RESUMEN
The [URE3] yeast prion is the self-propagating amyloid form of the Ure2 protein. [URE3] is cured by overexpression of several yeast proteins, including Ydj1, Btn2, Cur1, Hsp42, and human DnaJB6. To better understand [URE3] curing, we used real-time imaging with a yeast strain expressing a GFP-labeled full-length Ure2 construct to monitor the curing of [URE3] over time. [URE3] yeast cells exhibited numerous fluorescent foci, and expression of the GFP-labeled Ure2 affected neither mitotic stability of [URE3] nor the rate of [URE3] curing by the curing proteins. Using guanidine to cure [URE3] via Hsp104 inactivation, we found that the fluorescent foci are progressively lost as the cells divide until they are cured; the fraction of cells that retained the foci was equivalent to the [URE3] cell fraction measured by a plating assay, indicating that the foci were the prion seeds. During the curing of [URE3] by Btn2, Cur1, Hsp42, or Ydj1 overexpression, the foci formed aggregates, many of which were 0.5 µm or greater in size, and [URE3] was cured by asymmetric segregation of the aggregated seeds. In contrast, DnaJB6 overexpression first caused a loss of detectable foci in cells that were still [URE3] before there was complete dissolution of the seeds, and the cells were cured. We conclude that GFP labeling of full-length Ure2 enables differentiation among the different [URE3]-curing mechanisms, including inhibition of severing followed by seed dilution, seed clumping followed by asymmetric segregation between mother and daughter cells, and seed dissolution.
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Proteínas Fúngicas/metabolismo , Imagen Molecular , Priones/metabolismo , Levaduras/citología , Factores de Tiempo , Levaduras/metabolismoRESUMEN
Prions arise from proteins that have two possible conformations: properly folded and non-infectious or misfolded and infectious. The [PSI+] yeast prion, which is the misfolded and self-propagating form of the translation termination factor eRF3 (Sup35), can be cured of its infectious conformation by overexpression of Hsp104, which helps dissolve the prion seeds. This dissolution depends on the trimming activity of Hsp104, which reduces the size of the prion seeds without increasing their number. To further understand the relationship between trimming and curing, trimming was followed by measuring the loss of GFP-labeled Sup35 foci from both strong and weak [PSI+] variants; the former variant has more seeds and less soluble Sup35 than the latter. Overexpression of Saccharomyces cerevisiae Hsp104 (Sc-Hsp104) trimmed the weak [PSI+] variants much faster than the strong variants and cured the weak variants an order of magnitude faster than the strong variants. Overexpression of the fungal Hsp104 homologs from Schizosaccharomyces pombe (Sp-Hsp104) or Candida albicans (Ca-Hsp104) also trimmed and cured the weak variants, but interestingly, it neither trimmed nor cured the strong variants. These results show that, because Sc-Hsp104 has greater trimming activity than either Ca-Hsp104 or Sp-Hsp104, it cures both the weak and strong variants, whereas Ca-Hsp104 and Sp-Hsp104 only cure the weak variants. Therefore, curing by Hsp104 overexpression depends on both the trimming ability of the fungal Hsp104 homolog and the strength of the [PSI+] variant: the greater the trimming activity of the Hsp104 homolog and the weaker the variant, the greater the curing.
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Proteínas de Choque Térmico/metabolismo , Factores de Terminación de Péptidos/metabolismo , Priones/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/metabolismo , Adenosina Trifosfatasas/genética , Adenosina Trifosfatasas/metabolismo , Candida albicans/genética , Candida albicans/metabolismo , Prueba de Complementación Genética , Proteínas de Choque Térmico/genética , Factores de Terminación de Péptidos/genética , Priones/genética , Conformación Proteica , Saccharomyces cerevisiae/genética , Proteínas de Saccharomyces cerevisiae/genética , Schizosaccharomyces/genética , Schizosaccharomyces/metabolismoRESUMEN
Anti-HLA donor-specific antibodies are associated with worse outcomes after organ transplantation. Among sensitized pediatric heart candidates, requirement for negative donor-specific cytotoxicity crossmatch increases wait times and mortality. However, transplantation with positive crossmatch may increase posttransplantation morbidity and mortality. We address this clinical challenge in a prospective, multicenter, observational cohort study of children listed for heart transplantation (Clinical Trials in Organ Transplantation in Children-04 [CTOTC-04]). Outcomes were compared among sensitized recipients who underwent transplantation with positive crossmatch, nonsensitized recipients, and sensitized recipients without positive crossmatch. Positive crossmatch recipients received antibody removal and augmented immunosuppression, while other recipients received standard immunosuppression with corticosteroid avoidance. This first CTOTC-04 report summarizes study rationale and design and relates pretransplantation sensitization status using solid-phase technology. Risk factors for sensitization were explored. Of 317 screened patients, 290 were enrolled and 240 underwent transplantation. Core laboratory evaluation demonstrated that more than half of patients were anti-HLA sensitized. Greater than 80% of sensitized patients had class I (with or without class II) HLA antibodies, and one-third of sensitized patients had at least 1 HLA antibody with median fluorescence intensity of ≥8000. Logistic regression models demonstrated male sex, weight, congenital heart disease history, prior allograft, and ventricular assist device are independent risk factors for sensitization.
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Antígenos HLA/inmunología , Trasplante de Corazón/métodos , Isoanticuerpos/inmunología , Proyectos de Investigación , Donantes de Tejidos , Tolerancia al Trasplante/inmunología , Adolescente , Niño , Preescolar , Femenino , Estudios de Seguimiento , Prueba de Histocompatibilidad , Humanos , Terapia de Inmunosupresión , Lactante , Recién Nacido , Isoanticuerpos/sangre , Masculino , Pronóstico , Estudios Prospectivos , Factores de Riesgo , Trasplante HomólogoRESUMEN
Cardiovascular disease is increasingly recognized as a major cause of premature mortality among those with autoimmune disorders. There is an urgent need to identify those patients with autoimmune disease who are at risk for CVD so as to optimize therapeutic intervention and ultimately prevention. Accurate identification, monitoring and stratification of such patients will depend upon a panel of biomarkers of cardiovascular disease. This review will discuss some of the most recent biomarkers of cardiovascular diseases in autoimmune disease, including lipid oxidation, imaging biomarkers to characterize coronary calcium, plaque, and intima media thickness, biomarkers of inflammation and activated complement, genetic markers, endothelial biomarkers, and antiphospholipid antibodies. Clinical implementation of these biomarkers will not only enhance patient care but also likely accelerate the pharmaceutical pipeline for targeted intervention to reduce or eliminate cardiovascular disease in the setting of autoimmunity.
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Enfermedades Autoinmunes/inmunología , Autoinmunidad/inmunología , Biomarcadores/metabolismo , Enfermedades Cardiovasculares/inmunología , Enfermedades Autoinmunes/complicaciones , Enfermedades Autoinmunes/prevención & control , Calcinosis/complicaciones , Calcinosis/inmunología , Calcinosis/metabolismo , Calcio/inmunología , Calcio/metabolismo , Enfermedades Cardiovasculares/complicaciones , Enfermedades Cardiovasculares/prevención & control , Grosor Intima-Media Carotídeo , Enfermedad de la Arteria Coronaria/complicaciones , Enfermedad de la Arteria Coronaria/inmunología , Enfermedad de la Arteria Coronaria/metabolismo , Humanos , Lipoproteínas HDL/inmunología , Lipoproteínas HDL/metabolismoRESUMEN
Background: Neuropsychiatric sequelae of COVID-19 have been documented, including delusions, hallucinations, agitation, and disorganized behavior. Although the mechanisms for these symptoms remain unclear, there has been an increasing body of literature suggesting a correlation between COVID-19 infection and psychosis. Here, we illustrate the case of a 34-year-old female with no previous psychiatric history who contracted COVID-19 and subsequently developed severe symptoms of psychosis. After presenting to the emergency department with one month of worsening mood, auditory hallucinations, intrusive thoughts, and hyperreligiosity, she was admitted to the inpatient psychiatric unit. The patient was treated with multiple antipsychotic medications and was discharged in stable condition with resolution of her auditory hallucinations; however, her delusions, hyperreligiosity, and negative psychotic symptoms persisted, resulting in a second inpatient psychiatric admission eight days after discharge, during which she again did not reach full remission. Objectives: With this information, we hope to increase awareness of COVID-induced psychosis and further discuss the relationship between COVID-19 infection and neuropsychiatric symptoms. Conclusions: Although there has been increasing research about the COVID-19 pandemic, there is much to be elucidated regarding the neuropsychiatric symptoms related to these infections. Similar to previous studies, our case describes a patient with no previous psychiatric history who developed severe psychotic symptoms after COVID-19 infection and was admitted to the inpatient psychiatric unit. These symptoms resulting from infection can be severe or debilitating for the patient. Therefore, physicians should be aware of these potential neuropsychiatric sequelae when treating patients with active COVID-19 infections, and treatment with antipsychotics or acute inpatient psychiatric admission should be considered.
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Hsp104 propagates the yeast prion [PSI+], the infectious form of Sup35, by severing the prion seeds, but when Hsp104 is overexpressed, it cures [PSI+] in a process that is not yet understood but may be caused by trimming, which removes monomers from the ends of the amyloid fibers. This curing was shown to depend on both the N-terminal domain of Hsp104 and the expression level of various members of the Hsp70 family, which raises the question as to whether these effects of Hsp70 are due to it binding to the Hsp70 binding site that was identified in the N-terminal domain of Hsp104, a site not involved in prion propagation. Investigating this question, we now find, first, that mutating this site prevents both the curing of [PSI+] by Hsp104 overexpression and the trimming activity of Hsp104. Second, we find that depending on the specific member of the Hsp70 family binding to the N-terminal domain of Hsp104, both trimming and the curing caused by Hsp104 overexpression are either increased or decreased in parallel. Therefore, the binding of Hsp70 to the N-terminal domain of Hsp104 regulates both the rate of [PSI+] trimming by Hsp104 and the rate of [PSI+] curing by Hsp104 overexpression.
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Priones , Proteínas de Saccharomyces cerevisiae , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Proteínas de Saccharomyces cerevisiae/metabolismo , Proteínas HSP70 de Choque Térmico/genética , Proteínas HSP70 de Choque Térmico/metabolismo , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Priones/genética , Priones/metabolismo , Factores de Terminación de Péptidos/químicaRESUMEN
OBJECTIVE: Disease activity in systemic lupus erythematosus (SLE) is typically monitored by measuring serum C3 and C4. However, these proteins have limited utility as lupus biomarkers, because they are substrates rather than products of complement activation. The aim of this study was to evaluate the utility of measuring the erythrocyte-bound complement activation products, erythrocyte-bound C3d (E-C3d) and E-C4d, compared with that of serum C3 and C4 for monitoring disease activity in patients with SLE. METHODS: The levels of E-C3d and E-C4d were measured by flow cytometry in 157 patients with SLE, 290 patients with other diseases, and 256 healthy individuals. The patients with SLE were followed up longitudinally. Disease activity was measured at each visit, using the validated Systemic Lupus Activity Measure (SLAM) and the Safety of Estrogens in Lupus Erythematosus: National Assessment (SELENA) version of the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). RESULTS: At baseline, patients with SLE had higher median levels of E-C3d and E-C4d (P < 0.0001) in addition to higher within-patient and between-patient variability in both E-C3d and E-C4d when compared with the 2 non-SLE groups. In a longitudinal analysis of patients with SLE, E-C3d, E-C4d, serum C3, and anti-double-stranded DNA (anti-dsDNA) antibodies were each significantly associated with the SLAM and SELENA-SLEDAI. In a multivariable analysis, E-C4d remained significantly associated with these SLE activity measures after adjusting for serum C3, C4, and anti-dsDNA antibodies; however, E-C3d was associated with the SLAM but not with the SELENA-SLEDAI. CONCLUSION: Determining the levels of the erythrocyte-bound complement activation products, especially E-C4d, is an informative measure of SLE disease activity as compared with assessing serum C4 levels and should be considered for monitoring disease activity in patients with SLE.
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Complemento C3d/análisis , Eritrocitos/inmunología , Lupus Eritematoso Sistémico/fisiopatología , Fragmentos de Péptidos/sangre , Adolescente , Adulto , Anciano , Complemento C4b , Femenino , Citometría de Flujo , Humanos , Masculino , Persona de Mediana Edad , Índice de Severidad de la EnfermedadRESUMEN
OBJECTIVES: To evaluate the association between lupus severity and cell-bound complement activation products (CB-CAPs) or low complement proteins C3 and C4. METHODS: All subjects (n=495) fulfilled the American College of Rheumatology (ACR) classification criteria for SLE. Abnormal CB-CAPs (erythrocyte-bound C4d or B-lymphocyte-bound C4d levels >99th percentile of healthy) and complement proteins C3 and C4 were determined using flow cytometry and turbidimetry, respectively. Lupus severity was estimated using the Lupus Severity Index (LSI). Statistical analysis consisted of multivariable linear regression and groups comparisons. RESULTS: Abnormal CB-CAPs were more prevalent than low complement values irrespective of LSI levels (62% vs 38%, respectively, p<0.0001). LSI was low (median 5.44, IQR: 4.77-6.93) in patients with no complement abnormality, intermediate in patients with abnormal CB-CAPs (median 6.09, IQR: 5.31-8.20) and high in the group presenting with both abnormal CB-CAPs and low C3 and/or C4 (median 7.85, IQR: 5.51-8.37). Odds of immunosuppressant use was higher in subjects with LSI ≥5.95 compared with subjects with LSI <5.95 (1.60 vs 0.53, p<0.0001 for both). Multivariable regression analysis revealed that higher LSI scores associated with abnormal CB-CAPs-but not low C3/C4-after adjusting for younger age, race and longer disease duration (p=0.0001), which were also independent predictors of disease severity (global R2=0.145). CONCLUSION: Abnormalities in complement activation as measured by CB-CAPs are associated with increased LSI.
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Activación de Complemento/inmunología , Complemento C3/análisis , Complemento C4/análisis , Lupus Eritematoso Sistémico/inmunología , Adolescente , Adulto , Linfocitos B/química , Linfocitos B/inmunología , Estudios de Casos y Controles , Complemento C3/metabolismo , Complemento C4/metabolismo , Estudios Transversales , Eritrocitos/química , Eritrocitos/inmunología , Etnicidad , Femenino , Citometría de Flujo/métodos , Humanos , Inmunosupresores/uso terapéutico , Lupus Eritematoso Sistémico/diagnóstico , Lupus Eritematoso Sistémico/tratamiento farmacológico , Masculino , Persona de Mediana Edad , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
BACKGROUND AND PURPOSE: Platelets bearing complement C4d were recently reported to be 99% specific for a diagnosis of systemic lupus erythematosus (SLE) and associated with neuropsychiatric lupus. We compared the prevalence of platelet C4d and investigated the clinical associations of platelet C4d in patients with acute ischemic stroke. METHODS: We recruited 80 patients hospitalized for acute ischemic stroke. Stroke severity was measured by the National Institutes of Health Stroke Scale (NIH-SS). Infarct volume was determined by MRI. Platelet C4d was measured by flow cytometry. RESULTS: Mean age was 57.9 years (range: 24.6 to 86.8 years), 58% were male, and 91% were white. Eight patients (10%) with acute ischemic stroke were platelet C4d-positive, which was significantly higher in prevalence compared to healthy controls (0%, P<0.0001) and non-SLE patients with immune/inflammatory disease (2%, P=0.004). The median NIH-SS score and infarct volume for acute stroke patients were 6 (interquartile range [IQR]: 2 to 13) and 3.4 cc (IQR: 1.1 to 16.6), respectively. Platelet C4d-positive patients were more likely to have a severe stroke compared to those with negative platelet C4d (NIH-SS median: 17.5 versus 5, P=0.003). Positive platelet C4d was independently associated with stroke severity (P=0.03) after controlling for age, anticardiolipin antibody (aCL) status, and total anterior circulation of stroke involvement, and also with infarct volume (P=0.005) after controlling for age, aCL status, and old stroke by MRI. CONCLUSIONS: Platelet C4d is associated with severe acute ischemic stroke. Platelet C4d may be a biomarker as well as pathogenic clue that links cerebrovascular inflammation and thrombosis.
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Plaquetas/química , Isquemia Encefálica/sangre , Fragmentos de Péptidos/sangre , Enfermedad Aguda , Anciano , Anticuerpos Anticardiolipina/sangre , Artritis Reumatoide/sangre , Artritis Reumatoide/complicaciones , Artritis Reumatoide/inmunología , Biomarcadores , Isquemia Encefálica/tratamiento farmacológico , Isquemia Encefálica/inmunología , Isquemia Encefálica/patología , Enfermedades Cardiovasculares/sangre , Enfermedades Cardiovasculares/inmunología , Comorbilidad , Complemento C4b , Femenino , Fibrinolíticos/uso terapéutico , Humanos , Inflamación/sangre , Inflamación/inmunología , Lupus Eritematoso Sistémico/sangre , Lupus Eritematoso Sistémico/complicaciones , Lupus Eritematoso Sistémico/inmunología , Masculino , Persona de Mediana Edad , Inhibidores de Agregación Plaquetaria/uso terapéutico , Enfermedades Reumáticas/sangre , Enfermedades Reumáticas/inmunología , Factores de Riesgo , Índice de Severidad de la Enfermedad , Terapia TrombolíticaRESUMEN
Perivascular adipose tissue (PVAT) influences vascular function and pathology. We present a protocol using micro-computed tomography (microCT), a novel imaging technique typically used for hard biological tissue, to characterize the temporal and spatial development of aorta PVAT and luminal plaque soft tissue. Apolipoprotein E deficient (ApoE) and C57Bl/6J (control) mice were fed a high fat western diet up to 30 weeks. 3D microCT reconstructions were used to quantify: 1) vascular wall volume, a surrogate measure of remodeling, was greater in ApoE, 2) aorta PVAT volume was reduced in ApoE, 3) plaque volumes increased over time in ApoE, 4) plaque development co-localized with luminal ostia, origins of branching arteries, which traveled through areas of greatest PVAT volume, 5) qualitatively, the same arteries showed evidence of increased tortuosity in ApoE. This study reflects the potential of microCT analyses to assess vascular wall, PVAT and arterial trajectory modifications in relevant animal models. Abbreviations: PVAT: perivascular adipose tissue; ApoE: apolipoprotein E deficient mouse strain; Control: C57Bl/6J mouse strain; PTA: 0.3% phosphotungstic acid; microCT: micro-computed tomography; CV: cardiovascular; CVD: cardiovascular disease; IQR: interquartile range; PPARγ: peroxisome proliferator activated receptor - gamma; VV: vasa vasorum; 3D: three dimensional.
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Tejido Adiposo/patología , Aorta Torácica/patología , Apolipoproteínas/deficiencia , Apolipoproteínas/metabolismo , Modelos Animales de Enfermedad , Imagenología Tridimensional , Placa Aterosclerótica/patología , Microtomografía por Rayos X , Tejido Adiposo/metabolismo , Animales , Aorta Torácica/metabolismo , Apolipoproteínas/genética , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Placa Aterosclerótica/metabolismoRESUMEN
INTRODUCTION: Cell-bound complement activation products (CB-CAPs) were first reported in 2004, since which time multiple laboratories have demonstrated their value as biomarkers for diagnosis, monitoring, and stratification of patients with systemic lupus erythematosus. Areas covered: This review summarizes the highlights of these 14 years of CB-CAPs discovery and validation, concluding with a view toward their future potential for precision medicine. Expert commentary: The practice of medicine is both art and science and each physician can be considered both artist and scientist with a variable blend of the two skill sets. There is arguably no disease that presents a greater challenge, nor a greater opportunity, for implementation of precision medicine, as does lupus. The physician who is presented with diagnosis and/or management of a patient suspected of having lupus will need to augment artistic skills with scientific guidance, and that science will be delivered in the form of biomarkers. Ultimately, we will likely have a 'lupus liquid biopsy' that will be 100% sensitive and 100% specific for a diagnosis of lupus. This will undoubtedly be a panel of biomarkers rather than an individual laboratory test. Such a liquid biopsy could transform lupus diagnosis to an entirely scientific process.
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Biomarcadores/metabolismo , Proteínas del Sistema Complemento/metabolismo , Lupus Eritematoso Sistémico/diagnóstico , Activación de Complemento , Testimonio de Experto , Humanos , Monitoreo FisiológicoRESUMEN
BACKGROUND AND AIMS: Women with systemic lupus erythematosus (SLE) have an increased risk of cardiovascular disease (CVD). Perivascular adipose tissue (PVAT) surrounds the vascular wall, is associated with CVD risk factors, and may contribute to premature CVD in SLE. We previously found greater volumes of aortic PVAT associated with aortic calcification (AC) in female SLE patients. There is recent evidence that not only volume but adipose density may also be indicative of inflammation. We hypothesized that female SLE patients would have a difference in aPVAT quality associated with AC that is independent of volume. METHODS: Aorta PVAT quality was evaluated using the average radiodensity (density) of adipose tissue-specific Hounsfield Units (-190 to -30 HU) within each clinical CT scan of CVD-free, age-/race-matched SLE women (n = 143) and healthy controls (HC, n = 143). RESULTS: Aorta PVAT density was significantly higher in SLE (mean (SD): (-83.6 (1.9) HU) versus HC (-84.1 (1.8) HU), p=0.03). Increasing aPVAT volume was correlated with denser aPVAT in SLE (ρ, p-value: 0.75,<0.0001) and HC (0.74,<0.0001). Increasing AC score (log) was correlated with denser aPVAT for SLE (0.31, 0.0005) and HC (0.23, 0.008). In linear regression, denser aPVAT was more strongly associated with AC score in SLE (ß (SE): 0.445 (0.11), p<0.0001) versus HC (0.335 (0.12), p=0.006) independent of age, circulating inflammatory markers, CVD risk factors and BMI (p<0.05), but was attenuated with aPVAT volume (p=0.3). CONCLUSIONS: Denser aPVAT is associated with aPVAT volume and AC in SLE women. Adjusting for aPVAT volume attenuated the detected association between aPVAT density and AC, which may be indicative of adipose dysfunction.
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Tejido Adiposo/fisiopatología , Adiposidad , Aorta , Enfermedades de la Aorta/etiología , Lupus Eritematoso Sistémico/complicaciones , Calcificación Vascular/etiología , Tejido Adiposo/diagnóstico por imagen , Aorta/diagnóstico por imagen , Enfermedades de la Aorta/diagnóstico por imagen , Enfermedades de la Aorta/fisiopatología , Aortografía/métodos , Distribución de Chi-Cuadrado , Angiografía por Tomografía Computarizada , Estudios Transversales , Femenino , Humanos , Modelos Lineales , Lupus Eritematoso Sistémico/diagnóstico , Lupus Eritematoso Sistémico/fisiopatología , Persona de Mediana Edad , Análisis Multivariante , Estudios Retrospectivos , Factores de Riesgo , Calcificación Vascular/diagnóstico por imagen , Calcificación Vascular/fisiopatologíaRESUMEN
Premature, accelerated onset of atherothrombotic disease is prevalent in patients with systemic lupus erythematosus (SLE). Most, if not all, atherothrombotic diseases are likely to involve platelets and complement. Previously, we discovered that platelets bearing complement activation product C4d (P-C4d) are present in SLE patients, and are significantly associated with antiphospholipid (aPL) antibody positivity and stroke in SLE patients. The goal of the present study was to further elucidate the role of aPL and other platelet-reactive autoantibodies in the generation of P-C4d. To determine the association between P-C4d and aPL antibodies, the serum levels of aPL antibodies and P-C4d of 180 SLE patients were measured by enzyme-linked immunoassays and flow cytometry, respectively. To investigate the role of aPL antibodies, and possibly other autoantibodies as well, in mediating the generation of P-C4d, in vitro 2-step P-C4d induction experiments were performed. The results showed that the presence and levels of aPL antibodies in the serum were specifically elevated in SLE patients with positive P-C4d. The plasma and immunoglobulins purified from SLE patients who were positive for P-C4d and aPL were capable of inducing C4d deposition on normal platelets in vitro. The capacity of SLE plasma in inducing P-C4d appeared to correlate proportionately to the serum aPL levels. Collectively, the results demonstrate that both aPL and other platelet-reactive autoantibodies may participate in mediating the generation of P-C4d in SLE patients.
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PURPOSE: Inflammatory rheumatic diseases (IRD) are associated with accelerated coronary artery disease (CAD), which may result from both systemic and vascular wall inflammation. There are indications that complement may be involved in the pathogenesis of CAD in Systemic Lupus Erythematosus (SLE) and Rheumatoid Arthritis (RA). This study aimed to evaluate the associations between circulating complement and complement activation products with mononuclear cell infiltrates (MCI, surrogate marker of vascular inflammation) in the aortic media and adventitia in IRDCAD and non-IRDCAD patients undergoing coronary artery bypass grafting (CABG). Furthermore, we compared complement activation product deposition patterns in rare aorta adventitial and medial biopsies from SLE, RA and non-IRD patients. METHODS: We examined plasma C3 (p-C3) and terminal complement complexes (p-TCC) in 28 IRDCAD (SLE = 3; RA = 25), 52 non-IRDCAD patients, and 32 IRDNo CAD (RA = 32) from the Feiring Heart Biopsy Study. Aortic biopsies taken from the CAD only patients during CABG were previously evaluated for adventitial MCIs. The rare aortic biopsies from 3 SLE, 3 RA and 3 non-IRDCAD were assessed for the presence of C3 and C3d using immunohistochemistry. RESULTS: IRDCAD patients had higher p-TCC than non-IRDCAD or IRDNo CAD patients (p<0.0001), but a similar p-C3 level (p = 0.42). Circulating C3 was associated with IRD duration (ρ, p-value: 0.46, 0.03). In multiple logistic regression analysis, IRD remained significantly related to the presence and size of MCI (p<0.05). C3 was present in all tissue samples. C3d was detected in the media of all patients and only in the adventitia of IRD patients (diffuse in all SLE and focal in one RA). CONCLUSION: The independent association of IRD status with MCI and the observed C3d deposition supports the unique relationship between rheumatic disease, and, in particular, SLE with the complement system. Exaggerated systemic and vascular complement activation may accelerate CVD, serve as a CVD biomarker, and represent a target for new therapies.
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Biomarcadores/sangre , Proteínas del Sistema Complemento/metabolismo , Enfermedad de la Arteria Coronaria/sangre , Inflamación/sangre , Enfermedades Reumáticas/sangre , Anciano , Artritis Reumatoide/sangre , Complemento C3/metabolismo , Complemento C3d/metabolismo , Femenino , Humanos , Inmunohistoquímica , Técnicas In Vitro , Lupus Eritematoso Sistémico/sangre , Masculino , Persona de Mediana Edad , Factores de RiesgoRESUMEN
Cell death by apoptosis plays a significant and seemingly contradictory role in the development and pathogenesis of autoimmune diseases. Apoptosis is integral to the assembly and maintenance of a healthy, self-tolerant immune system. However, many of the molecular and cellular events specific to apoptosis generate a reservoir of self-antigens with the potential to initiate and possibly perpetuate autoimmune conditions. Recent findings that support this latter, more sinister role for apoptosis have shed light on a mystery that is common to many systemic autoimmune diseases, namely, why the majority of autoantibodies produced in patients with these diseases target proteins that are normally found inside the cell, often within the nucleus. This review will discuss how autoantigens are specifically altered during the apoptotic process, and how the complement system participates in recognizing and clearing these potentially immunogenic packages.
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Apoptosis/inmunología , Autoinmunidad , Tolerancia Inmunológica , Animales , Autoanticuerpos/inmunología , Autoantígenos/inmunología , Vía Clásica del Complemento , HumanosRESUMEN
Measurement of serum C3 and C4 has been used for several decades to monitor disease activity in patients with SLE. Despite the limited utility and recognized weaknesses of these assays, they have remained the gold standard during an era of unprecedented discovery in the complement field. The current urgent need for lupus biomarkers warrants efforts to mine the complement system for assays superior to serum C3 and C4. Recent studies of soluble and cell-bound complement activation products hold promise for achieving this goal.
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Biomarcadores/metabolismo , Activación de Complemento , Complemento C3/inmunología , Complemento C4/inmunología , Lupus Eritematoso Sistémico , Eritrocitos/metabolismo , Humanos , Lupus Eritematoso Sistémico/sangre , Lupus Eritematoso Sistémico/diagnóstico , Lupus Eritematoso Sistémico/inmunología , Reticulocitos/metabolismoRESUMEN
Deficiencies in the classical pathway of the complement system have been implicated in the etiology and pathogenesis of systemic lupus erythematosus (SLE) for several decades. Recent advances have suggested that this link is due to a critical role of complement in the recognition and clearance of the cellular remnants of apoptosis. In this review, we summarize the role of apoptosis in generation of an autoimmune response, and we integrate recent advances that link apoptosis, complement activation and the etiopathogenesis of SLE.
Asunto(s)
Apoptosis , Proteínas del Sistema Complemento/fisiología , Lupus Eritematoso Sistémico/etiología , Animales , Autoantígenos/metabolismo , Autoinmunidad , Humanos , Tolerancia Inmunológica , Lupus Eritematoso Sistémico/inmunología , FosforilaciónRESUMEN
The mystery that surrounds autoimmunity revolves around how the immune system of patients who have systemic autoimmune diseases becomes primed to recognize intracellular antigens, how the autoantibodies thus produced contribute to the pathogenesis of the disease, and how those autoantibodies access their target proteins. By examining the mechanisms that are involved in the normal cellular process of apoptosis, we are beginning to unravel this mystery. The intracellular autoantigen targets of many systemic autoimmune diseases become altered during apoptosis in ways that may change how they are perceived by the immune system. High concentrations of self-antigens, or in the case of viral infection, complexes of foreign and self-antigens, are packaged during generation of apoptotic cells. The packages also may contain altered fragments of self-antigens that have not been encountered previously by the immune system. Under normal circumstances, apoptotic cells are cleared rapidly by macrophages and DCs. The normal consequence of that clearance is that the apoptosis-altered self-antigens are either ignored by the immune system or tolerance to those antigens is maintained. Clearance is achieved through complex mechanisms that enable macrophages and DCs to recognize apoptotic cells as nonthreatening "self" particles. Defects in this process that cause a delay in clearance could change the appearance of apoptotic cells and cause them to be recognized as "foreign invaders," thereby stimulating an inflammatory response that, in turn, activates an immune response to self-antigens. By studying the mechanisms that are involved in recognition and clearance of apoptotic cells, we are uncovering clues to the defects that may underlie the development of systemic autoimmunity.
Asunto(s)
Apoptosis , Autoantígenos/inmunología , Enfermedades Autoinmunes/inmunología , Autoinmunidad/fisiología , Animales , HumanosRESUMEN
T cells bearing C4d, a complement activation product (CAP), have been shown to be highly sensitive and specific as diagnostic biomarkers for systemic lupus erythematosus (SLE). T cells bearing C4d are also functionally abnormal, suggesting a role for cell-bound CAPs in lupus pathogenesis. However, the mechanism responsible for generation of T-C4d has not been determined. The purpose of this cross-sectional and prospective study was to investigate the potential role of anti-T-cell autoantibodies in the generation of the T cell-bound C4d (T-C4d) signatures in SLE. Briefly, T cells from patients with SLE (n = 326), patients with other inflammatory diseases (n = 185), and healthy controls (n = 48) were characterized for surface deposition of either or both of C4d and immunoglobulin (Ig) by flow cytometry. In vitro phenotype transfer experiments were performed to characterize Ig from patients with SLE for the capacity to generate T-C4d signatures in vitro. The results demonstrate that individual patients with SLE harbor specific signatures reflecting the presence of either or both of C4d and Ig on their T cells and T-cell subsets. In addition, SLE patient-specific signatures can be transferred in vitro to normal T cells by exposure to Ig purified from the signature donor. Complement activation does not proceed through the generation of C5b-9 (membrane attack complex) or cellular lysis, and T-C4d does not correlate with lymphopenia. In conclusion, these results suggest that patient-specific T-C4d signatures are generated by anti-T-cell autoantibodies that trigger sublytic complement activation, a previously unrecognized pathway in lupus pathogenesis.
Asunto(s)
Suero Antilinfocítico/inmunología , Complemento C4b/inmunología , Lupus Eritematoso Sistémico/inmunología , Fragmentos de Péptidos/inmunología , Linfocitos T/inmunología , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD8-positivos/inmunología , Estudios de Casos y Controles , Humanos , Fenotipo , Donantes de TejidosRESUMEN
The last decade has witnessed an explosion in efforts to discover and validate lupus biomarkers. The currently steep trajectory of this progress is unprecedented. However, advances in the lupus biomarker field remain fewer and slower than physicians, patients, and pharmaceutical companies have hoped for. This chapter will review the challenges confronted by physicians and scientists in pursuit of lupus biomarkers and will present our experience on this path and specific efforts to surmount some of the obstacles in this endeavor. A comprehensive review of the current landscape in lupus biomarker research has recently been published elsewhere (Ahearn et al. Transl Res 159:326-342, 2012; Liu et al. Ther Adv Musculoskelet Dis 5:210-233, 2013; Liu and Ahearn Best Pract Res Clin Rheumatol 23:507-523, 2009; Liu et al. Curr Opin Rheumatol 17:543-549, 2005).