Determinación de la actividad alergénica de Blo t 2, un alérgeno del ácaro del polvo Blomia tropicalis

Mondol Miranda, Ernesto Carlos

dc.contributor.advisor	Caraballo Gracia, Luis
dc.contributor.author	Mondol Miranda, Ernesto Carlos
dc.date.accessioned	2021-08-03T15:10:22Z
dc.date.available	2021-08-03T15:10:22Z
dc.date.issued	2021
dc.identifier.citation	TM616.973 / M745	es
dc.identifier.uri	https://hdl.handle.net/11227/12318
dc.identifier.uri	http://dx.doi.org/10.57799/11227/1349
dc.description	Tesis (Magíster en Inmunología). -- Universidad de Cartagena, Instituto de Investigaciones Inmunológicas, 2021	es
dc.description.abstract	La incidencia de las enfermedades alérgicas, entre las que se encuentran el asma, la rinitis y la dermatitis atópica, ha aumentado en las últimas décadas, tanto en países desarrollados como en vía de desarrollo; ocasionando una gran morbilidad entre los pacientes y gastos para los organismos de seguridad social. La genética de los individuos alérgicos juega un papel importante en el desarrollo de estas enfermedades; además, a lo largo de la vida los individuos se exponen tanto a factores de riesgo como protectores, que en conjunto van a determinar la presentación de enfermedades alérgicas. La exposición a los ácaros del polvo como lo es Blomia tropicalis es un importante factor de riesgo para la presentación del asma alérgica, este ácaro posee proteínas alergénicas, con diferencias en su estructura, propiedades biológicas, estabilidad y nivel de expresión, por lo tanto, se espera que la respuesta inmune a los diferentes alergenos de B. tropicalis, sea heterogénea. La capacidad de producir IgE es una de las características más conocidas resultante de este proceso, lo que se ha asumido como una de las propiedades esenciales que hacen que un componente sea alergénico. Sin embargo, la inducción de IgE específica es necesaria, pero no suficiente para ser un alergeno, pues estos también deben inducir síntomas de alergia. Lo anterior ha obligado a replantearse los criterios para definir un verdadero alergeno y a sugerir pruebas que ayuden a definir si una molécula, además de tener alergenicidad (es decir que induce la síntesis de IgE y se une a dicho anticuerpo) tiene también actividad alergénica, es decir, que induce un proceso inflamatorio. Entre estas pruebas encontramos la evaluación de la frecuencia e intensidad de unión a la IgE; pruebas de provocación in vivo o in vitro, como la prueba de activación de basófilos o de liberación de histamina, ensayos de anafilaxia cutánea pasiva, o la evaluación de su capacidad de inducir cambios epigenéticos, entre otros. Lo anterior es esencial si se quiere sacar verdadero provecho al fundamento del diagnóstico basado en componentes (DBC, el cual apunta a la medicina de precisión) ya que no tendría sentido identificar supuestos alérgenos cuya participación real en la fisiopatología de la enfermedad es dudosa o por el contrario infravalorar alérgenos que por no tener una alta reactividad IgE son ignorados, aunque tengan una gran capacidad de inducir síntomas. Por lo anterior en este proyecto nos propusimos la caracterización del alérgeno del grupo 2 de Blomia tropicalis (Blo t 2) en términos de su actividad alergénica. Para lo cual, se obtuvo en primer lugar este alergeno en forma recombinante, con un alto nivel de pureza y bajos niveles de endotoxina para llevar a cabo ensayos de ELISA y pruebas de activación pasiva de basófilos, en pacientes y controles sanos; además para probar su actividad alergénica en dos modelos murinos, uno de alergia respiratoria y otro de anafilaxia cutánea pasiva. Se obtuvo proteína suficiente para el desarrollo de los experimentos con un nivel de pureza adecuado. Los resultados de los trabajos muestran que aunque el 30% de los asmáticos estaban sensibilizados a Blo t 2, este porcentaje no fue significativamente superior al de los sujetos sanos. Dentro de los sujetos sensibilizados a B. tropicalis el 40% reaccionó a rBlo t 2. Este alergeno demostró su capacidad de generar inflamación en el modelo murino de asma alérgica, evidenciado en el infiltrado inflamatorio pulmonar, aumento de eosinófilos en el lavado broncoalveolar, la generación de hiperreactividad en las vías respiratorias y el aumento de la IgE, IgG1 e IgG2a; en la prueba de anafilaxia cutánea pasiva también provocó el aumento de la extravasación de azul de Evans; además, indujo la sobre expresión del marcador CD203c en la membrana de basófilos sensibilizados con sueros de asmáticos. En conclusión, rBlo t 2 tiene actividad alergénica demostrada por ensayos in vitro e in vivo que podrían indicar que este es un alergeno clínicamente importante en enfermedades alérgicas como el asma.	es
dc.language.iso	spa	es
dc.publisher	Universidad de Cartagena	es
dc.rights.uri	https://creativecommons.org/licenses/by-nc-nd/4.0	es
dc.subject	Asma	es
dc.subject	Alergia - Enfermedades	es
dc.subject	Blomia tropicalis	es
dc.subject	Ácaros - Alergia	es
dc.subject	Alérgenos	es
dc.title	Determinación de la actividad alergénica de Blo t 2, un alérgeno del ácaro del polvo Blomia tropicalis	es
dc.type	Trabajo de grado - Maestría	spa
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