autoreactivity toward whole ß2GPI and corresponding peptides on different domains of ß2GPI and could lead to dysfunction of arterial endothelium by upregulation of chemokines.
Subject(s)
Endothelial Cells/cytology , Immunoglobulin G/isolation & purification , Oxidative Stress , Peptides/immunology , beta 2-Glycoprotein I/immunology , Antigen-Antibody Reactions , Autoantibodies/immunology , Cells, Cultured , Coronary Vessels/cytology , Cytokines/immunology , Dynamic Light Scattering , Healthy Volunteers , Humans , Immunoglobulin G/metabolismABSTRACT
Antiprothrombin antibodies can be measured by ELISA using either a prothrombin/phosphatidylserine complex (aPS/PT) or prothrombin alone (aPT) as antigen. We aimed to compare the clinical features of autoimmune patients with avidity of aPS/PT and determine the diagnostic efficiency of aPS/PT and aPT for assessing antiphospholipid syndrome (APS). aPS/PT were of low (n = 9), heterogeneous (n = 31) and high (n = 8) avidity out of 48 cases. None of the samples with low avidity were positive in aPT ELISA. Among patients with heterogeneous or high avidity aPS/PT, there was a significantly greater number of patients with APS as compared to patients with low avidity (38/39 vs. 7/9; p < 0.05). No SLE patients had high avidity antiprothrombin antibodies.
Subject(s)
Antiphospholipid Syndrome/diagnosis , Autoantibodies/blood , Phosphatidylserines/immunology , Prothrombin/immunology , Antibody Affinity , Antiphospholipid Syndrome/blood , Antiphospholipid Syndrome/immunology , Biomarkers/blood , Enzyme-Linked Immunosorbent Assay , Humans , Lupus Erythematosus, Systemic/blood , Lupus Erythematosus, Systemic/immunologyABSTRACT
Antibodies against ß(2)-glycoprotein I (anti-ß(2)GPI) are one of the hallmarks of the antiphospholipid syndrome (APS). However, they are heterogenic regarding their epitope specificity, pathogenic mechanisms and their avidity. In the current study we present some outstanding issues about avidity of anti-ß(2)GPI antibodies. Our results confirmed that high avidity anti-ß(2)GPI are associated with thrombosis and APS, while in low avidity anti-ß(2)GPI group non-APS (predominantly systemic lupus erythematosus) patients prevailed.
Subject(s)
Antibody Affinity , Antiphospholipid Syndrome/immunology , Autoantibodies/blood , beta 2-Glycoprotein I/immunology , Adult , Female , Humans , MaleABSTRACT
BACKGROUND: We analysed the outcomes of 726 cases of primary head and neck cancer patients managed between 1996 and 2008, including those managed in the multidisciplinary clinic or team setting (MDT) and those managed outside of an MDT by individual disciplines (non-MDT) in the same institution. METHODS: Data were collected from the Hospital Based Cancer Registry and a database within the Head and Neck Cancer Clinic. Univariable comparisons and multivariable analyses were performed using a logistic regression model. Survival by staging was analysed. Comparisons of management and outcomes were made between MDT and non-MDT patients. RESULTS: 395 patients (54%) had been managed in the MDT vs 331 patients (46%) non-MDT. MDT patients were more likely to have advanced disease (likelihood ratio χ(2)=44.7, P<0.001). Stage IV MDT patients had significantly improved 5-year survival compared with non-MDT patients (hazard ratio=0.69, 95% CI=0.51-0.88, P=0.004) and more synchronous chemotherapy and radiotherapy (P=0.004), and the non-MDT group had more radiotherapy as a single modality (P=0.002). CONCLUSIONS: The improved survival of MDT-managed stage IV patients probably represents both the selection of multimodality treatment and chemotherapeutic advances that these patients received in a multidisciplinary team setting by head and neck cancer specialists as opposed to cancer generalists in a non-MDT setting.
Subject(s)
Carcinoma/therapy , Combined Modality Therapy/methods , Head and Neck Neoplasms/therapy , Interdisciplinary Communication , Antineoplastic Combined Chemotherapy Protocols/therapeutic use , Carcinoma/mortality , Carcinoma/pathology , Combined Modality Therapy/trends , General Surgery/statistics & numerical data , Head and Neck Neoplasms/mortality , Head and Neck Neoplasms/pathology , Humans , Neoplasm Staging , Radiotherapy/statistics & numerical data , Retrospective Studies , Survival Analysis , Time Factors , Treatment OutcomeABSTRACT
OBJECTIVE: The objective of this study was to extend the findings of the preliminary study by measuring the avidity of IgG anti-ß2-glycoprotein I antibodies (anti-ß2-GPI) on a larger group of patients with primary or secondary antiphospholipid syndrome (APS) and anti-ß2-GPI positive patients without APS in the frame of the European Forum on antiphospholipid antibodies (aPL). METHODS: Serum from 137 patients with primary APS, APS associated with autoimmune diseases, and patients with autoimmune diseases other than APS from five EU rheumatology centres were tested for anti-ß2-GPI antibodies. The 109 patients who were sera positive for anti-ß2-GPI by the in-house anti-ß2-GPI enzyme-linked immunosorbent assay (ELISA) at the Immunology Laboratory, UMC Ljubljana were selected for further testing on avidity with chaotropic anti-ß2-GPI ELISA. RESULTS: High, low and heterogeneous avidity IgG anti-ß2-GPI was found in 32/109, 17/109 and 60/109 patients respectively. Significantly more patients with APS were in the high avidity than in the low avidity anti-ß2-GPI group, while the opposite was observed for non-APS (both p < 0.001). The most common clinical feature among patients with high avidity anti-ß2-GPI was thrombosis, mainly due to venous thrombosis (p < 0.01 and p < 0.001, versus low avidity anti-ß2-GPI group). CONCLUSION: Patients with or without APS had anti-ß2-GPI of high, low or heterogeneous avidity. High avidity anti-ß2-GPI was associated with thrombosis and APS, while in the low avidity anti-ß2-GPI group non-APS (predominantly SLE) patients prevailed. Determination of anti-ß2-GPI avidity should be considered in the analytical strategies for further differentiation of patients with anti-ß2-GPI antibodies.
Subject(s)
Antibodies, Antiphospholipid/blood , Antiphospholipid Syndrome/immunology , Autoantibodies/blood , beta 2-Glycoprotein I/immunology , Adolescent , Adult , Aged , Antibody Affinity , Child , Europe , Female , Humans , Immunoglobulin G/blood , Male , Middle Aged , Young AdultABSTRACT
High PTH levels have been reported in patients with chronic heart failure (CHF). Similarly, its levels increase with aging and are related to impaired survival in elderly adults. However, its relationship with neuroendocrine activation and endothelial dysfunction in CHF has not been previously studied. Seventy-three CHF males with New York Heart Association (NYHA) classes II and III and 20 control subjects aged ≥ 55 yr were recruited. PTH, 25-hydroxyvitamin D [25(OH)D], N-terminal pro-brain natriuretic peptide (NT-pro-BNP), adiponectin, and osteoprotegerin were measured. Endothelial function (brachial flow mediated dilation), echocardiography, physical performance, and quality of life were assessed, as well. CHF patients had markedly increased serum PTH (77 ± 33 vs 40 ± 11 pg/ml, p<0.0001), NT-pro-BNP [1809 (2742) vs 67 (74) pg/ml, p<0.0001], adiponectin (17 ± 9 vs 10 ± 2 µg/ml, p<0.0001), osteoprotegerin, whereas 25(OH)D levels were decreased compared to controls. Increased PTH is positively correlated with NTpro- BNP (r=0.399, p<0.0001), adiponectin (r=0.398, p<0.0001), and osteoprotegerin, whereas negatively with 25(OH)D in CHF patients. Additionally, increased serum PTH was associated with endothelial dysfunction, echocardiographic variables of heart failure progression, impaired physical performance, and deteriorated quality of life. In a multivariate linear regression analysis, increased serum PTH was independently associated with neuroendocrine activation (NT-pro-BNP, adiponectin) and endothelial dysfunction in elderly CHF men (R2=0.455). Additionally, demonstrated relations with other well-established variables of heart failure severity suggest the potential role of serum PTH in the pathogenesis and non-invasive monitoring of heart failure progression. Future studies are needed to evaluate the predictive value of serum PTH for clinical outcomes as well as beneficial potential of PTH suppression in CHF patients.
Subject(s)
Endothelium, Vascular/physiopathology , Heart Failure/blood , Heart Failure/physiopathology , Neurosecretory Systems/physiology , Parathyroid Hormone/blood , Adiponectin/blood , Aged , Chronic Disease , Humans , Male , Middle Aged , Natriuretic Peptide, Brain/blood , Osteoprotegerin/blood , Peptide Fragments/blood , ROC Curve , Vascular Diseases/blood , Vascular Diseases/physiopathology , Vitamin D/bloodABSTRACT
It is generally assumed that cerebrospinal fluid (CSF) is secreted in the brain ventricles, and so after an acute blockage of the aqueduct of Sylvius an increase in the ventricular CSF pressure and dilation of isolated ventricles may be expected. We have tested this hypothesis in cats. After blocking the aqueduct, we measured the CSF pressure in both isolated ventricles and the cisterna magna, and performed radiographic monitoring of the cross-sectional area of the lateral ventricle. The complete aqueductal blockage was achieved by implanting a plastic cannula into the aqueduct of Sylvius through a small tunnel in the vermis of the cerebellum in the chloralose-anesthetized cats. After the reconstitution of the occipital bone, the CSF pressure was measured in the isolated ventricles via a plastic cannula implanted in the aqueduct of Sylvius and in the cisterna magna via a stainless steel cannula. During the following 2 h, the CSF pressures in the isolated ventricles and cisterna magna were identical to those in control conditions. We also monitored the ventricular cross-sectional area by means of radiography for 2 h after the aqueductal blockage and failed to observe any significant changes. When mock CSF was infused into isolated ventricles to imitate the CSF secretion, the gradient of pressure between the ventricle and cisterna magna developed, and disappeared as soon as the infusion was terminated. However, when mock CSF was infused into the cisterna magna at various rates, the resulting increased subarachnoid CSF pressure was accurately transmitted across the brain parenchyma into the CSF of isolated ventricles. The lack of the increase in the CSF pressure and ventricular dilation during 2 h of aqueductal blockage suggests that aqueductal obstruction by itself does not lead to development of hypertensive acute hydrocephalus in cats.
Subject(s)
Cerebral Aqueduct/physiopathology , Cerebral Ventricles/pathology , Cerebrospinal Fluid Pressure/physiology , Cerebrospinal Fluid/physiology , Animals , Catheterization/adverse effects , Cats , Cerebral Ventricles/physiopathology , Cerebral Ventriculography/methods , Cisterna Magna/physiopathology , Dilatation, Pathologic/cerebrospinal fluid , Female , Flow Injection Analysis , Hydrocephalus/cerebrospinal fluid , Hydrocephalus/etiology , Male , Models, Animal , Time FactorsABSTRACT
Autoantibodies against dsDNA are utilized for the diagnosis and prognosis of SLE as they are highly specific and correlate with disease activity/renal involvement. However, different detection methods are used in routine diagnostic laboratories. Farr radioimmunoassay (Farr-RIA) has been designated as the preferred method, since it provides very specific and at the same time quantitative results, enabling follow-up of level variations over time. Using intercalating fluorescent dsDNA dye would enable all the benefits of Farr-RIA without the radioactive material and organic solvents. To develop a modified fluorescent Farr method (Farr-FIA) and compare it to the classical Farr-RIA in regard to laboratory parameters, as well as clinical utility. Assays were tested on sera of 70 SLE patients, 78 other autoimmune patients, and 145 healthy blood donors. DNA for Farr-FIA was isolated from healthy donor, for Farr-RIA, 14C-labeled dsDNA from E. coli was used and mixed with sera in borate-buffered saline, followed by precipitation with saturated ammonium sulfate solution and centrifugation. The supernatant (S) was separated from the precipitate (P), and content of dsDNA was measured with PicoGreen (Invitrogen) in Farr-FIA or radioactive isotope in scintillation solution in Farr-RIA. The results were calculated as a ratio (P-S)/(P+S). Farr-FIA has a diagnostic sensitivity of 53% and diagnostic specificity of 100% (ROC AUC 0.781). Good correlation and agreement were shown between Farr-RIA and Farr-FIA. Also, there is good correlation between Farr-FIA and SLEDAI, comparable to that of Farr-RIA. Farr-FIA differs from Farr-RIA in the changed detection system yielding comparable results and thus could represent a nonradioactive replacement for Farr-RIA.
Subject(s)
Antibodies, Antinuclear/blood , Lupus Erythematosus, Systemic/blood , Radioimmunoprecipitation Assay/methods , Adult , Antibodies, Antinuclear/analysis , Cross-Sectional Studies , DNA/immunology , Diagnostic Tests, Routine , Female , Humans , Linear Models , Lupus Erythematosus, Systemic/diagnosis , MaleABSTRACT
AIMS: Oxidation reactions can modify protein activity or specificity. Recently, a novel redox-reactive family of autoantibodies was described, which indicated involvement of altered antibodies (beside altered antigens) into autoimmune reactions. The aim of our study was to determine the binding capacity alterations of electro-oxidized blood donors' IgGs, and to evaluate their effects on released proinflammatory interleukin 6 in HUVEC. RESULTS: We found out that 1.) Isolated blood donor IgGs bound after electro-oxidation to beta2-glycoprotein I, cardiolipin, citrullinated cyclic peptide and protein 3 by enzyme-linked immunosorbent assay, extractable nuclear antigens by counterimmuno-electrophoresis, and cell antigens by indirect immunofluorescence; 2.) Alterations in immunoreactivity of IgGs due to oxidation highly depend on electric current, time of exposure and the presence of antioxidants, 3.) Treatment of HUVEC with oxidized IgGs resulted in changed cell morphology, accompanied by an increase in released interleukin-6. CONCLUSIONS: Our data suggest repeatable transformation of antibodies present in the blood of healthy persons and patients. Inter-individual differences in chemical stability of antibodies, patient's antioxidant status, and the microenvironmental changes at the cellular level may influence the range of antibody alterations and their involvement in pathophysiological autoimmune processes.
Subject(s)
Autoantibodies/immunology , Immunoglobulin G/immunology , Interleukin-6/biosynthesis , Antibody Specificity , Antioxidants/pharmacology , Autoantibodies/blood , Autoantibodies/metabolism , Blood Donors , Endothelium, Vascular/cytology , Endothelium, Vascular/immunology , Humans , Immunoglobulin G/blood , Immunoglobulin G/metabolism , Inflammation Mediators/metabolism , Interleukin-6/genetics , Kinetics , Oxidation-Reduction , RNA, Messenger/metabolismABSTRACT
The immune response may be changed due to altered proteins or modifications of immunoglobulins, including oxidative processes. The susceptibility to oxidative modifications depends greatly on amino-acid moiety composition due to chemical characteristics (instability) of their side-chains. Initial steps of oxidation may change the specificity and avidity of immunoglobulins due to chemical alteration of the hypervariable region. The oxidation of antibodies increases the hydrophilic nature of the paratopes and makes them more susceptible for the binding to cationic surfaces even without the strong surface-to-surface fitting. The electro-oxidation of IgG significantly changes the immunoreactivity and specificity of IgG fractions, regardless of the initial immunoreactivity to a specific autoantigen also in healthy persons. Data are presented on changes in the immunoreactivity as well as the avidity of antibodies against beta2-glycoprotein I after being exposed to direct current. ELISA measurements showed increased reactivity of anti-beta2-glycoprotein I antibodies at the beginning and various, fluctuating results after prolonged exposure to electro-oxidation. Inter-individual differences in chemical stability of immunoglobulins and patient's antioxidative status may influence the range of their alterations and their impact on health/disease balance.
Subject(s)
Antibody Affinity/immunology , Antibody Specificity/immunology , Autoantibodies/blood , Immunoglobulin G/immunology , beta 2-Glycoprotein I/immunology , Antigen-Antibody Reactions , Humans , Oxidation-ReductionABSTRACT
Vesicle shape transformations caused by decreasing the difference between the equilibrium areas of membrane monolayers were studied on phospholipid vesicles with small volume to membrane area ratios. Slow transformations of the vesicle shape were induced by lowering of the concentration of lipid monomers in the solution outside the vesicle. The complete sequence of shapes consisted of a string of pearls, and wormlike, starfish, discocyte and stomatocyte shapes. The transformation from discocyte to stomatocyte vesicle shapes was analyzed theoretically to see whether these observations accord with the area difference elasticity (ADE) model. The membrane shape equation and boundary conditions were derived for axisymmetrical shapes for low volume vesicles, part of whose membranes are in contact. Calculated shapes were arranged into a phase diagram. The theory predicts that the transition between discocyte and stomatocyte shapes is discontinuous for relatively high volumes and continuous for low volumes. The calculated shape sequences matched well with the observed ones. By assuming a linear decrease of the equilibrium area difference with time, the ratio between the nonlocal and local bending constants is in agreement with reported values.
Subject(s)
Membrane Lipids/chemistry , Phospholipids/chemistry , Elasticity , Lipid BilayersABSTRACT
The terms affinity and avidity are often used indiscriminately, despite clearly differing. Since affinity refers to monovalent binding of antibodies to a monovalent epitope, the majority of data on the binding of anti-beta2-glycoprotein I antibodies (anti-beta2-GPI) characterized their avidity rather than affinity. Anti-beta2-GPI were generally believed to be of low avidity, but heterogeneous avidity of patients' IgG anti-beta2-GPI has been demonstrated. High avidity anti-beta2-GPI monoclonals were reported to possess higher pathogenicity than low avidity anti-beta2-GPI. Polyclonal high avidity anti-beta2-GPI were found to be more common in patients with antiphospholipid syndrome (APS) and associated with thrombosis. Some conformational changes of beta2-GPI are required for the binding of polyclonal anti-beta2-GPI to the antigen: neither high density of the antigen nor high avidity of the anti-beta2-GPI alone is sufficient for the recognition. Avidity of anti-beta2-GPI should be considered in any attempt of inter-laboratory standardisation and/or evaluation of anti-beta2-GPI enzyme-linked immunosorbent assay (ELISA).
Subject(s)
Antibody Affinity , Autoantibodies/chemistry , Autoantibodies/metabolism , Glycoproteins/immunology , Animals , Antibody Affinity/genetics , Enzyme-Linked Immunosorbent Assay/standards , Glycoproteins/genetics , Humans , beta 2-Glycoprotein IABSTRACT
We aimed to evaluate avidity of anti-beta(2)-glycoprotein I antibodies (anti-beta(2)-GPIs) in patients with antiphospholipid syndrome (APS) at the time of acute thrombotic events or pregnancy loss as compared with clinical event-free periods. To do so, 69 sera samples from 16 patients (6 with primary APS and 10 with APS secondary to systemic lupus erythematosus ) were selected on the basis of anti-beta(2)-GPI positivity. Avidity of IgG anti-beta(2)-GPIs was determined by chaotropic enzyme-linked immunosorbent assay (ELISA), using increased NaCl concentration during antibody binding. High, heterogeneous, and low-avidity anti-beta(2)-GPIs were measured in APS patients, with no clear pattern regarding the time of thrombotic events or pregnancy failure. In general, anti-beta(2)-GPI avidity did not change substantially during disease course. We concluded that avidity of anti-beta(2)-GPIs appears to be a rather stable parameter in an individual APS patient. Considering the previously shown association of high-avidity anti-beta(2)-GPIs with venous thrombosis, avidity of anti-beta(2)-GPIs may be a better predictor of predisposition to thrombosis and unsuccessful pregnancy than levels of antiphospholipid antibodies, which may fluctuate over time owing to several factors.
Subject(s)
Abortion, Spontaneous/etiology , Antibody Affinity/immunology , Antiphospholipid Syndrome/immunology , Glycoproteins/immunology , Immunoglobulin G/blood , Thrombosis/etiology , Enzyme-Linked Immunosorbent Assay , Female , Humans , Pregnancy , beta 2-Glycoprotein IABSTRACT
The pathogenicity of antibodies against ß2-glycoprotein I (anti-ß2GPI) depends on multiple factors such as subclass type, epitope binding and avidity. Due to their large heterogeneity, their impact on antiphospholipid syndrome (APS) onset is still not fully clarified. We studied the binding characteristics of IgG anti-ß2GPI with known avidity from sera of 201 autoimmune patients (87 with APS, 67 with APS associated with systemic lupus erythematosus (SLE), 47 with only SLE) to six ß2GPI peptides corresponding to amino acid clusters on domains I-II, II, III and III-IV by indirect ELISA and evaluated their association with clinical features of APS. Peptides A (LKTPRV; domain I-II), B (KDKATF; domain IV) and C (TLRVYK; domain III) were derived from a hexapeptide phage display library previously shown to react with pathogenic monoclonal anti-ß2GPI. Peptides D (NGPANSK; domain III), E (YNPLWFV; domain II) and F (KMDGNHP; domain III-IV) represent surface amino acid clusters on ß2GPI. The percentage of patients positive for peptides were observed as follows: 30.3% for peptide D, 28.90% for B, 25.9% for C, 24.9% for E, 24.4% for F and 10.0% for A. The anti-peptide antibodies in studied serum samples were predominantly of heterogeneous avidity, followed by law avidity anti-peptide antibodies, whereas only a few were of high avidity. Positive and negative correlations were found between several anti-peptide antibodies and the rate of thrombosis. Our results indicated diverse reactivity of IgG anti-ß2GPI to different epitopes on ß2GPI. Classification of IgG anti-ß2GPI into subgroups regarding epitope specificity and avidity could represent an additional tool in understanding their pathogenicity in APS.
Subject(s)
Autoantibodies/immunology , Autoimmune Diseases/immunology , Immunoglobulin G/immunology , Peptides/immunology , beta 2-Glycoprotein I/immunology , Adolescent , Adult , Aged , Aged, 80 and over , Antibody Affinity/immunology , Autoantibodies/blood , Autoantibodies/metabolism , Autoimmune Diseases/blood , Autoimmune Diseases/metabolism , Child , Female , Humans , Immunoglobulin G/metabolism , Male , Middle Aged , Odds Ratio , Peptides/metabolism , Protein Binding/immunology , Young Adult , beta 2-Glycoprotein I/chemistry , beta 2-Glycoprotein I/metabolismABSTRACT
Elevated levels of antiphospholipid antibodies are associated with an increased risk of thrombosis. To establish the prevalence of these antibodies in deep vein thrombosis (DVT), IgG and IgM antibodies to cardiolipin (aCL) and phosphatidylserine (aPS) were determined by enzyme-linked immunosorbent assay in 118 patients with DVT either during an acute episode (N = 53) or at least 2 months after acute DVT (N = 65). Most patients (76%) had proximal leg DVT and no one had evident autoimmune disorder. aCL and aPS values higher than 4 standard deviations above the mean value of the control group (147 blood donors) were considered increased. Increased IgG aCL were observed in 10% of DVT patients (controls: 5%, not significant), increased IgG aPS in 16% of DVT patients (controls: 5%, p less than 0.005) and both types in 4% of DVT patients (controls: 3%, not significant). In the subgroup of 41 patients with previous idiopathic DVT, prevalence of increased IgG aPS was the highest: 27% (p less than 0.001). Increased antibodies of IgM isotype were observed in 3% (aCL) and 2% (aPS) of all DVT patients (controls: 8% and 4%, respectively, not significant). Elevated IgG aCL or aPS were not associated with significant changes in platelet count, antithrombin III and protein C. However, in patients with increased IgG aPS deficient fibrinolysis due to high plasminogen activator inhibitor activity was observed before and after 20 min upper arm venous occlusion. DVT patients with increased IgG aPS might be exposed to a greater risk of rethrombosis due to deficient fibrinolysis than DVT patients without these antibodies.