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1.
Bioconjug Chem ; 27(5): 1293-304, 2016 05 18.
Artigo em Inglês | MEDLINE | ID: mdl-27074387

RESUMO

We have synthesized and characterized a novel phosphorothioate CpG oligodeoxynucleotide (CpG ODN)-Ficoll conjugated nanoparticulate adjuvant, termed DV230-Ficoll. This adjuvant was constructed from an amine-functionalized-Ficoll, a heterobifunctional linker (succinimidyl-[(N-maleimidopropionamido)-hexaethylene glycol] ester) and the CpG-ODN DV230. Herein, we describe the evaluation of the purity and reactivity of linkers of different lengths for CpG-ODN-Ficoll conjugation, optimization of linker coupling, and conjugation of thiol-functionalized CpG to maleimide-functionalized Ficoll and process scale-up. Physicochemical characterization of independently produced lots of DV230-Ficoll reveal a bioconjugate with a particle size of approximately 50 nm and covalent attachment of more than 100 molecules of CpG per Ficoll. Solutions of purified DV230-Ficoll were stable for at least 12 months at frozen and refrigerated temperatures and stability was further enhanced in lyophilized form. Compared to nonconjugated monomeric DV230, the DV230-Ficoll conjugate demonstrated improved in vitro potency for induction of IFN-α from human peripheral blood mononuclear cells and induced higher titer neutralizing antibody responses against coadministered anthrax recombinant protective antigen in mice. The processes described here establish a reproducible and robust process for the synthesis of a novel, size-controlled, and stable CpG-ODN nanoparticle adjuvant suitable for manufacture and use in vaccines.


Assuntos
Adjuvantes Imunológicos/química , Antígenos de Bactérias/imunologia , Toxinas Bacterianas/imunologia , Desenho de Fármacos , Ficoll/química , Nanopartículas/química , Oligodesoxirribonucleotídeos/química , Oligodesoxirribonucleotídeos/imunologia , Animais , Estabilidade de Medicamentos , Humanos , Maleimidas/química , Metilação , Camundongos , Polietilenoglicóis/química
2.
J Immunol ; 196(1): 284-97, 2016 Jan 01.
Artigo em Inglês | MEDLINE | ID: mdl-26608924

RESUMO

Nanoparticulate delivery systems for vaccine adjuvants, designed to enhance targeting of secondary lymphoid organs and activation of APCs, have shown substantial promise for enhanced immunopotentiation. We investigated the adjuvant activity of synthetic oligonucleotides containing CpG-rich motifs linked to the sucrose polymer Ficoll, forming soluble 50-nm particles (DV230-Ficoll), each containing >100 molecules of the TLR9 ligand, DV230. DV230-Ficoll was evaluated as an adjuvant for a candidate vaccine for anthrax using recombinant protective Ag (rPA) from Bacillus anthracis. A single immunization with rPA plus DV230-Ficoll induced 10-fold higher titers of toxin-neutralizing Abs in cynomolgus monkeys at 2 wk compared with animals immunized with equivalent amounts of monomeric DV230. Monkeys immunized either once or twice with rPA plus DV230-Ficoll were completely protected from challenge with 200 LD50 aerosolized anthrax spores. In mice, DV230-Ficoll was more potent than DV230 for the induction of innate immune responses at the injection site and draining lymph nodes. DV230-Ficoll was preferentially colocalized with rPA in key APC populations and induced greater maturation marker expression (CD69 and CD86) on these cells and stronger germinal center B and T cell responses, relative to DV230. DV230-Ficoll was also preferentially retained at the injection site and draining lymph nodes and produced fewer systemic inflammatory responses. These findings support the development of DV230-Ficoll as an adjuvant platform, particularly for vaccines such as for anthrax, for which rapid induction of protective immunity and memory with a single injection is very important.


Assuntos
Adjuvantes Imunológicos/administração & dosagem , Vacinas contra Antraz/imunologia , Antraz/prevenção & controle , Antígenos de Bactérias/imunologia , Toxinas Bacterianas/imunologia , Oligonucleotídeos/imunologia , Infecções Respiratórias/prevenção & controle , Animais , Antraz/imunologia , Antraz/microbiologia , Vacinas contra Antraz/administração & dosagem , Antígenos de Bactérias/genética , Antígenos CD/biossíntese , Antígenos de Diferenciação de Linfócitos T/biossíntese , Linfócitos B/imunologia , Antígeno B7-2/biossíntese , Bacillus anthracis/imunologia , Bacillus anthracis/patogenicidade , Toxinas Bacterianas/genética , Células Dendríticas/imunologia , Ficoll/imunologia , Sequência Rica em GC/genética , Lectinas Tipo C/biossíntese , Macaca fascicularis , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Nanopartículas , Neutrófilos/imunologia , Oligonucleotídeos/genética , Proteínas Recombinantes/imunologia , Infecções Respiratórias/imunologia , Infecções Respiratórias/microbiologia , Linfócitos T/imunologia , Vacinação , Vacinas Sintéticas/imunologia
3.
J Clin Invest ; 119(9): 2564-76, 2009 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-19726873

RESUMO

CpG-containing immunostimulatory DNA sequences (ISS), which signal through TLR9, are being developed as a therapy for allergic indications and have proven to be safe and well tolerated in humans when administrated via the pulmonary route. In contrast, ISS inhalation has unexplained toxicity in rodents, which express TLR9 in monocyte/macrophage lineage cells as well as in plasmacytoid DCs (pDCs) and B cells, the principal TLR9-expressing cells in humans. We therefore investigated the mechanisms underlying this rodent-specific toxicity and its implications for humans. Mice responded to intranasally administered 1018 ISS, a representative B class ISS, with strictly TLR9-dependent toxicity, including lung inflammation and weight loss, that was fully reversible and pDC and B cell independent. Knockout mouse experiments demonstrated that ISS-induced toxicity was critically dependent on TNF-alpha, with IFN-alpha required for TNF-alpha induction. In contrast, human PBMCs, human alveolar macrophages, and airway-derived cells from Ascaris suum-allergic cynomolgus monkeys did not produce appreciable TNF-alpha in vitro in response to ISS stimulation. Moreover, sputum of allergic humans exposed to inhaled ISS demonstrated induction of IFN-inducible genes but minimal TNF-alpha induction. These data demonstrate that ISS induce rodent-specific TNF-alpha-dependent toxicity that is absent in humans and reflective of differential TLR9 expression patterns in rodents versus humans.


Assuntos
Oligodesoxirribonucleotídeos/toxicidade , Fator de Necrose Tumoral alfa/metabolismo , Adjuvantes Imunológicos/toxicidade , Administração por Inalação , Animais , Asma/genética , Asma/imunologia , Líquido da Lavagem Broncoalveolar/imunologia , Citocinas/metabolismo , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Humanos , Técnicas In Vitro , Pulmão/efeitos dos fármacos , Pulmão/imunologia , Pulmão/patologia , Macaca fascicularis , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Knockout , Oligodesoxirribonucleotídeos/administração & dosagem , Oligodesoxirribonucleotídeos/genética , Oligodesoxirribonucleotídeos/imunologia , Especificidade da Espécie , Receptor Toll-Like 9/agonistas , Receptor Toll-Like 9/deficiência , Receptor Toll-Like 9/genética , Receptor Toll-Like 9/metabolismo , Fator de Necrose Tumoral alfa/deficiência , Fator de Necrose Tumoral alfa/genética
4.
Clin Cancer Res ; 14(17): 5626-34, 2008 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-18765557

RESUMO

PURPOSE: CpG oligodeoxynucleotides (CpG-ODN) are being investigated as cancer vaccine adjuvants because they mature plasmacytoid dendritic cells (PDC) into potent antigen-presenting cells. CpG-ODN also induce PDC to secrete chemokines that alter lymphocyte migration. Whether CpG-ODN TLR signals enhance antigen-specific immunity and/or trafficking in humans is unknown. EXPERIMENTAL DESIGN: We conducted a phase I study of CpG-ODN (1018 ISS) given as a vaccine adjuvant with granulocyte-macrophage colony-stimulating factor (GM-CSF) to induce T-cell immunity to a peptide vaccine from the tumor-associated antigen hTERT. RESULTS: The adjuvant effect was limited; only 1 of 16 patients showed a high-frequency hTERT-specific tetramer CD8(+) T-cell response. However, CpG-ODN induced marked, transient peripheral blood lymphopenia. Biopsies showed dense lymphocytic infiltration at the vaccine site clustered around activated PDC. In vitro, CpG-ODN-treated PDC induced T-cell migration, showing that CpG-ODN stimulation of human PDC was sufficient to chemoattract T cells. CONCLUSIONS: Our results show that (a) CpG-ODN with GM-CSF may not be an effective adjuvant strategy for hTERT peptide vaccines but (b) GM-CSF/CpG-ODN causes a PDC-mediated chemokine response that recruits T-cell migration to the peripheral tissues. These findings suggest a novel therapeutic role for targeted injections of CpG-ODN to direct lymphocyte migration to specific sites such as the tumor bed.


Assuntos
Vacinas Anticâncer/uso terapêutico , Células Dendríticas/imunologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/imunologia , Oligodesoxirribonucleotídeos/uso terapêutico , Linfócitos T/imunologia , Telomerase/imunologia , Vacinas de Subunidades Antigênicas/uso terapêutico , Adjuvantes Imunológicos , Vacinas Anticâncer/efeitos adversos , Movimento Celular , Humanos , Oligodesoxirribonucleotídeos/imunologia , Receptor Toll-Like 9/antagonistas & inibidores
5.
Nat Med ; 13(5): 552-9, 2007 May.
Artigo em Inglês | MEDLINE | ID: mdl-17479101

RESUMO

The identification of the antigen recognition receptors for innate immunity, most notably the Toll-like receptors, has sparked great interest in therapeutic manipulation of the innate immune system. Toll-like receptor agonists are being developed for the treatment of cancer, allergies and viral infections, and as adjuvants for potent new vaccines to prevent or treat cancer and infectious diseases. As recognition grows of the role of inappropriate Toll-like receptor stimulation in inflammation and autoimmunity, significant efforts have begun to develop antagonists to Toll-like receptors as well.


Assuntos
Imunidade Inata , Receptores Toll-Like/imunologia , Animais , Antígenos/imunologia , Autoimunidade , Humanos , Hipersensibilidade/imunologia , Inflamação/imunologia , Modelos Imunológicos , Neoplasias/imunologia , Receptores Toll-Like/agonistas , Receptores Toll-Like/antagonistas & inibidores , Receptores Toll-Like/genética , Viroses/imunologia
6.
Immunity ; 25(4): 655-64, 2006 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-17000122

RESUMO

Although Toll-like Receptors (TLRs) play a major function in innate recognition of pathogens, their role in antigen processing and presentation in vivo is poorly understood. Here we establish that Toxoplasma gondii profilin, a TLR11 ligand present in the parasite, is an immunodominant antigen in the CD4(+) T cell response to the pathogen. The immunogenicity of profilin was entirely dependent on both TLR11 recognition and signaling through the adaptor myeloid differentiation factor 88 (MyD88). Selective responsiveness to this parasite protein was regulated at the level of antigen presentation by dendritic cells (DC) and required both TLR signaling and major histocompatibility complex (MHC) class II recognition acting in cis. These findings support a major influence of TLR recognition in antigen presentation by DC in vivo and establish a mechanism by which TLR ligand association regulates the immunogenicity of microbial antigens.


Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Antígenos de Protozoários/imunologia , Linfócitos T CD4-Positivos/imunologia , Profilinas/imunologia , Receptores Toll-Like/metabolismo , Toxoplasma/imunologia , Proteínas Adaptadoras de Transdução de Sinal/genética , Animais , Apresentação de Antígeno , Antígenos CD8/análise , Células Dendríticas/imunologia , Antígenos de Histocompatibilidade Classe II/metabolismo , Ligantes , Camundongos , Camundongos Mutantes , Fator 88 de Diferenciação Mieloide , Transdução de Sinais , Subpopulações de Linfócitos T/imunologia , Receptores Toll-Like/genética
7.
Blood ; 107(6): 2423-31, 2006 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-16293610

RESUMO

Recent studies suggest plasmacytoid predendritic cells (pDCs) and myeloid dendritic cells (mDCs) have the functional plasticity to produce similar amounts of type 1 interferons (IFNs) and interleukin-12 (IL-12), challenging the concept and existence of DC subsets with distinct function. In this study, we demonstrate that previous studies showed human pDCs produce large amounts of IL-12 because of contaminating mDCs. Using highly purified human DC subsets, we found that although pDCs make 300 times more IFN-alpha than mDCs and mDCs make 13 times more IL-12 p70 than pDCs in response to all the toll-like receptor ligands and CD40 ligands, pDCs rapidly make large amounts of IFN-alpha within the first 12 hours of activation and become refractory to further stimulation. pDCs preferentially expressed the transcriptional factors critical for type 1 IFN, but not for IL-12 transcription, and they dedicated 60% of new transcriptional activity to make 19 type 1 IFN subtypes. This study provides formal proof that the plasticity of DC subsets is limited and that different DC subsets evolve to perform distinct functions in linking innate and adaptive immunity.


Assuntos
Células Dendríticas/citologia , Células Dendríticas/imunologia , Imunidade , Interferon Tipo I/biossíntese , Ligante de CD40 , Separação Celular/métodos , Separação Celular/normas , Células Dendríticas/classificação , Humanos , Interferon-alfa/biossíntese , Interleucina-12/biossíntese , Cinética , Ligantes , Receptores Toll-Like , Transcrição Gênica
8.
J Leukoc Biol ; 78(4): 954-66, 2005 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-16081597

RESUMO

Depending on the activation status, plasmacytoid dendritic cells (PDC) and myeloid DC have the ability to induce CD4 T cell development toward T helper cell type 1 (Th1) or Th2 pathways. Thus, we tested whether different activation signals could also have an impact on the profile of chemokines produced by human PDC. Signals that induce human PDC to promote a type 1 response (i.e., viruses) and a type 2 response [i.e., CD40 ligand (CD40L)] also induced PDC isolated from tonsils to secrete chemokines preferentially attracting Th1 cells [such as interferon-gamma (IFN-gamma)-inducible protein (IP)-10/CXC chemokine ligand 10 (CXCL10) and macrophage inflammatory protein-1beta/CC chemokine ligand 4 (CCL4)] or Th2 cells (such as thymus and activation-regulated chemokine/CCL17 and monocyte-derived chemokine/CCL22), respectively. Activated natural killer cells were preferentially recruited by supernatants of virus-activated PDC, and supernatants of CD40L-activated PDC attracted memory CD4(+) T cells, particularly the CD4(+)CD45RO(+)CD25(+) T cells described for their regulatory activities. It is striking that CD40L and virus synergized to trigger the production of IFN-gamma by PDC, which induces another Th1-attracting chemokine monokine-induced by IFN-gamma/CXCL9 and cooperates with endogenous type I IFN for IP-10/CXCL10 production. In conclusion, our studies reveal that PDC participate in the selective recruitment of effector cells of innate and adaptive immune responses and that virus converts the CD40L-induced Th2 chemokine patterns of PDC into a potent Th1 mediator profile through an autocrine loop of IFN-gamma.


Assuntos
Ligante de CD40/farmacologia , Quimiocinas/biossíntese , Células Dendríticas/imunologia , Interferon gama/imunologia , Células Th1/imunologia , Células Th2/imunologia , Comunicação Autócrina/imunologia , Ligante de CD40/imunologia , Quimiocina CXCL9 , Quimiocinas/imunologia , Quimiocinas CXC/biossíntese , Quimiocinas CXC/imunologia , Quimiotaxia de Leucócito/imunologia , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/virologia , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/biossíntese , Peptídeos e Proteínas de Sinalização Intercelular/imunologia , Interferon gama/biossíntese , Interferon gama/efeitos dos fármacos , Interferon gama/farmacologia , Interleucina-3/farmacologia , Células Matadoras Naturais/citologia , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/virologia , Orthomyxoviridae/imunologia , Proteínas Recombinantes , Células Th1/efeitos dos fármacos , Células Th1/virologia , Células Th2/efeitos dos fármacos , Células Th2/virologia
9.
Arthritis Rheum ; 52(5): 1504-16, 2005 May.
Artigo em Inglês | MEDLINE | ID: mdl-15880822

RESUMO

OBJECTIVE: To investigate the activation and recruitment pathways of relevant leukocyte subsets during the initiation and amplification of cutaneous lupus erythematosus (LE). METHODS: Quantitative real-time polymerase chain reaction was used to perform a comprehensive analysis of all known chemokines and their receptors in cutaneous LE lesions, and the cellular origin of these chemokines and receptors was determined using immunohistochemistry. Furthermore, cytokine- and ultraviolet (UV) light-mediated activation pathways of relevant chemokines were investigated in vitro and in vivo. RESULTS: In the present study, we identified the CXCR3 ligands CXCL9 (interferon-gamma [IFNgamma]-induced monokine), CXCL10 (IFNgamma-inducible protein 10), and CXCL11 (IFN-inducible T cell alpha chemoattractant) as being the most abundantly expressed chemokine family members in cutaneous LE. Expression of these ligands corresponded with the presence of a marked inflammatory infiltrate consisting of mainly CXCR3-expressing cells, including skin-homing lymphocytes and blood dendritic cell antigen 2-positive plasmacytoid dendritic cells (PDCs). Within cutaneous LE lesions, PDCs accumulated within the dermis and were activated to produce type I IFN, as detected by the expression of the IFNalpha-inducible genes IRF7 and MxA. IFNalpha, in turn, was a potent and rapid inducer of CXCR3 ligands in cellular constituents of the skin. Furthermore, we demonstrated that the inflammatory CXCR3 ligands cooperate with the homeostatic chemokine CXCL12 (stromal cell-derived factor 1) during the recruitment of pathogenically relevant leukocyte subsets. Moreover, we showed that UVB irradiation induces the release of CCL27 (cutaneous T cell-attracting chemokine) from epidermal compartments into dermal compartments and up-regulates the expression of a distinct set of chemokines in keratinocytes. CONCLUSION: Taken together, our data suggest an amplification cycle in which UV light-induced injury induces apoptosis, necrosis, and chemokine production. These mechanisms, in turn, mediate the recruitment and activation of autoimmune T cells and IFNalpha-producing PDCs, which subsequently release more effector cytokines, thus amplifying chemokine production and leukocyte recruitment, finally leading to the development of a cutaneous LE phenotype.


Assuntos
Quimiocinas CXC/imunologia , Peptídeos e Proteínas de Sinalização Intercelular/imunologia , Leucócitos/imunologia , Lúpus Eritematoso Cutâneo/imunologia , Lesões por Radiação/imunologia , Raios Ultravioleta/efeitos adversos , Células Cultivadas , Quimiocina CXCL10 , Quimiocina CXCL11 , Quimiocina CXCL9 , Humanos , Lúpus Eritematoso Cutâneo/patologia , Ativação Linfocitária
10.
DNA Cell Biol ; 24(2): 63-72, 2005 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-15699627

RESUMO

CpG-C are a novel class of CpG motif-containing immunostimulatory sequences (ISS) that includes both a 5'-TCG element and a CpG-containing palindrome. CpG-C drive all known ISS activities and, in particular, are potent enhancers of IFN-alpha from plasmacytoid dendritic cells (PDCs). In our examination of CpG-C sequence requirements, we determined that optimal IFN-alpha-inducing activity could be achieved with longer palindromes. Longer palindromes also correlated with maintenance of the double-stranded (ds) form despite concentration and pH changes, indicating a preference for ds oligodeoxynucleotides (ODNs) by the ISS-induced signaling mechanism for IFN-alpha synthesis. This correlation did not hold for all arms of the ISS-induced immune response, since we did not observe increased B cell activity with the longer palindrome CpG-C ODNs. We further demonstrated that CpG-C retained activity in an in vitro primate system and induced the expression of several cytokines and IFN-alpha-inducible genes when CpG-C were administered in vivo to mice and primates. In conclusion, we have shown CpG-C to exert several types of immune functions across multiple species, and this novel class is thus an attractive candidate for ISS-based therapeutic strategies.


Assuntos
Adjuvantes Imunológicos/farmacologia , Linfócitos B/efeitos dos fármacos , Citocinas/biossíntese , Expressão Gênica/efeitos dos fármacos , Oligodesoxirribonucleotídeos/farmacologia , Adjuvantes Imunológicos/genética , Animais , Proteínas Reguladoras de Apoptose , Linfócitos B/imunologia , Proliferação de Células , Ilhas de CpG/genética , Feminino , Proteínas de Ligação ao GTP/biossíntese , Proteínas de Ligação ao GTP/genética , Expressão Gênica/fisiologia , Regulação da Expressão Gênica/fisiologia , Humanos , Interferon-alfa/metabolismo , Interferon gama/metabolismo , Interleucina-12/metabolismo , Interleucina-6/metabolismo , Linfonodos/química , Linfonodos/metabolismo , Camundongos , Proteínas de Resistência a Myxovirus , Oligodesoxirribonucleotídeos/genética , Papio , RNA Mensageiro/análise , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA , Fatores de Transcrição/biossíntese , Fatores de Transcrição/genética
11.
Blood ; 103(7): 2547-53, 2004 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-14670916

RESUMO

Type 1 interferon-producing cells (IPCs), also known as plasmacytoid dendritic cell (DC) precursors, represent the key effectors in antiviral innate immunity and triggers for adaptive immune responses. IPCs play important roles in the pathogenesis of systemic lupus erythematosus (SLE) and in modulating immune responses after hematopoietic stem cell transplantation. Understanding IPC development from hematopoietic progenitor cells (HPCs) may provide critical information in controlling viral infection, autoimmune SLE, and graft-versus-host disease. FLT3-ligand (FLT3-L) represents a key IPC differentiation factor from HPCs. Although hematopoietic cytokines such as interleukin-3 (IL-3), IL-7, stem cell factor (SCF), macrophage-colony-stimulating factor (M-CSF), and granulocyte M-CSF (GM-CSF) promote the expansion of CD34+ HPCs in FLT3-L culture, they strongly inhibit HPC differentiation into IPCs. Here we show that thrombopoietin (TPO) cooperates with FLT3-L, inducing CD34+ HPCs to undergo a 400-fold expansion in cell numbers and to generate more than 6 x 10(6) IPCs per 10(6) CD34+ HPCs within 30 days in culture. IPCs derived from HPCs in FLT3-L/TPO cultures display blood IPC phenotype and have the capacity to produce large amounts of interferon-alpha (IFN-alpha) and to differentiate into mature DCs. This culture system, combined with the use of adult peripheral blood CD34+ HPCs purified from G-CSF-mobilized donors, permits the generation of more than 10(9) IPCs from a single blood donor.


Assuntos
Células Dendríticas/citologia , Hematopoese/fisiologia , Células-Tronco Hematopoéticas/citologia , Proteínas de Membrana/fisiologia , Trombopoetina/fisiologia , Antígenos CD/análise , Antígenos CD34/análise , Técnicas de Cultura de Células/métodos , Diferenciação Celular/imunologia , Divisão Celular/fisiologia , Células Cultivadas , Células Dendríticas/imunologia , Feto , Idade Gestacional , Fator Estimulador de Colônias de Granulócitos/farmacologia , Mobilização de Células-Tronco Hematopoéticas/métodos , Humanos , Ativação Linfocitária , Plasmócitos/citologia , Plasmócitos/imunologia , Linfócitos T/imunologia
12.
Blood ; 102(13): 4487-92, 2003 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-12946990

RESUMO

Immunostimulatory sequences (ISS) are short oligonucleotides containing unmethylated cytosine-phosphate-guanine (CpG) dinucleotides that stimulate innate immune responses through Toll-like receptor-9 on B cells and plasmacytoid dendritic cell (PDC) precursors. The anti-inflammatory cytokine interleukin (IL)-10 is predicted to be a potent inhibitor of many of the activities described for ISS, and this may impact the use of ISS in disease states characterized by elevated IL-10. As the activities of ISS on PDCs are central to many clinical applications of ISS, we have studied the effects of IL-10 on PDC stimulation by 3 distinct classes of ISS. IL-10 inhibited cytokine production and survival of ISS-activated PDCs; however, IL-12 induction was much more sensitive to inhibition than interferon (IFN)-alpha induction. Within the PDC population are cells that respond to ISS by producing either IL-12 or IFN-alpha but not both cytokines. IL-12-producing PDCs require costimulation through CD40 and appear more mature than IFN-alpha-producing PDCs. The 3 distinct classes of ISS differed with respect to induction of PDC maturation and T-cell priming capacity. IL-10 regulated PDC activation but did not inhibit the subsequent T-cell-priming ability of PDCs already activated by ISS.


Assuntos
Adjuvantes Imunológicos/farmacologia , Ilhas de CpG/imunologia , Células Dendríticas/efeitos dos fármacos , Interleucina-10/farmacologia , Adjuvantes Imunológicos/classificação , Apresentação de Antígeno , Antígenos CD40/fisiologia , Células Cultivadas/efeitos dos fármacos , Células Dendríticas/imunologia , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Interferon-alfa/farmacologia , Interferon gama/biossíntese , Interferon gama/genética , Interleucina-10/fisiologia , Interleucina-12/biossíntese , Interleucina-12/genética , Interleucina-3/farmacologia , Ativação Linfocitária , Oligodesoxirribonucleotídeos/imunologia , Oligodesoxirribonucleotídeos/farmacologia , Proteínas Recombinantes/farmacologia , Linfócitos T/imunologia
13.
Immunity ; 16(6): 779-90, 2002 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12121660

RESUMO

An efficient Th1-driven adaptive immune response requires activation of the T cell receptor and secretion of the T cell stimulatory cytokine IL-12 by activated antigen-presenting cells. IL-12 triggers Th1 polarization of naive CD4(+) T cells and secretion of IFN-gamma. We describe a new heterodimeric cytokine termed IL-27 that consists of EBI3, an IL-12p40-related protein, and p28, a newly discovered IL-12p35-related polypeptide. IL-27 is an early product of activated antigen-presenting cells and drives rapid clonal expansion of naive but not memory CD4(+) T cells. It also strongly synergizes with IL-12 to trigger IFN-gamma production of naive CD4(+) T cells. IL-27 mediates its biologic effects through the orphan cytokine receptor WSX-1/TCCR.


Assuntos
Linfócitos T CD4-Positivos/fisiologia , Proteínas de Transporte/análise , Glutationa Transferase , Glicoproteínas/análise , Interleucinas/fisiologia , Células Th1/imunologia , Sequência de Aminoácidos , Células Apresentadoras de Antígenos/metabolismo , Divisão Celular , Dimerização , Interferon gama/biossíntese , Interleucina-12/fisiologia , Interleucinas/química , Antígenos de Histocompatibilidade Menor , Dados de Sequência Molecular , Conformação Proteica , Receptores de Citocinas/metabolismo , Receptores de Interleucina , Alinhamento de Sequência
14.
Nat Immunol ; 3(7): 673-80, 2002 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-12055625

RESUMO

Whether epithelial cells play a role in triggering the immune cascade leading to T helper 2 (T(H)2)-type allergic inflammation is not known. We show here that human thymic stromal lymphopoietin (TSLP) potently activated CD11c(+) dendritic cells (DCs) and induced production of the T(H)2-attracting chemokines TARC (thymus and activation-regulated chemokine; also known as CCL17) and MDC (macrophage-derived chemokine; CCL22). TSLP-activated DCs primed naïve T(H) cells to produce the proallergic cytokines interleukin 4 (IL-4), IL-5, IL-13 and tumor necrosis factor-alpha, while down-regulating IL-10 and interferon-gamma. TSLP was highly expressed by epithelial cells, especially keratinocytes from patients with atopic dermatitis. TSLP expression was associated with Langerhans cell migration and activation in situ. These findings shed new light on the function of human TSLP and the role played by epithelial cells and DCs in initiating allergic inflammation.


Assuntos
Antígenos CD11 , Citocinas/imunologia , Células Dendríticas/imunologia , Dermatite Atópica/imunologia , Interleucina-7/imunologia , Células Th2/imunologia , Timo/imunologia , Adulto , Biomarcadores , Linfócitos T CD4-Positivos/citologia , Linfócitos T CD4-Positivos/imunologia , Divisão Celular , Células Cultivadas , Técnicas de Cocultura , Citocinas/genética , Citocinas/farmacologia , Células Dendríticas/citologia , Células Dendríticas/efeitos dos fármacos , Dermatite Atópica/patologia , Células Epiteliais/citologia , Células Epiteliais/imunologia , Expressão Gênica , Humanos , Interleucina-7/farmacologia , Células de Langerhans/citologia , Células de Langerhans/imunologia , Tonsila Palatina/imunologia , RNA Mensageiro , Pele/imunologia , Pele/patologia , Células Th2/citologia , Linfopoietina do Estroma do Timo
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