RESUMEN
Trypanosome Lytic Factor (TLF) is a primate-specific high-density lipoprotein (HDL) complex that, through the cation channel-forming protein apolipoprotein L-1 (APOL1), provides innate immunity to select kinetoplastid parasites. The immunoprotective effects of TLF have been extensively investigated in the context of its interaction with the extracellular protozoan Trypanosoma brucei brucei, to which it confers sterile immunity. We previously showed that TLF could act against an intracellular pathogen Leishmania, and here we dissected the role of TLF and its synergy with host-immune cells. Leishmania major is transmitted by Phlebotomine sand flies, which deposit the parasite intradermally into mammalian hosts, where neutrophils are the predominant phagocytes recruited to the site of infection. Once in the host, the parasites are phagocytosed and shed their surface glycoconjugates during differentiation to the mammalian-resident amastigote stage. Our data show that mice producing TLF have reduced parasite burdens when infected intradermally with metacyclic promastigotes of L. major, the infective, fly-transmitted stage. This TLF-mediated reduction in parasite burden was lost in neutrophil-depleted mice, suggesting that early recruitment of neutrophils is required for TLF-mediated killing of L. major. In vitro we find that only metacyclic promastigotes co-incubated with TLF in an acidic milieu were lysed. However, amastigotes were not killed by TLF at any pH. These findings correlated with binding experiments, revealing that labeled TLF binds specifically to the surface of metacyclic promastigotes, but not to amastigotes. Metacyclic promastigotes of L. major deficient in the synthesis of surface glycoconjugates LPG and/or PPG (lpg1- and lpg5A-/lpg5B- respectively) whose absence mimics the amastigote surface, were resistant to TLF-mediated lysis. We propose that TLF binds to the outer surface glycoconjugates of metacyclic promastigotes, whereupon it kills the parasite in the acidic phagosome of phagocytes. We hypothesize that resistance to TLF requires shedding of the surface glycoconjugates, which occurs several hours after phagocytosis by immune cells, creating a relatively short-lived but effective window for TLF to act against Leishmania.
Asunto(s)
Interacciones Huésped-Parásitos/fisiología , Inmunidad Innata , Leishmaniasis Cutánea , Lipoproteínas HDL/metabolismo , Animales , Humanos , Leishmania major , Leishmaniasis Cutánea/inmunología , Leishmaniasis Cutánea/metabolismo , Leishmaniasis Cutánea/patología , Lipoproteínas HDL/inmunología , RatonesRESUMEN
Apolipoprotein A-I (apo A-I) represents the main component of the Trypanosome lytic factor (TLF) which contributes to the host innate immunity against Trypanosoma and Leishmania. These parasites use complex and multiple strategies such as molecular mimicry to evade or subvert the host immune system. Previous studies have highlighted the adaptation mechanisms of TLF-resistant Trypanosoma species. These data might support the hypothesis that Leishmania parasites (amastigote forms in macrophages) might express apo A-I to bypass and escape from TLF action as a component of the host innate immune responses. The anti-inflammatory property of apo A-I is another mechanism that supports our idea that apo A-I may play a role in Leishmania parasites allowing them to bypass the host innate immune system.
Asunto(s)
Apolipoproteína A-I/inmunología , Leishmania/inmunología , Leishmaniasis/inmunología , Proteínas Protozoarias/inmunología , Humanos , Evasión Inmune , Inmunidad Innata , Lipoproteínas HDL/inmunología , Macrófagos/inmunología , Macrófagos/parasitología , Imitación MolecularRESUMEN
High density lipoproteins (HDL) are heterogeneous particles composed by a vast array of proteins and lipids, mostly recognized for their cardiovascular (CV) protective effects. However, evidences from basic to clinical research have contributed to depict a role of HDL in the modulation of immune-inflammatory response thus paving the road to investigate their involvement in other diseases beyond those related to the CV system. HDL-C levels and HDL composition are indeed altered in patients with autoimmune diseases and usually associated to disease severity. At molecular levels, HDL have been shown to modulate the anti-inflammatory potential of endothelial cells and, by controlling the amount of cellular cholesterol, to interfere with the signaling through plasma membrane lipid rafts in immune cells. These findings, coupled to observations acquired from subjects carrying mutations in genes related to HDL system, have helped to elucidate the contribution of HDL beyond cholesterol efflux thus posing HDL-based therapies as a compelling interventional approach to limit the inflammatory burden of immune-inflammatory diseases.
Asunto(s)
Enfermedades Cardiovasculares/metabolismo , Enfermedades del Sistema Inmune/inmunología , Lipoproteínas HDL/inmunología , Animales , Humanos , Enfermedades del Sistema Inmune/genética , Inmunidad Innata , Linfocitos/inmunología , Macrófagos/inmunologíaRESUMEN
Inflammation and immune dysfunction have been increasingly recognized as crucial mechanisms in atherogenesis. Modifications in cell lipid metabolism, plasma dyslipidaemia and particularly low high-density lipoprotein (HDL) levels occur both in atherosclerosis and in autoimmune rheumatic diseases (which are strongly associated with an increased risk of atherosclerosis), suggesting the presence of a crucial link. HDL, the plasma lipoprotein responsible for reverse cholesterol transport, is known for its several protective effects in the context of atherosclerosis. Among these, HDL immunomodulatory effects are possibly the less understood. Through the efflux of cholesterol from plasma cell membranes with the consequent disruption of lipid rafts and the interaction with the cholesterol transporters present in the plasma membrane, HDL affects both the innate and adaptive immune responses. Animal and human studies have demonstrated a predominance of HDL anti-inflammatory effects, despite some pro-inflammatory actions having also been reported. The HDL role on the modulation of the immune response is further suggested by the detection of low levels together with a dysfunctional HDL in patients with autoimmune diseases. Here, we review the current knowledge of the immune mechanisms of atherosclerosis and the modulatory effects HDL may have on them.
Asunto(s)
Aterosclerosis/inmunología , Autoinmunidad/inmunología , Inmunidad/inmunología , Lipoproteínas HDL/inmunología , Animales , Enfermedades Autoinmunes/inmunología , Humanos , Inflamación/inmunologíaRESUMEN
The ability of HDL to inhibit inflammation in adipocytes and adipose tissue is reduced when HDL contains serum amyloid A (SAA) that is trapped by proteoglycans at the adipocyte surface. Because we recently found that the major extracellular matrix proteoglycan produced by hypertrophic adipocytes is versican, whereas activated adipose tissue macrophages produce mainly biglycan, we further investigated the role of proteoglycans in determining the antiinflammatory properties of HDL. The distributions of versican, biglycan, apolipoprotein A1 (the major apolipoprotein of HDL), and SAA were similar in adipose tissue from obese mice and obese human subjects. Colocalization of SAA-enriched HDL with versican and biglycan at the cell surface of adipocyte and peritoneal macrophages, respectively, was blocked by silencing these proteoglycans, which also restored the antiinflammatory property of SAA-enriched HDL despite the presence of SAA. Similar to adipocytes, normal HDL exerted its antiinflammatory function in macrophages by reducing lipid rafts, reactive oxygen species generation, and translocation of Toll-like receptor 4 and NADPH oxidase 2 into lipid rafts, effects that were not observed with SAA-enriched HDL. These findings imply that SAA present in HDL can be trapped by adipocyte-derived versican and macrophage-derived biglycan, thereby blunting HDL's antiinflammatory properties.
Asunto(s)
Adipocitos/inmunología , Biglicano/inmunología , Lipoproteínas HDL/inmunología , Macrófagos Peritoneales/inmunología , Obesidad/inmunología , Proteína Amiloide A Sérica/inmunología , Versicanos/inmunología , Adipocitos/patología , Adulto , Animales , Apolipoproteína A-I/genética , Apolipoproteína A-I/inmunología , Biglicano/antagonistas & inhibidores , Biglicano/genética , Dieta Alta en Grasa/efectos adversos , Femenino , Regulación de la Expresión Génica , Humanos , Resistencia a la Insulina/inmunología , Lipoproteínas HDL/genética , Macrófagos Peritoneales/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Persona de Mediana Edad , Obesidad/etiología , Obesidad/genética , Obesidad/patología , Unión Proteica , Transporte de Proteínas , ARN Interferente Pequeño/genética , ARN Interferente Pequeño/metabolismo , Especies Reactivas de Oxígeno/inmunología , Especies Reactivas de Oxígeno/metabolismo , Proteína Amiloide A Sérica/genética , Nitrato de Plata/administración & dosificación , Receptor Toll-Like 4/genética , Receptor Toll-Like 4/inmunología , Versicanos/antagonistas & inhibidores , Versicanos/genéticaRESUMEN
Trypanosomiasis is a devastating neglected tropical disease affecting livestock and humans. Humans are susceptible to two Trypanosoma brucei subspecies but protected from other trypanosomes by circulating high-density lipoprotein (HDL) complexes called trypanosome lytic factors (TLFs) 1 and 2. TLFs contain apolipoprotein L-1 contributing to lysis and haptoglobin-related protein (HPR), which can function as a ligand for a parasite receptor. TLF2 also uniquely contains non-covalently associated immunoglobin M (IgM) antibodies, the role and origin of which remain unclear. Here, we show that these TLF2-associated IgMs interact with both HPR and alternate trypanosome surface proteins, including variant surface glycoprotein, likely facilitating complex biogenesis and TLF uptake into parasites. TLF2-IgMs are germline antibodies that, while present at basal concentrations in healthy individuals, are elicited by trypanosome infection in both murine models and human sleeping sickness patients. These data suggest that poly- and self-reactive germline antibodies such as TLF2-associated IgMs play a role in antimicrobial immunity.
Asunto(s)
Anticuerpos Antiprotozoarios/inmunología , Antígenos de Neoplasias/inmunología , Apolipoproteína L1/inmunología , Haptoglobinas/inmunología , Inmunoglobulina M/inmunología , Lipoproteínas HDL/inmunología , Tripanosomiasis Africana/inmunología , Adolescente , Adulto , Anciano , Animales , Línea Celular , Niño , Femenino , Células Germinativas/inmunología , Interacciones Huésped-Parásitos , Humanos , Masculino , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Noqueados , Persona de Mediana Edad , Modelos Animales , Parásitos , Trypanosoma brucei brucei , Adulto JovenRESUMEN
BACKGROUND: Autoimmune diseases are closely associated with cardiovascular diseases (CVD). Over the last decades, the comprehension of atherosclerosis, the principal initiator of CVD, evolved from a lipidcentered disease to a predominant inflammatory and immune response-driven disease displaying features of autoimmunity against a broad range of auto-antigens, including lipoproteins. Among them, high density lipoproteins (HDL) are important actors of cholesterol transport and bear several anti-atherogenic properties, raising a growing interest as therapeutic targets to decrease atherosclerosis and CVD burden, with nevertheless rather disappointing results so far. Reflecting HDL composition complexity, autoimmune responses and autoantibodies against various HDL components have been reported. RESULTS: In this review, we addressed the important complexity of humoral autoimmunity towards HDL and particularly how this autoimmune response could help improving our understanding of HDL biological implication in atherosclerosis and CVD. We also discussed several issues related to specific HDL autoantibody subclasses characteristics, including etiology, prognosis and pathological mechanisms according to Rose criteria. CONCLUSION: Finally, we addressed the possible clinical value of using these antibodies not only as potential biomarkers of atherogenesis and CVD, but also as a factor potentially mitigating the benefit of HDL-raising therapies.
Asunto(s)
Aterosclerosis/inmunología , Enfermedades Cardiovasculares/inmunología , Inmunidad Humoral , Lipoproteínas HDL/inmunología , Autoanticuerpos/inmunología , Autoinmunidad , Biomarcadores/sangre , HDL-Colesterol , HumanosRESUMEN
High density lipoproteins (HDL) are considered cardio protective. Apolipoprotein A-I (apoA-I), a major component of HDL helps in reverse cholesterol transport, whose function is greatly affected during atherosclerosis due to oxidation by myeloperoxidase. Amino acid tyrosine residue of apoA-I at position 192 and 166 are sensitive to oxidation by myeloperoxidase resulting in the generation of chlorinated and nitrated apoA-I and they are believed to be present in atherosclerotic plaques and in circulation. These oxidized apoA-I have been suggested as potential indicator(s) of CVD risks in humans. To detect the levels of oxidized apoA-I there is a need for developing monoclonal antibodies (mAbs) with high specificity and sensitivity that could be utilized routinely in clinical immune based assays for blood plasma or for in vivo imaging. In this study, chemically chlorinated apoA-I (chlorinated 192tyrosine- apoA-I) and a short synthetic peptide, containing the corresponding chlorinated tyrosine residue, conjugated to keyhole limpet hemocyanin (KLH) carrier protein were used for immunization. Stable hybridoma clones F7D5 and G11E3 were found to be highly sensitive and reactive towards chlorinated 192tyrosine- apoA-I. Interestingly, these mAbs also displayed positive reaction with atherosclerotic plaques obtained from mouse and human biopsies. In vitro or in vivo diagnostic tests could be developed either by detecting oxidized apoA-I in human plasma or by directly imaging atheroma plaques as both mAbs were shown to stain human atheroma. The anti-chlorinated 192tyrosine- apoA-I mAbs described in this study may have a high diagnostic potential in predicting CVD risks.
Asunto(s)
Anticuerpos Monoclonales/inmunología , Apolipoproteína A-I/análisis , Aterosclerosis/diagnóstico , Enfermedad de la Arteria Coronaria/diagnóstico , Pruebas Inmunológicas , Lipoproteínas HDL/análisis , Animales , Especificidad de Anticuerpos , Apolipoproteína A-I/inmunología , Aterosclerosis/genética , Aterosclerosis/inmunología , Aterosclerosis/metabolismo , Biomarcadores/análisis , Enfermedad de la Arteria Coronaria/inmunología , Enfermedad de la Arteria Coronaria/metabolismo , Modelos Animales de Enfermedad , Halogenación , Humanos , Lipoproteínas HDL/inmunología , Ratones Noqueados para ApoE , Oxidación-Reducción , Placa Aterosclerótica , Valor Predictivo de las Pruebas , Reproducibilidad de los Resultados , TirosinaRESUMEN
Atherosclerosis, a chronic inflammatory disease of the arterial wall, is the leading cause of cardiac disorders and stroke. The onset and progression of these diseases are linked with the inflammatory response, especially NLRP3 inflammasome activation, inducing the production of proinflammatory cytokines, such as interleukin 1ß (IL-1ß). Because high-density lipoproteins (HDLs) have shown significant antiatherogenic and anti-inflammatory properties, we evaluated their immunomodulatory activity in response to cholesterol crystals and other innate immune activators. Human primary monocyte-derived macrophages, THP-1 cells, and murine macrophages were stimulated to activate NLRP3 inflammasome and other pattern recognition receptors, in the presence or absence of HDL. Then, HDL immunomodulatory effects were evaluated through IL-1ß and IL-6 production by enzyme-linked immunosorbent assay. Furthermore, in vivo HDL anti-inflammatory effects were evaluated in a murine model of peritoneal inflammatory infiltration. HDLs have an immunomodulatory effect on different cellular models, including peripheral blood mononuclear cells, THP-1 cells, and murine macrophages, by affecting the activity of innate immunity sensors, such as Toll-like receptors (TLRs), dectin-1, and inflammasomes. HDL reduces the proinflammatory role of cholesterol crystals, nigericin, and other NLRP3 and AIM2 inflammasome agonists, and several TLR agonists, leading to a decreased production of IL-1ß and IL-6. The results suggest that HDLs are highly important in the regulation of the innate immune response and may have a beneficial role in controlling diseases associated with the inflammatory response.
Asunto(s)
Inmunidad Innata/inmunología , Inflamación/inmunología , Lipoproteínas HDL/inmunología , Animales , Células Cultivadas , Humanos , RatonesRESUMEN
Despite strong evidence that high-density lipoproteins (HDLs) modulate the immune response, the role of HDL in allergies is still poorly understood. Many patients with allergic rhinitis (AR) develop a late-phase response, characterized by infiltration of monocytes and eosinophils into the nasal submucosa. Functional impairment of HDL in AR-patients may insufficiently suppress inflammation and cell infiltration, but the effect of AR on the composition and function of HDL is not understood. We used apolipoprotein (apo) B-depleted serum as well as isolated HDL from AR-patients (nâ¯=â¯43) and non-allergic healthy controls (nâ¯=â¯20) for detailed compositional and functional characterization of HDL. Both AR-HDL and apoB-depleted serum of AR-patients showed decreased anti-oxidative capacity and impaired ability to suppress monocyte nuclear factor-κB expression and pro-inflammatory cytokine secretion, such as interleukin (IL)-4, IL-6, IL-8, tumor necrosis factor alpha and IL-1 beta. Sera of AR-patients showed decreased paraoxonase and cholesteryl-ester transfer protein activities, increased lipoprotein-associated phospholipase A2 activity, while lecithin-cholesterol acyltransferase activity and cholesterol efflux capacity were not altered. Surprisingly, apoB-depleted serum and HDL from AR-patients showed an increased ability to suppress eosinophil effector responses upon eotaxin-2/CCL24 stimulation. Mass spectrometry and biochemical analyses showed reduced levels of apoA-I and phosphatidylcholine, but increased levels of apoA-II, triglycerides and lyso-phosphatidylcholine in AR-HDL. The changes in AR-HDL composition were associated with altered functional properties. In conclusion, AR alters HDL composition linked to decreased anti-oxidative and anti-inflammatory properties but improves the ability of HDL to suppress eosinophil effector responses.
Asunto(s)
Lipoproteínas HDL/inmunología , Rinitis Alérgica/inmunología , Adolescente , Adulto , Niño , Citocinas/análisis , Citocinas/inmunología , Femenino , Humanos , Inmunoglobulina E/inmunología , Lipoproteínas HDL/análisis , Masculino , Monocitos/inmunología , Adulto JovenRESUMEN
Presentation of exogenous antigens loaded on major histocompatibility complex class I molecules by antigen presenting cells, termed cross-presentation, is essential for the induction of CD8+ T cells and is performed mainly by specialized dendritic cell subsets. Research into this field has described two main mechanisms of cross-presentation, the cytosolic pathway and the vacuolar pathway. As the first step in cross-presentation, surface receptors relating to cross-presentation are required in the recognition and uptake of Ags, which include C-type lectin receptors, immunoglobulin γ Fc region receptor, chemokine receptor, scavenger receptor etc. After uptake by the cells, there are also many molecules that enable Ags to participate in cross-presentation pathways. By this approach, exogenous Ags can induce CD8+ T cells into cytotoxic T lymphocytes, which is of great significance to induce antitumor and antiviral immune responses, and the molecular mechanism would facilitate the development of related adjuvants. However, the detailed mechanisms of cross-presentation still remain unknown. In this paper, some latest researches, including two major pathways, DC surface receptors and application prospects are summarized.
Asunto(s)
Presentación de Antígeno , Antígenos Heterófilos/inmunología , Reactividad Cruzada , Isoantígenos/inmunología , Adyuvantes Inmunológicos/farmacología , Animales , Antígenos de Neoplasias/inmunología , Linfocitos T CD8-positivos/inmunología , Citosol/inmunología , Células Dendríticas/inmunología , Antígenos de Histocompatibilidad Clase I/inmunología , Humanos , Lipoproteínas HDL/inmunología , Ratones , Modelos Inmunológicos , Receptores Fc/inmunología , Receptores Inmunológicos/inmunología , Receptores de Lipoproteína/inmunología , Vacuolas/inmunologíaRESUMEN
The anti-inflammatory properties of high-density lipoproteins (HDL) are lost in uremia. These HDL may show pro-inflammatory features partially as a result of changed protein composition. Alterations of polymorphonuclear leukocytes (PMNLs) in chronic kidney disease (CKD) may contribute to chronic inflammation and high vascular risk. We investigated if HDL from uremic patients is related to systemic inflammation by interfering with PMNL function. PMNL apoptosis was investigated by assessing morphological features and DNA content. CD11b surface expression was quantified by flow cytometry. Oxidative burst was measured via cytochrome c reduction assay. Chemotaxis was assessed by using an under-agarose migration assay. We found that HDL from CKD and hemodialysis (HD) patients significantly attenuated PMNL apoptosis, whereas HDL isolated from healthy subjects had no effect on PMNL apoptosis. The use of signal transduction inhibitors indicated that uremic HDL exerts anti-apoptotic effects by activating pathways involving phosphoinositide 3-kinase and extracellular-signal regulated kinase. Healthy HDL attenuated the surface expression of CD11b, whereas HDL from CKD and HD patients had no effect. All tested isolates increased the stimulation of oxidative burst, but did not affect PMNL chemotactic movement. In conclusion, HDL may contribute to the systemic inflammation in uremic patients by modulating PMNL functions.
Asunto(s)
Lipoproteínas HDL/inmunología , Neutrófilos/inmunología , Insuficiencia Renal Crónica/inmunología , Uremia/inmunología , Adulto , Anciano , Apoptosis , Antígeno CD11b/inmunología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Diálisis RenalRESUMEN
PURPOSE OF REVIEW: Sepsis is a common syndrome of multiorgan system dysfunction caused by a dysregulated inflammatory response to an infection and is associated with high rates of mortality. Plasma lipid and lipoprotein levels and composition change profoundly during sepsis and have emerged as both biomarkers and potential therapeutic targets for this condition. The purpose of this article is to review recent progress in the understanding of the molecular regulation of lipid metabolism during sepsis. RECENT FINDINGS: Patients who experience greater declines in high-density lipoprotein during sepsis are at much greater risk of succumbing to organ failure and death. Although the causality of these findings remains unclear, all lipoprotein classes can sequester and prevent the excessive inflammation caused by pathogen-associated lipids during severe infections such as sepsis. This primordial innate immune function has been best characterized for high-density lipoproteins. Most importantly, results from human genetics and preclinical animal studies have suggested that several lipid treatment strategies, initially designed for atherosclerosis, may hold promise as therapies for sepsis. SUMMARY: Lipid and lipoprotein metabolism undergoes significant changes during sepsis. An improved understanding of the molecular regulation of these changes may lead to new opportunities for the treatment of sepsis.
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Proteínas de Transferencia de Ésteres de Colesterol/genética , Metabolismo de los Lípidos/genética , Lipoproteínas HDL/sangre , Insuficiencia Multiorgánica/genética , Proproteína Convertasa 9/genética , Sepsis/genética , Animales , Anticolesterolemiantes/uso terapéutico , Apolipoproteínas C/uso terapéutico , Biomarcadores/sangre , Proteínas de Transferencia de Ésteres de Colesterol/antagonistas & inhibidores , Proteínas de Transferencia de Ésteres de Colesterol/sangre , Proteínas de Transferencia de Ésteres de Colesterol/inmunología , Regulación de la Expresión Génica , Humanos , Inhibidores de Hidroximetilglutaril-CoA Reductasas/uso terapéutico , Inmunidad Innata , Metabolismo de los Lípidos/efectos de los fármacos , Metabolismo de los Lípidos/inmunología , Lipoproteína Lipasa/sangre , Lipoproteína Lipasa/genética , Lipoproteína Lipasa/inmunología , Lipoproteínas HDL/genética , Lipoproteínas HDL/inmunología , Insuficiencia Multiorgánica/sangre , Insuficiencia Multiorgánica/mortalidad , Insuficiencia Multiorgánica/prevención & control , Inhibidores de PCSK9 , Fragmentos de Péptidos/uso terapéutico , Proproteína Convertasa 9/sangre , Proproteína Convertasa 9/inmunología , Sepsis/sangre , Sepsis/tratamiento farmacológico , Sepsis/mortalidad , Análisis de SupervivenciaRESUMEN
The alternative pathway (AP) of complement is constantly active in plasma and can easily be activated on self surfaces and trigger local inflammation. Host cells are protected from AP attack by Factor H (FH), the main AP regulator in plasma. Although complement is known to play a role in atherosclerosis, the mechanisms of its contribution are not fully understood. Since FH via its domains 5-7 binds apoliporotein E (apoE) and macrophages produce apoE we examined how FH could be involved in the antiatherogenic effects of apoE. We used blood peripheral monocytes and THP-1 monocyte/macrophage cells which were also loaded with acetylated low-density lipoprotein (LDL) to form foam cells. Binding of FH and apoE on these cells was analyzed by flow cytometry. High-density lipoprotein (HDL)-mediated cholesterol efflux of activated THP-1 cells was measured and transcriptomes of THP-1 cells using mRNA sequencing were determined. We found that binding of FH to human blood monocytes and cholesterol-loaded THP-1 macrophages increased apoE binding to these cells. Preincubation of fluorescent cholesterol labeled THP-1 macrophages in the presence of FH increased cholesterol efflux and cholesterol-loaded macrophages displayed reduced transcription of proinflammatory/proatherogenic factors and increased transcription of anti-inflammatory/anti-atherogenic factors. Further incubation of THP-1 cells with serum reduced C3b/iC3b deposition. Overall, our data indicate that apoE and FH interact with monocytic cells in a concerted action and this interaction reduces complement activation and inflammation in the atherosclerotic lesions. By this way FH may participate in mediating the beneficial effects of apoE in suppressing atherosclerotic lesion progression.
Asunto(s)
Apolipoproteínas E/inmunología , Aterosclerosis/inmunología , Factor H de Complemento/inmunología , Células Espumosas/inmunología , Monocitos/inmunología , Aterosclerosis/patología , Complemento C3b/inmunología , Células Espumosas/patología , Humanos , Inflamación/inmunología , Inflamación/patología , Lipoproteínas HDL/inmunología , Monocitos/patología , Células THP-1 , Transcripción Genética/inmunologíaRESUMEN
Apolipoprotein A1 (ApoA1) of the high-density lipoprotein (HDL) plays a cardinal role in alleviating atherosclerosis in various ways. Its role in reverse cholesterol transport is preeminent. However, the ApoA1 undergoes oxidation under chronic inflammatory conditions and these oxidations are mediated by myeloperoxidase. It has been reported that the oxidation of the amino acids such as methionine, tyrosine, and tryptophan residues at specific sites of ApoA1 renders it not only dysfunctional but also proinflammatory and proatherogenic. Thus, assessing the quality of ApoA1 and, in turn, that of HDL in circulating blood can serve as an early diagnostic tool for cardiovascular diseases (CVDs). In this study, we developed monoclonal antibodies (mAbs) specific to modified ApoA1 with its tyrosine residue at the 166th position nitrated to 3-nitrotyrosine. A 20 amino acid peptide around the modification of interest was designed using an antigenicity prediction tool. The peptide was custom synthesized with ovalbumin as conjugate and used as an antigen to immunize BALB/c mice. Hybridomas were obtained by fusion of Sp2/0 mouse myeloma cells with spleen cells from the immunized mouse. A hybridoma clone 2E5B7, thus developed and characterized, was found to secrete mAb of the desired specificity and sensitivity against nitrated 166Tyrosine. The lowest concentration of the antigen that could be detected by the mAb with confidence was 15 ng. The mAb was able to detect nitrated 166Tyrosine peptide ovalbumin conjugate antigen spiked in human plasma with high specificity. The generated mAb could be potentially used in immuno-based diagnostic systems to screen the quality of HDL and in turn assess CVD risks in humans.
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Anticuerpos Monoclonales/biosíntesis , Apolipoproteína A-I/sangre , Aterosclerosis/sangre , Diagnóstico Precoz , Animales , Anticuerpos Monoclonales/inmunología , Apolipoproteína A-I/aislamiento & purificación , Aterosclerosis/inmunología , Aterosclerosis/patología , Humanos , Hibridomas/inmunología , Lipoproteínas HDL/sangre , Lipoproteínas HDL/inmunología , Ratones , Ratones Endogámicos BALB C , Oxidación-Reducción , Tirosina/análogos & derivados , Tirosina/inmunología , Tirosina/metabolismoRESUMEN
Quantitative and qualitative defects of high-density lipoprotein (HDL) are important in atherogenesis. In this study, we investigated whether antibodies against HDL components had additional value to conventional cardiovascular risk factors for the diagnosis of ischaemic stroke (IS) and coronary artery disease (CAD). Cross-sectional study was conducted on 53 patients with IS, 51 with CAD and 55 healthy controls, and in vitro studies to validate findings of the clinical study. We determined serum immunoglobulin G (IgG) antibodies against HDL (aHDL), apolipoproteins (aApoA-I, aApoA-II and aApoC-I) and paraoxonase-1 (aPON1) as well as PON1 activity (PON1a), total antioxidant capacity and biomarkers of endothelial activation (serum nitric oxide metabolites, 3-nitrotyrosine, VCAM-1 and ICAM-1); in vitro assays tested the capacity of IgG aHDL purified from high titer patients to inhibit PON1a and to reverse protective effect of HDL on endothelial cells. IgG aHDL, aApoA-I and aPON1 were higher in IS and CAD than controls (p < 0.001), predicted negatively PON1a and positively VCAM-1 and ICAM-1. By adding IgG aHDL and aApoA-I to a traditional cardiovascular risk factors model for IS and by adding IgG aHDL in a similar model for CAD, we obtained better discrimination of IS and CAD from healthy controls. IgG aHDL purified from IS and CAD inhibited PON1a by 38% (p < 0.01) and abrogated the protective effect of HDL on VCAM-1 expression by 126% compared with non-specific human IgG (p < 0.001). IgG against HDL components interfere with the antioxidant and anti-inflammatory properties of HDL and may represent novel biomarkers for vascular disease that need to be investigated in prospective studies.
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Biomarcadores/sangre , Enfermedad de la Arteria Coronaria/inmunología , Endotelio/fisiología , Inmunoglobulina G/sangre , Isquemia/inmunología , Lipoproteínas HDL/metabolismo , Accidente Cerebrovascular/inmunología , Anciano , Apolipoproteínas/inmunología , Arildialquilfosfatasa/inmunología , Arildialquilfosfatasa/metabolismo , Estudios de Casos y Controles , Enfermedad de la Arteria Coronaria/diagnóstico , Estudios Transversales , Femenino , Humanos , Isquemia/diagnóstico , Lipoproteínas HDL/inmunología , Masculino , Persona de Mediana Edad , Valor Predictivo de las Pruebas , Pronóstico , Riesgo , Accidente Cerebrovascular/diagnóstico , Molécula 1 de Adhesión Celular Vascular/metabolismoRESUMEN
The present study aimed to evaluate the possible role of immunoglobulin G (IgG) antibodies against high-density lipoproteins (HDL) and paraoxonase 1 (PON1) as possible biomarkers of cardiovascular disease (CVD) in systemic lupus erythematosus (SLE). To this end, levels of these autoantibodies, PON1 activity and total antioxidant capacity were quantified in serum samples from 198 SLE patients, 100 healthy controls (HC) and 42 non-autoimmune individuals with traditional cardiovascular risk factors. PON1 rs662 polymorphism was analysed in a subgroup of patients and controls. Subclinical CVD were determined by Doppler ultrasound in 118 SLE patients and 30 HC, analysing carotid intima-media thickness (IMT) and blood flow parameters in internal carotid, middle cerebral and basilar arteries. Serum levels of both anti-HDL and anti-PON1 antibodies were increased in SLE patients compared with HC (p < 0.001); however, only anti-PON1 antibodies, in addition to disease activity, were significant predictors of the impaired PON1 function in SLE (ß = -0.143, p = 0.045). Conversely, anti-HDL antibodies were associated with higher risk of CVD (odds ratio: 3.69; p = 0.012) and lower HDL levels at disease onset (ρ = -0.324, p = 0.044). Finally, anti-PON1 antibodies were associated with carotid IMT in SLE (ß = 0.201, p = 0.008) and inversely related to cranial arteries blood flow velocities in patients with clinical and subclinical CVD (all p < 0.001). In sum, these findings allowed us to propose serum levels of anti-PON1 and anti-HDL antibodies as potential early biomarkers of endothelial damage and premature atherosclerosis in SLE, thus constituting useful therapeutic targets for the prevention of future CVD in these patients.
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Arildialquilfosfatasa/inmunología , Autoanticuerpos/sangre , Enfermedades de las Arterias Carótidas/sangre , Inmunoglobulina G/sangre , Lipoproteínas HDL/inmunología , Lupus Eritematoso Sistémico/sangre , Adulto , Antioxidantes/análisis , Arildialquilfosfatasa/genética , Biomarcadores/sangre , Enfermedades de las Arterias Carótidas/diagnóstico por imagen , Enfermedades de las Arterias Carótidas/epidemiología , Enfermedades de las Arterias Carótidas/inmunología , Grosor Intima-Media Carotídeo , Estudios de Casos y Controles , Femenino , Humanos , Lupus Eritematoso Sistémico/diagnóstico , Lupus Eritematoso Sistémico/epidemiología , Lupus Eritematoso Sistémico/inmunología , Masculino , Persona de Mediana Edad , Polimorfismo de Nucleótido Simple , Valor Predictivo de las Pruebas , Prevalencia , Factores de Riesgo , España/epidemiología , Ultrasonografía DopplerRESUMEN
High-density lipoproteins (HDLs) can inhibit inflammatory cytokine expression on innate immune cells, but sometimes they promote cytokine production as suggested in a recent article in Cell Metabolism by van der Vorst et al. (2017). Kopecky et al. point out that the origin, handling, and storage conditions of HDL preparations dictate their functional properties and can specifically affect immune cells to evoke a pro-inflammatory response.
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Inmunidad Innata , Lipoproteínas HDL/inmunología , Animales , Citocinas/inmunología , Humanos , Inmunidad Celular , Inflamación/inmunología , Macrófagos/inmunologíaRESUMEN
Van der Vorst et al. underscore the relevance of HDL quality control, considering HDL source and processing, but argue that disease- or storage-associated structural modifications of HDL cannot explain the observed pro-inflammatory effects on macrophages. Discrepancies between reported effects of HDL in macrophages are probably related to methodological differences.
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Inflamación/inmunología , Lipoproteínas HDL/inmunología , Macrófagos/inmunología , Animales , Humanos , Inmunidad Innata , Macrófagos/química , Preservación Biológica , Proyectos de InvestigaciónRESUMEN
BACKGROUND: Both CD4+ T cells and macrophages are mainly involved in the autoimmune-mediated ß-cells destruction in type 1 diabetes (T1D). The aim of this study was to examine the effect of HDL on functional activities of macrophage and its ability to regulate the production of cytokines in autologous mixed macrophage/CD4+ T cells at the recent-onset human type 1 diabetes. METHODS: Cell samples were isolated from volunteers with recent-onset T1D or healthy controls. RESULTS: The levels of the production of IL-1ß, IL-2, IFN-γ, nitric oxide (NO), and hydrogen peroxide (H2O2) were significantly increased in the co-culture of T1D cells when compared to that of cells from healthy controls. Similarly, those of intracellular free calcium ions (ifCa2+) were slightly, but not significantly increased (p> 0.05). Conversely, macrophage exhibited significantly decreased levels of the relative tyrosine phosphorylation of STAT6 (p-STAT6, Tyr641) in culture of T1D cells than in that of cells from healthy controls; while those of p-STAT4 (Tyr693) were significantly increased. Likewise, the levels of IL-4 and IL-10 were significantly decreased in the co-culture of T1D cells compared to co-culture of cells from healthy controls. Additionally, HDL treatment significantly down-regulated the production of IL-1ß, IL-2, IFN-γ, NO, H2O2, phagocytosis, bacterial killing, the relative tyrosine phosphorylation of macrophage-expressed STAT4 (p-STAT4, Tyr693), as well as the ratio of IL-1ß/IL-10, NO production/arginase activity, p-STAT4/p-STAT6, IFN-γ/IL-4, IFN-γ/IL-10, and the combined proinflammatory (PICs)/anti-inflammatory (AICs) cytokines. Moreover, HDL treatment significantly up-regulated the production of IL-4, IL-10, arginase activity, and p-STAT6 (Tyr641) (for all comparisons, p< 0.001). CONCLUSIONS: We show for the first time that HDL may reverse both the functional activities of macrophages and immunoinflammatory response during reciprocal macrophage-CD4+ T cell crosstalk at the beginning of T1D. These findings should open the way for therapeutic trials in the short- and medium-term.