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1.
Reproduction ; 126(1): 49-59, 2003 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-12814347

RESUMEN

Mouse zona pellucida subunit 3 (mZP3) was tested for efficacy as an immunocontraceptive antigen by comparing the fertility of mice immunized with recombinant mZP3 proteins. Recombinant protein was expressed using either the vaccinia virus T7 mammalian (vmZP3 protein) or baculovirus insect cell (bmZP3 protein)-expression systems. Female BALB/c or wild mice were immunized by i.p. injection using Freund's complete adjuvant and boosted three times with affinity purified recombinant proteins in Freund's incomplete adjuvant. Most mice developed antibodies that crossreacted to the respective mZP3 antigens by ELISA or western blot. In BALB/c mice immunized with vmZP3, fertility and mean litter size were reduced transiently to 25% and 10%, respectively, of those of control mice. However, immunization with bmZP3 did not affect either the fertility or mean litter sizes in BALB/c or wild mice immunized with bmZP3. The results demonstrate that reduction in fertility can be achieved in female BALB/c mice immunized using Freund's adjuvants and recombinant mZP3 protein produced in a mammalian, but not an insect, cell-expression system. Arguments are presented for the likely role of glycosylation of the mZP3 antigen in inducing contraceptive immune responses.


Asunto(s)
Anticoncepción Inmunológica , Proteínas del Huevo/administración & dosificación , Glicoproteínas de Membrana/administración & dosificación , Receptores de Superficie Celular , Vacunas Anticonceptivas/administración & dosificación , Animales , Anticuerpos/sangre , Baculoviridae/genética , Secuencia de Bases , Reactores Biológicos , Western Blotting/métodos , Proteínas del Huevo/genética , Proteínas del Huevo/inmunología , Femenino , Adyuvante de Freund , Inyecciones Intraperitoneales , Tamaño de la Camada , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/inmunología , Ratones , Ratones Endogámicos BALB C , Datos de Secuencia Molecular , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/inmunología , Proteínas Recombinantes/aislamiento & purificación , Spodoptera , Transducción Genética , Vacunas Anticonceptivas/inmunología , Virus Vaccinia/genética , Glicoproteínas de la Zona Pelúcida
2.
J Invest Dermatol ; 117(5): 1193-9, 2001 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-11710932

RESUMEN

The ultraviolet radiation present in sunlight is the primary cause of nonmelanoma skin cancer and has been implicated in the development of cutaneous malignant melanoma. In addition, ultraviolet is immune suppressive and the suppression induced by ultraviolet radiation has been identified as a risk factor for skin cancer induction. Ultraviolet also suppresses the immune response to infectious agents. In most experimental models, ultraviolet is applied to immunologically naive animals prior to immunization. Of equal concern, however, is the ability of sunlight to suppress established immune reactions, such as the recall reaction in humans, which protects against microbial infections. Here we demonstrate that solar-simulated ultraviolet radiation, applied after immunization, suppresses immunologic memory and the elicitation of delayed-type hypersensitivity. Further, we found that wavelengths in the ultraviolet A region of the solar spectrum were critical for inducing immune suppression. Ultraviolet A (320-400 nm) radiation was as effective as solar-simulated ultraviolet A + B (290-400 nm) in suppressing the elicitation of an established immune response. Irradiation with ultraviolet AI (340-400 nm) had no effect. Supporting a critical role for ultraviolet A in ultraviolet-induced immune suppression was the observation that applying a sunscreen that contained an ultraviolet B only filter had no protective effect, whereas, a sunscreen containing both ultraviolet A and ultraviolet B filters totally blocked ultraviolet-induced immune suppression. These data suggest that sunlight may depress the protective effect of prior vaccination. In addition, the observation that ultraviolet A is immunosuppressive indicates the need for ultraviolet A protection when designing sun protection strategies.


Asunto(s)
Sistema Inmunológico/efectos de la radiación , Rayos Ultravioleta , Animales , Formación de Anticuerpos/efectos de los fármacos , Formación de Anticuerpos/efectos de la radiación , Química Farmacéutica , Hipersensibilidad Tardía/prevención & control , Sistema Inmunológico/efectos de los fármacos , Inmunización , Memoria Inmunológica/efectos de la radiación , Ratones , Ratones Endogámicos C3H , Protectores Solares/química , Protectores Solares/farmacología
3.
Immunol Cell Biol ; 79(1): 41-8, 2001 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-11168622

RESUMEN

Depletion of dendritic cells from UV-B-irradiated sheep skin was investigated by monitoring migration of these cells towards regional lymph nodes. By creating and cannulating pseudoafferent lymphatic vessels draining a defined region of skin, migrating cells were collected and enumerated throughout the response to UV-B irradiation. In the present study, the effects of exposing sheep flank skin to UV-B radiation clearly demonstrated a dose-dependent increase in the migration of Langerhans cells (LC) from the UV-B-exposed area to the draining lymph node. The range of UV-B doses assessed in this study included 2.7 kJ/m2, a suberythemal dose; 8 kJ/m2, 1 minimal erythemal dose (MED); 20.1 kJ/m2; 40.2 kJ/m2; and 80.4 kJ/m2, 10 MED. The LC were the cells most sensitive to UV-B treatment, with exposure to 8 kJ/m2 or greater reproducibly causing a significant increase in migration. Migration of gammadelta+ dendritic cells (gammadelta+ DC) from irradiated skin was also triggered by exposure to UV-B radiation, but dose dependency was not evident within the range of UV-B doses examined. This, in conjunction with the lack of any consistent correlation between either the timing or magnitude of migration peaks of these two cell types, suggests that different mechanisms govern the egress of LC and gammadelta+ DC from the skin. It is concluded that the depression of normal immune function in the skin after exposure to erythemal doses of UV-B radiation is associated with changes in the migration patterns of epidermal dendritic cells to local lymph nodes.


Asunto(s)
Movimiento Celular/efectos de la radiación , Células Dendríticas/fisiología , Células de Langerhans/fisiología , Piel/citología , Piel/efectos de la radiación , Animales , Movimiento Celular/fisiología , Células Dendríticas/inmunología , Células Dendríticas/ultraestructura , Relación Dosis-Respuesta en la Radiación , Cinética , Células de Langerhans/inmunología , Células de Langerhans/ultraestructura , Ganglios Linfáticos/citología , Ovinos , Piel/inmunología , Rayos Ultravioleta
4.
Immunol Cell Biol ; 79(6): 547-68, 2001 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-11903614

RESUMEN

This article reviews many of the complex events that occur after cutaneous ultraviolet (UV) exposure. The inflammatory changes of acute exposure of the skin include erythema (sunburn), the production of inflammatory mediators, alteration of vascular responses and an inflammatory cell infiltrate. Damage to proteins and DNA accumulates within skin cells and characteristic morphological changes occur in keratinocytes and other skin cells. When a cell becomes damaged irreparably by UV exposure, cell death follows via apoptotic mechanisms. Alterations in cutaneous and systemic immunity occur as a result of the UV-induced inflammation and damage, including changes in the production of cytokines by keratinocytes and other skin-associated cells, alteration of adhesion molecule expression and the loss of APC function within the skin. These changes lead to the generation of suppressor T cells, the induction of antigen-specific immunosuppression and a lowering of cell-mediated immunity. These events impair the immune system's capacity to reject highly antigenic skin cancers. This review gives an overview of the acute inflammatory and immunological events associated with cutaneous UV exposure, which are important to consider before dealing with the complex interactions that occur with chronic UV exposure, leading to photocarcinogenesis.


Asunto(s)
Epidermis/efectos de la radiación , Sistema Inmunológico/efectos de la radiación , Inflamación/etiología , Rayos Ultravioleta/efectos adversos , Animales , Epidermis/inmunología , Epidermis/metabolismo , Epidermis/patología , Terapia de Inmunosupresión , Inflamación/inmunología , Inflamación/patología , Células de Langerhans/inmunología , Células de Langerhans/efectos de la radiación , Macrófagos/inmunología , Macrófagos/efectos de la radiación , Mastocitos/inmunología , Mastocitos/efectos de la radiación , Ratones
5.
Nat Immunol ; 1(6): 521-5, 2000 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-11101875

RESUMEN

Ultraviolet (UV) radiation is carcinogenic and immunosuppressive. UV-induced immune suppression is mediated by antigen-specific T cells, which can transfer suppression to normal recipients. These cells are essential for controlling skin cancer development in the UV-irradiated host and in suppressing other immune responses, such as delayed-type hypersensitivity. Despite their importance in skin cancer development, their exact identity has remained elusive. We show here that natural killer T cells from UV-irradiated donor mice function as suppressor T cells and play a critical role in regulating the growth of UV-induced skin cancers and suppressing adaptive immune responses in vivo.


Asunto(s)
Células Asesinas Naturales/inmunología , Neoplasias Cutáneas/inmunología , Subgrupos de Linfocitos T/inmunología , Animales , Antígenos/administración & dosificación , Antígenos CD1/genética , Antígenos CD1/metabolismo , Femenino , Hipersensibilidad Tardía , Tolerancia Inmunológica , Ratones , Ratones Endogámicos C3H , Ratones Endogámicos C57BL , Ratones Noqueados , Neoplasias Inducidas por Radiación/inmunología , Bazo/inmunología , Rayos Ultravioleta/efectos adversos
6.
Mutat Res ; 422(1): 147-54, 1998 Nov 09.
Artículo en Inglés | MEDLINE | ID: mdl-9920439

RESUMEN

Depletion of epidermal Langerhans cells (LC) and the concomitant depression of the skin immune system after excessive exposure to ultraviolet B light (UVB) has been established in the international literature for some time. Our investigations were intended to determine whether or not these phenomena occurred as a direct result of increased LC migration being triggered by the UVB exposure. To test this hypothesis, a sheep model was established in which the lymphatic vessels draining a defined region of skin were cannulated and the cells migrating towards the regional lymph node continuously collected. Cell populations in these collections were identified and enumerated by indirect immunofluorescence and flow cytometry. These experiments showed there was a significant, dose-dependent increase in the rate of LC migration from sheep skin after exposure to doses of UVB light exceeding 1 minimal erythemal dose (MED). In a series of parallel experiments, the functional characteristics of dendritic cells (DC) migrating from normal or UVB irradiated sheep were studied. To assay them, enriched preparations of DC were collected via cannulated afferent lymphatic vessels and pulsed with antigen prior to incubation with autologous peripheral blood lymphocytes. The relative efficiency of antigen presentation was determined by the ability of DC to induce T cell proliferation. Our data clearly demonstrate that there is a profound loss of normal antigen-presenting cell function after exposure to UVB light. Various experiments were undertaken to determine the mechanism(s) associated with these changes in migration kinetics and cellular function. Electron microscopic examinations of LC migrating from normal or UVB irradiated skin have demonstrated a profound loss of dendritic processes after UVB exposure. This provides a possible explanation for the changes in skin immunity after UVB exposure.


Asunto(s)
Células de Langerhans/fisiología , Piel/inmunología , Piel/efectos de la radiación , Rayos Ultravioleta , Animales , Movimiento Celular/efectos de la radiación , Epidermis/inmunología , Epidermis/efectos de la radiación , Citometría de Flujo , Células de Langerhans/efectos de la radiación , Células de Langerhans/ultraestructura , Ganglios Linfáticos/fisiología , Ganglios Linfáticos/efectos de la radiación , Sistema Linfático/fisiología , Microscopía Electrónica , Ovinos , Factores de Tiempo
7.
J Pharm Sci ; 86(1): 135-41, 1997 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-9002473

RESUMEN

Molecular modeling techniques using attachment energy calculations have been applied, for the first time to our knowledge, to simulate the morphology of an organic hydrate: alpha-lactose monohydrate. Calculation of the strong intermolecular forces using the atom-atom approximation and the potential parameters of Nemethy et al. (Némethy, G.; Pottle, M. S.; Scheraga, H. A. J. Phys. Chem. 1983, 87, 1883-1887) reveals the crystallization to be dominated by intermolecular interactions between lactose molecules rather than lactose-water interactions, suggesting that water of hydration plays a space-filling role in the growth process. The simulated crystal shows a tomahawk-like morphology with the polar effect of the monoclinic space group P21 correctly reproduced by the occurrence of only one of the [010] forms. Confrontation of these results with experimental work of our own, involving crystals precipitated from aqueous solutions at various degrees of undercooling examined by scanning electron microscopy, and that of Visser and Bennema (Visser, R. A.; Bennema, P. Neth. Milk Dairy J. 1983, 37,109-137), who assigned faces to a crystal, gives good agreement, suggesting the suitability of the force field and atom-atom approaches to model the crystallization of organic hydrates.


Asunto(s)
Lactosa/química , Cristalografía por Rayos X , Excipientes , Modelos Moleculares
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