RESUMO
The purpose of this study was to evaluate the efficacy of imiquimod-containing nanovesicles prepared with lipids extracted from the hyperhalophile archaebacterium Halorubrum tebenquichense (nanoARC-IMQ) to induce protection against Trypanosoma cruzi infection. The therapeutic efficacy of archaeolipid nanovesicles was assessed in an experimental murine model of acute infection with T. cruzi. The administration of nanoARQ-IMQ prevented mortality as compared to infected untreated animals, reduced parasitemia levels and diminished myocardial and musculoskeletal lesions in mice infected with a lethal strain of T. cruzi. Our findings suggest that the immunotherapy with nanoARC-IMQ has potential to limit the progression of Chagas disease.
Assuntos
Adjuvantes Imunológicos/uso terapêutico , Doença de Chagas/terapia , Imiquimode/uso terapêutico , Imunoterapia , Nanopartículas/uso terapêutico , Adjuvantes Imunológicos/química , Animais , Doença de Chagas/patologia , Imiquimode/química , Lipídeos/química , Masculino , Camundongos , Camundongos Endogâmicos C3H , Nanopartículas/química , Tamanho da Partícula , Propriedades de SuperfícieRESUMO
Hyperhalophilic archaebacteria exclusively produce sn2,3 diphytanylglycerol diether archaeolipids, unique structures absent in bacteria and eukaryotes. Nanovesicles made of archaeolipids known as nanoarchaeosomes (nanoARC), possess highly stable bilayers, some of them displaying specific targeting ability. Here we hypothesize that nanoARC made from Halorubrum tebenquichense archaebacteria, may constitute efficient carriers for the TLR7 agonist imiquimod (IMQ). NanoARC-IMQ takes advantage of the intense interaction between IMQ and the highly disordered, poorly fluid branched archaeolipid bilayers, rich in archaeol analog of methyl ester of phosphatidylglycerophosphate (PGP-Me), a natural ligand of scavenger receptor A1 (SR-A1). This approach lacks complex manufacture steps required for bilayers labeling, enabling future analytical characterization, batch reproducibility, and adaptation to higher scale production. SR-A1 mediated internalization of particulate material is mostly targeted to macrophages and is extensive because it is not submitted to a negative feedback. A massive and selective intracellular delivery of IMQ may concentrate its effect specifically into the endosomes, where the TLR7 is expressed, magnifying its immunogenicity, at the same time reducing its systemic bioavailability, and therefore it's in vivo adverse effects. NanoARC-IMQ (600-900 nm diameter oligolamellar vesicles of ~-43 mV Z potential) were heavily loaded with IMQ at ~44 µg IMQ/mg phospholipids [~20 folds higher than the non-SR-A1 ligand soyPC liposomes loaded with IMQ (LIPO-IMQ)]. In vitro, nanoARC-IMQ induced higher TNF-α and IL-6 secretion by J774A1 macrophages compared to same dose of IMQ and same lipid dose of LIPO-IMQ. In vivo, 3 subcutaneous doses of nanoARC-IMQ+ 10 µg total leishmania antigens (TLA) at 50 µg IMQ per Balb/C mice, induced more pronounced DTH response, accompanied by a nearly 2 orders higher antigen-specific systemic IgG titers than IMQ+TLA and LIPO-IMQ. The isotype ratio of nanoARC-IMQ+TLA remained ~0.5 indicating, the same as IMQ+TLA, a Th2 biased response distinguished by a pronounced increase in antibody titers, without negative effects on splenocytes lymphoproliferation, with a potential CD8+LT induction 10 days after the last dose. Overall, this first approach showed that highly SR-A1 mediated internalization of heavily loaded nanoARC-IMQ, magnified the effect of IMQ on TLR7 expressing macrophages, leading to a more intense in vivo immune response.
RESUMO
INTRODUCTION AND PURPOSE: Laparoscopic sleeve gastrectomy (LSG) in patients with a BMI between 30 and 35 kg/m2 plus comorbidities has shown to be safe and effective. The purpose of this study is to describe our outcomes in this group of patients after 3 years of follow-up. MATERIALS AND METHODS: Retrospective descriptive analysis of patients with initial BMI between 30 and 35 kg/m2 plus comorbidities were submitted to LSG between 2006 and 2013. We analyzed gender, age, comorbidities, BMI, total weight loss (%TWL), excess weight loss (%EWL), comorbidity resolution, morbidity, and mortality. Postoperative success was defined as %TWL over 20% and EWL% over 50% maintained for at least 1 year and comorbidity remission with no need of medication. RESULTS: Of the patients, 477 underwent a LSG in the above period and 252 met inclusion criteria; 188 (75%) were female and 64 (25%) were male. Median age was 39 years (15-70). Three-year follow-up was 43.9% (111 patients). Median preoperative BMI was 32.3 kg/m2 (30-34.3). Median postoperative %TWL was 12.9, 23.2, 28.2, 24.3, and 22.1% at 1, 6, 12, 24, and 36 months, respectively. %EWL was 42.88, 77.44, 98.42, 83.2, and 75.8%. Median surgical time was 86.9 min (40-120). There was comorbidity remission at 36 months. Insulin resistance was remitted in 89.4%, dyslipidemia 52%, non-alcoholic fatty liver disease 84.6%, hypertension 75%, and GERD 65%. T2DM had 60% of complete remission and 40% improvement. There were morbidity in six patients (2.4%), two reoperations, no leaks, and no mortality. CONCLUSIONS: Performing LSG in patients with grade I obesity is safe and effective. BMI should not be the only indicator to consider bariatric and metabolic surgery. We still require further studies and longer follow-up.
Assuntos
Índice de Massa Corporal , Gastrectomia , Obesidade Mórbida/epidemiologia , Obesidade Mórbida/cirurgia , Adolescente , Adulto , Idoso , Feminino , Seguimentos , Gastrectomia/efeitos adversos , Gastrectomia/métodos , Humanos , Laparoscopia/efeitos adversos , Laparoscopia/métodos , Masculino , Pessoa de Meia-Idade , Obesidade Mórbida/patologia , Complicações Pós-Operatórias/epidemiologia , Complicações Pós-Operatórias/etiologia , Período Pós-Operatório , Estudos Retrospectivos , Resultado do Tratamento , Adulto JovemRESUMO
Ultradeformable archaeosomes (UDA) are nanovesicles made of total polar archaeolipids (TPA) from the archaea Halorubrum tebenquichense, soybean phosphatidylcholine and sodium cholate (3:3:1w/w). Fresh dispersions of UDA including different type of antigens are acknowledged as efficient topical vaccination agents. UDA dispersions however, if manufactured for pharmaceutical use, have to maintain colloidal stability upon liposomicidal processes such as sterilization and lyophilization (SLRUDA), needed to extend shelf life during storage. The remaining capacity of SLRUDA to act as adjuvants was therefore tested here for the first time. Another unexplored issue addressed here, is the outcome of replacing classical antigen inclusion into nanovesicles by their physical mixture. Our results showed that UDA behaved as super-stable nanovesicles because of its high endurance during heat sterilization and storage for 5 months at 40°C. The archaeolipid content of UDA however, was insufficient to protect it against lyophilization, which demanded the addition of 2.5% v/v glycerol plus 0.07% w/v glucose. No significant differences were found between serum anti-ovalbumin (OVA) IgG titers induced by fresh or SLRUDA upon topical application of 4 weekly doses at 600µg lipids/75µg OVA to Balb/c mice. Finally, SLRUDA mixed with OVA elicited the same Th2 biased plus a non-specific cell mediated response than OVA encapsulated within UDA. Concluding, we showed that TPA is key component of super-stable nanovesicles that confers resistance to heat sterilization and to storage under cold-free conditions. The finding of SLRUDA as ready-to-use topical adjuvant would lead to simpler manufacture processing and cheaper products. .