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Evaluación de la capacidad de hemocianinas de moluscos para activar la vía clásica del sistema del complemento humano
(Universidad de Chile, 2016)
continuar con esta línea de investigación, para proveer
información adicional para un uso seguro de las hemocianinas en biomedicina.
xvi
ABSTRACT
Evaluation of the Capacity of Molluskan Hemocyanins to Activate the Classical
Pathway of the Human...
en biomedicina. Descritas a comienzos del siglo XX; han sido proteínas modelo en bioquímica y, unas décadas después, herramientas instrumentales en inmunología básica y aplicada, especialmente para el caso de la hemocianina de la Lapa Californiana...
en biomedicina. Descritas a comienzos del siglo XX; han sido proteínas modelo en bioquímica y, unas décadas después, herramientas instrumentales en inmunología básica y aplicada, especialmente para el caso de la hemocianina de la Lapa Californiana...
Derecho a la autodeterminación informativa respecto a los datos genéticos
(Universidad de Chile, 2018)
Participación de receptores lectina tipo C en el reconocimiento de hemocianinas de moluscos por células presentadoras de antígeno y consecuencias en la presentación antigénica
(2016)
biomedicina como proteína carrier de antígenos tumorales y como inmunoestimulantes no específicos en el tratamiento de ciertos cáncer. Para explicar esta propiedad se han invocado características tales como su tamaño, xenogenicidad, estructura compleja y...
Hemocyanins from the gastropod Megathura crenulata (KLH), Concholepas concholepas (CCH) and Fissurella latimarginata (FLH) have mainly been used as non-specific immunostimulants with beneficial clinical outcomes, to prevent the progression of recurrent superficial bladder cancer or as carriers/adjuvants for producing antibodies against tumor-associated antigens in therapeutic vaccines against cancer. However, the mechanism involved in the positive immunomodulatory effects of these proteins has been scarcely studied. An important feature of these hemocyanins is its carbohydrate content, which can reach up to 3% (w/w), being mannose as the most abundant sugar. These oligosaccharides can be recognized by C-type lectin receptors (CLRs) of innate immunity, such as Mannose Receptor (MR), DC-SIGN (Dendritic Cell-Specific Intercellular adhesion molecule-3-Grabbing Non-integrin) or MGL (macrophage galactose lectine), present on antigen presenting cells (APCs). Furthermore, these innate receptors have been involved in classical and alternative antigen presentation by MHC I and MHC II to CD8+ and CD4+ lymphocytes, respectively. Hence, we proposed that CLRs-mediated endocytosis of hemocyanins in antigen presenting cells triggers the cross-presentation of these proteins. In this work, we studied the interaction between CLRs and hemocyanins using indirect ELISA and SPR (surface plasmon resonance). We found that CCH, FLH, and KLH bind to MR and DC-SIGN with high affinity constants; additionally FLH and KLH also bind to MGL. Our results demonstrate that the interaction between MR and hemocyanins trigger their endocytosis, analysed by an artificial CHO-MR system. Furthermore, human and murine APCs-expressing CLRs can uptake hemocyanins and ovalbumin. We also found that hemocyanins uptake by murine macrophages and dendritic cells can be partially inhibited by EDTA and D-Mannose. These results suggest an important role of CLRs in endocytosis and signalling transduction of hemocyanins. On the other hand, we studied the cellular destination of hemocyanins in bone marrow derived dendritic cells (BMDCs) using confocal microscopy. FLH and KLH showed a similar localization in Rab5+, Rab7+ and Lamp-1+ compartments. However, CCH was not found in Rab5+ early endosomes, suggesting differential compartmentalization between the hemocyanins. Thus, FLH and KLH could be involved in an alternative processing pathway such as cross-presentation. Finally, we determined if murine APCs could present hemocyanins on MHC I context. Our results showed that in vitro stimulation of macrophages with FLH, but not with CCH or KLH, increased MHC I and MHC II expression, supporting our hypothesis. Hemocyanins cross-presentation could explain one of the mechanisms involved in the antitumoral effect of these proteins, in addition to the classic presentation by MHC II. Moreover, CD8+ lymphocytes activation could promote cytokines and humoral factors release that indirectly trigger other latent immune responses against the tumoral cells, effect known as bystander effect...
Hemocyanins from the gastropod Megathura crenulata (KLH), Concholepas concholepas (CCH) and Fissurella latimarginata (FLH) have mainly been used as non-specific immunostimulants with beneficial clinical outcomes, to prevent the progression of recurrent superficial bladder cancer or as carriers/adjuvants for producing antibodies against tumor-associated antigens in therapeutic vaccines against cancer. However, the mechanism involved in the positive immunomodulatory effects of these proteins has been scarcely studied. An important feature of these hemocyanins is its carbohydrate content, which can reach up to 3% (w/w), being mannose as the most abundant sugar. These oligosaccharides can be recognized by C-type lectin receptors (CLRs) of innate immunity, such as Mannose Receptor (MR), DC-SIGN (Dendritic Cell-Specific Intercellular adhesion molecule-3-Grabbing Non-integrin) or MGL (macrophage galactose lectine), present on antigen presenting cells (APCs). Furthermore, these innate receptors have been involved in classical and alternative antigen presentation by MHC I and MHC II to CD8+ and CD4+ lymphocytes, respectively. Hence, we proposed that CLRs-mediated endocytosis of hemocyanins in antigen presenting cells triggers the cross-presentation of these proteins. In this work, we studied the interaction between CLRs and hemocyanins using indirect ELISA and SPR (surface plasmon resonance). We found that CCH, FLH, and KLH bind to MR and DC-SIGN with high affinity constants; additionally FLH and KLH also bind to MGL. Our results demonstrate that the interaction between MR and hemocyanins trigger their endocytosis, analysed by an artificial CHO-MR system. Furthermore, human and murine APCs-expressing CLRs can uptake hemocyanins and ovalbumin. We also found that hemocyanins uptake by murine macrophages and dendritic cells can be partially inhibited by EDTA and D-Mannose. These results suggest an important role of CLRs in endocytosis and signalling transduction of hemocyanins. On the other hand, we studied the cellular destination of hemocyanins in bone marrow derived dendritic cells (BMDCs) using confocal microscopy. FLH and KLH showed a similar localization in Rab5+, Rab7+ and Lamp-1+ compartments. However, CCH was not found in Rab5+ early endosomes, suggesting differential compartmentalization between the hemocyanins. Thus, FLH and KLH could be involved in an alternative processing pathway such as cross-presentation. Finally, we determined if murine APCs could present hemocyanins on MHC I context. Our results showed that in vitro stimulation of macrophages with FLH, but not with CCH or KLH, increased MHC I and MHC II expression, supporting our hypothesis. Hemocyanins cross-presentation could explain one of the mechanisms involved in the antitumoral effect of these proteins, in addition to the classic presentation by MHC II. Moreover, CD8+ lymphocytes activation could promote cytokines and humoral factors release that indirectly trigger other latent immune responses against the tumoral cells, effect known as bystander effect...
Efecto inmunomodulador de hemocianinas de moluscos combinadas con adyuvantes de vacuna en cáncer oral murino
(Universidad de Chile, 2015)
: polysorbate 20 / polisorbato 20
1
RESUMEN
Las hemocianinas de algunos moluscos son ampliamente usadas en
biomedicina, entre ellas, la de Megathura crenulata (KLH), de Concholepas
concholepas (CCH) y de Fissurella latimarginata (FLH), éstas dos...
. Luego de más de 35 años de uso en biomedicina, los investigadores han provisto evidencia que sustenta este atributo, al tener la habilidad para incrementar la respuesta inmune del hospedero, a través de su interacción con diferentes células inmunes...
. Luego de más de 35 años de uso en biomedicina, los investigadores han provisto evidencia que sustenta este atributo, al tener la habilidad para incrementar la respuesta inmune del hospedero, a través de su interacción con diferentes células inmunes...
Síntesis y caracterización de nanopartículas de níquel, cobalto y mezclas níquel-cobalto dispersadas en diferentes matrices
(Universidad de Chile, 2015)
literatura para catálisis homogénea con
metales de transición.
11
ÍNDICE DE TABLAS
1. Aplicaciones de las propiedades magnéticas en biomedicina
2. Materiales magnéticos y sus aplicaciones
3. Resumen de parámetros magnéticos...
aplicaciones presentes y futuras, las más interesantes quizás en el campo de la biomedicina [3]. El origen en el cambio de estas propiedades comparadas con el material macroscópico o “bulk”, se debe a que la reducción de tamaño, 19 produce cambios en los...
aplicaciones presentes y futuras, las más interesantes quizás en el campo de la biomedicina [3]. El origen en el cambio de estas propiedades comparadas con el material macroscópico o “bulk”, se debe a que la reducción de tamaño, 19 produce cambios en los...
Compromiso renal en vasculitis asociadas a anticuerpos anticitoplasma de neutrófilos. Recomendaciones de consenso de las sociedades chilenas de nefrología y reumatología
(Sociedad Médica de Santiago, 2018-02)
. santiago, chile.
7Departamento de nefrología,
Facultad de medicina, Pontificia
universidad católica de chile.
santiago, chile.
8centro de Biología celular y
Biomedicina. Facultad de medicina.
universidad san sebastián.
santiago, chile.
9Departamento...
Utilidad de Tinción de Tricrómico de Masson en la Cuantificación de Densidad Media Vascular en Mucosa Oral Normal, Displasia Epitelial y Carcinoma Oral de Células Escamosas
(Sociedad Chilena de Anatomía, 2017)
Departamento de Cirugía y Patología, Facultad de Odontología, Universidad Andrés Bello, Santiago, Chile.
2 Programa de Doctorado en BioMedicina, Facultad de Medicina, Universidad de los Andes, Santiago, Chile.
3 BioMAT’X, Centro de Investigación Biomédica...
Estudio del rol de la caspasa 8 en la infección por Trypanosoma cruzi en una linea celular de trofoblasto humano (Be Wo)
(Universidad de Chile, 2016)
Vanessa V. Carrillo Werner
TESIS PARA OPTAR AL GRADO DE MAGISTER EN BIOMEDICINA CELULAR Y
MOLECULAR
Director de Tesis: Dra. Ulrike Kemmerling
2016
AGRADECIMIENTOS
Quisiera dar mi más profundo agradecimiento a mi tutora, la Dra...
Manifiesto Florence para unos mejores cuidados basados en la evidencia
(Ediciones Rol, 2020)
in Evidence-Based Health Care.
University of Oxford. Oxford. UK.
josepmariagarciaa@gmail.com
RAMÓN ESCURIET PEIRÓ
PhD Biomedicina. Director del Programa
de formación teórica de matronas. Departament
de Salut. Generalitat de Catalunya. Barcelona.
NÚRIA...
La neuroinflamación como factor detonante del desarrollo de la enfermedad de Alzheimer
(Sociedad Chilena de Neurolología, Psiquiatría y Neurocirugía, 2010-01)
/03/2010
Laboratorio de Neurociencias Celulares y Moleculares, Facultad de Ciencias, Universidad de Chile y Centro Internacional de
Biomedicina (ICC):
1 Tesista Ingeniería en Biotecnología Molecular, Universidad de Chile.
2 M.D.
3 Ph.D.
ARTÍCULO DE REVISIÓN...