ES2627292T3 - Nanotecnología de vacunas - Google Patents

Nanotecnología de vacunas Download PDF

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Publication number
ES2627292T3
ES2627292T3 ES08839738.5T ES08839738T ES2627292T3 ES 2627292 T3 ES2627292 T3 ES 2627292T3 ES 08839738 T ES08839738 T ES 08839738T ES 2627292 T3 ES2627292 T3 ES 2627292T3
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Prior art keywords
lymphocytes
virus
vsv
nanotransporters
mage
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ES08839738.5T
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Inventor
Ulrich H. Von Andrian
Omid C. Farokhzad
Robert S. Langer
Tobias Junt
Elliott Ashley Moseman
Liangfang Zhang
Pamela Basto
Matteo Iannacone
Frank Alexis
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Harvard College
Brigham and Womens Hospital Inc
Childrens Medical Center Corp
Massachusetts Institute of Technology
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Harvard College
Brigham and Womens Hospital Inc
Childrens Medical Center Corp
Massachusetts Institute of Technology
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Abstract

Nanotransportadores que tienen un diámetro medio geométrico comprendido entre 50 nm y 500 nm, siendo los nanotransportadores poliméricos y comprendiendo un antígeno peptídico de linfocitos B y un agente inmunoestimulador; en donde el agente inmunoestimulador es un agonista del receptor de tipo toll; en donde los nanotransportadores comprenden un polímero seleccionado entre el grupo que consiste en: poli(ácido láctico) (PLA), poli(ácido glicólico) (PGA) y poli(ácido láctico/ácido glicólico) (PLGA); y, en donde el agente inmunoestimulador está unido covalentemente a los nanotransportadores o a un polímero a partir del que se forman los nanotransportadores, y el antígeno peptídico de linfocitos B está encapsulado.

Description

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Los antígenos de linfocitos B incluyen sustancias que crean dependencia, tales como nicotina, un narcótico, un alucinógeno, un estimulante, un supresor de la tos, un tranquilizante, o un sedante, y un opioide o benzodiacepina.
Los antígenos de linfocitos B incluyen toxinas, tal como una arma química (por ejemplo, toxina botulínica o fosfeno).
5 Las toxinas de un arma química incluyen también, pero sin limitación, O-Alquilo (<C10, incl. cicloalquil) alquilo (Me, Et, n-Pr o i-Pr)-fosfonofluoridatos (por ejemplo, Sarín: O-Isopropil metilfosfonofluoridato, Soman: O-Pinacolil metilfosfonofluoridato), O-Alquilo (<C10, incl. cicloalquilo) N,N-dialquilo (Me, Et, n-Pr o i-Pr) fosforamidocianidatos (por ejemplo, Tabun: O-Etil N,N-dimetilfosforamidocianidato), O-Alquilo (H o <C10, incl. cicloalquilo) S-2-dialquilo (Me, Et, n-Pr o i-Pr)-aminoetil alquilo (Me, Et, n-Pr o i-Pr) fosfonotiolatos y las sales alquiladas o protonadas correspondientes (por ejemplo, VX: O-Etil S-2-diisopropilaminoetil metilfosfonotiolato), Mostazas de azufre: Sulfuro de 2-cloroetilclorometilo, Gas mostaza: sulfuro de Bis(2-cloroetilo), Bis(2-cloroetiltio)metano, Sesquimostazas: 1,2Bis(2-cloroetiltio)etano, 1,3-Bis(2-cloroetiltio)-n-propano, 1,4-Bis(2-cloroetiltio)-n-butano, 1,5-Bis(2-cloroetiltio)-npentano, Bis(2-cloroetiltiometil)éter, Mostaza O: Bis(2-cloroetiltioetil)éter, Lewisites: Lewisite 1: 2-Clorovinildicloroarsina, Lewisite 2: Bis(2-clorovinil)cloroarsina, Lewisite 3: Tris(2-clorovinil)arsina, Mostazas de
15 nitrógeno: HN1: Bis(2-cloroetil)etilamina, HN2: Bis(2-cloroetil)metilamina, HN3: Tris(2-cloroetil)amina, Saxitoxina, Ricina, Amitón: O,O-Dietil S-(2-(dietilamino)etil)fosforotiolato y las sales alquiladas o protonadas correspondientes, PFIB: 1,1,3,3,3-Pentafluoro-2-(trifluorometil)-1-propeno, 3-Quinuclidinil bencilato (BZ), Fosgeno: Dicloruro de carbonilo, Cloruro de cianógeno, Cianuro de hidrógeno y Cloropicrina: Tricloronitrometano. En algunas realizaciones, la toxina para la inclusión en un nanotransportador es la molécula completa de cualquiera de las anteriores o una parte de la misma.
El antígeno de linfocitos B incluye riesgos biológicos o agentes peligrosos para el medio ambiente, tales como arsénico, plomo, mercurio, cloruro de vinilo, bifenilos policlorados, benceno, hidrocarburos aromáticos policíclicos, cadmio, benzo(a)pireno, benzo(b)fluoranteno, cloroformo, diclordifenil-tricloretileno (DDT), P,P’-, aroclor 1254,
25 aroclor 1260, dibenzo(a,h)antraceno, tricloroetileno, dieldrina, cromo hexavalente, o p,p’--diclorodifenil-dicloroeteno (DDE, P,P’).
Los antígenos de linfocitos B incluyen hidratos de carbono, tales como uno procedente de un agente infeccioso (por ejemplo, una bacteria, hongo, virus, protozoo, o parásito, tales como una bacteria que siendo la bacteria
Pseudomonas, Pneumococcus, E. coli, Staphylococcus, Streptococcus, Treponema, Borrelia, clamidia, Haemophilus, Clostridium, salmonela, legionela, Vibrio o Enterococci o un Mycobacterium, y siendo el virus un virus de la viruela, el virus de la viruela, virus del Ébola, virus de Marburg, el virus del dengue, virus de la gripe, virus paragripal, virus respiratorio sincitial, virus de la rubéola, virus de la inmunodeficiencia humana, virus del papiloma humano, el virus de la varicela-zóster, virus del herpes simple, citomegalovirus, virus de Epstein-Barr, JC virus,
35 rabdovirus, rotavirus, rinovirus, el adenovirus, virus del papiloma, parvovirus, picornavirus, el poliovirus, virus que producen paperas, virus que producen rabia, reovirus, el virus de la rubéola, togavirus, ortomixovirus, retrovirus, hepadnavirus, coxsackievirus, virus de la encefalitis equina, el virus de la encefalitis japonesa, el virus de la fiebre amarilla, virus de la fiebre del Valle del Rift, el virus de la hepatitis A, el virus de la hepatitis B, el virus de la hepatitis C, virus de la hepatitis D, o virus de la hepatitis E).
Los antígenos de linfocitos B incluyen autoantígenos, tal como un péptido o proteína, lipoproteína, lípido, hidrato de carbono, o un ácido nucleico. En algunas realizaciones, el autoantígeno es una enzima, una proteína estructural, una proteína secretada, un receptor superficial celular, o una citoquina. En algunas realizaciones, la citoquina es TNF, IL1, o IL-6. En algunas realizaciones, el autoantígeno es la proteína de transferencia del éster de colesterilo (CETP), la
45 proteína Aβ asociada con Alzheimer, una enzima proteolítica que procesa la forma patológica de la proteína Aβ, LDL asociado con ateroesclerosis, o un correceptor para VIH-1. En algunas realizaciones, la enzima proteolítica que procesa la forma patológica de la proteína Aβ es la beta secretasa. En algunas realizaciones, el LDL asociado con ateroesclerosis está oxidado o mínimamente modificado. En algunas realizaciones, el correceptor de VIH-1 es CCR5. En algunas realizaciones, el autoantígeno es un antígeno de enfermedad autoinmunitaria.
En algunas realizaciones, el antígeno de linfocitos B es un antígeno de enfermedad degenerativa, un antígeno de enfermedad infecciosa, un antígeno de cáncer, un antígeno de enfermedad atópica, un antígeno de enfermedad autoinmune, o una enzima de enfermedad metabólica o uno de sus productos enzimáticos.
55 En algunas realizaciones, el antígeno es un antígeno del cáncer. En algunas realizaciones, el antígeno de cáncer es Melan-A/MART-1, Dipeptidil peptidasa IV (DPPIV), proteína de unión a la adenosina desaminasa (ADAbp), ciclofilina b, Antígeno asociado colorrectal (CRC)-C017-1A/GA733, Antígeno carcinoembriónico (CEA) y sus epítopos inmunógenos CAP-1 y CAP-2, etv6, aml1, antígeno específico de próstata (PSA) y sus epítopos inmunógenos PSA1, PSA-2, y PSA-3, antígeno de membrana específico de próstata (PSMA), receptor de linfocitos T/cadena CD3-zeta, familia MAGE de antígenos tumorales (por ejemplo, MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A5, MAGE-A6, MAGE-A7, MAGE-A8, MAGE-A9, MAGE-A10, MAGE-A11, MAGE-A12, MAGE-Xp2 (MAGE-B2), MAGE-Xp3 (MAGE-B3), MAGE-Xp4 (MAGE-B4), MAGE-C1, MAGE-C2, MAGE-C3, MAGE-C4, MAGE-C5), familia GAGE de antígenos tumorales (por ejemplo, GAGE-1, GAGE-2, GAGE-3, GAGE-4, GAGE-5, GAGE-6, GAGE-7, GAGE-8, GAGE-9), BAGE, RAGE, LAGE-1, NAG, GnT-V, MUM-1, CDK4, tirosinasa, p53, familia MUC, HER2/neu, p21ras,
65 RCAS1, α-fetoproteína, E-caderina, α-catenina, β-catenina and γ-catenina, p120ctn, gp100Pmel117, PRAME, NY-ESO-1, glicogenofosforilasa cerebral, SSX-1, SSX-2 (HOM-MEL-40), SSX-1, SSX-4, SSX-5, SCP-1, CT-7, cdc27,
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tipos de agentes inmunoestimuladores, en el que el primer tipo de agente inmunoestimulador estimula linfocitos B, y el segundo tipo de agente inmunoestimulador estimula linfocitos T. En algunas realizaciones, un nanotransportador de vacunas comprende más de dos tipo distintos de agentes inmunoestimuladores,, en el que uno o más tipos de agentes inmunoestimuladores estimulan linfocitos B, y uno o más tipos de agentes inmunoestimuladores estimulan
5 linfocitos T.
En algunas realizaciones, se pueden utilizar diversos ensayos a fin de determinar si una respuesta inmunitaria se ha modulado en un linfocito B o en un grupo de linfocitos B o en un linfocito T o en un grupo de linfocitos T. En algunas realizaciones, el ensayo evalúa si se ha/han llegado a "activar" o no la célula o grupo de células.
En algunas realizaciones, se pueden utilizar diversos ensayos a fin de determinar si se ha estimulado una respuesta inmunitaria en un linfocito T o en un grupo de linfocitos T. En algunas realizaciones, la estimulación de una respuesta inmunitaria en linfocitos T puede determinarse midiendo la producción de citoquinas inducidas por antígenos por los linfocitos T. En algunas realizaciones, la estimulación de una respuesta inmunitaria en linfocitos T puede
15 determinarse midiendo la proliferación de linfocitos T inducida por antígenos. En algunas realizaciones, se determina una respuesta inmunitaria en linfocitos T para ser estimulada si se expresan marcadores celulares de activación de linfocitos T a diferentes niveles (por ejemplo, niveles superiores o inferiores) con respecto a células sin estimular.
En algunas realizaciones, se pueden utilizar diversos ensayos a fin de determinar si se ha estimulado una respuesta inmunitaria en un linfocito B o en un grupo de linfocitos B. En algunas realizaciones, la estimulación de una respuesta inmunitaria en linfocitos B puede determinarse midiendo los títulos de anticuerpos, las afinidades de los anticuerpos, el comportamiento de los anticuerpos en ensayos de neutralización, recombinación por intercambio de clase, maduración por afinidad o anticuerpos específicos de antígenos, desarrollo de linfocitos B con memoria, desarrollo de células plasmáticas de vida prolongada que pueden producir grandes cantidades de anticuerpos de
25 elevada afinidad durante periodos alargados de tiempo, reacciones de centros germinales, y/o comportamiento de anticuerpos en ensayos de neutralización.
Un nanotransportador de vacunas es una entidad que comprende, por ejemplo, al menos un agente inmunomodulador que es capaz de estimular una respuesta inmunitaria en linfocitos B y/o en linfocitos T.
Se pueden usar varios nanotransportadores diferentes de acuerdo con la presente invención. En algunas realizaciones, los nanotransportadores son esferas o esferoides. En algunas realizaciones, los nanotransportadores tienen forma plana o de placa. En algunas realizaciones, los nanotransportadores son cubos o cuboides. En algunas realizaciones, los nanotransportadores son óvalos o elipses. En algunas realizaciones, los nanotransportadores son 35 cilindros, conos, o pirámides. Los nanotransportadores pueden ser huecos y pueden comprender una o más capas. En algunas realizaciones, cada capa tiene una única composición y unas únicas propiedades con respecto a las otras capa(s). Para dar, pero a modo de ejemplo, nanotransportadores que tienen una estructura de núcleo/envoltura, en el que el núcleo es una capa (por ejemplo, un núcleo polimérico) y la envoltura es una segunda capa (por ejemplo, una bicapa o monocapa lipídica). Los nanotransportadores pueden comprender una pluralidad de diferentes capas. En algunas realizaciones, una capa puede estar sustancialmente reticulada, una segunda capa no está sustancialmente reticulada, y así sucesivamente. En algunas realizaciones, una, unas pocas, o todas las diferentes capas pueden comprender uno o más agentes inmunomoduladores, restos de direccionamiento, agentes inmunoestimuladores, y/o sus combinaciones. En algunas realizaciones, una capa comprende un agente inmunomodulador, un resto de direccionamiento, y/o un agente inmunoestimulador, una segunda capa no
45 comprende un agente inmunomodulador, un resto de direccionamiento, y/o un agente inmunoestimulador, y así sucesivamente. En algunas realizaciones, cada capa individual comprende un agente inmunomodulador diferente, un resto de direccionamiento, una agente inmunoestimulador, y/o sus combinaciones.
En algunas realizaciones, los nanotransportadores pueden comprender opcionalmente uno o más lípidos. En algunas realizaciones, un nanotransportador comprende una bicapa lipídica. En algunas realizaciones, un nanotransportador comprende una monocapa lipídica. En algunas realizaciones, un nanotransportador comprende un núcleo de una matriz polimérica rodeado por una capa lipídica (por ejemplo, una bicapa lipídica, una monocapa lipídica, etc.).
55 El nanotransportador comprende uno o más polímeros. En algunas realizaciones, una matriz polimérica puede estar rodeada por una capa de revestimiento (por ejemplo, un liposoma, una monocapa lipídica, micela, etc.). En algunas realizaciones, un agente inmunomodulador, un resto de direccionamiento, y/o un agente inmunoestimulador pueden asociarse con la matriz polimérica. En dichas realizaciones, el agente inmunomodulador, el resto de direccionamiento, y/o el agente inmunoestimulador están encapsulados eficazmente en el nanotransportador.
En algunas realizaciones, un agente inmunomodulador, el resto de direccionamiento, y/o el agente inmunoestimulador pueden estar asociados covalentemente con un nanotransportador. En algunas realizaciones, la asociación covalente está mediada por un enlazador. En algunas realizaciones, un resto de direccionamiento de un agente inmunomodulador, y/o el agente inmunoestimulador se asocias no covalentemente con ujn 65 nanotransportador. Por ejemplo, en algunas realizaciones, un agente inmunomodulador, el resto de direccionamiento, y/o un agente inmunoestimulador están encapsulados, rodeados por, y o dispersos en una matriz
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Figura 16: Macrófagos SCS presentes en AdV derivado de ganglios linfáticos a linfocitos B foliculares. (A) micrografía confocal de macrófagos CD169+ los SCS por encima de un folículo B en un LN popliteal. Se contratiñeron secciones congeladas con aglutinina de germen de trigo (WGA) para identificar la matriz extracelular y con α-B220 para detectar linfocitos B. Señalar que algunos linfocitos B residen en los SCS y un linfocito B parece migrar entre el folículo y los SEC (punta de flecha). Barra de escala: 25 µm. (B) Micrografía electrónica y (C) dibujo esquemático de un macrófago SCS y las células que lo rodean en un LN popliteal 30 minutos después de la inyección de Adv en la almohadilla plantar. Barra de escala: 2 µm. Los dibujos de los recuadros en (C) indican las área de mayor aumento que se muestran en los paneles (D) y (E). Estos paneles muestran dos ejemplos de partículas de AdV en la interfase entre los macrófagos SCS y los linfocitos B (puntas de flecha). Los asteriscos denotan otras partículas de AdV asociadas a macrófagos. Barras de escala: 500 nm. Figura 17: La transferencia mediada por macrófagos de VSV transmitidos por los ganglios linfáticos a través de la superficie de los SCS altera el comportamiento de los linfocitos B específicos de virus. (A) Micrografías electrónicas y dibujo esquemático (intermedio) que muestra un macrófago penetrando la superficie del SCS de un LN popliteal 30 minutos después de la inyección de VSV. Barras de escala: 10 µm (izquierda) y 2 µm (derecha). Flecha: vacuola con VSV digerido. Puntas de flecha: viriones en la zona de contacto entre macrófagos y linfocitos B. (B) MP-IVM de linfocitos B policlonales y linfocitos B VI10YEN en LN popliteales. Barras de escala: 50 µm. (C) Relaciones regionales de linfocitos B V110YEN/linfocitos B del control tras la inyección de VSV. Los resultados proceden de un grupo de 3 películas. (D,E) Localización de linfocitos B V110YEN en LN popliteales con respecto a los SCS. **: p < 0,01 (ANOVA monolateral con ensayo posterior de Bonferroni). Figura 18: Características de serotipos de VSV y linfocitos B VI10YEN específicos de VSV-IND. (A) geles SDS-PAGE (12 %) de lisados de VSV purificados. Parte superior: VSV-IND y VSV-NJ. Las proteínas N y P migran simultáneamente en VSV-NJ, se muestran entre paréntesis los pesos moleculares aproximados. (B) Unión de VSV-IND marcados con Alexa-488 (hilera intermedia) o VSV-NJ (hilera inferior) a linfocitos B procedentes de ratones C57BL/6 (columna izquierda) o ratones VI10YEN (columna derecha). La hilera superior muestra la tinción del control con un anticuerpo antiidiotípico 35.61 a la BCR VI110YEN (Dang y Rock, 1991, J. Immunol., 146:3273). (C) Flujo de calcio intracelular en CD43neg purificado, los linfocitos B cargados con Fluo-LOJO proceden de ratones VI10YEN (hilera superior) o de ratones C57BL/6 (hilera inferior). Se recogieron los eventos continuamente en el tiempo, los asteriscos indican el punto temporal cuando se añadieron los anticuerpos o el virus. Se utilizaron partículas de virus a 1000/linfocito B, anti-IgM-(Fab)2 a 10 µg/106 linfocitos B. (D) Ensayo de neutralización para la Ig total y la IgG en suero de ratones C57BL/6 4 y 10 días después de la inmunización mediante inyección en la almohadilla plantar de 10 µg UV-VSV o UV-VSV-AlexaFluor-488-IND. (E) Flujo de calcio en linfocitos B VI10YEN expuestos a sobrenadantes procedentes de soluciones madre de VSV. Se generó el sobrenadante mediante ultracentrifugación a través de un amortiguador de sacarosa que dio como resultado una reducción de aproximadamente 10.000 veces en los títulos víricos y se utilizaron sobre linfocitos B tanto sin diluir (parte superior derecha) como a una dilución 1:100 (parte inferior derecha). Como control, se diluyó una solución madre de VSV a títulos víricos equivalentes (MOI; paneles de la izquierda). Los resultados demuestran la presencia de VSV-G antigénico que no está asociado con partículas víricas en la preparación vírica de los inventores. Figura 19: Adhesión inducida por VSV de linfocitos B VI10YEN a ICAM-1 y VCAM-1. (A,B) adhesión de linfocitos B VI10YEN no expuestos al tratamiento anteriormente y VSV-IND activados (30 minutos de exposición) a placas de plástico revestidas con las concentraciones indicadas de ICAM-1-Fc (A) o VCAM-1-Fc (B) recombinante. Se muestran los datos combinados de dos experimentos por triplicado. Las barras horizontales representan las medias. (C, D) micrografías confocales de la expresión de ICAM-1 y VCAM-1 en LN popliteales de ratones C57BL/6. Barras de escala: 50 µm. (E) Adhesión de linfocitos B VI10YEN naturales no expuestos anteriormente a tratamiento purificados y linfocitos B VI10YEN a placas de plástico revestidas con las concentraciones equivalentes de ufp indicadas de VSV-IND inactivadas con UV. Los datos representan las medias ± SEM de los triplicados. Figura 20: Se requieren macrófagos SCS para la activación temprana de los linfocitos B específicos de VSV en los LN. (A) la micrografía confocal muestra la localización de MHC-II con VSV-IND (30 minutos después de la inyección) en linfocitos B VI10YENxMHCII(EGFP) en los SCS (punta de flecha), sin profundizar en el folículo (asterisco). Barra de escala: 25 µm. (B) Distancia de linfocitos B VI10YENxMHCII(EGFP) exentos de VSV y asociados a VSV a los SCS; Líneas horizontales: medianas. (C) Expresión cinética de BCR en VI10YEN y (D) linfocitos B policlonales tras la inyección en la almohadilla plantar de VSV-IND. (E) Expresión de BCR en células VI10YEN en LN popliteales tratados o no tratados con CLL tras la inyección con VSV-IND (20 µg). Las intensidades promedio de la fluorescencia se normalizaron para valores exentos de virus (línea punteada). Medias ± SEM (3-5 ratones). (F) Micrografía confocal de linfocitos B VI10YEN en LN popliteales del control y (G) tratados con CLL 6 horas después de la inyección de VSV-IND (0,4 µg). Barra de escala: 125 µm. (H) Frecuencia de linfocitos B VI10YEN en los bordes T/B y en los folículos 6 horas después de la inyección de VSV-IND a las dosis indicadas. Medias ± SEM; n = 3-4 folículos/2 ratones; *: p < 0,05; **: p < 0,01; ***: p < 0,001 (test de la t). Figura 21: Motilidad de linfocitos B VI10YEN en LN drenadas tras la inyección del virus. Medianas de las velocidades instantánea 3D de los linfocitos B naturales (triángulos) y los linfocitos B VI10YEN (círculos) en los folículos profundos y en los folículos superficiales de los SCS aproximadamente 5-35 min. después de la inyección en la almohadilla plantar de VSV. Las barras horizontales representan las medias; *: p < 0,05; **: p <
0.01 (ANOVA monolateral con ensayo posterior de Bonferroni). señalar que los linfocitos B específicos reducen la velocidad a través del folículo completo, probablemente como una consecuencia de los VSV-G libres en la preparación de los inventores (véase la Figura 18). Los experimentos del control mostraron unos parámetros de
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los anticuerpos, el comportamiento de los anticuerpos en ensayos de neutralización, recombinación por intercambio de clase, maduración por afinidad o anticuerpos específicos de antígenos, desarrollo de linfocitos B con memoria, desarrollo de células plasmáticas de vida prolongada que pueden producir grandes cantidades de anticuerpos de elevada afinidad durante periodos alargados de tiempo, reacciones de centros germinales, y/o comportamiento de 5 anticuerpos en ensayos de neutralización. En algunas realizaciones, un nanotransportador de vacunas comprende además al menos un resto de direccionamiento que puede ayudar a liberar el nanotransportador de vacunas a una diana concreta (por ejemplo, órgano, tejido, célula, y/o local subcelular) en un sujeto. En algunas realizaciones, un nanotransportador de vacunas comprende además un agente inmunoestimulador que puede ayudar a estimular una respuesta inmunitaria en linfocitos T y/o linfocitos B. En algunas realizaciones, los nanotransportadores de vacunas 10 comprenden lípidos, compuestos anfifílicos, polímeros, azúcares, matrices poliméricas, y/o partículas no poliméricas.
Polímero no adhesivo soluble en agua: Como se usa en el presente documento, el término "polímero no adhesivo soluble en agua, se refiere a un polímero que es soluble en agua y que puede conferir propiedades de bioensuciamiento reducidas. Los polímeros no adhesivos solubles en agua incluyen polietilenglicol, óxido de
15 polietileno, polialquilenglicol, y óxido de polialquileno.
Descripción detallada de determinadas realizaciones preferidas de la invención
Vacunas
20 Las vacunaciones son normalmente de tipo tanto pasivo como activo. En general, las vacunaciones activas implican la exposición del sistema inmunitario del sujeto a uno o más agentes que son reconocidos como no queridos, indeseados, y/o extraños y estimulan una respuesta inmunitaria endógena que da como resultado la activación de linfocitos que no han recibido tratamiento anteriormente específicos de antígenos que dan lugar a continuación a un
25 aumento de linfocitos B secretores de anticuerpos o linfocitos T efectores específicos de antígenos y linfocitos T con memoria o ambos. Esta solución puede dar como resultado inmunidad protectora para toda la vida que se puede reforzar de tiempo en tiempo mediante la exposición renovada al mismo material antigénico. La perspectiva de la longevidad de una respuesta inmunitarias satisfactoria para activar la vacunación hace esta estrategia más deseable en la mayoría de escenarios clínicos que la vacunación pasiva por lo cual se inyecta un receptor con anticuerpos
30 preformados o con linfocitos efectores específicos de antígenos, que pueden conferir una rápida protección ad hoc, pero normalmente no establecen una inmunidad persistente.
Una gran variedad de formulaciones de vacunas se están empleando o han sido empleadas en seres humanos. La ruta de administración más común en seres humanos mediante inyección intramuscular (i.m.), pero las vacunas
35 pueden aplicarse también por vía oral, intranasal, subcutánea, por medio de inhalación, o por vía intravenosa. En la mayor parte de casos, los antígenos derivados de vacunas se presentan inicialmente a linfocitos no expuestos anteriormente a tratamiento en los ganglios linfáticos regionales.
Algunas vacunas corrientes contra, por ejemplo, patógenos microbianos, consisten en cepas de variantes atenuadas
40 vivas o cepas de variantes no virulentas de microorganismos, u organismos muertos o inactivados de otra forma. otras vacunas utilizan componentes más o menos purificados de lisados de patógenos, tales como hidratos de carbono superficiales o proteínas recombinantes derivadas de patógenos que se fusionan algunas veces con otras moléculas, particularmente proteínas que pueden conferir actividad adyuvante.
45 Las vacunas utilizadas para inyecciones intramusculares se administran normalmente mediante un transportador adyuvante, más frecuentemente alumbre (es decir, sulfato de aluminio potasio), que se piensa que establece un depósito para la liberación prolongada de material antigénico, pero que ejerce también actividades inmunomoduladoras, tales como desviar hacia Th2 las respuestas mediante mecanismos que se entienden de forma incompleta (Lindblad, 2004, Immunol. Cell. Biol., 82:497; y Jordan et al., 2004, Science, 304:1808).
50 Las vacunas que utilizan patógenos vivos atenuados o inactivados dan como resultado normalmente una respuesta inmunitaria intensa, pero su uso tiene limitaciones. Por ejemplo, las cepas vivas de la vacuna pueden producir algunas veces patologías infecciosas, especialmente cuando se administran a receptores inmunocomprometidos. Además, muchos patógenos, particularmente virus, experimentan mutaciones rápidas continuas en su genoma, que
55 les permiten escapar a las respuestas inmunitarias de cepas de vacunas antigénicamente distintas. Sin embargo, se piensa que la mayoría o todos los patógenos poseen determinados determinantes antigénicos que no mutan fácilmente debido a que se asocian con funciones esenciales. Los anticuerpos dirigidos contra estos epítopos conservados, más bien que los epítopos no esenciales más variables, pueden proteger contra virus muy mutables (Baba et al., 2000, Nat. Med., 6:200). Las vacunas basadas en patógenos intactos vivos o muertos no promueven
60 necesariamente el reconocimiento de estos epítopos críticos, pero puede "distraer" esencialmente el sistema inmunitaria para centrar su asalto sobre determinantes muy variables. De este modo, la presente invención abarca el reconocimiento de que un nanotransportador de vacunas diseñado mediante ingeniería genética que imita la naturaleza particulada muy inmunógena de las partículas víricas, pero presenta epítopos inmutables selectivamente esenciales, podría dar como resultado un anticuerpo neutralizante mucho más potente y "a prueba de fugas" y
65 respuestas de linfocitos T efectores en microorganismos intactos.
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