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AIMS: Narrowing or occlusion of arteries that supply the limbs can evolve to critical limb ischemia. M-CSF promotes proliferation, differentiation and survival of monocytes and macrophages, and polarization of macrophages to M2-subtype, which are essential elements for vessel formation and tissue repair. Based on these properties of M-CSF, we hypothesize that transfection of M-CSF into ischemic limbs may promote vessel formation and repair of ischemic limbs. MAIN METHODS: Hindlimb ischemia was surgically induced in 10-12 weeks old Balb/c and gene therapy was performed with intramuscular application of either uP-MCSF or uP plasmids (100 µg). Macrophage and monocyte subpopulations were assessed by flow cytometry and blood flow was monitored by Laser Doppler Perfusion Imaging (LDPI). Thirty days after transfection, we assessed gastrocnemius mass and muscle force, subsequently collecting the muscle for histology. KEY FINDINGS: We successfully developed the uP-MCSF plasmid, which increases M-CSF expression in the muscle transiently. Thirty days after uP-MCSF gene therapy in ischemic muscles, the treated group presented: improved muscle force, reduced fibrosis and increased arteriogenesis, although LDPI analysis did not show any significant difference in blood flow among groups. Noteworthy, we observed a temporary increase in MHCIIhighCD206high macrophages after uP-MCSF transfection. SIGNIFICANCE: M-CSF gene therapy improved ischemic muscle functionality by promoting arteriogenesis and decreasing fibrosis, likely through increased MHCIIhighCD206high macrophages and not via classically known M2-macrophages.
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Fator Estimulador de Colônias de Macrófagos , Macrófagos , Animais , Humanos , Macrófagos/metabolismo , Monócitos/metabolismo , Músculo Esquelético/patologia , Isquemia/metabolismo , Membro Posterior/irrigação sanguíneaRESUMO
OBJECTIVES: To evaluate the impact of invasive mechanical ventilation associated with two serum inflammatory cytokines and clinical indicators, on the second day of life, as predictors of bronchopulmonary dysplasia in very low birth weight preterm infants. It was hypothesized that the use of invasive mechanical ventilation in the first hours of life is associated with biomarkers that may predict the chances of preterm infants to develop bronchopulmonary dysplasia. METHODS: Prospective cohort of 40 preterm infants with gestational age <34 weeks and birth weight <1500â¯g. The following were analyzed: clinical variables; types of ventilator support used (there is a higher occurrence of bronchopulmonary dysplasia when oxygen supplementation is performed by long periods of invasive mechanical ventilation); hospitalization time; quantification of two cytokines (granulocyte and macrophage colony stimulating factor [GM-CSF] and eotaxin) in blood between 36 and 48â¯h of life. The preterm infants were divided in two groups: with and without bronchopulmonary dysplasia. RESULTS: The GM-CSF levels presented a significantly higher value in the bronchopulmonary dysplasia group (pâ¯=â¯0.002), while eotaxin presented higher levels in the group without bronchopulmonary dysplasia (pâ¯=â¯0.02). The use of continuous invasive mechanical ventilation was associated with increased ratios between GM-CSF and eotaxin (100% sensitivity and 80% specificity; receiver operating characteristic areaâ¯=â¯0.9013, CIâ¯=â¯0.7791-1.024, pâ¯<â¯0.0001). CONCLUSIONS: The duration of invasive mechanical ventilation performed in the first 48â¯h of life in the very low birth weight infants is a significant clinical predictor of bronchopulmonary dysplasia. The use of continuous invasive mechanical ventilation was associated with increased ratios between GM-CSF and eotaxin, suggesting increased lung injury and consequent progression of the disease.
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Displasia Broncopulmonar , Biomarcadores , Displasia Broncopulmonar/diagnóstico , Humanos , Lactente , Recém-Nascido , Recém-Nascido Prematuro , Estudos Prospectivos , Respiração ArtificialRESUMO
[This corrects the article DOI: 10.1155/2020/4086929.].
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BACKGROUND: After traumatic skeletal muscle injury, muscle healing is often incomplete and produces extensive fibrosis. The sequence of M1 and M2 macrophage accumulation and the duration of each subtype in the injured area may help to direct the relative extent of fibrogenesis and myogenesis during healing. We hypothesized that increasing the number of M1 macrophages early after traumatic muscle injury would produce more cellular and molecular substrates for myogenesis and fewer substrates for fibrosis, leading to better muscle healing. METHODS: To test this hypothesis, we transfected skeletal muscle with a plasmid vector to transiently express GM-CSF shortly after injury to drive the polarization of macrophages towards the M1 subset. C57BL/6 mouse tibialis anterior (TA) muscles were injured by contusion and electroporated with uP-mGM, which is a plasmid vector that transiently expresses GM-CSF. Myogenesis, angiogenesis, and fibrosis were evaluated by histology, immunohistochemistry, and RT-qPCR; subpopulations of macrophages by flow cytometry; and muscle functioning by the maximum running speed on the treadmill and the recovery of muscle mass. RESULTS: Muscle injury increased the number of local M1-like macrophages and decreased the number of M2-like macrophages on day 4, and uP-mGM treatment enhanced this variation. uP-mGM treatment decreased TGF-ß1 protein expression on day 4, and the Sirius Red-positive area decreased from 35.93 ± 15.45% (no treatment) to 2.9% ± 6.5% (p < 0.01) on day 30. uP-mGM electroporation also increased Hgf, Hif1α, and Mtor gene expression; arteriole density; and muscle fiber number during regeneration. The improvement in the quality of the muscle tissue after treatment with uP-mGM affected the increase in the TA muscle mass and the maximum running speed on a treadmill. CONCLUSION: Collectively, our data show that increasing the number of M1-like macrophages immediately after traumatic muscle injury promotes muscle recovery with less fibrosis, and this can be achieved by the transient expression of GM-CSF.
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Fator Estimulador de Colônias de Granulócitos e Macrófagos , Macrófagos , Animais , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético , CicatrizaçãoRESUMO
The present study is aimed at evaluating serological method using scFv anti-Strongyloides sp. and reporting the frequencies of the results with conventional parasitological technique (faeces) in elderly individuals. Among 112 elderly individuals (≥60 years of age), 14.28% were positive for at least one enteroparasite, with one individual positive for S. stercoralis. Sera were evaluated for the presence of anti-Strongyloides sp. antibodies using total or detergent fraction extracts of Strongyloides venezuelensis, which presented positivity rates of 19.64% and 10.71%, respectively. An anti-HSP60 single-chain variable fragment from Strongyloides sp. was used to detect parasite antigens, with 5.36% (6 individuals) of ELISA-positive individuals returning a positive result. While the serological test indicates previous or recent infection and may be limited by antigen purification, the anti-HSP60 method reflects the presence of Strongyloides sp. immune complexes and exhibits greater sensitivity and specificity. Our results demonstrate the variable occurrence of enteroparasites in elderly individuals residing in long-term nursing homes and validate a novel epidemiological tool to describe infection cases by Strongyloides sp.
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Anticorpos Anti-Helmínticos/sangue , Complexo Antígeno-Anticorpo/sangue , Antígenos de Helmintos/sangue , Chaperonina 60/sangue , Anticorpos de Cadeia Única/sangue , Estrongiloidíase/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Animais , Anticorpos Anti-Helmínticos/imunologia , Antígenos de Helmintos/imunologia , Brasil , Chaperonina 60/imunologia , Fezes/parasitologia , Feminino , Instituição de Longa Permanência para Idosos , Humanos , Masculino , Pessoa de Meia-Idade , Casas de Saúde , Sensibilidade e Especificidade , Anticorpos de Cadeia Única/imunologia , Strongyloides/crescimento & desenvolvimento , Strongyloides/imunologia , Strongyloides/patogenicidade , Estrongiloidíase/sangue , Estrongiloidíase/imunologia , Estrongiloidíase/parasitologiaRESUMO
Leprosy reactions are acute immunological events that occur during the evolution of chronic infectious disease causing neural damage and disabilities. A study using blood samples of 17 leprosy reaction patients and 17 reaction-free was carried out by means of associations between antigens, receptors, and expression of cytokines, using path analysis providing new insights into the immunological mechanisms involved in triggering leprosy reactions. Toll-like receptors (TLR) such as TLR1 and TLR2, presented balanced expression in the reaction-free multibacillary (MB) group (TLR1: 1.01 ± 0.23, TLR2: 1.22 ± 0.18; p = 0.267). On the other hand, downgrading type 1 reaction (T1R) (TLR1: 1.24 ± 0.17, TLR2: 2.88 ± 0.37; p = 0.002) and erythema nodosum leprosum (ENL) (TLR1: 1.93 ± 0.17, TLR2: 2.81 ± 0.15; p = 0.004) revealed an unbalance in relation to the expression of these receptors. When the path analysis was approached, it was noted that interleukin 10 (IL-10) expression showed a dependence relation with phenolic glycolipid I (PGL-I) in downgrading T1R (direct effect = 0.503 > residual effect = 0.364), whereas in ENL, such relationship occurred with lipoarabinomannan (LAM) (direct effect = 0.778 > residual effect = 0.280). On the contrary, in the reaction-free leprosy group, interferon-gamma (IFN-γ) levels were dependent on the association between TLR2 and TLR1 (0.8735). The high TLR2 expression associated with IL-10 levels, in the leprosy reaction groups, may be hypothetically related to the formation of TLR2/2 homodimers and/or TLR2/6 heterodimers linked to evasion mechanisms in downgrading reactions and pathophysiology of ENL.
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Eritema Nodoso/etiologia , Regulação da Expressão Gênica , Interferon gama/genética , Interleucina-10/genética , Hanseníase/etiologia , Receptor 1 Toll-Like/genética , Receptor 2 Toll-Like/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores , Suscetibilidade a Doenças , Eritema Nodoso/diagnóstico , Eritema Nodoso/epidemiologia , Eritema Nodoso/metabolismo , Feminino , Humanos , Interferon gama/metabolismo , Interleucina-10/metabolismo , Hanseníase/diagnóstico , Hanseníase/epidemiologia , Hanseníase/metabolismo , Masculino , Pessoa de Meia-Idade , Modelos Biológicos , Ligação Proteica , Transdução de Sinais , Receptor 1 Toll-Like/metabolismo , Receptor 2 Toll-Like/metabolismo , Adulto JovemRESUMO
This work shows the antitumor and antimetastatic effects of BthTX-II, an Asp-49 PLA2 from Bothrops jararacussu venom, on MDA-MB-231 human triple negative breast cancer cells. BthTX-II caused a dose-dependent cell death of MDA-MB-231 cells when compared with the non-tumorigenic breast cells by inducing apoptosis and autophagy. BthTX-II was also able to decrease the proliferation and to inhibit cell cycle progression. We also observed an upregulation of the ATM gene, which is responsible for cell-cycle arrest and DNA repair such as CCND1, CCNE1, CDC25A, E2F1, AKT1 and AKT3. Interestingly, BthTX-II inhibited invasion, migration and 3D cell growth of MDA-MB-231 cells, as well as inhibited the epithelial-mesenchymal transition (EMT) of this cell by increasing E-cadherin (CDH-1) and decreasing TWIST1, CTNNB1, vimentin and cytokeratin-5 expression. In conclusion, these results showed that BthTX-II displays antitumor and antimetastatic effects on MDA-MB-231 cells and may be useful for the development of new approaches and therapeutic strategies to manage triple negative breast cancer.
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Antineoplásicos/química , Antineoplásicos/farmacologia , Bothrops , Venenos de Crotalídeos/química , Venenos de Crotalídeos/farmacologia , Fosfolipases A2 do Grupo II/química , Fosfolipases A2 do Grupo II/farmacologia , Animais , Antineoplásicos/isolamento & purificação , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Biomarcadores Tumorais , Adesão Celular/efeitos dos fármacos , Pontos de Checagem do Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Venenos de Crotalídeos/isolamento & purificação , Fosfolipases A2 do Grupo II/isolamento & purificação , Humanos , Venenos de Serpentes/química , Venenos de Serpentes/farmacologiaRESUMO
Safety and biocompatibility assessment of biomaterials are themes of constant concern as advanced materials enter the market as well as products manufactured by new techniques emerge. Within this context, this review provides an up-to-date approach on current methods for the characterization and safety assessment of biomaterials and biomedical devices from a physical-chemical to a biological perspective, including a description of the alternative methods in accordance with current and established international standards.
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Materiais Biocompatíveis/efeitos adversos , Materiais Biocompatíveis/normas , Teste de Materiais/normas , Pesquisa Biomédica/normas , HumanosRESUMO
The present work reports the effects of a C-type lectin (BpLec) isolated from Bothrops pauloensis snake venom upon in vitro and in vivo angiogenesis models. Initially, we noted that BpLec was not cytotoxic to endothelial cells (tEnd) in doses up to 40µg/mL, but lower doses (2.5µg/mL, 5µg/mL, 10µg/mL and 20µg/mL) reduced tEnd cells adhesion to some extracellular matrix proteins and inhibited the in vitro vessel formation in Matrigel assay stimulated by bFGF. ß-galactosides (d-lactose, N-acetyl-d-galactosamine and d-galactose) at 400mM reversed the effect of BpLec on tEnd cells adhesion, whereas d-galactose (400mM) partially reversed BpLec property of inhibiting vessel formation by tEnd cells in Matrigel. In vivo assays showed that BpLec increased hemoglobin content and capillary vessels number in polyether-polyurethane sponge discs subcutaneously implanted into dorsal skin mice. Additionally, BpLec also reduced collagen deposition and did not induce a pro-inflammatory response, as demonstrated by the decreased the secretion of some inflammatory cytokines, whereas myeloperoxidase (MPO) and N-acetylglucosaminidase (NAG) activities were not altered by BpLec. Taken together, our results indicate that BpLec might represent an interesting angiogenesis and inflammatory modulator that could also be used for searching possible therapeutic targets involved in these processes.
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Inibidores da Angiogênese/farmacologia , Bothrops , Venenos de Crotalídeos/química , Lectinas Tipo C/metabolismo , Acetilglucosaminidase/metabolismo , Inibidores da Angiogênese/isolamento & purificação , Inibidores da Angiogênese/toxicidade , Animais , Adesão Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Células Endoteliais/citologia , Células Endoteliais/efeitos dos fármacos , Lectinas Tipo C/isolamento & purificação , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Neovascularização Fisiológica/efeitos dos fármacos , Peroxidase/metabolismoRESUMO
BACKGROUND: The development of a vaccine for the prevention of visceral leishmaniasis (VL) still represents a significant unmet medical need. A human vaccine can be found if one takes into consideration that many people living in endemic areas of disease are infected but do not develop active VL, including those subjects with subclinical or asymptomatic infection. METHODS: In this study, a phage display was used to select phage-exposed peptides that were specific to immunoglobulin G (IgG) antibodies from asymptomatic and symptomatic VL patients, separating them from non-infected subjects. Phage clones presenting valid peptide sequences were selected and used as stimuli of peripheral blood mononuclear cells (PBMCs) obtained from both patients' groups and controls. Those with higher interferon-gamma (IFN-γ)/interleukin (IL)-10 ratios were further selected for vaccination tests. RESULTS: Among 17 evaluated clones, two were selected, B1 and D11, and used to immunize BALB/c mice in an attempt to further validate their in vivo protective efficacy against Leishmania infantum infection. Both clones induced partial protection against the parasite challenge, which was evidenced by the reduction of parasitism in the evaluated organs, a process mediated by a specific T helper (Th)1 immune response. CONCLUSIONS: To the best of our knowledge, this study is the first to use a rational strategy based on in vitro stimulation of human PBMCs with selected phage-displayed clones to obtain new immunogens against VL.
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Anticorpos Antiprotozoários/sangue , Antígenos de Protozoários/imunologia , Leishmania infantum/imunologia , Leishmaniose Visceral/prevenção & controle , Vacinas Protozoárias/imunologia , Vacinas Protozoárias/isolamento & purificação , Células Th1/imunologia , Animais , Humanos , Imunoensaio , Interferon gama/metabolismo , Interleucina-10/metabolismo , Leishmaniose Visceral/imunologia , Programas de Rastreamento , Camundongos Endogâmicos BALB C , Biblioteca de PeptídeosRESUMO
The cyclopentenone prostaglandin 15-deoxy-Δ(12,14)-prostaglandin J2 (15d-PGJ2) is a natural ligand of peroxisome proliferator-activated receptor gamma (PPAR-γ) and a potential mediator of apoptosis in cancer cells. In the present study, we evaluated the effect of 15d-PGJ2 in human thyroid papillary carcinoma cells (TPC-1) using different doses of 15d-PGJ2 (0.6 to 20 µM) to determine IC50 (9.3 µM) via the MTT assay. The supernatant culture medium of the TPC-1 cells that was treated either with 15d-PGJ2 or with vehicle (control) for 24 hours was assessed for IL-6 secretion via CBA assay. RT-qPCR was used to evaluate mRNA expression of IL-6, SOCS1, SOCS3, and STAT3. TPC-1 cells treated with 15d-PGJ2 decreased the secretion and expression of IL-6 and STAT3, while it increased SOCS1 and SOCS3. Overall, we demonstrated that 15d-PGJ2 downregulated IL-6 signaling pathway and led TPC-1 cells into apoptosis. In conclusion, 15d-PGJ2 shows the potential to become a new therapeutic approach for thyroid tumors.
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Background: Metallothioneins (MTs) gene polymorphisms have been associated with the ability of free radicalscavenging and detoxification of heavy metals leading to cancer development. Our aim was to revisit, in a Brazil-ian population, single-nucleotide polymorphisms (SNPs) of the MT gene family previously associated with oralsquamous cell carcinoma (OSCC).Material and Methods: A case-control investigation with 28 OSCC patients and 45 controls was conducted, usingconventional risk factors (tobacco use and alcohol consumption) as covariates. SNPs genotyping for rs8052334(MT1B), rs964372 (MT1B), and rs1610216 (MT2A) was performed by PCR-RFLP, and SNPs for rs11076161(MT1A) were analyzed by TaqMan assay.Results: The only SNP associated with increased risk for OSCC was the MT-1A AA genotype (OR = 4.7; p = 0.01).We have also evidenced for the first time a significant linkage disequilibrium between the SNPs of MT-2A andMT-1A in this population with the highest frequency (30%) of the unfavorable haplotype G/A/C/T (rs1610216 /rs11076161 / rs964372 / rs8052334) of MT gene polymorphisms (OR = 6.2; p = 0.04). Interestingly, after removingthe effects of conventional risk factors, we have uncovered the significance of the AA genotype of the rs11076161with increased odds of 19-fold higher towards OSCC development.Conclusions: This is the first demonstration that a significant linkage disequilibrium among gene polymor-phisms of the MT family may affect susceptibility to oral cancer, which is conditioned by the G/A/C/T haplotype(rs1610216/rs11076161/rs964372/ rs8052334) and the MT-1A gene polymorphism has a potential clinical utility forthe OSCC risk assessment.(AU)
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Humanos , Masculino , Feminino , Metalotioneína , Polimorfismo Genético , Neoplasias de Cabeça e Pescoço , Carcinoma de Células Escamosas , Carcinoma de Células Escamosas de Cabeça e Pescoço , Neoplasias Bucais/genética , Brasil , Estudos de Casos e Controles , Fatores de Risco , Saúde BucalRESUMO
This work shows the antitumoral effects of BnSP-6, a Lys 49 PLA2 isolated from Bothrops pauloensis venom, on human breast cancer MDA-MB-231 cells. BnSP-6 caused a dose-dependent cytotoxicity and inhibited cell adhesion. Interestingly, cytotoxic activity of BnSP-6 was significantly lower against MCF10A, a non-tumorigenic breast cell line, suggesting that this PLA2 presented a possible preference for targets in cancer cells. Analysis of cell death on MDA-MB-231 cells showed that BnSP-6 stimulated the autophagy process, as evidenced by labeling of autophagic vacuoles. Moreover, apoptosis assays showed that BnSP-6 induced both early and late apoptosis. Apoptosis of MDA-MB-231 cells was also confirmed by up-regulation of different genes related to the apoptosis pathway, such as TNF, TNFRSF10B, TNFRSF1A and CASP8 and decreased expression of anti-apoptotic genes (BCL2 and BCL2L). In addition, BnSP-6 caused a remarkable increase in gene expression of BRCA2 and TP53 tumor suppressors. Finally, BnSP-6 induced down-regulation of Angiopoetin 1 gene (potent pro-angiogenic factor) and inhibited adhesion and migration of MDA-MB-231 cells suggesting pharmaceutical applications of this PLA2 as an antiangiogenic and anti-metastatic agent. Taken together, our results show that the PLA2 BnSP-6 presents anticancer potential that can be exploited as prototype for the design of new therapies.
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Bothrops/metabolismo , Venenos de Crotalídeos/química , Venenos de Crotalídeos/toxicidade , Fosfolipases A2/química , Fosfolipases A2/toxicidade , Animais , Apoptose/efeitos dos fármacos , Autofagia/efeitos dos fármacos , Neoplasias da Mama , Adesão Celular/efeitos dos fármacos , Morte Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular , Venenos de Crotalídeos/isolamento & purificação , Feminino , Expressão Gênica , Humanos , Fosfolipases A2/isolamento & purificação , Domínios e Motivos de Interação entre ProteínasRESUMO
After oral infection, Toxoplasma gondii invades intestinal cells, induces breakdown of intestinal physiology and barrier functions, and causes intestinal pathology in some animal species. Although parasites' invasion into host cells is a known phenomenon, the effects of T. gondii infection in the intestinal barrier are still not well established. To evaluate morphological and physiological modifications on the colorectal adenocarcinoma-derived Caco-2 cell line during T. gondii infection, microvilli, tight junction integrity, and transepithelial electrical resistance (TEER) were investigated under infection. It was observed that the dextran uptake (endocytosis) and distribution were smaller in infected than in noninfected Caco-2 cells. The infection leads to the partial loss of microvilli at the cell surface. Claudin-1, zonula occludens-1 (ZO-1), and occludin expressions were colocalized by immunofluorescence and presented discontinuous net patterns in infected cells. Immunoblotting analysis at 24 hr postinfection revealed decreasing expression of occludin and ZO-1 proteins, whereas claudin-1 presented similar expression level compared with noninfected cells. T. gondii decreased TEER in Caco-2 cells 24 hr after infection. Our results suggest that T. gondii infection may lead to the loss of integrity of intestinal mucosa, resulting in impaired barrier function.
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Mucosa Intestinal/parasitologia , Toxoplasma/fisiologia , Citoesqueleto de Actina/ultraestrutura , Células CACO-2 , Polaridade Celular , Claudina-1/metabolismo , Dextranos/metabolismo , Impedância Elétrica , Endocitose , Humanos , Mucosa Intestinal/metabolismo , Mucosa Intestinal/ultraestrutura , Microvilosidades/metabolismo , Microvilosidades/parasitologia , Microvilosidades/ultraestrutura , Ocludina/metabolismo , Junções Íntimas/metabolismo , Junções Íntimas/parasitologia , Junções Íntimas/ultraestrutura , Proteína da Zônula de Oclusão-1/metabolismoRESUMO
The cluster of differentiation antigen 14 (CD14) is a key molecule of the innate immunity. This pattern recognition receptor binds mainly to lipopolysaccharide (LPS), lipotechoic acid (LTA), arachidonic acid, and thus induces the releases various cytokines, as a defense mechanism. Several studies suggest that different regions of the amino-terminal portion of the molecule may be involved in the LPS binding; however, controversial results on the recognition sequence still persist. In this work, functional epitopes of the CD14 molecule were mapped through Phage Display by using a 7-mer conformational constrained random peptide library against a monoclonal antibody anti-soluble CD14-fraction ST and a polyclonal anti-CD14. In silico and empirical analyses were performed to map the selected peptides into the CD14 3D structure. Immunoreactivity tests of peptides against bacterial components of Gram+ and Gram- bacteria were performed in order to demonstrate their functional recognition. All peptides strongly reacted against all bacteria, and besides the recognition of the amino-terminal region, we were able to demonstrate a second epitope site in the middle of the receptor. Additional in silico analysis suggests a possible role of CD14 epitopes as natural antimicrobial peptides.