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1.
Oncol Rep ; 42(2): 521-532, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31173268

RESUMO

Breakpoint cluster region (BCR)­Abelson murine leukemia (ABL)1+ acute B­lymphoblastic leukemia (B­ALL) is a disease associated with a dismal prognosis and a high incidence of central nervous system (CNS) metastasis. However, BCR­ABL1+ B­ALL with CNS infiltration has not been previously characterized, at least to the best of our knowledge. In the present study, a murine model of BCR­ABL1+ B­ALL with CNS metastasis was established using retroviral transduction. The vast majority of BCR­ABL1+ leukemic cells were found to be immature B cells with a variable proportion of pro­B and pre­B populations. The present results indicated that the BCR­ABL1+ B­leukemic cells expressed high levels integrin subunit alpha 6 (Itga6) and L­selectin adhesion molecules, and have an intrinsic ability to disseminate and accumulate in CNS tissues, predominantly in meninges. On the whole, these results provide an approach for addressing the mechanisms of BCR­ABL1+ B­ALL with CNS metastasis and may guide the development of novel therapeutic strategies.


Assuntos
Neoplasias do Sistema Nervoso Central/secundário , Modelos Animais de Doenças , Proteínas de Fusão bcr-abl/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras B/patologia , Animais , Transplante de Medula Óssea , Neoplasias do Sistema Nervoso Central/metabolismo , Neoplasias do Sistema Nervoso Central/terapia , Feminino , Proteínas de Fusão bcr-abl/genética , Integrinas/metabolismo , Selectina L/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Leucemia-Linfoma Linfoblástico de Células Precursoras B/metabolismo , Leucemia-Linfoma Linfoblástico de Células Precursoras B/terapia
2.
Pharm Res ; 36(7): 97, 2019 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-31076925

RESUMO

PURPOSE: The aim of this research was to design dexamethasone palmitate (DP) loaded sialic acid modified liposomes, with the eventual goal of using peripheral blood neutrophils (PBNs) that carried drug-loaded liposomes to improve the therapeutic capacity for rheumatoid arthritis (RA). METHODS: A sialic acid - cholesterol conjugate (SA-CH) was synthesized and anchored on the surface of liposomal dexamethasone palmitate (DP-SAL). The physicochemical characteristics and in vitro cytotoxicity of liposomes were evaluated. Flow cytometry and confocal laser scanning microscopy were utilized to investigate the accumulation of liposomes in PBNs. The adjuvant-induced arthritis was adopted to investigate the targeting ability and anti-inflammatory effect of DP loaded liposomes. RESULTS: Both DP-CL and DP-SAL existed an average size less than 200 nm with remarkably high encapsulation efficiencies more than 90%. In vitro and in vivo experiments manifested SA-modified liposomes provided a reinforced accumulation of DP in PBNs. As well, DP-SAL displayed a greater degree of accumulation in the joints and a stronger anti-inflammatory effect in terms of RA suppression. CONCLUSIONS: SA-modified liposomal DP was a promising candidate for RA-targeting treatment through the neutrophil-mediated drug delivery system.


Assuntos
Artrite Reumatoide/tratamento farmacológico , Dexametasona/farmacocinética , Lipossomos/química , Ácido N-Acetilneuramínico/química , Neutrófilos/metabolismo , Animais , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/farmacocinética , Anti-Inflamatórios/toxicidade , Artrite Experimental/tratamento farmacológico , Artrite Experimental/patologia , Artrite Reumatoide/patologia , Colesterol/química , Dexametasona/administração & dosagem , Dexametasona/toxicidade , Liberação Controlada de Fármacos , Articulações/efeitos dos fármacos , Articulações/patologia , Selectina L/metabolismo , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/patologia , Masculino , Neutrófilos/patologia , Palmitatos/química , Ratos Wistar , Distribuição Tecidual
3.
PLoS Pathog ; 15(3): e1007633, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30875408

RESUMO

Memory CD8+ T cells in the circulation rapidly infiltrate non-lymphoid tissues following infection and provide protective immunity in an antigen-specific manner. However, the subsequent fate of memory CD8+ T cells after entering non-lymphoid tissues such as the skin during a secondary infection is largely unknown. Furthermore, because expression of CD62L is often used to identify the central memory (TCM) CD8+ T cell subset, uncoupling the physical requirement for CD62L-mediated lymph node homing versus other functional attributes of TCM CD8+ T cells remains unresolved. Here, we show that in contrast to naïve CD8+ T cells, memory CD8+ T cells traffic into the skin independent of CD62L-mediated lymph node re-activation and provide robust protective immunity against Vaccinia virus (VacV) infection. TCM, but not effector memory (TEM), CD8+ T cells differentiated into functional CD69+/CD103- tissue residents following viral clearance, which was also dependent on local recognition of antigen in the skin microenvironment. Finally, we found that memory CD8+ T cells expressed granzyme B after trafficking into the skin and utilized cytolysis to provide protective immunity against VacV infection. Collectively, these findings demonstrate that TCM CD8+ T cells become cytolytic following rapid infiltration of the skin to protect against viral infection and subsequently differentiate into functional CD69+ tissue-residents.


Assuntos
Linfócitos T CD8-Positivos/metabolismo , Memória Imunológica/fisiologia , Pele/imunologia , Animais , Antígenos CD/metabolismo , Antígenos CD/fisiologia , Antígenos de Diferenciação de Linfócitos T/metabolismo , Antígenos de Diferenciação de Linfócitos T/fisiologia , Linfócitos T CD8-Positivos/virologia , Feminino , Selectina L/metabolismo , Lectinas Tipo C/metabolismo , Lectinas Tipo C/fisiologia , Linfonodos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Pele/virologia , Subpopulações de Linfócitos T/metabolismo , Subpopulações de Linfócitos T/fisiologia , Vírus Vaccinia/imunologia , Vírus Vaccinia/patogenicidade
4.
Int J Mol Sci ; 20(6)2019 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-30897843

RESUMO

Viral infections and reactivations remain a serious obstacle to successful hematopoietic stem cell transplantation (HSCT). When antiviral drug treatment fails, adoptive virus-specific T-cell transfer provides an effective alternative. Assuming that naive T cells (TN) are mainly responsible for GvHD, methods were developed to generate naive T-cell-depleted products while preserving immune memory against viral infections. We compared two major strategies to deplete potentially alloreactive T cells: CD45RA and CD62L depletion and analyzed phenotype and functionality of the resulting CD45RA-/CD62L- naive T-cell-depleted as well as CD45RA⁺/CD62L⁺ naive T-cell-enriched fractions in the CMV pp65 and IE1 antigen model. CD45RA depletion resulted in loss of terminally differentiated effector memory T cells re-expressing CD45RA (TEMRA), and CD62L depletion in loss of central memory T cells (TCM). Based on these differences in target cell-dependent and target cell-independent assays, antigen-specific T-cell responses in CD62L-depleted fraction were consistently 3⁻5 fold higher than those in CD45RA-depleted fraction. Interestingly, we also observed high donor variability in the CD45RA-depleted fraction, resulting in a substantial loss of immune memory. Accordingly, we identified donors with expected response (DER) and unexpected response (DUR). Taken together, our results showed that a naive T-cell depletion method should be chosen individually, based on the immunophenotypic composition of the T-cell populations present.


Assuntos
Imunoterapia/métodos , Selectina L/metabolismo , Antígenos Comuns de Leucócito/metabolismo , Subpopulações de Linfócitos T/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/imunologia , Linfócitos T CD8-Positivos/metabolismo , Citomegalovirus/imunologia , Doença Enxerto-Hospedeiro/imunologia , Doença Enxerto-Hospedeiro/metabolismo , Humanos , Imunofenotipagem , Microesferas , Linfócitos T Reguladores/imunologia , Linfócitos T Reguladores/metabolismo
5.
Immunol Invest ; 48(6): 632-643, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-30887869

RESUMO

It is well documented that age-related impaired functioning of immunocompetent cells is associated with an increase in the rates of chronic inflammatory diseases. Recently, an ability of melatonin to modulate inflammatory processes by regulating leucocyte recruitment has been demonstrated. However, to date, no studies have attempted to determine the impact of melatonin on the expression of CD62L by lymphocytes. CD62L, also known as L-selectin, is required for the entry of lymphocytes into secondary lymphoid organs, sites of tumor growth and chronic inflammation through high endothelial venules. Here, we investigated the effect of melatonin at physiological concentrations on the expression of CD62L by T and NK cells in vivo and in vitro. We demonstrated that NK and CD3+ T cells obtained from the spleen of aged mice were characterized by decreased expression of CD62L compared to young mice. Melatonin administration up-regulated the levels of surface CD62L on NK and T cell populations in aged mice under non-inflammatory conditions and on CD8+ T cells in aged mice with chronic inflammation. Pre-incubation with melatonin prevented the reduction in CD62L expression by CD8+ T cells induced by the co-cultivation of peripheral blood mononuclear cells with human pancreatic adenocarcinoma cell line (MiaPaCa-2). The obtained results suggest that melatonin can modulate lymphocyte homing into lymph nodes and sites of chronic inflammation and, therefore, can stimulate immune responses in chronic inflammatory conditions associated with aging.


Assuntos
Linfócitos T CD8-Positivos/fisiologia , Inflamação/imunologia , Células Matadoras Naturais/fisiologia , Selectina L/metabolismo , Melatonina/metabolismo , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/microbiologia , Envelhecimento , Animais , Humanos , Inflamação/tratamento farmacológico , Selectina L/genética , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos , Neoplasias Pancreáticas/patologia , Células Tumorais Cultivadas , Regulação para Cima
6.
Innate Immun ; 25(1): 46-59, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30782043

RESUMO

Myeloid-derived suppressor cells (MDSCs) inhibit T cell responses and are relevant to cancer, autoimmunity and transplant biology. Anti-thymocyte globulin (ATG) is a commonly used T cell depletion agent, yet the effect of ATG on MDSCs has not been investigated. MDSCs were generated in Lewis Lung Carcinoma 1 tumor-bearing mice. MDSC development and function were assessed in vivo and in vitro with and without ATG administration. T cell suppression assays, RT-PCR, flow cytometry and arginase activity assays were used to assess MDSC phenotype and function. MDSCs increased dramatically in tumor-bearing mice and the majority of splenic MDSCs were of the polymorphonuclear subset. MDSCs potently suppressed T cell proliferation. ATG-treated mice developed 50% fewer MDSCs and these MDSCs were significantly less suppressive of T cell proliferation. In vitro, ATG directly bound 99.6% of MDSCs. CCR7, L-selectin and LFA-1 were expressed by both T cells and MDSCs, and binding of LFA-1 was inhibited by ATG pre-treatment. Arg-1 and PD-L1 transcript expression were reduced 30-40% and arginase activity decreased in ATG-pretreated MDSCs. MDSCs were bound and functionally inhibited by ATG. T cells and MDSCs expressed common Ags which were also targets of ATG. ATG may be helpful in tumor models seeking to suppress MDSCs. Alternatively, ATG may inadvertently inhibit important T cell regulatory events in autoimmunity and transplantation.


Assuntos
Soro Antilinfocitário/farmacologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Carcinoma Pulmonar de Lewis/patologia , Células Supressoras Mieloides/efeitos dos fármacos , Animais , Antígeno B7-H1/genética , Antígeno B7-H1/metabolismo , Proliferação de Células , Células Cultivadas , Regulação para Baixo , Imunossupressão , Selectina L/genética , Selectina L/metabolismo , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos C57BL , Células Supressoras Mieloides/fisiologia , Receptores CCR7/metabolismo
7.
Gastroenterology ; 156(6): 1775-1787, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30710527

RESUMO

BACKGROUND & AIMS: Crohn's disease (CD) is characterized by an imbalance of effector and regulatory T cells in the intestinal mucosa. The efficacy of anti-adhesion therapies led us to investigate whether impaired trafficking of T-regulatory (Treg) cells contributes to the pathogenesis of CD. We also investigated whether proper function could be restored to Treg cells by ex vivo expansion in the presence of factors that activate their regulatory activities. METHODS: We measured levels of the integrin α4ß7 on Treg cells isolated from peripheral blood or lamina propria of patients with CD and healthy individuals (controls). Treg cells were expanded ex vivo and incubated with rapamycin with or without agonists of the retinoic acid receptor-α (RARA), and their gene expression profiles were analyzed. We also studied the cells in cytokine challenge, suppression, and flow chamber assays and in SCID mice with human intestinal xenografts. RESULTS: We found that Treg cells from patients with CD express lower levels of the integrin α4ß7 than Treg cells from control patients. The pathway that regulates the expression of integrin subunit α is induced by retinoic acid (RA). Treg cells from patients with CD incubated with rapamycin and an agonist of RARA (RAR568) expressed high levels of integrin α4ß7, as well as CD62L and FOXP3, compared with cells incubated with rapamycin or rapamycin and all-trans retinoic acid. These Treg cells had increased suppressive activities in assays and migrated under conditions of shear flow; they did not produce inflammatory cytokines, and RAR568 had no effect on cell stability or lineage commitment. Fluorescently labeled Treg cells incubated with RAR568 were significantly more likely to traffic to intestinal xenografts than Treg cells expanded in control medium. CONCLUSIONS: Treg cells from patients with CD express lower levels of the integrin α4ß7 than Treg cells from control patients. Incubation of patients' ex vivo expanded Treg cells with rapamycin and an RARA agonist induced expression of α4ß7 and had suppressive and migratory activities in culture and in intestinal xenografts in mice. These cells might be developed for treatment of CD. ClinicalTrials.gov, Number: NCT03185000.


Assuntos
Doença de Crohn/imunologia , Integrinas/metabolismo , Receptor alfa de Ácido Retinoico/agonistas , Linfócitos T Reguladores/efeitos dos fármacos , Linfócitos T Reguladores/metabolismo , Adulto , Animais , Antineoplásicos/farmacologia , Estudos de Casos e Controles , Técnicas de Cultura de Células , Movimento Celular/efeitos dos fármacos , Células Cultivadas , Feminino , Fatores de Transcrição Forkhead/metabolismo , Expressão Gênica/efeitos dos fármacos , Xenoenxertos , Humanos , Imunossupressores/farmacologia , Integrinas/genética , Mucosa Intestinal/imunologia , Mucosa Intestinal/transplante , Selectina L/metabolismo , Ativação Linfocitária , Masculino , Camundongos , Camundongos SCID , Pessoa de Meia-Idade , Compostos Orgânicos/farmacologia , Sirolimo/farmacologia , Linfócitos T Reguladores/imunologia , Transcriptoma/efeitos dos fármacos , Tretinoína/farmacologia
8.
J Infect Dis ; 219(9): 1474-1482, 2019 04 16.
Artigo em Inglês | MEDLINE | ID: mdl-30452713

RESUMO

BACKGROUND: Eosinophils are a prominent cell type in the host response to helminths, and some evidence suggests that neutrophils might also play a role. However, little is known about the activation status of these granulocytes during helminth infection. METHODS: We analyzed the expression of eosinophil and neutrophil activation markers in peripheral blood by flow cytometry and measured serum levels of eosinophil granule proteins in 300 subjects residing in an area endemic for soil-transmitted helminths (STH). The data generated are on samples before and after 1 year of 3-monthly albendazole treatment. RESULTS: Anthelmintic treatment significantly reduced the prevalence of STH. While eosinophil numbers were significantly higher in STH-infected compared to uninfected subjects and significantly decreased following albendazole treatment, there was no effect exerted by the helminths on either eosinophil nor neutrophil activation. Although at baseline eosinophil granule protein levels were not different between STH-infected and uninfected subjects, treatment significantly reduced the levels of eosinophil-derived neurotoxin (EDN) in those infected at baseline. CONCLUSIONS: These results show that besides decreasing eosinophil numbers, anthelmintic treatment does not significantly change the activation status of eosinophils, nor of neutrophils, and the only effect seen was a reduction in circulating levels of EDN. CLINICAL TRIALS REGISTRATION: http://www.isrctn.com/ISRCTN75636394.


Assuntos
Albendazol/uso terapêutico , Anti-Helmínticos/uso terapêutico , Proteínas Granulares de Eosinófilos/sangue , Eosinófilos/metabolismo , Helmintíase/sangue , Adulto , Antígenos CD/metabolismo , Antígenos de Diferenciação de Linfócitos T/metabolismo , Grupo com Ancestrais do Continente Asiático , Biomarcadores/sangue , Antígeno CD11b/metabolismo , Estudos de Casos e Controles , Moléculas de Adesão Celular/metabolismo , Proteína Catiônica de Eosinófilo/sangue , Proteína Básica Maior de Eosinófilos/sangue , Neurotoxina Derivada de Eosinófilo/sangue , Eosinófilos/imunologia , Grupo com Ancestrais do Continente Europeu , Feminino , Proteínas Ligadas por GPI/metabolismo , Helmintíase/tratamento farmacológico , Helmintíase/imunologia , Humanos , Indonésia , Selectina L/metabolismo , Lectinas Tipo C/metabolismo , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Ativação de Neutrófilo , Neutrófilos/imunologia , Neutrófilos/metabolismo , Receptores de Complemento 3b/metabolismo
9.
Am J Pathol ; 189(3): 604-618, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30593820

RESUMO

Necrotizing enterocolitis (NEC) is a devastating disease affecting premature infants with intestinal inflammation and necrosis. The neonatal intestinal inflammatory response is rich in macrophages, and blood monocyte count is low in human NEC. We previously found that NF-κB mediates the intestinal injury in experimental NEC. However, the role of NF-κB in myeloid cells during NEC remains unclear. Herein, inhibitor of kappaB kinase ß (IKKß), a critical kinase mediating NF-κB activation, was deleted in lysozyme M (Lysm)-expressing cells, which were found to be Cd11b+Ly6c+ monocytes but not Cd11b+Ly6c- macrophages in the dam-fed neonatal mouse intestine. NEC induced differentiation of monocytes into intestinal macrophages and up-regulation of monocyte recruitment genes (eg, L-selectin) in the macrophage compartment in wild-type mice, but not in pups with IKKß deletion in Lysm+ cells. Thus, NF-κB is required for NEC-induced monocyte activation, recruitment, and differentiation in neonatal intestines. Furthermore, pups with Lysm-IKKß deletion had improved survival and decreased incidence of severe NEC compared with littermate controls. Decreased NEC severity was not associated with an improved intestinal barrier. In contrast, NEC was unabated in mice with IKKß deletion in intestinal epithelial cells. Together, these data suggest that recruitment of Ly6c+ monocytes into the intestine, NF-κB activation in these cells, and differentiation of Ly6c+ monocytes into macrophages are critical cellular and molecular events in NEC development to promote disease.


Assuntos
Antígenos Ly/metabolismo , Enterocolite Necrosante/metabolismo , Células Epiteliais/metabolismo , Macrófagos/metabolismo , Monócitos/metabolismo , NF-kappa B/metabolismo , Animais , Antígenos Ly/genética , Enterocolite Necrosante/genética , Enterocolite Necrosante/patologia , Células Epiteliais/patologia , Deleção de Genes , Quinase I-kappa B/genética , Quinase I-kappa B/metabolismo , Selectina L/genética , Selectina L/metabolismo , Macrófagos/patologia , Camundongos , Camundongos Transgênicos , NF-kappa B/antagonistas & inibidores , NF-kappa B/genética , Regulação para Cima
11.
J Immunol ; 202(1): 171-182, 2019 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-30504420

RESUMO

Innate lymphoid cells (ILCs) guard epithelial tissue integrity during homeostasis, but can be potent immune effector cells during inflammation. Precursors to all ILC subsets (ILC precursors [ILCP]) have been identified in human peripheral blood (PB). We found that during homeostasis, ILCP in PB of mouse and human expressed homing receptors for secondary lymphoid organs, mainly CD62L. These ILCP entered mouse lymph nodes in a CD62L-dependent way and relied on S1P receptors for their exit. Importantly, CD62L expression was absent on human ILCs expressing NKp44 in tonsils and PB of Crohn disease patients, and relatively fewer CD62L+ ILCP were present in PB of Crohn disease patients. These data are in agreement with selective expression of CD62L on nonactivated ILCP. As such, we conclude that CD62L not only serves as a functional marker of ILCP, but has potential to be used in the clinic as a diagnostic marker in inflammatory disorders.


Assuntos
Células Sanguíneas/imunologia , Doença de Crohn/imunologia , Selectina L/metabolismo , Linfonodos/imunologia , Linfócitos/imunologia , Células Progenitoras Linfoides/fisiologia , Animais , Células Cultivadas , Feminino , Homeostase , Humanos , Imunidade Inata , Selectina L/genética , Ativação Linfocitária , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Receptor 2 Desencadeador da Citotoxicidade Natural/metabolismo , Receptores de Lisoesfingolipídeo/metabolismo
12.
J Dermatol ; 46(1): 65-69, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30450688

RESUMO

We report the case of an 88-year-old Japanese man with erythrodermic involvement of T-cell prolymphocytic leukemia (T-PLL). He had a history of pharyngeal diffuse large B-cell lymphoma successfully treated with polychemotherapy including cyclophosphamide and epirubicin, 6 years before the current illness. He presented with numerous reddish, coalescing, flat-topped papules on the trunk and extremities, sparing the skin folds of the abdomen, the features of which mimicked those of papuloerythroderma. Immunohistochemistry showed perivascular and epidermotropic infiltration of CD3+ CD4+ T cells in the cutaneous lesion. However, flow cytometric analysis revealed that the skin infiltrating T cells were negative for surface CD4, and that CD3+ CD4- CD8- cells made up 92% of the T-cell fraction of peripheral blood. The circulating atypical T cells had a round or oval nucleus and prominent nucleoli, and the deletion of chromosomes 6q, 13 and 17. These cytological profiles were consistent with those of T-PLL and distinct from those of Sézary cells. The same T-cell clone was detected in the cutaneous lesion and peripheral blood, but the expression of CD62L was absent in the skin infiltrates and present in the circulating cells. No specific mutation was detected in STAT3 or STAT5B. Although low-dose oral etoposide had a beneficial effect on the skin rash, a fatal crisis of marked leukocytosis (169 × 103 /µL) occurred 19 months after the illness onset. CD62L-leukemic cells of T-PLL may infiltrate the skin to form papuloerythroderma-like cutaneous lesions.


Assuntos
Dermatite Esfoliativa/patologia , Selectina L/metabolismo , Leucemia Prolinfocítica de Células T/patologia , Neoplasias Cutâneas/patologia , Idoso de 80 Anos ou mais , Biópsia , Linfócitos T CD4-Positivos/metabolismo , Dermatite Esfoliativa/sangue , Dermatite Esfoliativa/diagnóstico , Evolução Fatal , Citometria de Fluxo , Humanos , Leucemia Prolinfocítica de Células T/sangue , Leucemia Prolinfocítica de Células T/diagnóstico , Masculino , Testes Sorológicos , Pele/patologia , Neoplasias Cutâneas/sangue , Neoplasias Cutâneas/diagnóstico
13.
Inflammation ; 42(1): 276-289, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30251217

RESUMO

Recent data have demonstrated that chronic inflammation is a crucial component of tumor initiation and progression. We previously reported that immature myeloid-derived suppressor cells (MDSCs) with immunosuppressive activity toward effector T cells were expanded in experimental chronic inflammation. We hypothesized that elevated levels of MDSCs, induced by chronic inflammation, may contribute to the progression of tumor growth. Using the Ehrlich carcinoma animal model, we found increased tumor growth in mice with chronic adjuvant arthritis, which was accompanied by a persistent increase in the proportion of splenic monocytic and granulocytic MDSCs expressing CD62L (L-selectin), when compared to tumor mice without adjuvant arthritis. Depletion of inflammation-induced MDSCs resulted in decreased tumor growth. In vitro studies demonstrated that increased expression of CD62L by MDSCs was mediated by TNFα, elevated concentrations of which were found in tumor mice subjected to chronic inflammation. Moreover, the addition of exogenous TNFα markedly enhanced the suppressive activity of bone marrow-derived MDSCs, as revealed by the ability to impair the proliferation of CD8+ T cells in vitro. This study provides evidence that chronic inflammation may promote tumor growth via induction of CD62L expression by MDSCs that can facilitate their migration to tumor and lymph nodes and modulation of their suppressor activity.


Assuntos
Artrite Experimental/complicações , Inflamação/complicações , Selectina L/metabolismo , Células Supressoras Mieloides/metabolismo , Carga Tumoral , Animais , Movimento Celular , Doença Crônica , Camundongos , Fator de Necrose Tumoral alfa/farmacologia
14.
Int J Cancer ; 144(5): 1128-1134, 2019 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-30178523

RESUMO

Reprogramming of neutrophils by malignant cells is well-described for many types of solid tumors, but data remain scarce for hematological diseases. Chronic lymphocytic leukemia (CLL) is characterized for a deep immune dysregulation mediated by leukemic cells that compromises patient's outcome. Murine models of CLL highlight the relevance of myeloid cells as tumor-driven reprogramming targets. In our study, we evaluated neutrophil reprogramming by CLL cells. We first show that the proportion of the CD16high CD62Ldim neutrophil subset in peripheral blood of CLL patients is increased compared to age-matched healthy donors (HD). In vitro, neutrophils from HD cultured in the presence of CLL cells or conditioned media (CM) from CLL cells exhibited a longer lifespan. Depletion of G-CSF and GM-CSF from CM partially reversed the protective effect. In addition, the proportion of viable neutrophils that displayed a CD16high CD62Ldim phenotype was increased in the presence of CM from CLL cells, being TGF-ß/IL-10 responsible for this effect. Altogether, our results describe a novel mechanism through which CLL cells can manipulate neutrophils.


Assuntos
Diferenciação Celular/fisiologia , Tolerância Imunológica/fisiologia , Selectina L/metabolismo , Leucemia Linfocítica Crônica de Células B/metabolismo , Leucemia Linfocítica Crônica de Células B/patologia , Neutrófilos/patologia , Receptores de IgG/metabolismo , Idoso , Linhagem Celular Tumoral , Feminino , Proteínas Ligadas por GPI/metabolismo , Fator Estimulador de Colônias de Granulócitos/metabolismo , Fator Estimulador de Colônias de Granulócitos e Macrófagos/metabolismo , Humanos , Masculino , Pessoa de Meia-Idade , Neutrófilos/metabolismo , Fator de Crescimento Transformador beta/metabolismo
15.
FASEB J ; 33(3): 3225-3236, 2019 03.
Artigo em Inglês | MEDLINE | ID: mdl-30383448

RESUMO

Previously we reported that the sensitivity of CD4+ T cells to ATP does not depend on P2X7 receptor (P2X7R) expression levels but on their activation and differentiation stages. Therefore, here we have investigated a potential relationship between the sensitivity of CD8+ T cells to ATP and their stages of differentiation. Thus, the CD8+ subpopulation exhibits a drastically reduced sensitivity to ATP with aging, which parallels the strong increase of an effector/memory CD8+ subset expressing high levels of CD44 cell adhesion molecule and CD45RB transmembrane phosphatase (CD44hiCD45RBhi). Using l-selectin/CD62L, CC-chemokine receptor 7, and CD127/IL-7 receptor-α markers, we showed that effector/memory CD8+ T cells belong to a central or effector memory subset. In contrast, the CD44hiCD45RBhi effector/memory subset is absent or poorly expressed in the CD4+ T subpopulation regardless of age. While ATP treatment can trigger channel and pore formation, CD62L shedding, phosphatidylserine exposure, and cell death in the CD44loCD45RBhi-naive CD8+ subset, it is unable to induce these cellular activities in the CD44hiCD45RBhi effector/memory CD8+ subset. Importantly, both CD44loCD45RBhi-naive and CD44hiCD45RBhi effector/memory subsets express similar low levels of P2X7R, demonstrating that the sensitivity of CD8+ T cells to ATP depends on the stage of differentiation instead of P2X7R expression levels.-Mellouk, A., Bobé, P. CD8+, but not CD4+ effector/memory T cells, express the CD44highCD45RBhigh phenotype with aging, which displays reduced expression levels of P2X7 receptor and ATP-induced cellular responses.


Assuntos
Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD8-Positivos/metabolismo , Receptores Purinérgicos P2X7/metabolismo , Trifosfato de Adenosina/metabolismo , Envelhecimento/imunologia , Envelhecimento/metabolismo , Animais , Linfócitos T CD4-Positivos/classificação , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/classificação , Linfócitos T CD8-Positivos/imunologia , Sinalização do Cálcio , Diferenciação Celular/imunologia , Receptores de Hialuronatos/metabolismo , Memória Imunológica , Selectina L/metabolismo , Antígenos Comuns de Leucócito/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Fenótipo , Receptores Purinérgicos P2X7/deficiência , Receptores Purinérgicos P2X7/genética , Subpopulações de Linfócitos T/citologia , Subpopulações de Linfócitos T/imunologia , Subpopulações de Linfócitos T/metabolismo
16.
J Immunol Res ; 2018: 8175810, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30364054

RESUMO

Introduction: It has been previously shown that immunoregulatory DX5+NKT cells are able to prevent colitis induced by CD4+CD62Lhigh T lymphocytes in a SCID mouse model. The aim of this study was to further investigate the underlying mechanism in vitro. Methods: CD4+CD62Lhigh and DX5+NKT cells from the spleen of Balb/c mice were isolated first by MACS, followed by FACS sorting and cocultured for up to 96 h. After polyclonal stimulation with anti-CD3, anti-CD28, and IL-2, proliferation of CD4+CD62Lhigh cells was assessed using a CFSE assay and activity of proapoptotic caspase-3 was determined by intracellular staining and flow cytometry. Extrinsic apoptotic pathway was blocked using an unconjugated antibody against FasL, and activation of caspase-3 was measured. Results: As previously shown in vivo, DX5+NKT cells inhibit proliferation of CD4+CD62Lhigh cells in vitro after 96 h coculture compared to a CD4+CD62Lhigh monoculture (proliferation index: 1.39 ± 0.07 vs. 1.76 ± 0.12; P = 0.0079). The antiproliferative effect of DX5+NKT cells was likely due to an induction of apoptosis in CD4+CD62Lhigh cells as evidenced by increased activation of the proapoptotic caspase-3 after 48 h (38 ± 3% vs. 28 ± 3%; P = 0.0451). Furthermore, DX5+NKT cells after polyclonal stimulation showed an upregulation of FasL on their cell surface (15 ± 2% vs. 2 ± 1%; P = 0.0286). Finally, FasL was blocked on DX5+NKT cells, and therefore, the extrinsic apoptotic pathway abrogated the activation of caspase-3 in CD4+CD62Lhigh cells. Conclusion: Collectively, these data confirmed that DX5+NKT cells inhibit proliferation of colitis-inducing CD4+CD62Lhigh cells by induction of apoptosis. Furthermore, DX5+NKT cells likely mediate their cytotoxic and proapoptotic potentials via FasL, confirming recent reports about iNKT cells. Further studies will be necessary to evaluate the therapeutical potential of these immunoregulatory cells in patients with colitis.


Assuntos
Linfócitos T CD4-Positivos/imunologia , Colite/imunologia , Células T Matadoras Naturais/imunologia , Animais , Apoptose , Caspase 3/metabolismo , Proliferação de Células , Células Cultivadas , Técnicas de Cocultura , Citotoxicidade Imunológica , Modelos Animais de Doenças , Proteína Ligante Fas/metabolismo , Integrina alfa2/metabolismo , Selectina L/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos SCID
17.
PLoS One ; 13(10): e0206175, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30376575

RESUMO

Endurance exercise is associated with a transient increase in neutrophil counts in the peripheral blood. Here we investigate the impact of intensified endurance exercise on the neutrophil compartment. We hypothesized that intensified endurance exercise leads to mobilization of neutrophil subsets, which are normally absent in the blood. Furthermore, we followed the potential build-up of neutrophil activation and the impact on overnight recovery of the neutrophil compartment during a seven-day cycling tour. The neutrophil compartment was studied in 28 healthy amateur cyclists participating in an eight-day strenuous cycling tour. Blood samples were taken at baseline, after 4 days and after 7 days of cycling. The neutrophil compartment was analyzed in terms of numbers and its phenotype by deep phenotyping of flow cytometry data with the multi-dimensional analysis method FLOOD. Repeated endurance exercise led to a gradual increase in total neutrophil counts over the days leading to a 1.26 fold-increase (95%CI 1.01-1.51 p = 0.0431) in the morning of day 8. Flow cytometric measurements revealed the appearance of 2 additional neutrophil subsets: CD16brightCD62Ldim and CD16dimCD62Lbright. A complex change in neutrophil phenotypes was present characterized by decreased expression of both CD11b and CD62L and marked increased expression of LAIR-1, VLA-4 and CBRM1/5. The changes in expression were found on all neutrophils present in the blood. Strikingly, in strong contrast to our findings during acute inflammation evoked by LPS challenge, these neutrophils did not upregulate classical degranulation markers. In fact, our FLOOD analysis revealed that the exercise induced neutrophil phenotype did not overlap with the neutrophil subsets arising upon acute inflammation. In conclusion, during multiple days of endurance exercise the neutrophil compartment does not regain homeostasis overnight. Thereby our study supports the concept of a build-up of inflammatory cues during repeated endurance exercise training, causing a prolonged change of the systemic neutrophil compartment.


Assuntos
Citometria de Fluxo/métodos , Neutrófilos/citologia , Neutrófilos/imunologia , Resistência Física/fisiologia , Adulto , Ciclismo , Contagem de Células Sanguíneas , Antígeno CD11b/metabolismo , Feminino , Proteínas Ligadas por GPI/metabolismo , Regulação da Expressão Gênica , Voluntários Saudáveis , Humanos , Integrina alfa4beta1/metabolismo , Selectina L/metabolismo , Masculino , Pessoa de Meia-Idade , Fenótipo , Receptores de IgG/metabolismo , Receptores Imunológicos/metabolismo
18.
Chem Biodivers ; 15(11): e1800269, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30193010

RESUMO

In an initial screening, the dichloromethane extract from the leaves of Melodorum fruticosum showed distinct inhibitory effects on the release of interleukin-8 (IL-8) in human neutrophils. Therefore, the aim of the present study was the phytochemical and pharmacological investigation of this extract, to better understand which compounds might be responsible for the anti-inflammatory effect. Phytochemical analysis led to the isolation of 12 known compounds and two new natural products, 5-hydroxy-6-(2-hydroxybenzyl)-4',7-dimethoxyflavanone (13) and 2',4'-dihydroxy-3'-(2-hydroxybenzyl)-4,6'-dimethoxychalcone (14). The influence of the isolated compounds on the production and release of the pro-inflammatory factors IL-8, tumor necrosis factor alpha (TNF-α), reactive oxygen species (ROS), and adhesion molecules (CD62L and CD11b) in human neutrophils was evaluated. Three constituents, melodamide A, 2',4'-dihydroxy-4,6'-dimethoxychalcone, and 2',6'-dihydroxy-4'-methoxychalcone, showed significant inhibition of IL-8 release (IC50 =6.6, 8.6, and 11.6 µm, respectively) and TNF-α production (IC50 =4.5, 13.3, and 6.2 µm, respectively).


Assuntos
Annonaceae/química , Anti-Inflamatórios não Esteroides/farmacologia , Neutrófilos/efeitos dos fármacos , Anti-Inflamatórios não Esteroides/química , Anti-Inflamatórios não Esteroides/isolamento & purificação , Antígeno CD11b/antagonistas & inibidores , Antígeno CD11b/metabolismo , Relação Dose-Resposta a Droga , Humanos , Inflamação/tratamento farmacológico , Inflamação/metabolismo , Interleucina-8/antagonistas & inibidores , Interleucina-8/metabolismo , Selectina L/antagonistas & inibidores , Selectina L/metabolismo , Neutrófilos/metabolismo , Folhas de Planta/química , Espécies Reativas de Oxigênio/antagonistas & inibidores , Espécies Reativas de Oxigênio/metabolismo , Relação Estrutura-Atividade , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/metabolismo
19.
eNeuro ; 5(4)2018.
Artigo em Inglês | MEDLINE | ID: mdl-30225356

RESUMO

L-selectin, a lectin-like receptor on all leukocyte classes, functions in adhesive and signaling roles in the recruitment of myeloid cells from the blood to sites of inflammation. Here, we consider L-selectin as a determinant of neurological recovery in a murine model of spinal cord injury (SCI). Spinal cord-injured, L-selectin knock-out (KO) mice (male) showed improved long-term recovery with greater white matter sparing relative to wild-type (WT) mice and reduced oxidative stress in the injured cord at 72 h post-SCI. There was a partial and transient reduction in accumulation of neutrophils in the injured spinal cords of KOs at 24 h post-injury. To complement these findings with KO mice, we sought a pharmacologic means for lowering L-selectin levels. We found that diclofenac, a nonsteroidal anti-inflammatory drug (NSAID), induced the shedding of L-selectin from the cell surface of myeloid subsets, specifically neutrophils and non-classical monocytes, in the blood and the injured spinal cord. Diclofenac administration to injured WT mice enhanced neurological recovery to a level comparable to that of KOs but did not improve recovery in KOs. While diclofenac treatment had no effect on myeloid cell accumulation, there was a reduction in oxidative stress at 72 h post-SCI. These findings implicate L-selectin in secondary pathogenesis beyond a role in leukocyte recruitment and raise the possibility of repurposing diclofenac for the treatment of SCI.


Assuntos
Anti-Inflamatórios não Esteroides/farmacologia , Diclofenaco/farmacologia , Inflamação , Selectina L/metabolismo , Leucócitos/metabolismo , Células Mieloides/metabolismo , Estresse Oxidativo/fisiologia , Traumatismos da Medula Espinal , Animais , Modelos Animais de Doenças , Inflamação/tratamento farmacológico , Inflamação/imunologia , Inflamação/metabolismo , Selectina L/efeitos dos fármacos , Leucócitos/efeitos dos fármacos , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Células Mieloides/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Traumatismos da Medula Espinal/tratamento farmacológico , Traumatismos da Medula Espinal/imunologia , Traumatismos da Medula Espinal/metabolismo
20.
Med Hypotheses ; 120: 4-6, 2018 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-30220337

RESUMO

In either natural fertilization or artificial insemination, hundreds of millions of sperm are ejaculated or inseminated and then deposited in the female reproductive tract, but only a few sperm reach the ampulla or the site of fertilization. This dramatic reduction in numbers clearly highlights the obstacles that sperm must overcome in order to reach the destination for egg fertilizing. Phagocytosis of sperm by leukocytes are repeatedly observed and generally considered to be one of the most important barriers, but the mechanism of the phagocytosis of sperm remains unclear. L-selectin is constitutively expressed on leukocytes, which can influence the behaviors of leukocytes when bind to ligands. Sialic acids are found as the outer most monosaccharide, capping the majority of glycans at the sperm surface, can act as recognition molecule. Since the sialic acids are considered as ligands of L-selectin, we propose that leukocytes bind to the sperm through the L-selectin on leukocytes and sialic acid on sperm surface during the sperm phagocytosis in female reproductive tract.


Assuntos
Genitália Feminina/fisiologia , Selectina L/metabolismo , Leucócitos/citologia , Fagocitose , Ácidos Siálicos/metabolismo , Espermatozoides/fisiologia , Animais , Feminino , Fertilização , Humanos , Infertilidade , Inseminação Artificial , Lectinas/metabolismo , Masculino , Camundongos , Polissacarídeos/metabolismo , Transdução de Sinais , Superóxidos/metabolismo
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