RESUMO
Stimulator of interferon genes (STING) is an endoplasmic reticulum-membrane protein that plays important roles in cancer immunotherapy by activating innate immune responses. We designed and synthesized STING modulators and characterized compounds 4a and 4c that share a crucial amidobenzimidazole moiety. In vitro STING binding and cell-based activity assays demonstrated the potency and efficacy of the compounds that function as direct STING agonists by stimulating STING downstream signaling and promoting type I interferon immune responses. In vitro metabolic studies and the pharmacokinetic properties of the compounds led us to investigate their anticancer activity in an in vivo syngeneic model. Intravenous injection of compounds 4a and 4c significantly decreased tumor volume in a CT26 murine colorectal carcinoma model, and the immunological memory-derived cancer inhibition was observed in 4c-treated mouse models. The present results suggest the therapeutic potential of the compounds for cancer immunotherapy via STING-mediated immune activation.
Assuntos
Neoplasias , Receptores de Interferon , Animais , Fatores Imunológicos/uso terapêutico , Imunoterapia/métodos , Interferons , Proteínas de Membrana/metabolismo , Camundongos , Neoplasias/tratamento farmacológico , Receptores de Interferon/uso terapêuticoRESUMO
BACKGROUND: Hepatitis E viruses (HEV) are an underestimated global cause of enterically transmitted viral hepatitis, which may persist in immunocompromised hosts, posing a risk for progressive liver fibrosis with limited treatment options. We previously established liver-humanized mice as a model for chronic HEV infections, which can be cleared by a 2-week pegylated (peg)-Interferon(IFN)α treatment course. However, severe side effects may hamper the use of IFNα in immunocompromised transplant recipient patients. IFNλ may be a valuable alternative, as its receptor is less ubiquitously expressed. AIMS: In this study, we assess the in vitro and in vivo potency of pegIFNλ to induce innate immune signalling in liver cells and to clear a persistent HEV infection in liver-humanized mice. METHODS & RESULTS: We found that human liver cells expressed the IFNλ receptor (IFNLR1) and are responsive to pegIFNλ. Treatment with pegIFNλ of liver-humanized mice persistently infected with HEV genotype 3 showed that pegIFNλ was well tolerated. Dose escalation studies showed that although HEV was not cleared at pegIFNλ doses up to 0.12 mg/kg for a maximum of 8 weeks, a dose of 0.3 mg/kg pegIFNλ treatment resulted in complete clearance of HEV antigen and HEV RNA from the liver in 8 out of 9 liver-humanized mice. CONCLUSIONS: PegIFNλ is well tolerated in mice and leads to clearance of a persistent HEV infection in liver-humanized mice.
Assuntos
Vírus da Hepatite E , Hepatite E , Animais , Antivirais/uso terapêutico , Vírus da Hepatite E/genética , Humanos , Interferon-alfa/farmacologia , Interferon-alfa/uso terapêutico , Camundongos , Receptores de Interferon/uso terapêuticoRESUMO
Thrombotic occlusion of inflammatory plaque in coronary arteries causes myocardial infarction. Treatment with emergent balloon angioplasty (BA) and stent implant improves survival, but restenosis (regrowth) can occur. Periodontal bacteremia is closely associated with inflammation and native arterial atherosclerosis, with potential to increase restenosis. Two virus-derived anti-inflammatory proteins, M-T7 and Serp-1, reduce inflammation and plaque growth after BA and transplant in animal models through separate pathways. M-T7 is a broad spectrum C, CC and CXC chemokine-binding protein. Serp-1 is a serine protease inhibitor (serpin) inhibiting thrombotic and thrombolytic pathways. Serp-1 also reduces arterial inflammation and improves survival in a mouse herpes virus (MHV68) model of lethal vasculitis. In addition, Serp-1 demonstrated safety and efficacy in patients with unstable coronary disease and stent implant, reducing markers of myocardial damage. We investigate here the effects of Porphyromonas gingivalis, a periodontal pathogen, on restenosis after BA and the effects of blocking chemokine and protease pathways with M-T7 and Serp-1. ApoE-/- mice had aortic BA and oral P. gingivalis infection. Arterial plaque growth was examined at 24 weeks with and without anti-inflammatory protein treatment. Dental plaques from mice infected with P. gingivalis tested positive for infection. Neither Serp-1 nor M-T7 treatment reduced infection, but IgG antibody levels in mice treated with Serp-1 and M-T7 were reduced. P. gingivalis significantly increased monocyte invasion and arterial plaque growth after BA (P<0.025). Monocyte invasion and plaque growth were blocked by M-T7 treatment (P<0.023), whereas Serp-1 produced only a trend toward reductions. Both proteins modified expression of TLR4 and MyD88. In conclusion, aortic plaque growth in ApoE-/- mice increased after angioplasty in mice with chronic oral P. gingivalis infection. Blockade of chemokines, but not serine proteases significantly reduced arterial plaque growth, suggesting a central role for chemokine-mediated inflammation after BA in P. gingivalis infected mice.
Assuntos
Angioplastia com Balão/efeitos adversos , Aorta/cirurgia , Placa Aterosclerótica/tratamento farmacológico , Receptores de Interferon/uso terapêutico , Proteínas Virais/uso terapêutico , alfa 1-Antitripsina/uso terapêutico , Animais , Aorta/efeitos dos fármacos , Infecções por Bacteroidaceae/complicações , Infecções por Bacteroidaceae/tratamento farmacológico , Placa Dentária/tratamento farmacológico , Placa Dentária/microbiologia , Camundongos , Placa Aterosclerótica/microbiologia , Porphyromonas gingivalis/patogenicidade , Proteínas Virais/farmacologia , alfa 1-Antitripsina/farmacologiaRESUMO
Interferon-beta (IFN-beta) is biologically unstable under physiologic conditions in vitro and is cleared rapidly from the bloodstream on administration in vivo. In the present study, we demonstrate that a soluble recombinant form of the type I IFN receptor subunit, sIFNAR-2, can neutralize the bioactivity of type I IFNs at high concentrations and, at lower concentrations, causes an enhancement of IFN-beta-mediated antiviral activity. The in vitro enhancement is due to the specific interaction of IFN-beta with sIFNAR-2, followed by dissociation of IFN-beta from the complex over time in culture. In vivo, the serum half-life of IFN-beta is extended from minutes to hours when administered intravenously in mice as a sIFNAR-2-associated complex. Moreover, the antitumor effect of IFN-beta is increased by between 9-fold and 27-fold when injected as an sIFNAR-2-associated complex, as demonstrated by an increase in the mean survival time of immunodeficient mice challenged with human Burkitt lymphoma cell (Daudi) xenografts (sIFNAR-2-complexed vs. free IFN-beta treatment). These results show that on association with sIFNAR-2, IFN-beta is more stable in vitro and exhibits increased efficacy when administered in vivo. Administration as a complex with sIFNAR-2 may, therefore, provide a method of enhancing the delivery and effectiveness of type I IFNs.
Assuntos
Interferon Tipo I/uso terapêutico , Interferon beta/farmacologia , Receptores de Interferon/uso terapêutico , Animais , Linfoma de Burkitt/imunologia , Células CHO , Linhagem Celular Tumoral , Cricetinae , Feminino , Meia-Vida , Humanos , Imunoterapia , Injeções Intravenosas , Interferon Tipo I/farmacocinética , Interferon Tipo I/farmacologia , Interferon beta/farmacocinética , Interferon beta/uso terapêutico , Proteínas de Membrana , Camundongos , Camundongos Endogâmicos , Camundongos SCID , Transplante de Neoplasias/imunologia , Receptor de Interferon alfa e beta , Proteínas Recombinantes/farmacocinética , Proteínas Recombinantes/farmacologia , Proteínas Recombinantes/uso terapêutico , Transplante Heterólogo/imunologiaRESUMO
PURPOSE OF REVIEW: Interferons are pleiotropic cytokines that exhibit important biologic activities, including antiviral, antitumor, and immunomodulatory effects. These cytokines have found important applications in clinical medicine, including the treatment of certain malignancies. The purpose of this review is to provide an update on basic and clinical research in the interferon field. RECENT FINDINGS: Significant advances have recently occurred in the field of type I interferon signal transduction. It is well known that the interferons transduce signals via activation of multiple signaling cascades, involving Jak kinases, signal transducer and activator of transcription proteins, Map kinases, and IRS and Crk proteins. Recent evidence indicates that the p38 Map kinase pathway plays an important role in type I interferon signaling in malignant cells and that its function is required for type I interferon-dependent gene transcription and generation of the antiproliferative of type I interferons. In clinical oncology, interferon-alpha remains an active and useful agent in the treatment of several malignant disorders, and efforts are underway to improve its efficacy by using different schedules and combinations with other agents. SUMMARY: This review summarizes the mechanisms of signal transduction of interferons and the emerging new concepts in this area. An update on the clinical applications of interferons in oncology is also provided, and potential translational applications, reflecting recent advances in the field, are discussed.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal , Interferons/metabolismo , Interferons/uso terapêutico , Proteínas Adaptadoras de Transporte Vesicular/efeitos dos fármacos , Proteínas Adaptadoras de Transporte Vesicular/metabolismo , Apoptose/efeitos dos fármacos , Humanos , Proteínas Quinases Ativadas por Mitógeno/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Neoplasias/tratamento farmacológico , Neoplasias/metabolismo , Proteínas Proto-Oncogênicas c-crk , Receptor de Insulina/efeitos dos fármacos , Receptor de Insulina/metabolismo , Receptores de Interferon/metabolismo , Receptores de Interferon/uso terapêutico , Transdução de Sinais/efeitos dos fármacosRESUMO
M-T7 is a myxoma virus-encoded protein that has been found to bind and disrupt human chemokine gradients. This study examined whether purified M-T7 could prevent chronic rejection in a rat renal allograft model. Fisher F344 renal allografts were transplanted into Lewis rats. Recipients were randomly grouped into two groups: control animals treated with cyclosporine alone and animals treated with cyclosporine combined with low-, medium- and high-dose M-T7 viral protein. The survival rate was not significantly different between allograft groups. Renal allografts treated with high-dose M-T7 demonstrated a significant reduction in tubular atrophy, glomerular atrophy, vascular hyalinization, cortical scarring, and lymphocyte infiltration. Morphometric analyses demonstrated that the high-dose M-T7 group also showed a significantly decreased amount of glomerulosclerosis and transplant arteriosclerosis. These data demonstrate for the first time that the immunoregulatory viral protein M-T7 can effectively attenuate chronic rejection in rat renal allografts.
Assuntos
Rejeição de Enxerto/prevenção & controle , Transplante de Rim/imunologia , Receptores de Interferon/uso terapêutico , Proteínas Virais/uso terapêutico , Animais , Quimiocinas/imunologia , Doença Crônica , Sobrevivência de Enxerto/efeitos dos fármacos , Sobrevivência de Enxerto/imunologia , Ratos , Transplante Homólogo/imunologiaRESUMO
We describe three patients with multifocal osteomyelitis caused by Mycobacterium avium and a family history of one or more first degree family members diagnosed with various clinical presentations of infections with nontuberculous mycobacteria. There was a significant delay in the diagnosis and they had a protracted course of their illness, which responded only slowly to prolonged multi-drug treatment. In one patient, additional treatment with interferon-gamma (IFN-gamma) was necessary. Macrophages of these patients had decreased in vitro responsiveness to IFN-gamma. Genomic sequencing revealed that these patients and their affected family members were heterozygous for a previously described dominant negative mutation in the gene encoding the IFN-gamma binding receptor-1 chain. The clinical presentations of the infections with nontuberculous mycobacteria in these families, with spread limited to skin, bone and lymph nodes, is discussed in the light of the immune mechanisms that are responsible for the clearance of otherwise poorly pathogenic environmental mycobacteria.
Assuntos
Predisposição Genética para Doença/genética , Infecção por Mycobacterium avium-intracellulare/genética , Infecção por Mycobacterium avium-intracellulare/microbiologia , Osteomielite/genética , Osteomielite/microbiologia , Receptores de Interferon/deficiência , Receptores de Interferon/genética , Adulto , Feminino , Genes Dominantes/genética , Testes Genéticos , Humanos , Masculino , Mutação/genética , Infecção por Mycobacterium avium-intracellulare/diagnóstico , Infecção por Mycobacterium avium-intracellulare/tratamento farmacológico , Osteomielite/diagnóstico , Osteomielite/tratamento farmacológico , Linhagem , Receptores de Interferon/uso terapêutico , Fatores de Tempo , Receptor de Interferon gamaRESUMO
Estudamos o papel da degradação do triptofano pela indoleamina 2,3-dioxigenase (INDO) no controle da replicação do T. cruzzi ou T. gondii em fibroblastos e macrófagos humanos estimulados com rIFN-gama e/ou rTNF-alfa. O T. gondii foi utlizado como controle, por ser sensível à degradação do triptofano induzida pelo rIFN-gama. A estimulação de fibroblastos humanos com rIFN-gama e o rTNF-alfa inibiu consideravelmente o desenvolvimento do T. gondii, mas não influenciou o desenvolvimento do T. cruzi. O desenvolvimento do T. gondii foi triptofano dependente, enquanto que o do T. cruzi foi triptofano independente. Ao contrário dos fibroblastos parentais, os fibroblastos defectivos na via de transdução de sinal do IFN-gama (JAKI, JAK2 e STAT1alfa)não modularam o desenvolvimento intracelular de T. gondii e nem expressaram mRNA da INDO quando estimulados pelo rIFN-gama. Houve uma pequena indução de mRNA da INDO em fibroblastos parentais e mutantes (JAK2) estimulados com rTNF-alfa, que provavelmente foi estimulado através da indução da síntese de IFN-es em fibroblastos humanos. O rIFN-gama e/ou rTNF-alfa induziram expressiva atividade tripanosomicida em macrófagos humanos.
A estimulação de macrófagos humanos com rIFN-gama ou rIFN-gama + rTNF-alfa, produziu uma considerável expressão do mRNA da INDO, e pouca expressão foi detectada quando as células foram estimuladas apenas com rTNF-alfa. A adição de triptofano, ou do inibidor da INDO, norharmane, à cultura de macrófagos humanos ativados com rIFN-gama e/ou rTNF-alfa, não reverteu a inibição do crescimento do parasita , mostrando que a degradação do triptofano não é um mecanismo responsável pelo efeito tripanosomicida de macrófagos humanos ativados com rIFN-gama e/ou rTNF-alfa. Os catabólitos do triptofano (ácido quinolínico, ácido quinurênico e L-quinurenina)inibiram parcialmente o desenvolvimento do T. cruzi em macrófagos humanos. A amplificação por PCR do DNA T. cLsintese de Escherichia coli ou Neurospora crassa e Saccharomyces cerevisae e a hibridização cruzada via dot blot utilizando como sondas os produtos de PCR, sugerem a possibilidade da existência da enzima triptofano sintetase em T. cruzi, o que explicaria a insensibilidade do parasita à depleção do triptofano. Alternativamente reconhecemos que mais estudos devam ser feitos para a comprovação da existência da enzima em T. cruzi
Assuntos
Plantas Medicinais/química , Toxoplasma/parasitologia , Toxoplasma/patogenicidade , Trypanosoma cruzi/enzimologia , Trypanosoma cruzi/parasitologia , Fibroblastos/parasitologia , Interferon gama/uso terapêutico , Macrófagos/parasitologia , Receptores de Interferon/uso terapêutico , Triptofano/metabolismo , Triptofano/uso terapêuticoRESUMO
Estudamos o papel da degradação do triptofano pela indoleamina 2,3-dioxigenase (INDO) no controle da replicação do T. cruzzi ou T. gondii em fibroblastos e macrófagos humanos estimulados com rIFN-gama e/ou rTNF-alfa. O T. gondii foi utlizado como controle, por ser sensível à degradação do triptofano induzida pelo rIFN-gama. A estimulação de fibroblastos humanos com rIFN-gama e o rTNF-alfa inibiu consideravelmente o desenvolvimento do T. gondii, mas não influenciou o desenvolvimento do T. cruzi. O desenvolvimento do T. gondii foi triptofano dependente, enquanto que o do T. cruzi foi triptofano independente. Ao contrário dos fibroblastos parentais, os fibroblastos defectivos na via de transdução de sinal do IFN-gama (JAKI, JAK2 e STAT1alfa) não modularam o desenvolvimento intracelular de T. gondii e nem expressaram mRNA da INDO quando estimulados pelo rIFN-gama. Houve uma pequena indução de mRNA da INDO em fibroblastos parentais e mutantes (JAK2) estimulados com rTNF-alfa, que provavelmente foi estimulado através da indução da síntese de IFN-es em fibroblastos humanos. O rIFN-gama e/ou rTNF-alfa induziram expressiva atividade tripanosomicida em macrófagos humanos. A estimulação de macrófagos humanos com rIFN-gama ou rIFN-gama + rTNF-alfa, produziu uma considerável expressão do mRNA da INDO, e pouca expressão foi detectada quando as células foram estimuladas apenas com rTNF-alfa. A adição de triptofano, ou do inibidor da INDO, norharmane, à cultura de macrófagos humanos ativados com rIFN-gama e/ou rTNF-alfa, não reverteu a inibição do crescimento do parasita, mostrando que a degradação do triptofano não é um mecanismo responsável pelo efeito tripanosomicida de macrófagos humanos ativados com rIFN-gama e/ou rTNF-alfa. Os catabólitos do triptofano (ácido quinolínico, ácido quinurênico e L-quinurenina) inibiram parcialmente o desenvolvimento do T. cruzi em macrófagos humanos. A amplificação por PCR do DNA T. cLsintese de Escherichia coli ou Neurospora crassa e Saccharomyces cerevisae e a hibridização cruzada via dot blot utilizando como sondas os produtos de PCR, sugerem a possibilidade da existência da enzima triptofano sintetase em T. cruzi, o que explicaria a insensibilidade do parasita à depleção do triptofano. Alternativamente reconhecemos que mais estudos devam ser feitos para a comprovação da existência da enzima em T. cruzi.
Assuntos
Doença de Chagas/diagnóstico , Toxoplasma/parasitologia , Toxoplasma/patogenicidade , Trypanosoma cruzi/enzimologia , Trypanosoma cruzi/parasitologia , Fibroblastos/parasitologia , Interferon gama/uso terapêutico , Macrófagos/parasitologia , Receptores de Interferon/uso terapêutico , Triptofano/metabolismo , Triptofano/uso terapêuticoRESUMO
NZB x NZW F1 hybrid (B/W) mice develop altered behavior in the elevated plus maze and novel object tasks between 6 and 12 weeks of age in parallel with lupus-like autoimmune disease. To confirm the relationship between disease progression and development of behavioral abnormalities, B/W and nonautoimmune NZW mice received chronic treatment with a soluble IFN gamma receptor (sIFN gamma R), a treatment known to retard autoimmune disease progression, or vehicle, beginning at 6 weeks of age. After 6 weeks of treatment, elevated plus maze and novel object testing revealed that although sIFN gamma R treated B/W mice still differed from NZW mice, chronic sIFN gamma R treatment significantly retarded the development of behavioral abnormalities in the B/W mice, while the NZW mice were not affected by this treatment. sIFN gamma R treated B/W mice were more active in both the plus maze and novel object tasks, and displayed less plus maze anxiety behavior and more exploratory activity in the novel object task compared to vehicle treated B/W mice. To clarify the role of acute action of the sIFN gamma R on the elevated IFN gamma levels of B/W mice, a second experiment examined the effects of a single injection of sIFN gamma R on B/W and NZW mice. Unlike chronic treatment, acute treatment with the same dose of sIFN gamma R did not affect plus maze or novel object behavior in 12-week-old mice. These results add to the growing evidence that lupus-associated behavioral abnormalities are a direct effect of the autoimmune disease.