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INTRODUCTION: The expansion of tumor-infiltrating lymphocytes (TIL) for adoptive cellular therapy is under investigation in many solid tumors of adulthood. Marrow-infiltrating lymphocytes (MIL) have demonstrated antitumor reactivity preclinically. Successful expansion of TIL/MIL has not been reported across pediatric solid tumor histologies. The objective of this study was to demonstrate successful expansion of TIL from pediatric solid tumors for translation in an adoptive cell therapy (ACT) treatment strategy. METHODS: A prospective study of TIL/MIL expansion was performed on solid tumors of pediatric patients undergoing standard-of-care procedures. TIL/MIL expansions were performed in the presence of high-dose interleukin 2. To demonstrate a full-scale expansion to clinically-relevant cell doses for TIL therapy, initial TIL culture was followed by a rapid expansion protocol for select patients. Expanded specimens were analyzed for phenotype by flow cytometry and for anti-tumor reactivity by the interferon-gamma release assay. RESULTS: Eighteen tumor samples were obtained. Initial TIL cultures were successfully generated from 14/18 samples (77.7%). A median of 5.52 × 107 (range: 2.5 × 106-3.23 × 108) cells were produced from initial cultures, with 46.9% expressing a CD3 phenotype (46.9%). Eight samples underwent rapid expansion, demonstrating a median 458-fold expansion and a CD3 phenotype of 98%. Initial MIL cultures were successfully generated from five samples, with a predominantly CD3 phenotype (45.2%). Sufficient tumor tissue was only available for seven TIL samples to be tested for reactivity; none demonstrated responsiveness to autologous tumor. CONCLUSIONS: TIL and MIL expansion from pediatric solid tumors was successful, including the full-scale expansion process. This data supports translation to an ACT-TIL treatment strategy in the pediatric population and thus a Phase I trial of ACT-TIL in pediatric high-risk solid tumors is planned.
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Pseudomonas aeruginosa and Klebsiella pneumoniae are two common gram-negative pathogens that are associated with bacterial pneumonia and can often be isolated from the same patient. We used a mixed-pathogen pneumonia infection model in which mice were infected with sublethal concentrations of P. aeruginosa and K. pneumoniae, resulting in significant lethality, outgrowth of both bacteria in the lung, and systemic dissemination of K. pneumoniae. Inflammation, induced by P. aeruginosa activation of Toll-like receptor 5, results in prolonged neutrophil recruitment to the lung and increased levels of neutrophil elastase in the airway, resulting in lung damage and epithelial barrier dysfunction. Live P. aeruginosa was not required to potentiate K. pneumoniae infection, and flagellin alone was sufficient to induce lethality when delivered along with Klebsiella. Prophylaxis with an anti-Toll-like receptor 5 antibody or Sivelestat, a neutrophil elastase inhibitor, reduced neutrophil influx, inflammation, and mortality. Furthermore, pathogen-specific monoclonal antibodies targeting P. aeruginosa or K. pneumoniae prevented the outgrowth of both bacteria and reduced host inflammation and lethality. These findings suggest that coinfection with P. aeruginosa may enable the outgrowth and dissemination of K. pneumoniae, and that a pathogen- or host-specific prophylactic approach targeting P. aeruginosa may prevent or limit the severity of such infections by reducing neutrophil-induced lung damage.
Assuntos
Coinfecção/imunologia , Infecções por Klebsiella/imunologia , Klebsiella pneumoniae/imunologia , Neutrófilos/imunologia , Pneumonia/imunologia , Infecções por Pseudomonas/imunologia , Pseudomonas aeruginosa/imunologia , Animais , Células Cultivadas , Coinfecção/microbiologia , Coinfecção/patologia , Feminino , Infecções por Klebsiella/microbiologia , Infecções por Klebsiella/patologia , Camundongos , Camundongos Endogâmicos C57BL , Infiltração de Neutrófilos , Neutrófilos/microbiologia , Neutrófilos/patologia , Pneumonia/microbiologia , Pneumonia/patologia , Infecções por Pseudomonas/microbiologia , Infecções por Pseudomonas/patologia , Receptor 5 Toll-Like/metabolismoRESUMO
Purpose: To report the ocular and systemic adverse events in eyes receiving ≥10 intravitreal ziv-aflibercept (IVZ) injections.Methods: Medical records of patients who received ≥10 IVZ for various chorioretinal conditions with minimum follow up period of 12 months were retrospectively analysed. These eyes received standard dose of IVZ (1.25 mg/0.05 ml) on pro-re-nata (PRN) or treat and extend (T&E) protocol. The primary study outcome was ocular and systemic adverse events related to IVZ injections whereas secondary outcomes were change in best corrected visual acuity (BCVA) and central macular thickness (CMT) on optical coherence tomography (OCT) at last visit compared to baseline. Comparison of BCVA and CMT at baseline and final visit was done using paired t-test.Results: A total of 94 eyes which received a mean ± standard deviation (mean±SD = 14.4 ± 4.6) IVZ injections were studied. A total of 41 eyes were treatment naïve whereas 53 eyes received intravitreal injections in the past with last injection at least 3 months prior. Mean (±SD) follow up period was 26.7 ± 8.7 months. Ocular adverse events were limited with a case each of acute iridocyclitis, endophthalmitis, cataract progression and early epiretinal membrane formation. No systemic events were recorded within a month of IVZ injection. There was a significant improvement in BCVA (p = 0.001) and change in CMT (p = 0.001) at last visit.Conclusion: Ocular use of ziv-aflibercept is safe with limited ocular and systemic side effects. Multiple injections of IVZ can be used in various chorioretinal diseases over the long term.
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Coriorretinite/tratamento farmacológico , Corioide/patologia , Angiofluoresceinografia/métodos , Receptores de Fatores de Crescimento do Endotélio Vascular/administração & dosagem , Proteínas Recombinantes de Fusão/administração & dosagem , Retina/patologia , Tomografia de Coerência Óptica/métodos , Idoso , Inibidores da Angiogênese/administração & dosagem , Coriorretinite/diagnóstico , Feminino , Seguimentos , Fundo de Olho , Humanos , Injeções Intravítreas , Masculino , Receptores de Fatores de Crescimento do Endotélio Vascular/antagonistas & inibidores , Estudos Retrospectivos , Resultado do TratamentoRESUMO
Efficacious use of therapeutic gene delivery via nanoparticles is hampered by the challenges associated with targeted delivery to tissues of interest. Systemic administration of lipid nanoparticle (LNP)-encapsulated mRNA leads to a protein expressed predominantly in the liver and spleen. Here, LNP encapsulating mRNA was covalently conjugated to an antibody, specifically binding plasmalemma vesicle-associated protein (PV1) as a means to target lung tissue. Systemic administration of PV1-targeted LNPs demonstrated significantly increased delivery of mRNA to the lungs and a 40-fold improvement in protein expression in the lungs, compared with control LNPs. We also investigated the effect of LNP size to determine optimal tissue distribution and transfection. Larger-size PV1-targeted LNPs not only have the elasticity to target the PV1 expressed in the caveolae but also enable robust mRNA expression in the lungs. Targeted delivery of mRNA to the lungs is a promising approach in the treatment of lung diseases.
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Portadores de Fármacos/química , Lipídeos/química , Pulmão/metabolismo , Nanopartículas/química , RNA Mensageiro/farmacologia , Animais , Anticorpos Imobilizados/química , Anticorpos Imobilizados/imunologia , Cavéolas/imunologia , Feminino , Técnicas de Transferência de Genes , Proteínas de Membrana/imunologia , Camundongos Endogâmicos BALB CRESUMO
Analizamos el uso de las drogas en una perspectiva histórica y cultural de caracter necesariamente antropológico, de como los diversos pueblos antiguos usaban las drogas, que valores, contextos y formas de control social poseian en relación al conocimiento y uso de varios tipos de sustancias psicoactivas o fármacos de origem vegetal, y que sentido esas prácticas teníam. Contrastando con la epidemia de drogadicción en las sociedades modernas com su volumen de problemas de salud, de legislación y desestructuración familiar: la droga como característica de fenómeno cultural de la modernidad: crisis y ruptura, la busca de felicidad, consuelo y confort, como también formas de interacción y convivio social; tal vez una apelación por desespero de la soledad, un remedio para el vazio y tedio, caracteristicos del hombre moderno. En suma, se trata de estabelecer la posible relación entre individuo y sociedad, su contexto y ¿por que sus desvios por la droga? (AU)