RESUMEN
We have developed a transgenic female goat harboring goat beta-casein promoter/human granulocyte colony stimulating factor (G-CSF) fusion gene by microinjection into fertilized one-cell goat zygotes. Human G-CSF was produced at levels of up to 50 microg/ml in transgenic goat milk. Its biological activity was equivalent to recombinant human G-CSF expressed from Chinese hamster ovary (CHO) cell when assayed using in vitro HL-60 cell proliferation. Human G-CSF from transgenic goat milk increased the total number of white blood cells in C57BL/6N mice with leucopenia induced by cyclophosphamide (CPA). The secreted human G-CSF was glycosylated although the degree of O-glycosylation was lower compared to CHO cell-derived human G-CSF.
Asunto(s)
Animales Modificados Genéticamente , Cabras/fisiología , Factor Estimulante de Colonias de Granulocitos/biosíntesis , Leche/metabolismo , Proteínas Recombinantes de Fusión/biosíntesis , Animales , Western Blotting , Caseínas/genética , División Celular , Ciclofosfamida/metabolismo , Cartilla de ADN/química , Ensayo de Inmunoadsorción Enzimática , Femenino , Vectores Genéticos , Factor Estimulante de Colonias de Granulocitos/genética , Factor Estimulante de Colonias de Granulocitos/aislamiento & purificación , Humanos , Ratones , Ratones Endogámicos C57BL , Reacción en Cadena de la Polimerasa , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/aislamiento & purificaciónRESUMEN
Cellular and humoral immunity was studied in pigmented guinea pigs after one injection in the two hind footpads of extract of bovine retina or crude suspensions of rod outer segments (ROS) with complete Freund's adjuvant. In vitro macrophage migration inhibition experiments showed that the antigens which give rise to cellular immunity are mainly located in the pigment epithelium-photoreceptor layers and are devoid of species specificity. With the complement fixation method, the evolution of serum antibodies in anti-ROS guinea pigs was compared with clinical and histological data during the course of the ocular disease. Antibodies appeared in the 2nd week, slighly before the lesions, reaching their maximum level at the 4th week and decreased pari passu with the clinical ocular inflammation, while choroidal cellular infiltration and retinal alterations persisted for several months. Attempts to transfer the disease passively with concentrated serum from enucleated donors were successful. The immune sera were injected locally by different routes. Repeated subconjunctival injections - the less traumatic way - induced severe chorio-retinal lesions.
