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1.
Front Immunol ; 15: 1358800, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38803493

RESUMO

During inflammation and tissue regeneration, the alarmin High Mobility Group Box 1 (HMGB1), in its reduced isoform, enhances the activity of the chemokine CXCL12, forming a heterocomplex that acts via the chemokine receptor CXCR4. Despite the established roles of both HMGB1 and CXCL12 in tumor progression and metastatic spread to distal sites, the role of the CXCL12/HMGB1 heterocomplex in cancer has never been investigated. By employing a newly established mass spectrometry protocol that allows an unambiguous distinction between reduced (red-HMGB1) and oxidized (ox-HMGB1) HMGB1 isoforms in cell lysates, we demonstrate that human epithelial cells derived from breast (MCF-7 and MDA-MB-231) and prostate (PC-3) cancer predominantly express red-HMGB1, while primary CD3+ T lymphocytes from peripheral blood express both HMGB1 isoforms. All these cancer cells release HMGB1 in the extracellular microenvironment together with varying concentrations of thioredoxin and thioredoxin reductase. The CXCL12/HMGB1 heterocomplex enhances, via CXCR4, the directional migration and invasiveness of cancer cells characterized by high metastatic potential that possess a fully active thioredoxin system, contributing to maintain red-HMGB1. On the contrary, cancer cells with low metastatic potential, lack thioredoxin reductase, promptly uptake CXCL12 and fail to respond to the heterocomplex. Our study demonstrates that the responsiveness of cancer cells to the CXCL12/HMGB1 heterocomplex, resulting in enhanced cell migration and invasiveness, depends on the maintenance of HMGB1 in its reduced isoform, and suggests disruption of the heterocomplex as a potential therapeutic target to inhibit invasion and metastatic spread in cancer therapies.


Assuntos
Movimento Celular , Quimiocina CXCL12 , Proteína HMGB1 , Isoformas de Proteínas , Receptores CXCR4 , Humanos , Proteína HMGB1/metabolismo , Proteína HMGB1/genética , Receptores CXCR4/metabolismo , Receptores CXCR4/genética , Quimiocina CXCL12/metabolismo , Linhagem Celular Tumoral , Feminino , Microambiente Tumoral , Masculino , Neoplasias/metabolismo , Neoplasias/patologia
2.
RMD Open ; 10(1)2024 Feb 23.
Artigo em Inglês | MEDLINE | ID: mdl-38395454

RESUMO

OBJECTIVES: Ankylosing spondylitis (AS) is a chronic inflammatory rheumatic disease affecting mainly the axial skeleton. Peripheral involvement (arthritis, enthesitis and dactylitis) and extra-musculoskeletal manifestations, including uveitis, psoriasis and bowel inflammation, occur in a relevant proportion of patients. AS is responsible for chronic and severe back pain caused by local inflammation that can lead to osteoproliferation and ultimately spinal fusion. The association of AS with the human leucocyte antigen-B27 gene, together with elevated levels of chemokines, CCL17 and CCL22, in the sera of patients with AS, led us to study the role of CCR4+ T cells in the disease pathogenesis. METHODS: CD8+CCR4+ T cells isolated from the blood of patients with AS (n=76) or healthy donors were analysed by multiparameter flow cytometry, and gene expression was evaluated by RNA sequencing. Patients with AS were stratified according to the therapeutic regimen and current disease score. RESULTS: CD8+CCR4+ T cells display a distinct effector phenotype and upregulate the inflammatory chemokine receptors CCR1, CCR5, CX3CR1 and L-selectin CD62L, indicating an altered migration ability. CD8+CCR4+ T cells expressing CX3CR1 present an enhanced cytotoxic profile, expressing both perforin and granzyme B. RNA-sequencing pathway analysis revealed that CD8+CCR4+ T cells from patients with active disease significantly upregulate genes promoting osteogenesis, a core process in AS pathogenesis. CONCLUSIONS: Our results shed light on a new molecular mechanism by which T cells may selectively migrate to inflammatory loci, promote new bone formation and contribute to the pathological ossification process observed in AS.


Assuntos
Espondilite Anquilosante , Humanos , Espondilite Anquilosante/genética , Espondilite Anquilosante/metabolismo , Osteogênese/genética , Subpopulações de Linfócitos T/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Inflamação
3.
J Med Chem ; 64(18): 13439-13450, 2021 09 23.
Artigo em Inglês | MEDLINE | ID: mdl-34510899

RESUMO

During inflammatory reactions, the production and release of chemotactic factors guide the recruitment of selective leukocyte subpopulations. The alarmin HMGB1 and the chemokine CXCL12, both released in the microenvironment, can form a heterocomplex, which exclusively acts on the chemokine receptor CXCR4, enhancing cell migration, and in some pathological conditions such as rheumatoid arthritis exacerbates the immune response. An excessive cell influx at the inflammatory site can be diminished by disrupting the heterocomplex. Here, we report the computationally driven identification of the first peptide (HBP08) binding HMGB1 and selectively inhibiting the activity of the CXCL12/HMGB1 heterocomplex. Furthermore, HBP08 binds HMGB1 with the highest affinity reported so far (Kd of 0.8 ± 0.4 µM). The identification of this peptide represents an important step toward the development of innovative pharmacological tools for the treatment of severe chronic inflammatory conditions characterized by an uncontrolled immune response.


Assuntos
Quimiocina CXCL12/antagonistas & inibidores , Proteína HMGB1/antagonistas & inibidores , Peptídeos/farmacologia , Ligação Proteica/efeitos dos fármacos , Sequência de Aminoácidos , Animais , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Quimiocina CXCL12/metabolismo , Proteína HMGB1/metabolismo , Humanos , Camundongos , Simulação de Acoplamento Molecular , Peptídeos/metabolismo , Receptores CXCR4/metabolismo
4.
Front Immunol ; 9: 2118, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30283452

RESUMO

Chemokine synergy-inducing molecules are emerging as regulating factors in cell migration. The alarmin HMGB1, in its reduced form, can complex with CXCL12 enhancing its activity on monocytes via the chemokine receptor CXCR4, while the form containing a disulfide bond, by binding to TLR2 or TLR4, initiates a cascade of events leading to production of cytokines and chemokines. So far, the possibility that the CXCL12/HMGB1 heterocomplex could be maintained in chronic inflammation was debated, due to the release of reactive oxygen species. Therefore, we have assessed if the heterocomplex could remain active in Rheumatoid Arthritis (RA) and its relevance in the disease assessment. Monocytes from RA patients with active disease require a low concentration of HMGB1 to enhance CXCL12-induced migration, in comparison to monocytes from patients in clinical remission or healthy donors. The activity of the heterocomplex depends on disease activity, on the COX2 and JAK/STAT pathways, and is determined by the redox potential of the microenvironment. In RA, the presence of an active thioredoxin system correlates with the enhanced cell migration, and with the presence of the heterocomplex in the synovial fluid. The present study highlights how, in an unbalanced microenvironment, the activity of the thioredoxin system plays a crucial role in sustaining inflammation. Prostaglandin E2 stimulation of monocytes from healthy donors is sufficient to recapitulate the response observed in patients with active RA. The activation of mechanisms counteracting the oxidative stress in the extracellular compartment preserves HMGB1 in its reduced form, and contributes to fuel the influx of inflammatory cells. Targeting the heterocomplex formation and its activity could thus be an additional tool for dampening the inflammation sustained by cell recruitment, for those patients with chronic inflammatory conditions who poorly respond to current therapies.


Assuntos
Artrite Reumatoide/metabolismo , Movimento Celular/efeitos dos fármacos , Quimiocina CXCL12/farmacologia , Proteína HMGB1/farmacologia , Monócitos/efeitos dos fármacos , Adulto , Idoso , Artrite Reumatoide/imunologia , Artrite Reumatoide/patologia , Movimento Celular/imunologia , Células Cultivadas , Dinoprostona/farmacologia , Sinergismo Farmacológico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Monócitos/imunologia , Monócitos/metabolismo , Oxirredução , Ligação Proteica/efeitos dos fármacos , Receptores CXCR4/imunologia , Receptores CXCR4/metabolismo , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia
5.
Arthritis Res Ther ; 20(1): 114, 2018 06 07.
Artigo em Inglês | MEDLINE | ID: mdl-29880013

RESUMO

BACKGROUND: B cells exert their pathogenic action in rheumatoid arthritis (RA) locally in the synovium. This study was undertaken to elucidate the chemokines responsible for the recruitment of B cells in the inflamed synovium, taking into account that the rich chemokine milieu present in the synovial tissue can fine-tune modulate discrete chemokine receptors. METHODS: Expression levels of chemokine receptors from the CC and CXC family, as well as CD27, were assessed by flow cytometry in CD20+ mononuclear cells isolated from the peripheral blood (PB) and synovial fluid (SF) of RA and psoriatic arthritis patients. Transwell experiments were used to study migration of B cells in response to a chemokine or in the presence of multiple chemokines. RESULTS: B cells from the SF of arthritis patients showed a significant increase in the surface expression of CCR1, CCR2, CCR4, CCR5 and CXCR4 with respect to PB. Conversely, SF B cells expressed consistently lower amounts of CXCR5, CXCR7 and CCR6, independent of CD27 expression. Analysis of permeabilized B cells suggested internalization of CXCR5 and CCR6 in SF B cells. In Transwell experiments, CCL20 and CXCL13, ligands of CCR6 and CXCR5, respectively, caused a significantly higher migration of B cells from PB than of those from SF of RA patients. Together, these two chemokines synergistically increased B-cell migration from PB, but not from SF. CONCLUSIONS: These results suggest that CXCL13 and CCL20 might play major roles in RA pathogenesis by acting singly on their selective receptors and synergistically in the accumulation of B cells within the inflamed synovium.


Assuntos
Artrite Reumatoide/metabolismo , Linfócitos B/metabolismo , Movimento Celular/fisiologia , Quimiocina CCL20/fisiologia , Quimiocina CXCL13/fisiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Artrite Reumatoide/imunologia , Artrite Reumatoide/patologia , Linfócitos B/imunologia , Células Cultivadas , Feminino , Humanos , Inflamação/imunologia , Inflamação/metabolismo , Inflamação/patologia , Masculino , Pessoa de Meia-Idade , Líquido Sinovial/citologia , Líquido Sinovial/imunologia , Líquido Sinovial/metabolismo , Membrana Sinovial/imunologia , Membrana Sinovial/metabolismo , Membrana Sinovial/patologia
6.
J Immunol ; 198(1): 184-195, 2017 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-27895171

RESUMO

CD4+ T cell repopulation of the gut is rarely achieved in HIV-1-infected individuals who are receiving clinically effective antiretroviral therapy. Alterations in the integrity of the mucosal barrier have been indicated as a cause for chronic immune activation and disease progression. In this study, we present evidence that persistent immune activation causes impairment of lymphocytes to respond to chemotactic stimuli, thus preventing their trafficking from the blood stream to peripheral organs. CCR6+ and CXCR3+ Th cells accumulate in the blood of aviremic HIV-1-infected patients on long-term antiretroviral therapy, and their frequency in the circulation positively correlates to levels of soluble CD14 in plasma, a marker of chronic immune activation. Th cells show an impaired response to chemotactic stimuli both in humans and in the pathogenic model of SIV infection, and this defect is due to hyperactivation of cofilin and inefficient actin polymerization. Taking advantage of a murine model of chronic immune activation, we demonstrate that cytoskeleton remodeling, induced by okadaic acid, restores lymphocyte migration in response to chemokines, both in vitro and in vivo. This study calls for novel pharmacological approaches in those pathological conditions characterized by persistent immune activation and loss of trafficking of T cell subsets to niches that sustain their maturation and activities.


Assuntos
Actinas/metabolismo , Quimiotaxia de Leucócito/imunologia , Infecções por HIV/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Animais , Separação Celular , Citoesqueleto/metabolismo , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , HIV-1 , Humanos , Imuno-Histoquímica , Macaca mulatta , Camundongos , Camundongos Endogâmicos C57BL , Polimerização , Reação em Cadeia da Polimerase em Tempo Real , Receptores CCR6/imunologia , Receptores CXCR3/imunologia , Síndrome de Imunodeficiência Adquirida dos Símios/imunologia
7.
Eur J Immunol ; 39(4): 1118-28, 2009 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-19291700

RESUMO

The migration of monocytes to sites of inflammation is largely determined by their response to chemokines. Although the chemokine specificities and expression patterns of chemokine receptors are well defined, it is still a matter of debate how cells integrate the messages provided by different chemokines that are concomitantly produced in physiological or pathological situations in vivo. We present evidence for one regulatory mechanism of human monocyte trafficking. Monocytes can integrate stimuli provided by inflammatory chemokines in the presence of homeostatic chemokines. In particular, migration and cell responses could occur at much lower concentrations of the CCR2 agonists, in the presence of chemokines (CCL19 and CCL21) that per se do not act on monocytes. Binding studies on CCR2(+) cells showed that CCL19 and CCL21 do not compete with the CCR2 agonist CCL2. Furthermore, the presence of CCL19 or CCL21 could influence the degradation of CCL2 and CCL7 on cells expressing the decoy receptor D6. These findings disclose a new scenario to further comprehend the complexity of chemokine-based monocyte trafficking in a vast variety of human inflammatory disorders.


Assuntos
Movimento Celular/imunologia , Quimiotaxia de Leucócito/imunologia , Inflamação/imunologia , Monócitos/imunologia , Receptores CCR2/imunologia , Receptores CCR7/imunologia , Sequência de Aminoácidos , Movimento Celular/efeitos dos fármacos , Quimiocina CCL19/química , Quimiocina CCL19/imunologia , Quimiocina CCL19/farmacologia , Quimiocina CCL2/imunologia , Quimiocina CCL2/farmacologia , Quimiocina CCL21/química , Quimiocina CCL21/imunologia , Quimiocina CCL21/farmacologia , Quimiocina CCL7/imunologia , Quimiocina CCL7/farmacologia , Quimiotaxia de Leucócito/efeitos dos fármacos , MAP Quinases Reguladas por Sinal Extracelular/imunologia , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Glicosaminoglicanos/imunologia , Glicosaminoglicanos/metabolismo , Humanos , Inflamação/metabolismo , Ligantes , Dados de Sequência Molecular , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Fosforilação/imunologia , Estrutura Terciária de Proteína , Receptores CCR10/imunologia , Receptores CCR10/metabolismo , Receptores CCR2/agonistas , Receptores CCR2/química , Receptores CCR7/agonistas , Receptores CCR7/química , Receptor D6 de Quimiocina
8.
Eur J Immunol ; 35(3): 746-56, 2005 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-15714581

RESUMO

In an attempt to clarify how cells integrate the signals provided by multiple chemokines expressed during inflammation, we have uncovered a novel mechanism regulating leukocyte trafficking. Our data indicate that the concomitant exposure to CCR4 agonists and CXCL10/IP-10 strongly enhances the chemotactic response of human T lymphocytes. This enhancement is synergistic rather than additive and occurs via CCR4 since it persists after CXCR3 blockade. Besides chemotaxis, other cellular responses are enhanced upon stimulation of CCR4-transfected cells with CCL22/MDC plus CXCL10. Several other chemokines in addition to CXCL10 were able to increase CCL22-mediated chemotaxis. The first beta-strand of the chemokine structure is highly and specifically implicated in this phenomenon, as established using synergy-inducing and non-synergy-inducing chimeric chemokines. As shown in situ for skin from atopic and allergic contact dermatitis patients, this organ becomes the ideal environment in which skin-homing CCR4(+) T lymphocytes can accumulate under the stimulus offered by CCR4 agonists, together with the synergistic chemokines that are concomitantly expressed. Overall, our results indicate that chemokine-induced synergism strengthens leukocyte recruitment towards tissues co-expressing several chemokines.


Assuntos
Quimiocinas CC/imunologia , Quimiocinas/química , Quimiotaxia de Leucócito/imunologia , Receptores de Quimiocinas/imunologia , Pele/imunologia , Sequência de Aminoácidos , Linhagem Celular , Quimiocina CCL22 , Quimiocina CXCL10 , Quimiocinas/imunologia , Quimiocinas CXC/imunologia , Dermatite Alérgica de Contato/imunologia , Dermatite Atópica/imunologia , Humanos , Imuno-Histoquímica , Imunoprecipitação , Dados de Sequência Molecular , Técnicas de Cultura de Órgãos , Estrutura Secundária de Proteína , Receptores CCR4 , Linfócitos T/imunologia
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