RESUMO
BACKGROUND: Natural killer (NK) cell therapy is considered an attractive and safe strategy for anticancer therapy. Nevertheless, when autologous or allogenic NK cells are used alone, the clinical benefit has been disappointing. This is partially due to the lack of target specificity. Recently, CD19-specific chimeric antigen receptor (CAR)-NK cells have proven to be safe and potent in patients with B-cell tumors. However, the generation of CAR-NK cells is a complicated manufacturing process. We aim at developing a targeted NK cell therapy without the need for cellular genetic modifications. We took advantage of the natural expression of the IgG Fc receptor CD16a (FcγRIIIa) to induce strong antigen-specific effector functions through antibody-dependent cell-mediated cytotoxicity (ADCC). We have generated the new technology "Pin", which enables the arming of modified monoclonal antibodies (mAbs) onto the CD16a of ex vivo expanded NK (eNK) cells. Methods Ex vivo eNK were prepared from umbilical cord blood cells and expanded using interleukin (IL)-2/IL-15 and Epstein-Barr virus (EBV)-transformed B-lymphoblastoid feeder cells. mAbs were engineered with four substitutions called Pin mutations to increase their affinity to CD16a. eNK were incubated with anti-CD20 or anti-CD19 Pin-mAbs to generate "armed" eNK and were used to assess effector functions in vitro on cancer cell lines, lymphoma patient cells and in vivo. RESULTS: CD16a/Pin-mAb interaction is stable for several days and Pin-mAb eNK inherit the mAb specificity and exclusively induce ADCC against targets expressing the cognate antigen. Hence, Pin-mAbs confer long-term selectivity to eNK, which allows specific elimination of the target cells in several in vivo mouse models. Finally, we showed that it is possible to arm eNK with at least two Pin-mAbs simultaneously, to increase efficacy against heterogenous cancer cell populations. CONCLUSIONS: The Pin technology provides an off-the-shelf NK cell therapy platform to generate CAR-like NK cells, without genetic modifications, that easily target multiple tumor antigens.
Assuntos
Células Matadoras Naturais , Receptores de IgG , Células Matadoras Naturais/imunologia , Células Matadoras Naturais/metabolismo , Humanos , Animais , Camundongos , Receptores de IgG/metabolismo , Receptores de IgG/imunologia , Imunoterapia Adotiva/métodos , Linhagem Celular Tumoral , Antígenos CD19/imunologia , Citotoxicidade Celular Dependente de Anticorpos , Receptores de Antígenos Quiméricos/imunologia , Receptores de Antígenos Quiméricos/genética , Receptores de Antígenos Quiméricos/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto , Anticorpos Monoclonais/uso terapêutico , Anticorpos Monoclonais/farmacologiaRESUMO
COVID-19 is caused by the infection of the lungs by SARS-CoV-2. Monoclonal antibodies, such as sotrovimab, showed great efficiency in neutralizing the virus before its internalization by lung epithelial cells. However, parenteral routes are still the preferred route of administration, even for local infections, which requires injection of high doses of antibody to reach efficacious concentrations in the lungs. Lung administration of antibodies would be more relevant requiring lower doses, thus reducing the costs and the side effects. But aerosolization of therapeutic proteins is very challenging, as the different processes available are harsh and trigger protein aggregation and conformational changes. This decreases the efficiency of the treatment, and can increase its immunogenicity. To address those issues, we developed a series of new excipients composed of a trehalose core, a succinyl side chain and a hydrophobic carbon chain (from 8 to 16 carbons). Succinylation increased the solubility of the excipients, allowing their use at relevant concentrations for protein stabilization. In particular, the excipient with 16 carbons (C16TreSuc) used at 5.6 mM was able to preserve colloidal stability and antigen-binding ability of sotrovimab during the nebulization process. It could also be used as a cryoprotectant, allowing storage of sotrovimab in a lyophilized form during weeks. Finally, we demonstrated that C16TreSuc could be used as an excipient to stabilize antibodies for the treatment against COVID-19, by in vitro and in vivo assays. The presence of C16TreSuc during nebulization preserved the neutralization capacity of sotrovimab against SARS-CoV-2 in vitro; an increase of its efficacy was even observed, compared to the non-nebulized control. The in vivo study also showed the wide distribution of sotrovimab in mice lungs, after nebulization with 5.6 mM of excipient. This work brings a solution to stabilize therapeutic proteins during storage and nebulization, making pulmonary immunotherapy possible in the treatment of COVID-19 and other lung diseases.
Assuntos
COVID-19 , Excipientes , Camundongos , Animais , Excipientes/química , Trealose/química , SARS-CoV-2 , Anticorpos AntiviraisRESUMO
BACKGROUND AND OBJECTIVES: The aim of this work was to test the hypothesis that patients with temporal lobe epilepsy (TLE) with subjective initial memory complaints (not confirmed by an objective standard assessment) and various phenotypes also show objective very long-term memory deficit with accelerated long-term forgetting. We tested patients with TLE with 2 surprise memory tests after 3 weeks: the standard Free and Cued Selective Reminding Test (FCSRT) and Epireal, a new test specifically designed to capture more ecologic aspects of autobiographical memory. METHODS: Forty-seven patients with TLE (12 with hippocampal sclerosis, 12 with amygdala enlargement, 11 with extensive lesions, 12 with normal MRI) who complained about their memory, but for whom the standard neuropsychological assessment did not reveal any memory impairment after a standard delay of 20 minutes, underwent 2 surprise memory tests after 3 weeks. They were compared to 35 healthy controls. RESULTS: After 3 weeks, FCSRT and Epireal recall scores were significantly lower in patients than in controls (p < 0.001). There was no significant correlation between FCSRT and Epireal scores (p = 0.99). Seventy-six percent of patients with TLE had objective impairment on at least 1 of these very long-term memory tests, regardless of the existence and type of lesion or response to antiseizure medication. Easily applicable, Epireal had a higher effect size, detected deficits in 28% more patients, and is a useful addition to the standard workup. DISCUSSION: Assessing long-term memory should be broadened to a wide spectrum of patients with TLE with a memory complaint, regardless of the epileptic syndrome, regardless of whether it is associated with a lesion. This could lead to rethinking TLE nosology associated with memory.
Assuntos
Epilepsia do Lobo Temporal , Memória Episódica , Epilepsia do Lobo Temporal/complicações , Epilepsia do Lobo Temporal/diagnóstico por imagem , Epilepsia do Lobo Temporal/patologia , Humanos , Transtornos da Memória/complicações , Transtornos da Memória/etiologia , Rememoração Mental , Testes NeuropsicológicosRESUMO
Modulation of gut microbiome composition seems to be a promising therapeutic strategy for a wide range of pathologic states. However, these microbiota-targeted interventions may affect production of microbial metabolites, circulating factors in the gut-liver axis influencing hepatic drug metabolism with possible clinical relevance. Butyrate, a short-chain fatty acid produced through microbial fermentation of dietary fibers in the colon, has well established anti-inflammatory role in the intestine, while the effect of butyrate on the liver is unknown. In this study, we have evaluated the effect of butyrate on hepatic AhR activity and AhR-regulated gene expression. We have showed that AhR and its target genes were upregulated by butyrate in dose-dependent manner in HepG2-C3 as well as in primary human hepatocytes. The involvement of AhR has been proved using specific AhR antagonists and siRNA-mediated AhR silencing. Experiments with AhR reporter cells have shown that butyrate regulates the expression of AhR target genes by modulating the AhR activity. Our results suggest also epigenetic action by butyrate on AhR and its repressor (AHRR) presumably through mechanisms based on HDAC inhibition in the liver. Our results demonstrate that butyrate may influence the drug-metabolizing ability of liver enzymes e.g., through the interaction with AhR-dependent pathways.
Assuntos
Butiratos , Microbioma Gastrointestinal , Butiratos/metabolismo , Butiratos/farmacologia , Colo/metabolismo , Ácidos Graxos Voláteis/metabolismo , Humanos , Fígado/metabolismo , Receptores de Hidrocarboneto Arílico/genética , Receptores de Hidrocarboneto Arílico/metabolismoRESUMO
The consistency of cerebrospinal fluid amyloid-ß (Aß)42/40 ratio and Aß42 has not been assessed in the AT(N) classification system. We analyzed the classification changes of the dichotomized amyloid status (A+/A-) in 363 patients tested for Alzheimer's disease biomarkers after Aß42 was superseded by the Aß42/40 ratio. The consistency of Aß42 and the Aß42/40 ratio was very low. Notably, the proportions of "false" A+T-patients were considerable (74-91%) and corresponded mostly to patients not clinically diagnosed with Alzheimer's disease. Our results suggest that the interchangeability of Aß42/40 ratio and Aß42 is limited for classifying patients in clinical setting using the AT(N) scheme.
Assuntos
Peptídeos beta-Amiloides/líquido cefalorraquidiano , Biomarcadores/líquido cefalorraquidiano , Disfunção Cognitiva/líquido cefalorraquidiano , Fragmentos de Peptídeos/líquido cefalorraquidiano , Terminologia como Assunto , Idoso , Doença de Alzheimer/classificação , Humanos , Proteínas tau/líquido cefalorraquidianoRESUMO
The large experience accumulated with the therapeutic use of monoclonal antibodies has revealed undesirable effects, among which opportunistic infections when prescribed in inflammatory or hematological diseases. This deleterious effect is a direct consequence of the immunosuppression induced by these antibodies through the blockade of several key pathways involved in both innate and adaptative immune responses, including migration of effector cells, depletion of B or T lymphocytes, inhibition of key cell-cell interactions. Four antibodies are concerned, targeting CD52, CD20, TNF-a and VLA-4, and major risks include activation of latent tuberculosis, or of normally silent viruses. Precise evaluation of these risks and understanding of their mechanisms have now led to the improvement of clinical safety, based on the detection of patients at risk, weighting of the benefit/risk ratio, and a very rigorous detection of latent infections before the onset of treatment by monoclonal antibodies know to induce immunosuppression.
Assuntos
Anticorpos Monoclonais/efeitos adversos , Infecções Oportunistas/etiologia , Alemtuzumab , Anticorpos Monoclonais/uso terapêutico , Anticorpos Monoclonais Humanizados , Anticorpos Monoclonais Murinos , Anticorpos Antineoplásicos/efeitos adversos , Anticorpos Antineoplásicos/uso terapêutico , Antígenos CD/imunologia , Antígenos CD20/imunologia , Antígenos de Neoplasias/imunologia , Antígeno CD52 , Estudos de Coortes , Glicoproteínas/imunologia , Humanos , Terapia de Imunossupressão/efeitos adversos , Integrina alfa4beta1/imunologia , Metanálise como Assunto , Infecções Oportunistas/epidemiologia , Recidiva , Estudos Retrospectivos , Rituximab , Tuberculose/etiologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/imunologia , Ativação ViralRESUMO
INTRODUCTION: 20-30% of depressed patients experience Treatment Resistant Depression (TRD). Electroconvulsive Therapy (ECT) remains the treatment of choice for TRD. However, the exact mechanism of ECT remains unclear. We aim to assess grey matter changes in patients with TRD undergoing bilateral ECT treatment at different points during and after treatment. METHODS: Patients are recruited at the University Hospital of Toulouse. Eligibility criteria include a diagnosis of TRD and an age between 50 and 70 years old. Patients received clinical assessments (Hamilton Depression Rating Scale) and structural scans (MRI) at three points: baseline (within 48â¯h before the first ECT); V2 (after the first ECT considered effective); and V3 (within 1 week of completing ECT). RESULTS: At baseline, controls had significantly higher cortical thickness than patients in the fusiform gyrus, the inferior, middle and superior temporal gyrus, the parahippocampal gyrus and the transverse temporal gyrus (respectively: t(35)=2.7, pâ¯=â¯0.02; t(35)=2.89, pâ¯=â¯0.017; t(35)=3.1, pâ¯=â¯0.015; t(35)=3.6, pâ¯=â¯0.009; t(35)=2.37, pâ¯=â¯0.031; t(35)=2.46, pâ¯=â¯0.03). This difference was no longer significant after ECT. We showed an increase in cortical thickness in superior temporal gyrus between (i) baseline and V3 (t(62)=-3.43 pâ¯=â¯0.009) and (ii) V2 and V3 (t(62)=-3.42 pâ¯=â¯0.009). We showed an increase in hippocampal volume between (i) baseline and V3 (t(62)=-5.23 p < 0.001) and (ii) V2 and V3 (t(62)=-5.3 p < 0.001). CONCLUSION: We highlight that there are grey matter changes during ECT treatment in a population with TRD compared to a healthy control population. These changes seem to occur after several rounds of ECT.
Assuntos
Transtorno Depressivo Resistente a Tratamento/patologia , Transtorno Depressivo Resistente a Tratamento/terapia , Eletroconvulsoterapia , Substância Cinzenta/patologia , Adulto , Idoso , Encéfalo/diagnóstico por imagem , Encéfalo/patologia , Transtorno Depressivo Resistente a Tratamento/diagnóstico por imagem , Feminino , Substância Cinzenta/diagnóstico por imagem , Hipocampo/diagnóstico por imagem , Hipocampo/patologia , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Giro Para-Hipocampal/diagnóstico por imagem , Giro Para-Hipocampal/patologia , Lobo Temporal/diagnóstico por imagem , Lobo Temporal/patologiaRESUMO
OBJECTIVE: To evaluate resilience and frequency of behavioral symptoms in Haitian children internationally adopted before and after the earthquake of January 12, 2010. METHODS: We conducted a retrospective quantitative study in 40 Haitian children. Families were also asked to participate in a qualitative study (individual interview at 18-24 months after the earthquake) and to complete State-Trait Anxiety Inventory (STAI) and STAI for children (STAI-C) questionnaires. RESULTS: Demographic and clinical characteristics were similar in the group who experienced the earthquake (n=22) and in the group who did not (n=18). The families of 30 adoptees were interviewed. There was no statistical difference between the two groups for the STAI (P=0.53) and STAI-C (P=0.75) or for the frequency of behavioral problems. Plenary adoption was pronounced for 84.6% and 33.3% of the children adopted in the pre- and post-earthquake group, respectively (P=0.02). Children rarely talked about the experience of the earthquake, which, by contrast, was a stressful experience for the adoptive families. CONCLUSIONS: Haitian children adopted after the earthquake did not express more stress or behavioral problems than those adopted before it. However, the possibility of a resurgence of mental disorders after age 10 should be borne in mind. (Disaster Med Public Health Preparedness. 2018;12:450-454).
Assuntos
Adoção/etnologia , Terremotos/estatística & dados numéricos , Internacionalidade , Adulto , Criança , Pré-Escolar , Feminino , França , Haiti/etnologia , Humanos , Masculino , Pessoa de Meia-Idade , Psicometria/instrumentação , Psicometria/métodos , Pesquisa Qualitativa , Resiliência Psicológica , Estudos Retrospectivos , Inquéritos e Questionários , Sobreviventes/estatística & dados numéricosRESUMO
Internationally adopted children often present diseases contracted in the country of origin. Skin diseases are common in new arrivals, and diagnosis may prove challenging for GPs or even dermatologists if they are inexperienced in the extensive geographic and ethnic diversity of international adoptees. To analyse the frequency and characteristics of skin diseases in international adoptees. In total, 142 adoptees were evaluated for a cross-sectional cohort study. The most frequent diseases observed at arrival were dermatological conditions. Of the adoptees, 70% presented at least one skin disease, of which 57.5% were infectious; Tinea capitis being the most frequent (n = 42). The recovery rate of Tinea capitis was 89% (n = 32/36). Ten cases of scabies were diagnosed. Other diseases included viral skin infection (n = 22), with 16 cases of Molluscum contagiosum and bacterial infection. Skin diseases are very common in internationally adopted children. There is a need for close collaboration between dermatologists and paediatricians to diagnose such infections, as well as clear guidelines to treat them.
Assuntos
Adoção , Dermatopatias/diagnóstico , Dermatopatias/etnologia , África/etnologia , Ásia/etnologia , Brasil/etnologia , Criança , Pré-Escolar , Colômbia/etnologia , Estudos Transversais , Emigração e Imigração , Feminino , França , Haiti/etnologia , Humanos , Lactente , Letônia/etnologia , Lituânia/etnologia , Masculino , Estudos Retrospectivos , Escabiose/diagnóstico , Escabiose/etnologia , Dermatopatias Bacterianas/diagnóstico , Dermatopatias Bacterianas/etnologia , Dermatopatias Virais/diagnóstico , Dermatopatias Virais/etnologia , Tinha do Couro Cabeludo/diagnóstico , Tinha do Couro Cabeludo/etnologiaRESUMO
Recurrent staphylococcal skin and soft tissue infections may recur despite decontamination and multiple courses of antibiotic therapy and may dramatically impair the patient's quality of life. We report successful use of long-term azithromycin prophylaxis in a recurrent laryngitis and a scalp folliculitis due to methicillin-susceptible Staphylococcus aureus.
Assuntos
Antibacterianos/administração & dosagem , Antibioticoprofilaxia/métodos , Azitromicina/administração & dosagem , Staphylococcus aureus Resistente à Meticilina/efeitos dos fármacos , Infecções Estafilocócicas/tratamento farmacológico , Idoso , Antibacterianos/farmacologia , Azitromicina/farmacologia , Foliculite/tratamento farmacológico , Foliculite/microbiologia , Foliculite/prevenção & controle , Humanos , Laringite/tratamento farmacológico , Laringite/microbiologia , Laringite/prevenção & controle , Masculino , Pessoa de Meia-Idade , Recidiva , Infecções Estafilocócicas/microbiologia , Infecções Estafilocócicas/prevenção & controleRESUMO
Monoclonal antibodies (mAbs), especially those that interact with immune or hematologic leukocyte membrane targets, have changed the outcome of numerous diseases. However, mAbs can block or reduce immune cells and cytokines, and can lead to increased risk of infection. Some of these risks are predictable and can be explained by their mechanisms of action. Others have been observed only after the mAbs were licensed and used extensively in patients. In this review, we focus on infectious complications that occur upon treatment with mAbs or Fc-containing fusion proteins targeting leukocyte membrane proteins, including CD52, CD20, tumor necrosis factor, VLA4, CD11a and CTLA4. We report their known infectious risks and the recommendations for their use. Although most of these drugs are clinically safe when the indications are respected, we emphasize the need for regular updating of pharmacovigilance data.