RESUMEN
Plant-made vaccines have been the subject of intense interest because they can be produced economically in large scale without the use of animal-derived components. Plant-made therapeutic vaccines against challenging chronic diseases, such as cancer, have received little research attention, and no previous human clinical trials have been conducted in this vaccine category. We document the feasibility of using a plant viral expression system to produce personalized (patient-specific) recombinant idiotype vaccines against follicular B cell lymphoma and the results of administering these vaccines to lymphoma patients in a phase I safety and immunogenicity clinical trial. The system allowed rapid production and recovery of idiotypic single-chain antibodies (scFv) derived from each patient's tumor and immunization of patients with their own individual therapeutic antigen. Both low and high doses of vaccines, administered alone or co-administered with the adjuvant GM-CSF, were well tolerated with no serious adverse events. A majority (>70%) of the patients developed cellular or humoral immune responses, and 47% of the patients developed antigen-specific responses. Because 15 of 16 vaccines were glycosylated in plants, this study also shows that variation in patterns of antigen glycosylation do not impair the immunogenicity or affect the safety of the vaccines. Collectively, these findings support the conclusion that plant-produced idiotype vaccines are feasible to produce, safe to administer, and a viable option for idiotype-specific immune therapy in follicular lymphoma patients.
Asunto(s)
Vacunas contra el Cáncer/uso terapéutico , Linfoma de Células B/terapia , Linfoma Folicular/terapia , Adulto , Anciano , Anticuerpos Antineoplásicos/sangre , Anticuerpos Antineoplásicos/química , Anticuerpos Antineoplásicos/genética , Anticuerpos Antineoplásicos/uso terapéutico , Vacunas contra el Cáncer/administración & dosificación , Vacunas contra el Cáncer/efectos adversos , Vacunas contra el Cáncer/inmunología , Femenino , Factor Estimulante de Colonias de Granulocitos y Macrófagos/administración & dosificación , Humanos , Inmunidad Celular , Idiotipos de Inmunoglobulinas/química , Idiotipos de Inmunoglobulinas/genética , Idiotipos de Inmunoglobulinas/uso terapéutico , Inyecciones Subcutáneas , Linfoma de Células B/genética , Linfoma de Células B/inmunología , Linfoma Folicular/genética , Linfoma Folicular/inmunología , Masculino , Persona de Mediana Edad , Plantas Modificadas Genéticamente , Proteínas Recombinantes , Seguridad , Vacunas Sintéticas/administración & dosificación , Vacunas Sintéticas/efectos adversos , Vacunas Sintéticas/inmunología , Vacunas Sintéticas/uso terapéuticoAsunto(s)
Sacos Aéreos/metabolismo , Peces/metabolismo , Acetatos/metabolismo , Sacos Aéreos/ultraestructura , Animales , Colesterol/metabolismo , Metabolismo de los Lípidos , Microscopía Electrónica , Fosfolípidos/metabolismo , Agua de Mar , Especificidad de la Especie , Escualeno/metabolismo , Vacuolas/metabolismo , Vacuolas/ultraestructuraAsunto(s)
Adenosina Trifosfatasas/metabolismo , Cloroplastos/enzimología , Células Vegetales , Calcio , Carbodiimidas , Anhidrasas Carbónicas/metabolismo , Carotenoides/análisis , Fraccionamiento Celular , Clorofila/análisis , Cloroplastos/análisis , Reductasas del Citocromo/metabolismo , Ferredoxinas , Fructosa-Bifosfatasa/metabolismo , Glicolatos , Magnesio , Microscopía Electrónica , Microsomas , Mitocondrias , NADH NADPH Oxidorreductasas/metabolismo , Fosfatidilcolinas/análisis , Fosfolípidos/análisis , Monoéster Fosfórico Hidrolasas/metabolismo , Proteínas de Plantas/análisis , Plantas/enzimologíaAsunto(s)
Sacos Aéreos/anatomía & histología , Peces , Sacos Aéreos/análisis , Sacos Aéreos/citología , Animales , Colesterol/análisis , Cromatografía de Gases , Cromatografía en Capa Delgada , Ácidos Grasos/análisis , Ácidos Grasos Insaturados/análisis , Membranas/análisis , Microscopía Electrónica , Fosfatidilcolinas/análisis , Fosfatidiletanolaminas/análisis , Fosfolípidos/análisis , Agua de Mar , Especificidad de la Especie , Esfingomielinas/análisisAsunto(s)
Membrana Celular/metabolismo , Crustáceos/metabolismo , Ceras/biosíntesis , Adenosina Trifosfatasas/metabolismo , Animales , Isótopos de Carbono , Membrana Celular/enzimología , Centrifugación por Gradiente de Densidad , Crustáceos/citología , Crustáceos/enzimología , Glucosa-6-Fosfatasa/metabolismo , Microscopía Electrónica , Nucleotidasas , Ácidos Palmíticos/metabolismo , Potasio , Sodio , Especificidad de la Especie , Succinato Deshidrogenasa/metabolismoRESUMEN
The membranes of sporophore cap tissue from the cultivated mushroom, Agaricus bisporus (Lange) Sing., were isolated using discontinuous sucrose gradient ultracentrifugation of a tissue homogenate. A membrane-rich fraction was concentrated at the 1.16/1.18 g/cc interface and a mitochondria-rich fraction at the 1.18/1.20 g/cc interface. The membrane fraction was judged to be greater than 90% membrane vesicles by electron microscopy. The protein to lipid ratio of the membrane fraction was 1.1; the molar ratio of sterol to phospholipid was 0.77. The specific radioactivity of a Mg-activated ATPase was 2.5 times greater in the membrane fraction than in the homogenate. No 5'-nucleotidase or Na-K-Mg-activated ATPase activity was observed.
RESUMEN
One of the major contaminants of reagent grade chloroform is identified as a diester of sebacic acid. The contaminant is not present in redistilled chloroform.