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Antígenos

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Antígenos

  1. 1. Antígenos reacción Ag-Ab Pruebas inmunológicas
  2. 2. ANTIGENOS
  3. 3. Conceptos <ul><li>Un antígeno es cualquier sustancia (molécula) que reacciona con productos de la respuesta inmune. </li></ul><ul><li>Epitopo o determinante antigénico. Es la porción de un Ag que se combina con los productos de una respuesta inmune especifica. </li></ul>
  4. 4. Características de los Ag <ul><li>Inmunogenicidad: capacidad de inducir respuesta (celular o humoral). </li></ul><ul><li>Cel B + Ag </li></ul><ul><li>Cel plasmaticas + Cel B de memoria </li></ul><ul><li>Anticuerpos </li></ul><ul><li>Antigenicidad: capacidad de combinarse de forma especifica a los productos de la respuesta inmune </li></ul><ul><li>Mol. Inmunogenicas = antigenicas </li></ul><ul><li>Mol. antigenicas ≠inmunogenicas </li></ul><ul><li>(ej. haptenos) </li></ul>
  5. 5. Que es un Hapteno <ul><li>Son moléculas pequeñas que no son inmunogénicas pero que pueden reaccionar con los productos de una respuesta inmune. </li></ul><ul><li>No inducen respuesta inmune por si solas, pero si cuando son transportadas por otras moléculas. </li></ul>
  6. 6. Propiedades de los Antígenos <ul><li>ALTERIDAD : entre mas extraña es la molécula es mejor la respuesta inmune. Depende de la distancia filogenética. </li></ul><ul><li>Albumina bovina ≠ albumina de ratón </li></ul><ul><li>Excepciones moléculas conservadas como el citocromo C </li></ul><ul><li>COMPLEJIDAD QQ : entre mas compleja es la molécula será mas inmunogenica ( estructura 1 ª ,2 ª ,3 ª ,4 ª. heteropolimeros>>homopolimeros ). </li></ul><ul><li>prots>polisacaridos>lipidos>acidos nucleicos </li></ul><ul><li>Ag particulados>>solubles </li></ul><ul><li>Ag desnaturalizados>>forma nativa </li></ul>
  7. 7. Propiedades de los Antígenos <ul><li>TAMAÑO MOLECULAR : entre mas grande es una molécula es mas inmunogenica. </li></ul><ul><li>100,000 Da >> inmunogenicidad </li></ul><ul><li>DEGRADABILIDAD : capacidad de procesarse y presentarse junto a MHC. </li></ul><ul><li>Ag fagocitados >> inmunogenicos </li></ul>
  8. 8. Tipos de antígenos T independientes <ul><li>Son los que pueden estimular directamente a las células B para producir ab. </li></ul><ul><li>No requieren de la ayuda de los linfocitos T </li></ul><ul><li>Generalmente polisacáridos </li></ul>
  9. 9. Ag T independientes Propiedades <ul><li>Estructura polimérica. Repetición del mismo determinante antigénico </li></ul><ul><li>Activación policlonal de células B . </li></ul><ul><li>Tipo 1: activadores policlonales </li></ul><ul><li>Tipo 2: no </li></ul><ul><li>Resistencia a la degradación . Persisten por mas tiempo estimulando el SI. </li></ul>
  10. 10. Tipos de antígenos T dependientes <ul><li>Son aquellos que no estimulan directamente la producción de ab, sin la ayuda de las células T. </li></ul><ul><li>Estructuralmente se caracterizan por pocas copias de un mismo epitopo. </li></ul><ul><li>Las Proteínas son un buen ejemplo </li></ul>
  11. 11. Superantígenos <ul><li>Son antígenos capaces de activar una gran población de linfocitos T (25 %) </li></ul>Monoclonal/Oligoclonal T cell response 1:10 4 - 1:10 5 Ag convencional Superantigeno Polyclonal T cell response 1:4 - 1:10
  12. 12. REACCION Ag-Ab
  13. 13. Reconocimiento Ag-Ac <ul><li>Depende de los receptores de T y B </li></ul><ul><ul><li>B: Igs </li></ul></ul><ul><ul><li>T: TCR </li></ul></ul><ul><li>T y B reconocen los ags de diferente manera </li></ul><ul><li>Ambos tiene regiones variables y constantes, </li></ul><ul><li>El proceso de recombinación en genes permite la variabilidad </li></ul>
  14. 14. Reconocimiento Ag-Ac
  15. 15. Reconocimiento Ag-Ac <ul><li>Los ab reconocen los ag en solución o en la superficie de células, en su forma nativa. </li></ul>
  16. 16. Reconocimiento Ag-Ac <ul><li>El TCR sólo reconoce ags asociado con el MHC en la superficie de células, este antígeno ha sido degradado en fragmentos </li></ul>
  17. 17. Reconocimiento Ag-Ac
  18. 18. Naturaleza de las Rx Ag-Ab <ul><li>Afinidad </li></ul><ul><li>es la fuerza de la Rx entre un determinante Ag y un sitio de union de un Ab . </li></ul><ul><li>Es la suma de las fuerzas de atracción y repulsión </li></ul>Ab Ag High Affinity Ab Ag Low Affinity
  19. 19. Naturaleza de las Rx Ag-Ab <ul><li>Avidez </li></ul><ul><li>Es la medida de la fuerza de unión de un Ag con muchos determinantes Ag y Ab multivalentes. </li></ul><ul><li>Esta influenciada por las valencias del Ag y del Ab. </li></ul>
  20. 20. Fuerzas que intervienen <ul><li>Puentes de hidrogeno </li></ul><ul><li>Enlaces hidrofobicos </li></ul><ul><li>Enlaces electrostaticos </li></ul><ul><li>Fuerzas de Van der Wals </li></ul><ul><li>No son enlaces covalentes por lo tanto la unión es REVERSIBLE </li></ul>
  21. 22. Especificidad y Rx cruzada <ul><li>Especificidad: se refiere a la habilidad de un Ab individual de reaccionar con solo un determinante Ag. Un Ab puede distinguir entre: </li></ul><ul><li>La estructura 1 ª de un ag. </li></ul><ul><li>Formas isomericas </li></ul><ul><li>Estructura 2 ª y 3 ª </li></ul>
  22. 23. Especificidad y Rx cruzada <ul><li>Rx cruzada: se refiere a la habilidad de uno o varios Ab de reaccionar con mas de un determinante Ag. </li></ul><ul><li>Estas Rx se dan porque los Ag comparten epitopos en común o epitopos con estructuras similares </li></ul>
  23. 25. Pruebas para la detección de la reacción Ag-Ab
  24. 26. Factores que afectan la medición de la Rx Ag-Ab <ul><li>Afinidad </li></ul><ul><li>Avidez </li></ul><ul><li>Radio Ag/Ab </li></ul><ul><li>Forma física del Ag. </li></ul>Ab excess Ag excess Equivalence – Lattice formation
  25. 27. Pruebas de aglutinación <ul><li>Son pruebas que tienen como punto de visualización la aglutinación de un Ag particulado. La prueba puede ser cualitativa o cuantitativa. </li></ul>Y Y Y + 
  26. 28. Hemaglutinación Usos: tipeaje sanguíneo, infecciones es fácil de realizar y es semicuantitativa 1/2 1/4 1/8 1/16 1/32 1/64 1/128 1/256 1/512 1/1024 Pos. Neg. Titer 64 8 512 <2 32 128 32 4 Patient 1 2 3 4 5 6 7 8
  27. 29. Aglutinación pasiva <ul><li>Prueba de aglutinación donde una partícula (latex) se recubre con el Ag. </li></ul><ul><li>Aplicación: medida de Ab para Ag solubles </li></ul>Y Y Y + 
  28. 30. Inhibición de la aglutinación/hemaglutinación Y Y Y +  Prior to Test Y +  Y Y + Test Patient’s sample
  29. 31. Pruebas de precipitación <ul><li>Inmunodifusión radial </li></ul><ul><li>inmunoelectroforesis </li></ul>Ag Ab - +
  30. 32. Ensayos inmunoenzimaticos (ELISA) radioinmunoensayos (RIA) Utiliza un contador de radiación La determinación puede ser visual o por un lector de ELISA
  31. 33. ELISA y RIA competitiva para Ag Y Y +  Prior to Test Labeled Ag Y Y +  Test + Patient’s sample Labeled Ag + Y Y +  Test + Patient’s sample Labeled Ag +
  32. 34. ELISA y RIA no competitiva para Ag y Ab Solid Phase Y Ag Immobilized Y Ab in Patient’s sample Labeled Anti-Ig Solid Phase Y Ag Immobilized Y Ag in Patient’s sample Labeled Ab
  33. 35. Inmunofluorescencia Utiliza un microscopio de fluorescencia <ul><li>Directa: detecta Ag en tejidos, usando Ab marcados con fluorocromo </li></ul><ul><li>Indirecta: detecta Ab. El Ab para el Ag no esta marcado. Luego se usa un Ab marcado con fluorocromo </li></ul>Ag Y Fluorochrome Labeled Ab Tissue Section Ag Y Y Fluorochrome Labeled Anti-Ig Tissue Section Unlabeled Ab
  34. 37. Inmunocromatografía
  35. 39. Western blot

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