RESUMEN
Mallory-Denk bodies (MDBs) are aggresomes composed of undigested ubiqutinated short lived proteins which have accumulated because of a decrease in the rate of their degradation by the 26s proteasome. The decrease in the activity of the proteasome is due to a shift in the activity of the 26s proteasome to the immunoproteasome triggered by an increase in expression of the catalytic subunits of the immunoproteasome which replaces the catalytic subunits of the 26s proteasome. This switch in the type of proteasome in liver cells is triggered by the binding of IFNγ to the IFNγ sequence response element (ISRE) located on the FAT10 promoter. To determine if either FAT10 or IFNγ are essential for the formation of MDBs we fed both IFNγ and FAT10 knock out (KO) mice DDC added to the control diet for 10weeks in order to induce MDBs. Mice fed the control diet and Wild type mice fed the DDC or control diet were compared. MDBs were located by immunofluorescent double stains using antibodies to ubiquitin to stain MDBs and FAT10 to localize the increased expression of FAT10 in MDB forming hepatocytes. We found that MDB formation occurred in the IFNγ KO mice but not in the FAT10 KO mice. Western blots showed an increase in the ubiquitin smears and decreases ß 5 (chymotrypsin-like 26S proteasome subunit) in the Wild type mice fed DDC but not in the FAT10 KO mice fed DDC. To conclude, we have demonstrated that FAT10 is essential to the induction of MDB formation in the DDC fed mice.
Asunto(s)
Modelos Animales de Enfermedad , Hepatocitos/efectos de los fármacos , Hígado/efectos de los fármacos , Cuerpos de Mallory/patología , Ubiquitinas/fisiología , Animales , Enfermedad Hepática Inducida por Sustancias y Drogas/patología , Dicarbetoxidihidrocolidina/toxicidad , Silenciador del Gen/fisiología , Hepatocitos/metabolismo , Hepatocitos/patología , Humanos , Interferón gamma/metabolismo , Hígado/metabolismo , Hígado/patología , Masculino , Ratones , Ratones Endogámicos C3H , Ratones Noqueados , Tamaño de los Órganos/efectos de los fármacos , Complejo de la Endopetidasa Proteasomal/efectos de los fármacos , Unión Proteica , Elementos de Respuesta/efectos de los fármacos , Elementos de Respuesta/fisiología , Especificidad de la Especie , Ubiquitina/metabolismoRESUMEN
The papillomavirus E2 protein plays an important role in viral transcriptional regulation and replication. We chose to study the cottontail rabbit papillomavirus (CRPV) E2 protein as a transcriptional regulator because of the availability of an animal model for papilloma formation, which may be relevant for human papillomavirus (HPV) infection and replication. We studied the effect of expression levels of E2 on the long control region, which contains transcriptional promoter and enhancer elements, and synthetic E2-dependent artificial promoters in which the E2 was the dominant factor in the transcriptional activation. These experiments indicated that high levels of E2 were inhibitory and low levels were stimulatory for transactivation. In addition, we showed that the complex formed between CRPV E2 and the cognate binding site was less stable than the complex formed between HPV E2 and the same cognate binding site. Furthermore, we showed that CRPV E2 binding to its transcriptional regulatory sequence was stabilized by other proteins such as E1, which produced increments in transcriptional activation of E2-dependent genes. The data may be used to define conditions in which the rabbit model can be used for the screening of drugs which are inhibitory to the HPV and CRPV replication and gene expression.
Asunto(s)
Papillomavirus del Conejo de Rabo Blanco/genética , Regulación Viral de la Expresión Génica , Proteínas Oncogénicas/genética , Proteínas Oncogénicas/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Proteínas Virales/genética , Proteínas Virales/metabolismo , Animales , Secuencia de Bases , Sitios de Unión , Línea Celular , Papillomavirus del Conejo de Rabo Blanco/fisiología , Genes Reporteros , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Regiones Promotoras Genéticas , Conejos , Proteínas Recombinantes/metabolismo , Secuencias Reguladoras de Ácidos Nucleicos , Spodoptera , Transcripción Genética , Activación Transcripcional , TransfecciónRESUMEN
B-chronic lymphocytic leukemia (B-CLL) is characterized by the clonal accumulation of CD5+ B cells. It has been suggested that CLL cells may be regulated by inhibitory and growth-promoting signals exerted by autologous T cells. We have recently described a model for human B-CLL in which peripheral blood mononuclear cells (PBMCs) are transplanted into the peritoneal cavity of lethally irradiated mice radioprotected with bone marrow from mice with severe combined immunodeficiency. In this model, adoptive transfer of low-stage PBMCs leads to marked engraftment of T cells or combined T and CLL cell engraftment, whereas infusion of high-stage PBMCs leads to dominance of CLL cells with a miniscule level of T-cell engraftment. This mutual exclusive pattern of engraftment indicated that T cells might control the expansion of tumor cells in the peritoneum of recipient BALB/c mice. In the present study, we further investigated this question and we demonstrate that in vivo T-cell depletion, using OKT3 antibody, markedly enhances the engraftment of B-CLL cells from patients with early-stage disease. In mice receiving PBMCs from 11 donors with advanced-stage disease, the results were more heterogeneous. In five patients the results were similar to those observed in early stage, whereas in two cases no CLL cell engraftment was found in the absence of T cells. The addition of purified T cells to PBMCs led to a substantial decrease of CLL engraftment in three advanced-stage cases. These results strengthen the working hypothesis that autologous T cells can actively suppress the expansion of the pathological cells in human-->mouse radiation chimera. This effect is prominent in early-stage disease, whereas in advanced stage suppressive and/or stimulatory effects may occur in different patients. The interaction of T cells with tumor cells and the potential of autologous T cell/immune-therapy in CLL can be further explored in this model.
Asunto(s)
Leucemia Linfocítica Crónica de Células B/inmunología , Transfusión de Linfocitos , Quimera por Radiación/inmunología , Linfocitos T/inmunología , Anciano , Anciano de 80 o más Años , Animales , Progresión de la Enfermedad , Femenino , Humanos , Depleción Linfocítica , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos NOD , Ratones Desnudos , Ratones SCID , Persona de Mediana Edad , Muromonab-CD3/farmacología , Estadificación de Neoplasias , Formación de Roseta , Trasplante HeterólogoRESUMEN
Radiation chimeras, generated by transplantation of SCID bone marrow into C3H/HeJ mice, show lethal susceptibility to staphylococcal enterotoxin B (SEB), thus constituting a valid murine model for SEB shock. This SEB sensitivity is due to the ability of the irradiated host to restore residual T cell populations, since the SCID donor bone marrow is unable to generate T cells. SCID bone marrow transplanted into irradiated nude mice does not generate SEB-sensitive chimeras, as a consequence of the inability of the recipient nude mice to develop mature T cells. Thymectomy of normal recipient mice prior to bone marrow transplantation does not affect the development of susceptibility to SEB, suggesting that postthymic, residual T cells of the host probably mediate this SEB sensitivity. In vivo depletion experiments show that CD4+ T cells are required for the SEB-triggered shock, while CD8+ cells suppress it. A further examination of the T helper subpopulations in the SEB-sensitive mice reveals a prevalence of CD4+ CD45RB(dim) cells over CD4+ CD45RB(bright) cells. This T helper balance was statistically significant when correlated with SEB-induced mortality. Our model provides a possible explanation for the SEB resistance of normal mice: they have a prevalence of CD4+ CD45RB(dim) over CD4+ CD45RB(bright) cells.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Enterotoxinas/toxicidad , Antígenos Comunes de Leucocito/análisis , Staphylococcus aureus/patogenicidad , Animales , Femenino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones SCID , Quimera por RadiaciónRESUMEN
We have previously shown that lethally irradiated normal strains of mice, radioprotected with severe combined immunodeficient (SCID) bone marrow, can be engrafted with human peripheral blood mononuclear cells (PBMC). The human/mouse radiation chimera can mount marked humoral and cellular responses to recall antigens, as well as primary responses. In the present study, we adoptively transferred splenocytes from patients with chronic immune thrombocytopenic purpura (ITP) into lethally irradiated BALB/c mice, radioprotected with SCID bone marrow. High titres of total human immunoglobulin appeared as early as 2 weeks post-transplant and declined after 6 weeks, while human anti-human platelet antibodies were detected 2-8 weeks after the transfer of splenocytes. The immunoglobulin G (IgG) fraction contained antibodies against glycoprotein (GP) IIb/IIIa (CD41) or GPIb/IX (CD42). The human platelet antibodies showed a low level of cross-reactivity with mouse platelets, and thrombocytopenia in the animals was not observed. Splenocytes from individual ITP patients differed in their capacity to produce either human platelet antibodies or total human immunoglobulin. Furthermore, antibodies produced in the murine system were not always identical to the original antibodies present in the serum of the patients. The study of the serological aspects of autoantibodies against human platelets in an animal model might be useful for the investigation of potential therapeutics in ITP.
Asunto(s)
Traslado Adoptivo , Autoanticuerpos/sangre , Plaquetas/inmunología , Púrpura Trombocitopénica Idiopática/inmunología , Quimera por Radiación/inmunología , Bazo/citología , Animales , Trasplante de Médula Ósea , Citometría de Flujo , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones SCID , Modelos Inmunológicos , Bazo/trasplanteRESUMEN
Photodynamic therapy (PDT) using the photosensitizer BPD-Verteporfin (liposomal benzoporphyrin derivative-monoacid ring A) has been shown in previous studies to be effective in the amelioration of inflammatory arthritis in both the MRL-lpr mouse and the New Zealand White (NZW) rabbit models, and could potentially offer alleviation of certain inflammation-related symptoms of rheumatoid arthritis. Time and dose dependency of BPD-MA tissue uptake was carried out in the inflamed synovium and other articular and peri-articular tissues following intravenous and intra-articular administration in the NZW rabbit model. As some articular and peri-articular tissues are difficult to extract, this study uses a rapid fluorimetric sampling of tissues following dissolution in Soluene 350. Our results showed that i.v. injected BPD-MA preferentially distributed in the inflamed synovium, and in tissues with a high degree of vascularization. Little or no association was found with avascular tissues such as cartilage and tendons. Clearance from the synovium was rapid, supporting earlier rather than late light treatment. Much higher association of BPD-MA with the synovium was achieved using intra-articular injection, and BPD-MA concentrations were maintained at relatively steady levels for several hours. These observations support the possibility that PDT could offer a safe, highly versatile clinical option for the management of inflamed joints in autoimmune disorders.
Asunto(s)
Antineoplásicos/farmacocinética , Fármacos Fotosensibilizantes/farmacocinética , Porfirinas/farmacocinética , Líquido Sinovial/metabolismo , Animales , Antineoplásicos/administración & dosificación , Artritis/inducido químicamente , Artritis/metabolismo , Inyecciones Intraarticulares , Inyecciones Intravenosas , Liposomas , Ovalbúmina , Fotoquimioterapia , Fármacos Fotosensibilizantes/administración & dosificación , Porfirinas/administración & dosificación , Conejos , Distribución Tisular , VerteporfinaAsunto(s)
Rechazo de Injerto/inmunología , Trasplante de Riñón/inmunología , Inmunodeficiencia Combinada Grave , Animales , Modelos Animales de Enfermedad , Humanos , Transfusión de Linfocitos , Quimera por Radiación , Ratas , Ratas Endogámicas Lew , Ratas Desnudas , Ensayo de Capsula Subrrenal , Trasplante Heterólogo , Trasplante HomólogoRESUMEN
Previous studies performed in our laboratory have shown that B-CLL cells are involved in the production of anti-red cell auto-antibodies, providing a possible mechanism for the autoimmune hemolytic anemia occurring during the course of B-CLL. In order to confirm this hypothesis, we attempted to transfer human B-CLL with AIHA to immunodeficient mice. Peripheral blood mononuclear cells (PBMC) from 11 B-CLL patients suffering from AIHA were transplanted into the peritoneal cavity of lethally irradiated Balb/c mice reconstituted with SCID bone marrow. Chimeric mice generated from PBMC of these patients (in stage III-IV of the disease) exhibited an engraftment profile with dominance of tumor cells and minuscule levels of T cells. Eighty-five percent of the chimeric mice generated from 10 out of the 11 B-CLL patients with Coombs'-positive AIHA, produced human Ig with anti-human red cell specificity as detected by indirect anti-globulin test. In addition, anti-red cell auto-antibodies were produced in 36% of chimeric mice generated from PBMC of Coombs'-negative B-CLL. In contrast, control experiments in which splenic cells from idiopathic AIHA or PBMC from normal donors were transplanted, failed to produce anti-RBC. This in vivo model further supports the relationship between the B cell expansion and the autoimmune hemolytic process.
Asunto(s)
Anemia Hemolítica/complicaciones , Linfocitos B/inmunología , Linfoma de Burkitt/inmunología , Transfusión de Linfocitos , Quimera por Radiación , Anciano , Anciano de 80 o más Años , Anemia Hemolítica/sangre , Anemia Hemolítica/inmunología , Anemia Hemolítica/patología , Animales , Formación de Anticuerpos , Células de la Médula Ósea , Linfoma de Burkitt/sangre , Linfoma de Burkitt/complicaciones , Linfoma de Burkitt/patología , Humanos , Inmunofenotipificación , Depleción Linfocítica , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos NOD , Ratones SCID , Persona de Mediana Edad , Estadificación de Neoplasias , Valores de Referencia , Bazo/inmunología , Linfocitos T/inmunología , Trasplante HeterólogoRESUMEN
BACKGROUND: We recently described a new approach that enables the generation of human/mouse chimera by adoptive transfer of human peripheral blood mononuclear cells into lethally irradiated normal strains of mice or rats, radioprotected with bone marrow from donors with severe combined immune deficiency. In such human/mouse chimera, a marked humoral response to recall antigens, as well as a significant primary response to keyhole limpet hemocyanin, has been generated. METHODS: In the present study, the organ distribution of the engrafted human cells in the human/mouse and human/rat chimera was investigated by immunohistochemistry. RESULTS: Our results show that the T cells seem to be distributed throughout the reticular endothelial system, almost behaving like particles without any homing specificity. The B cells, however, can barely be found in internal organs, such as the liver or the pancreas, and are concentrated in the secondary lymphoid system (e.g., spleen, lymph node, and nonencapsulated lymphoid tissue). The B cells, together with the engrafted human T cells, form mixed lymphoid follicles. CONCLUSIONS: The different homing patterns exhibited by the T and B lymphocytes indicate that the homing receptors on human B cells might be cross-reactive with their mouse counterparts, in contrast to the human T cells, which seem to be unable to interact with the mouse homing receptors. The presence of human B and T lymphocytes in close proximity to each other in the lymphoid tissues is in accordance with the ability of human/BALB radiation chimera to mount significant primary human antibody responses.
Asunto(s)
Linfocitos B/trasplante , Tejido Linfoide/citología , Linfocitos T/trasplante , Traslado Adoptivo , Animales , Linfocitos B/inmunología , Humanos , Inmunoglobulina G/análisis , Cadenas Pesadas de Inmunoglobulina/análisis , Cadenas Ligeras de Inmunoglobulina/análisis , Inmunoglobulina M/análisis , Inmunohistoquímica , Ganglios Linfáticos/citología , Ratones , Ratones Endogámicos BALB C , Ratones SCID , Quimera por Radiación , Ratas , Ratas Endogámicas Lew , Receptores de Antígenos de Linfocitos B/análisis , Bazo/citología , Coloración y Etiquetado , Linfocitos T/inmunología , Factores de TiempoRESUMEN
B-chronic lymphocytic leukemia (BCLL) is a lymphoproliferative disease that is characterized by clonal expansion of CD5+ B cells. BCLL is associated with secondary immunodeficiency and hypogammaglobulinemia. It has been suggested that T-cell dysregulation may play a role in the hypogammaglobulinemia and in the increased incidence of autoimmunity in BCLL patients. We attempted to transfer human peripheral blood mononuclear cells (PBMC) from BCLL patients in different stages of the disease into immunodeficient mice. PBMC from BCLL patients in stage 0, stages I to II, and stages III to IV were transplanted into the peritoneal cavity of lethally irradiated Balb/c or beige/nude/Xid (BNX) mice radioprotected with bone marrow (BM) from severe combined immunodeficiency (SCID) mice. Different engraftment profiles were found in the chimeric mice 2 weeks after transplantation of PBMC according to the disease stage of the BCLL donors. Infusion of PBMC from donors in stage 0 led to marked engraftment of human T cells, whereas the human tumor cells could hardly be detected. In contrast, chimeric mice receiving PBMC from patients in stage III to IV disease exhibited engraftment with a dominance of tumor cells, compared with a miniscule level of T cells. The ability of the engrafted cells to produce human Ig was also found to be correlated with the disease stage of the donor, although all donors had the same magnitude of hypogammaglobulinemia. Total human Ig production in the chimeric mice was normal in mice receiving PBMC from donors in stage 0, whereas in chimeric mice engrafted with PBMC from donors in stages III to IV almost no human Igs could be detected. This differential reconstitution of antibody production in the mouse model according to the stage of the patient's disease will allow further studies on possible cellular interactions between malignant and immune cells in BCLL.
Asunto(s)
Traslado Adoptivo , Refuerzo Inmunológico de Injertos , Inmunoglobulinas/biosíntesis , Leucemia Linfocítica Crónica de Células B/genética , Leucemia Linfocítica Crónica de Células B/inmunología , Quimera por Radiación , Anciano , Anciano de 80 o más Años , Animales , Modelos Animales de Enfermedad , Femenino , Humanos , Transfusión de Linfocitos , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Endogámicos NOD , Ratones Desnudos , Ratones SCID , Persona de Mediana Edad , Especificidad de Órganos/inmunologíaRESUMEN
BACKGROUND: We have recently shown that lethally irradiated normal strains of mice and rats, reconstituted with bone marrow from severe combined immune deficiency (SCID) mice, can be engrafted with human peripheral blood mononuclear cells (PBMC). METHODS: The feasibility of transplanting human renal tissue under the kidney capsule of the SCID/Lewis and SCID/nude radiation chimera and the effects of intraperitoneal infusion of allogeneic human PBMC on the human renal implants were investigated by histology, electron microscopy, immunohistochemistry, and fluorescence-activated cell sorter analysis. RESULTS: Sequential evaluation of the human renal implants from 10 days to 2 months after transplantation showed that human parenchymal elements survive in the implants up to 2 months after transplantation. The overall architecture of the transplanted kidney tissue and the normal structure of individual cells in the glomeruli and tubuli were preserved. Infusion of allogeneic human PBMC after kidney implantation resulted in patchy cellular infiltrates, composed mainly of activated human T cells, and led to prompt rejection of the human renal tissue, whereas no signs of inflammation were observed in human renal implants of chimeric rats that did not receive human PBMC. Treatment with OKT3 antibody, anti-human CD25 antibody, or CTLA4Ig fusion protein in vivo ameliorated the rejection process. CONCLUSIONS: Human adult kidney fragments transplanted into SCID-like rats transiently retain competent parenchymal structures. When these grafts are combined with allogeneic human PBMC, acute cellular rejection develops. We suggest that this chimeric model might be useful for the investigation of the effects of experimental manipulation on the kinetics of the inflammatory response during human renal allograft rejection.
Asunto(s)
Rechazo de Injerto , Inmunoconjugados , Trasplante de Riñón , Quimera por Radiación , Abatacept , Adulto , Animales , Antígenos CD , Antígenos de Diferenciación/farmacología , Antígeno CTLA-4 , Modelos Animales de Enfermedad , Citometría de Flujo , Rechazo de Injerto/inmunología , Supervivencia de Injerto/efectos de los fármacos , Supervivencia de Injerto/inmunología , Humanos , Fragmentos Fc de Inmunoglobulinas/farmacología , Inmunosupresores/farmacología , Trasplante de Riñón/inmunología , Transfusión de Leucocitos , Leucocitos Mononucleares/inmunología , Ratones , Ratones SCID , Microscopía Electrónica , Muromonab-CD3/farmacología , Quimera por Radiación/inmunología , Ratas , Ratas Endogámicas Lew , Receptores de Interleucina-2/inmunología , Proteínas Recombinantes de Fusión/farmacologíaRESUMEN
Severe combined immunodeficient (SCID) mice are increasingly used as hosts for the adoptive transfer of human lymphocytes. Human antibody responses can be obtained in these xenogeneic chimeras, but information about the functionality of the human T cells in SCID mice is limited and controversial. Studies using human peripheral blood lymphocytes (PBL) injected intraperitoneally (IP) into SCID mice (hu-PBL-SCID mice) have shown that human T cells from these chimeras are anergic and have a defective signaling via the T-cell receptor. In addition, their antigenic repertoire is limited to xenoreactive clones. In the present study, we tested the functionality of human T cell in a recently described chimeric model. In this system, BALB/c mice are conditioned by irradiation and then transplanted with SCID bone marrow, followed by IP injection of human PBL. Our experiments demonstrated that human T cells, recovered from these hu-PBL-BALB mice within 1 month posttransplant, proliferated and expressed activation markers upon stimulation with anti-CD3 monoclonal antibody. A vigorous antiallogeneic human cytotoxic T-lymphocyte (CTL) response could be generated in these mice by immunizing them with irradiated allogeneic cells. Moreover, anti-human immunodeficiency virus type 1 (HIV-1) Net-specific human CTLs could be generated in vivo from naive lymphocytes by immunization of mouse-human chimeras with a recombinant vaccinia-nef virus. This model may be used to evaluate potential immunomodulatory drugs or cytokines, and could provide a relevant model for testing HIV vaccines, for production of antiviral T-cell clones for adoptive therapy, and for studying human T-cell responses in vivo.
Asunto(s)
Productos del Gen nef/inmunología , VIH-1/inmunología , Isoantígenos/inmunología , Ratones SCID/inmunología , Quimera por Radiación/inmunología , Linfocitos T Citotóxicos/inmunología , Linfocitos T/trasplante , Trasplante Heterólogo/inmunología , Vacunas contra el SIDA/inmunología , Animales , Trasplante de Médula Ósea , Linfoma de Burkitt/inmunología , Linfoma de Burkitt/patología , Citotoxicidad Inmunológica , Rechazo de Injerto/inmunología , Humanos , Inmunización , Inmunoterapia Adoptiva , Inyecciones Intraperitoneales , Activación de Linfocitos , Ratones , Ratones Endogámicos BALB C , Muromonab-CD3/inmunología , Trasplante de Neoplasias/inmunología , Receptores de Interleucina-2/análisis , Trasplante Homólogo , Células Tumorales Cultivadas , Vacunas Sintéticas/inmunología , Virus Vaccinia/inmunología , Productos del Gen nef del Virus de la Inmunodeficiencia HumanaRESUMEN
It has been shown that engraftment of human peripheral blood lymphocytes (PBL) from Epstein-Barr virus (EBV) seropositive donors in C.B-17/SCID mice is associated with a high incidence of human B cell tumors. More recently, we described a new approach enabling engraftment of human PBL in normal strains of mice or rats receiving lethal split-dose radiation and radioprotected with SCID bone marrow. We now demonstrate that, in contrast to SCID recipients of human PBL, Balb/c and C3H/HeJ recipients of 50-100 x 10(6) human PBL did not develop any EBV lymphoma during a 7-month follow-up period, but were successfully engrafted with human B and T cells. On the other hand, lymphoma developed in 90% of the C.B-17/SCID mice infused with 70 x 10(6) human PBL from the same donor. Likewise, 36% of beige/nude/xid (BNX) mice, exposed to 12 Gy TBI, radioprotected with SCID bone marrow and then transplanted with human PBL developed lymphoma. Similar results were obtained when different strains were infused with PBL of the same donor. Immunohistochemical analysis indicated that the tumor cells were of human B cell origin and expressed the EBV-encoded latent membrane protein-1 and nuclear antigen 2. While further studies are required to understand the mechanisms which suppressed outgrowth of EBV lymphoma in human --> mouse radiation chimera, compared to human --> C.B-17/SCID or human --> BNX chimera, this marked resistance offers new possibilities for transplantation of hematopoietic tissues or cells from EBV-positive donors.
Asunto(s)
Linfocitos B/fisiología , Infecciones por Herpesviridae/inmunología , Herpesvirus Humano 4/patogenicidad , Linfoma/etiología , Infecciones Tumorales por Virus/etiología , Animales , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones SCID , Quimera por Radiación , Especificidad de la Especie , Bazo/citología , Linfocitos T/fisiología , Factores de TiempoRESUMEN
Benzoporphyrin derivative monoacid ring A (BPD), a hydrophobic chlorin-like porphyrin derivative, which fluoresces strongly at 690 nm, may have potential for both oncologic and nononcologic applications in photodynamic therapy (PDT). To study the influence of cellular characteristics on the uptake of BPD, the murine tumor cell line (P815), and in vitro and in vivo concanavalin A (Con A) -stimulated and unstimulated murine splenic lymphocytes were incubated with 2 micrograms/mL BPD at 37 degrees C for 0-60 min. At various times, cells were lysed and the amount of BPD taken up by cells was quantified by fluorescence measurements. The subsets of cells taking up BPD were analyzed using a panel of monoclonal antibodies and the Coulter XL fluorescence-activated cell sorter. Furthermore, Con A-stimulated and unstimulated spleen cells were incubated with 0-50 ng/mliter of BPD for 1 h prior to exposure to red light (7.2 J/cm2). Cell survival 24 h post-PDT was measured by the MTT assay. We found that the rapidly dividing tumor cell line and mitogen-stimulated murine T cells (mainly CD4+/IL-2R+) took up significantly more BPD (5-10-fold) than do unstimulated splenic lymphocytes. Increased BPD uptake correlated with greater photoinactivation when these cells were exposed to light at a wavelength of 690 nm. These findings suggest that activated cells of the immune system may be a target for photoinactivation by BPD.
Asunto(s)
Activación de Linfocitos , Linfocitos/efectos de los fármacos , Fármacos Fotosensibilizantes/farmacología , Porfirinas/farmacología , Bazo/citología , Animales , Médula Ósea/efectos de los fármacos , Células de la Médula Ósea , Citometría de Flujo , Ratones , Mitógenos/farmacología , Fotoquimioterapia , Células Tumorales CultivadasRESUMEN
Lubin et al recently described a new approach that enables the generation of human/mouse chimera by adoptive transfer of human peripheral blood mononuclear cells (PBMC) into lethally irradiated normal strains of mice, radioprotected with bone marrow (BM) from donors with severe combined immune deficiency (SCID). In the present study, we demonstrate in such human/mouse chimera a marked humoral response to recall antigen, such as tetanus toxoid (TT) or hepatitis B surface antigen (HBsAg), as well as a significant primary response to keyhole limpet hemocyanin (KLH). Maximal anti-KLH response in human/Balb chimera was attained 2 to 4 weeks after the immunization and declined thereafter. One week after transplantation, the predominant anti-KLH subtype was IgM, while after 2 weeks, the dominance had shifted to IgG. Similar primary antibody response was also demonstrated against the human immunodeficiency virus (HIV) Nef protein. Comparison between human/Balb and human/SCID chimera showed a major difference in their ability to mount a primary response against KLH. In Balb/c recipients, more than half of the mice exhibited marked IgM titers against KLH, while there was hardly any anti-KLH IgM response in the SCID recipients. From the earliest time point onwards, when anti-KLH antibodies were found in the latter chimera, they were predominantly of the IgG type. We have previously shown that in human/Balb chimera, unlike in SCID recipients, dissemination of transplanted PBMC into the spleen and other internal organs occurs within 24 hours. Therefore, it is likely that the early seeding in the appropriate microenvironment of the lymphoid tissues, is crucial for the maintenance of virgin human B cells.
Asunto(s)
Formación de Anticuerpos , Memoria Inmunológica , Inmunoterapia Adoptiva , Transfusión de Leucocitos , Quimera por Radiación/inmunología , Animales , Especificidad de Anticuerpos , Trasplante de Médula Ósea , Movimiento Celular , Hemocianinas/inmunología , Antígenos de Superficie de la Hepatitis B/inmunología , Humanos , Inmunoglobulina G/biosíntesis , Inmunoglobulina G/inmunología , Inmunoglobulina M/biosíntesis , Inmunoglobulina M/inmunología , Depleción Linfocítica , Tejido Linfoide/inmunología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones SCID , Toxoide Tetánico/inmunología , Trasplante HeterólogoRESUMEN
Normal strains of mice are rendered sensitive to small amounts (3-10 micrograms) of staphylococcal enterotoxin B (SEB) by transplanting bone marrow cells of SCID donor mice to lethally irradiated recipients. Four to 12 weeks post-transplantation, SEB induces 56-100% lethality. Transplantation of normal mouse bone marrow cells, either alone or with the SCID mouse selected bone marrow cells, does not confer SEB sensitivity. These data imply that either irradiation ablates certain cell population(s), that confer resistance to SEB in normal mice (populations that are absent in the SCID donor mice) or that the donor cells selectively repopulate recipients with SEB-sensitive cells. This model will help elucidate the cells, cytokines and the SEB peptide fragments responsible for SEB toxicity and will be useful in identifying promising vaccine candidates and in developing preventive medicines to protect against this potent toxin.
Asunto(s)
Trasplante de Médula Ósea/efectos adversos , Enterotoxinas/toxicidad , Quimera por Radiación/inmunología , Traumatismos Experimentales por Radiación/inmunología , Staphylococcus aureus/inmunología , Irradiación Corporal Total/efectos adversos , Animales , Relación Dosis-Respuesta Inmunológica , Inmunidad Innata , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C3H , Ratones SCID , Datos de Secuencia Molecular , Traumatismos Experimentales por Radiación/patología , Traumatismos Experimentales por Radiación/prevención & controlRESUMEN
In vitro experiments with benzoporphyrin derivative monoacid ring A (BPD) confirmed earlier studies that it was taken up rapidly (within 30 min) to maximum concentrations by all cells tested. It was also confirmed that rapidly dividing tumor cell lines and mitogen-activated murine T lymphocytes took up significantly more (5-10-fold) BPD than did normal splenic lymphocytes. Further experiments were undertaken to determine whether BPD could be activated by whole-body irradiation with red light in the blood of animals, shortly after intravenous (i.v.) administration, in the absence of skin photosensitivity. It was found that shaved and depilated mice injected i.v. 60 min earlier with BPD at between 0.5 and 1.0 mg/kg could tolerate 160 J/cm2 of broad-band red light (560-900 nm) delivered, at a relatively low rate, over a 90 min time interval without developing skin photosensitivity or general phototoxicity. During the treatment time, plasma levels of BPD were between 0.7 and 1.0 micrograms/mL. The light treatment resulted in between 70 and 80% photoinactivation of circulating BPD. When L1210 tumor cells were preincubated with BPD and injected i.v. into mice immediately before total-body light treatment (160 J/cm2 of 590-900 nm light delivered over 90 min), significant reductions in circulating clonogenic tumor cells were observed in blood samples taken immediately following treatment. This indicated that sufficient light was being delivered to BPD in the blood flowing in the peripheral vasculature to effect cytotoxicity to cells containing the photosensitizer without causing either vascular or skin photosensitivity.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Porfirinas/sangre , Porfirinas/efectos de la radiación , Animales , Transporte Biológico Activo , Línea Celular , Técnicas In Vitro , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos DBA , Fotoquimioterapia/métodos , Porfirinas/farmacocinética , Piel/metabolismo , Piel/efectos de la radiación , Células Tumorales Cultivadas/efectos de los fármacosRESUMEN
Efectuamos un seguimiento de 138 pacientes a largo plazo durante un período mínimo de 1,5 años y máximo de 11 con el objeto de determinar en nuestra población de pacientes con AR clásica o definida la morbimortalidad y obtener información respecto a su evolución . 112 mujeres y 26 hombres edad media 54 (+15 años)(rango :10 a 90 años). 20 pacientes ( 14,5 por ciento )fallecieron durante el seguimiento y las causas fueron:sépticas:5 pacientes;cardíacas:5;neoplasias:4;digestivas:2; insuficiencia respiratoria:2;neurológicas:1;endócrinas:1.La mortalidad fue mayor en los hombres (27 por ciento) que en las mujeres (11,6 por ciento) (p<0,001).40 por ciento de los muertos tenían clase funcional III-IV y el 85 por ciento de ellos tuvieron 2 o más admisiones hospitalarias (p<0,005).El 80por ciento de los fallecidos y el 41 por ciento de los vivos presentaron infecciones graves (p<0,005).El 55 por ciento de los que fallecieron tenían cardiopatías y fribosis pulmonar y el 30 por ciento de ellos desarrollaron neoplasias. El 85 por ciento de los muertos tuvo queratoconjuntivitis sicca y el 45,8por ciento de los vivos (p<0,005).El 89 por ciento de los fallecidos tuvieron Factor Reumatoideo en títulos superiores a 1/1280 y el 43,5 por ciento de los vivos e hipergammaglobulinemia mayor de 2g por ciento el 58,8 por ciento y 21,3 por ciento respectivamente (p<0,005).Conclusiones:La mortalidad global fue del 14,5 por ciento y las enfermedades infecciosas y cardiovasculares fueron las principales causas.Parámetros de mayor morbimortalidad resultaron :sexo masculino,clase funcional III-IV,presencia de QC sicca,fibrosis pulmonar,cardiopatías,infecciones graves,altos títulos de Factor Reumatoideo y marcada hipergammaglobulinemia.
Asunto(s)
Artritis Reumatoide/epidemiología , MortalidadRESUMEN
Efectuamos un seguimiento de 138 pacientes a largo plazo durante un período mínimo de 1,5 años y máximo de 11 con el objeto de determinar en nuestra población de pacientes con AR clásica o definida la morbimortalidad y obtener información respecto a su evolución . 112 mujeres y 26 hombres edad media 54 (+15 años)(rango :10 a 90 años). 20 pacientes ( 14,5 por ciento )fallecieron durante el seguimiento y las causas fueron:sépticas:5 pacientes;cardíacas:5;neoplasias:4;digestivas:2; insuficiencia respiratoria:2;neurológicas:1;endócrinas:1.La mortalidad fue mayor en los hombres (27 por ciento) que en las mujeres (11,6 por ciento) (p<0,001).40 por ciento de los muertos tenían clase funcional III-IV y el 85 por ciento de ellos tuvieron 2 o más admisiones hospitalarias (p<0,005).El 80por ciento de los fallecidos y el 41 por ciento de los vivos presentaron infecciones graves (p<0,005).El 55 por ciento de los que fallecieron tenían cardiopatías y fribosis pulmonar y el 30 por ciento de ellos desarrollaron neoplasias. El 85 por ciento de los muertos tuvo queratoconjuntivitis sicca y el 45,8por ciento de los vivos (p<0,005).El 89 por ciento de los fallecidos tuvieron Factor Reumatoideo en títulos superiores a 1/1280 y el 43,5 por ciento de los vivos e hipergammaglobulinemia mayor de 2g por ciento el 58,8 por ciento y 21,3 por ciento respectivamente (p<0,005).Conclusiones:La mortalidad global fue del 14,5 por ciento y las enfermedades infecciosas y cardiovasculares fueron las principales causas.Parámetros de mayor morbimortalidad resultaron :sexo masculino,clase funcional III-IV,presencia de QC sicca,fibrosis pulmonar,cardiopatías,infecciones graves,altos títulos de Factor Reumatoideo y marcada hipergammaglobulinemia.
Asunto(s)
Artritis Reumatoide/epidemiología , MortalidadRESUMEN
Biodistribution studies were carried out on 14C-labeled benzoporphyrin derivative monoacid ring A (BPD), which had been formulated as a unilamellar liposome or taken from a stock solution in dimethyl sulfoxide diluted into phosphate-buffered saline immediately before intravenous injection into DBA/2 mice. By and large the general distribution of BPD to various organs and tissues was comparable for both formulations. It was noted, however, that liposomal material appeared to enter tissues more rapidly and to be cleared more rapidly, as demonstrated by shorter half-lives for a number of tissues including skin, lung and fat, and generally lower levels in most tissues 24 h following administration. Accumulation in tumor tissue was slightly higher with liposomal BPD, and clearance rates for this tissue were equivalent (half-lives 16.1 h for liposomal BPD and 16.9 h for aqueous BPD). When the two preparations were tested in a bioassay in tumor-bearing mice, photodynamic therapy (PDT) with liposomal BPD proved to be superior to the aqueous preparation when PDT was administered 3 h following intravenous administration of BPD. Plasma distribution studies in vitro demonstrated that 91.1 +/- 0.3% of the liposomal BPD distributed to the lipoprotein fraction within the first hour of mixing, whereas only 49.1 +/- 2.6% of nonliposomal BPD was associated with lipoprotein under the same conditions. Furthermore, while lipoprotein-associated liposomal BPD distributed evenly between all three types of lipoprotein (high, low and very low density), a majority of nonliposomal BPD associated with the high-density lipoprotein fraction.