RESUMEN
The development of medicinal products often continues throughout the different phases of a clinical study and may require challenging changes in raw and starting materials at later stages. Comparability between the product properties pre- and post-change thus needs to be ensured. Here, we describe and validate the regulatory compliant change of a raw material using the example of a nasal chondrocyte tissue-engineered cartilage (N-TEC) product, initially developed for treatment of confined knee cartilage lesions. Scaling up the size of N-TEC as required for the treatment of larger osteoarthritis defects required the substitution of autologous serum with a clinical-grade human platelet lysate (hPL) to achieve greater cell numbers necessary for the manufacturing of larger size grafts. A risk-based approach was performed to fulfill regulatory requirements and demonstrate comparability of the products manufactured with the standard process (autologous serum) already applied in clinical indications and the modified process (hPL). Critical attributes with regard to quality, purity, efficacy, safety and stability of the product as well as associated test methods and acceptance criteria were defined. Results showed that hPL added during the expansion phase of nasal chondrocytes enhances proliferation rate, population doublings and cell numbers at passage 2 without promoting the overgrowth of potentially contaminant perichondrial cells. N-TEC generated with the modified versus standard process contained similar content of DNA and cartilaginous matrix proteins with even greater expression levels of chondrogenic genes. The increased risk for tumorigenicity potentially associated with the use of hPL was assessed through karyotyping of chondrocytes at passage 4, revealing no chromosomal changes. Moreover, the shelf-life of N-TEC established for the standard process could be confirmed with the modified process. In conclusion, we demonstrated the introduction of hPL in the manufacturing process of a tissue engineered product, already used in a late-stage clinical trial. Based on this study, the national competent authorities in Switzerland and Germany accepted the modified process which is now applied for ongoing clinical tests of N-TEC. The described activities can thus be taken as a paradigm for successful and regulatory compliant demonstration of comparability in advanced therapy medicinal products manufacturing.
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Condrocitos , Ingeniería de Tejidos , Humanos , Cariotipificación , Articulación de la RodillaRESUMEN
During acute graft-versus-host disease (aGVHD) in mice, autoreactive T cells can be generated de novo in the host thymus implying an impairment in self-tolerance induction. As a possible mechanism, we have previously reported that mature medullary thymic epithelial cells (mTEC(high)) expressing the autoimmune regulator are targets of donor T-cell alloimmunity during aGVHD. A decline in mTEC(high) cell pool size, which purges individual tissue-restricted peripheral self-antigens (TRA) from the total thymic ectopic TRA repertoire, weakens the platform for central tolerance induction. Here we provide evidence in a transgenic mouse system using ovalbumin (OVA) as a model surrogate TRA that the de novo production of OVA-specific CD4(+) T cells during acute GVHD is a direct consequence of impaired thymic ectopic OVA expression in mTEC(high) cells. Our data, therefore, indicate that a functional compromise of the medullary mTEC(high) compartment may link alloimmunity to the development of autoimmunity during chronic GVHD.
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Autoinmunidad , Linfocitos T CD4-Positivos/patología , Enfermedad Injerto contra Huésped/patología , Autotolerancia , Timo/patología , Animales , Autoantígenos/análisis , Autoantígenos/inmunología , Linfocitos T CD4-Positivos/inmunología , Femenino , Enfermedad Injerto contra Huésped/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Ovalbúmina/inmunología , Timo/inmunologíaRESUMEN
BACKGROUND: With increasing demand for umbilical cord blood units (CBUs) with total nucleated cell (TNC) counts of more than 150 × 10(7) , preshipping assessment is mandatory. Umbilical cord blood processing requires aseptic techniques and laboratories with specific air quality and cleanliness. Our aim was to establish a fast and efficient method for determining TNC counts at the obstetric ward without exposing the CBU to the environment. STUDY DESIGN AND METHODS: Data from a total of 151 cord blood donations at a single procurement site were included in this prospective study. We measured TNC counts in cord blood aliquots taken from the umbilical cord (TNCCord ), from placenta (TNCPlac ), and from a tubing segment of the sterile collection system (TNCTS ). TNC counts were compared to reference TNC counts in the CBU which were ascertained at the cord blood bank (TNCCBU ). RESULTS: TNCTS counts (173 ± 33 × 10(7) cells; calculated for 1 unit) correlated fully with the TNCCBU reference counts (166 ± 33 × 10(7) cells, Pearson's r = 0.97, p < 0.0001). In contrast, TNCCord and TNCPlac counts were more disparate from the reference (r = 0.92 and r = 0.87, respectively). CONCLUSIONS: A novel method of measuring TNC counts in tubing segments from the sterile cord blood collection system allows rapid and correct identification of CBUs with high cell numbers at the obstetric ward without exposing cells to the environment. This approach may contribute to cost efficacy as only CBUs with satisfactory TNC counts need to be shipped to the cord blood bank.
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Donantes de Sangre , Recuento de Células , Sangre Fetal/citología , Cordón Umbilical/irrigación sanguínea , Almacenamiento de Sangre/métodos , Femenino , Humanos , Placenta/irrigación sanguínea , Sistemas de Atención de Punto , Embarazo , Estudios ProspectivosRESUMEN
Development of acute graft-versus-host disease (aGVHD) predisposes to chronic GVHD with autoimmune manifestations. A characteristic of experimental aGVHD is the de novo generation of autoreactive T cells. Central tolerance is dependent on the intrathymic expression of tissue-restricted peripheral self-antigens (TRA), which is in mature medullary thymic epithelial cells (mTEC(high)) partly controlled by the autoimmune regulator (Aire). Because TECs are targets of donor T-cell alloimmunity, we tested whether murine aGVHD interfered with the capacity of recipient Aire(+)mTEC(high) to sustain TRA diversity. We report that aGVHD weakens the platform for central tolerance induction because individual TRAs are purged from the total repertoire secondary to a decline in the Aire(+)mTEC(high) cell pool. Peritransplant administration of an epithelial cytoprotective agent, fibroblast growth factor-7, maintained a stable pool of Aire(+)mTEC(high), with an improved TRA transcriptome despite aGVHD. Taken together, our data provide a mechanism for how autoimmunity may develop in the context of antecedent alloimmunity.
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Antígenos/genética , Células Epiteliales/inmunología , Enfermedad Injerto contra Huésped/genética , Enfermedad Injerto contra Huésped/inmunología , Timo/metabolismo , Enfermedad Aguda , Animales , Antígenos/metabolismo , Autoantígenos/genética , Autoantígenos/metabolismo , Femenino , Expresión Génica/fisiología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Endogámicos DBA , Ratones Transgénicos , Timo/patologíaRESUMEN
Cytoreductive conditioning regimens used in the context of allogeneic hematopoietic cell transplantation (HCT) elicit deficits in innate and adaptive immunity, which predispose patients to infections. As such, transplantation outcomes depend vitally on the successful reconstruction of immune competence. Restoration of a normal peripheral T-cell pool after HCT is a slow process that requires the de novo production of naive T cells in a functionally competent thymus. However, there are several challenges to this regenerative process. Most notably, advanced age, the cytotoxic pretransplantation conditioning, and posttransplantation alloreactivity are risk factors for T-cell immune deficiency as they independently interfere with normal thymus function. Here, we discuss preclinical allogeneic HCT models and clinical observations that have contributed to a better understanding of the transplant-related thymic dysfunction. The identification of the cellular and molecular mechanisms that control regular thymopoiesis but are altered in HCT patients is expected to provide the basis for new therapies that improve the regeneration of the adaptive immune system, especially with functionally competent, naive T cells.
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Trasplante de Células Madre Hematopoyéticas , Linfocitos T/citología , Timo/inmunología , Animales , Enfermedad Injerto contra Huésped/inmunología , Trasplante de Células Madre Hematopoyéticas/métodos , Humanos , Linfocitos T/inmunología , Timo/citología , Acondicionamiento Pretrasplante/métodos , Trasplante HomólogoRESUMEN
BACKGROUND: Scientific progress in the biology of hematopoietic stem cells (HSCs) provides opportunities for advances in therapy for different diseases. While stem cell sources such as umbilical cord blood (UCB) are unproblematic, other sources such as human embryonic stem cells (hESCs) raise ethical concerns. STUDY DESIGN AND METHODS: In a prospective survey we established the ethical acceptability of collection, research, and therapy with UCB HSCs versus hESCs among health care professionals, pregnant women, patients undergoing in vitro fertilization therapy, parents, and HSC donors and recipients in Switzerland. RESULTS: There was overall agreement about an ethical justification for the collection of UCB for research and therapy in the majority of participants (82%). In contrast, research and therapy with hESCs was acceptable only by a minority (38% of all responders). The collection of hESCs solely created for HSC collection purposes met overall with the lowest approval rates. Hematologists displayed among the participants the highest acceptance rates for the use of hESCs with 55% for collection, 63% for research, and 73% for therapy. CONCLUSIONS: This is the first study assessing the perception of hESCs for research and therapy in comparison with UCB HSCs in different target groups that are exposed directly, indirectly, or not at all to stem cell-based medicine. Our study shows that the debate over the legitimacy of embryo-destructive transplantation medicine is far from over as particularly hESC research continues to present an ethical problem to an overwhelming majority among laypersons and even among health care professionals.
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Investigación Biomédica , Trasplante de Células Madre de Sangre del Cordón Umbilical , Células Madre Embrionarias/trasplante , Sangre Fetal/citología , Personal de Salud , Adulto , Separación Celular , Femenino , Humanos , Técnicas de Transferencia Nuclear , Embarazo , Estudios Prospectivos , SuizaRESUMEN
BACKGROUND: Two competitive concepts of umbilical cord blood (UCB) banking are currently available: either allogeneic UCB is donated to a public bank or autologous cells are stored in a private bank. Allogeneic-autologous hybrid banking is a new concept that combines these two approaches. However, acceptance of hybrid UCB banking among potential donors is unknown to date. STUDY DESIGN AND METHODS: In a prospective survey, we aimed to establish the acceptance of the hybrid banking model among actual and potential UCB donors in Switzerland. The study groups consisted of parents and pregnant women with or without children. As control group, women at reproductive ages were investigated. RESULTS: The majority of participants agreed fundamentally with UCB donation, and overall acceptance of private banking was 47%. If a possibility for hybrid banking were to be made available, 49% would opt for such a public-private model and only 13% would choose private banking alone. Among the proponents of hybrid banking, a majority of participants chose donor cell splitting over the sequential banking mode. Fifty-six percent of responders wished prior notification before the release of their donated UCB to a foreign recipient. CONCLUSIONS: This is the first study which compared the acceptance of allogeneic, autologous, and hybrid allogeneic-autologous UCB banking in different target groups. We demonstrated that hybrid cord blood banking is the preferred model of banking among actual and potential UCB donors. With increasing demand for UCB in the future, health care providers should therefore consider offering hybrid banking as a viable storage option.
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Bancos de Sangre , Sangre Fetal , Aceptación de la Atención de Salud/estadística & datos numéricos , Adulto , Bancos de Sangre/economía , Bancos de Sangre/legislación & jurisprudencia , Femenino , Humanos , Embarazo , Estudios Prospectivos , Trasplante Autólogo , Trasplante HomólogoRESUMEN
Thymic T cell development is dependent on a specialized epithelial microenvironment mainly composed of cortical and medullary thymic epithelial cells (TECs). The molecular programs governing the differentiation and maintenance of TECs remain largely unknown. Wnt signaling is central to the development and maintenance of several organ systems but a specific role of this pathway for thymus organogenesis has not yet been ascertained. In this report, we demonstrate that activation of the canonical Wnt signaling pathway by a stabilizing mutation of beta-catenin targeted exclusively to TECs changes the initial commitment of endodermal epithelia to a thymic cell fate. Consequently, the formation of a correctly composed and organized thymic microenvironment is prevented, thymic immigration of hematopoietic precursors is restricted, and intrathymic T cell differentiation is arrested at a very early developmental stage causing severe immunodeficiency. These results suggest that a precise regulation of canonical Wnt signaling in thymic epithelia is essential for normal thymus development and function.
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Diferenciación Celular/inmunología , Células Epiteliales/metabolismo , Inhibidores de Crecimiento/genética , Inhibidores de Crecimiento/metabolismo , Timo/inmunología , Timo/metabolismo , beta Catenina/genética , beta Catenina/metabolismo , Animales , Diferenciación Celular/genética , Línea Celular , Células Cultivadas , Endodermo/citología , Endodermo/inmunología , Endodermo/metabolismo , Células Epiteliales/inmunología , Células Epiteliales/patología , Factores de Transcripción Forkhead/genética , Marcación de Gen , Inhibidores de Crecimiento/biosíntesis , Inhibidores de Crecimiento/fisiología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Transducción de Señal/genética , Transducción de Señal/inmunología , Timo/citología , Timo/patología , Proteínas Wnt/genética , Proteínas Wnt/metabolismo , beta Catenina/biosíntesis , beta Catenina/fisiologíaRESUMEN
A favorable outcome of allogeneic hematopoietic stem-cell transplantation (HSCT) depends on the complete reconstitution of the host's immune system. While recovery of peripheral T cells occurs in transplant recipients via both thymus-dependent and thymus-independent pathways, the regeneration of a population of phenotypically naive T cells with a broad T-cell receptor (TCR) repertoire relies entirely on the de novo generation of T cells in the thymus. However, preclinical models and clinical studies of allogeneic HSCT have identified the thymus as a target of graft-versus-host disease (GVHD). The present review focuses on recent insight into how GVHD affects thymic function and how this knowledge aides the design of new strategies to improve immune reconstitution following allogeneic HSCT.
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Enfermedad Injerto contra Huésped/fisiopatología , Reacción Injerto-Huésped/inmunología , Timo/fisiopatología , Células Epiteliales/fisiología , Enfermedad Injerto contra Huésped/inmunología , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Humanos , Péptidos y Proteínas de Señalización Intercelular/fisiología , Timo/inmunologíaRESUMEN
Haematopoietic precursors have to undergo a complex series of maturational steps in the thymus before they exit into the periphery as functional T lymphocytes. Thymic stroma cells, the majority being of epithelial origin, provide the functional partners for the maturational progression along this differentiation pathway. Here we review some of the molecular and cellular mechanisms that account for thymus organogenesis and discuss a strategy to use thymic epithelial precursor cells for the regeneration of the thymic microenvironment.
RESUMEN
The significance of the peripheral immune system in Alzheimer's disease pathogenesis remains controversial. To study the CNS invasion of hematopoietic cells in the course of cerebral amyloidosis, we used a green fluorescence protein (GFP)-bone marrow chimeric amyloid precursor protein transgenic mouse model (APP23 mice). No difference in the number of GFP-positive invading cells was observed between young APP23 mice and nontransgenic control mice. In contrast, in aged, amyloid-depositing APP23 mice, a significant increase in the number of invading ameboid-like GFP-positive cells was found compared with age-matched nontransgenic control mice. Interestingly, independent of the time after transplantation, only a subpopulation of amyloid deposits was surrounded by invading cells. This suggests that not all amyloid plaques are a target for invading cells or, alternatively, all amyloid plaques attract invading cells but only for a limited time, possibly at an early stage of plaque evolution. Immunological and ultrastructural phenotyping revealed that macrophages and T-cells accounted for a significant portion of these ameboid-like invading cells. Macrophages did not show evidence of amyloid phagocytosis at the electron microscopic level, and no obvious signs for T-cell-mediated inflammation or neurodegeneration were observed. The observation that hematopoietic cells invade the brain in response to cerebral amyloidosis may hold an unrecognized therapeutic potential.
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Precursor de Proteína beta-Amiloide/metabolismo , Células de la Médula Ósea , Trasplante de Médula Ósea , Encéfalo/metabolismo , Movimiento Celular , Hematopoyesis , Amiloide , Neuropatías Amiloides/metabolismo , Neuropatías Amiloides/patología , Neuropatías Amiloides/fisiopatología , Animales , Células de la Médula Ósea/patología , Encefalopatías/metabolismo , Encefalopatías/patología , Encefalopatías/fisiopatología , Proteínas Fluorescentes Verdes , Macrófagos , Masculino , Ratones , Ratones Transgénicos , Fagocitosis , Linfocitos T/patología , Quimera por TrasplanteRESUMEN
Haematopoietic precursors have to undergo a complex series of maturational steps in the thymus before they exit into the periphery as functional T lymphocytes. Thymic stroma cells, the majority being of epithelial origin, provide the functional partners for the maturational progression along this differentiation pathway. Here we review some of the molecular and cellular mechanisms that account for thymus organogenesis and discuss a strategy to use thymic epithelial precursor cells for the regeneration of the thymic microenvironment.
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Organogénesis , Regeneración , Linfocitos T/inmunología , Timo/citología , Autoantígenos , Células Epiteliales/citología , Células Madre Hematopoyéticas/citología , Humanos , Transducción de Señal , Células del Estroma/citología , Linfocitos T/citología , Timo/inmunologíaRESUMEN
BACKGROUND: The thyroid is composed of endocrine epithelial cells, blood vessels, and mesenchyme. However, no data exist thus far on absolute cell numbers, relative distribution, and proliferation of the different cell populations in the developing and mature thyroid. The aim of this study was therefore to establish a flow cytometry protocol that allows detection and quantification of discrete cell populations in embryonic and adult murine thyroid tissues. METHODS: Cell-type anti-mouse specific antibodies were used for erythroid cells (Ter119), hematopoietic cells (CD45), epithelial cells (EpCam/CD326, E-cadherin/CD324), thyroid follicular cells and C-cells (Nkx2-1), endothelial cells (Pecam/CD31, Icam-1/CD54), and fibroblasts (PDGFRa/CD140a). Proliferating cells were detected after labeling with 5-bromo-2'-deoxyuridine (BrdU). For flow cytometry analyses, micro-dissected embryonic (E) and adult thyroids were pooled (E13.5, n = 25; E15.5, n = 15; E17.5, n = 15; adult, n = 4) in one sample. RESULTS: The absolute parenchymal cell numbers per mouse thyroid (M ± SD), excluding the large number of CD45(+) and Ter119(+) cells, increased from 7425 ± 1338 at E13.5 to 271,561 ± 22,325 in adult tissues. As expected, Nkx2-1(+) cells represented the largest cell population in adult tissues (61.2 ± 1.1%). Surprisingly, at all three embryonic stages analyzed, thyroid follicular cells and C-cells accounted only for a small percentage of the total thyroid cell mass (between 4.7 ± 0.4% and 9.4 ± 1.6%). In contrast, the largest cell population at all three embryonic stages was identified as PDGFRa/CD140a(+) fibroblasts (61.4 ± 0.4% to 77.3 ± 1.1%). However, these cells represented the smallest population in adult tissues (5.2 ± 0.8%). Pecam/CD31(+) endothelial cells increased from E13.5 to E15.5 from 3.7 ± 0.8% to 8.5 ± 3.0%, then remained stable at E17.5 and adult tissues. Proliferation rates were sizable during the entire organogenesis but differed between cell populations, with distinct proliferative peaks at E13.5 in epithelial cells (32.7 ± 0.6% BrdU(+) cells), and at E15.5 in endothelial cells (22.4 ± 2.4% BrdU(+) cells). Fibroblasts showed a constant proliferation rate in embryonic tissues. In adult tissues, BrdU(+) cells were between 0.1% and 0.4% in all cell types. CONCLUSIONS: Using a novel flow cytometry-based method, a previously unobserved highly dynamic growth pattern of thyroid cell populations during embryogenesis was uncovered. This approach will provide a useful new tool for cell function analyses in murine thyroid disease models.
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Glándula Tiroides/citología , Glándula Tiroides/embriología , Animales , Anticuerpos/química , Diferenciación Celular , Proliferación Celular , Separación Celular , Células Epiteliales/citología , Femenino , Fibroblastos/citología , Citometría de Flujo , Mesodermo/citología , RatonesRESUMEN
INTRODUCTION: Circulating endothelial progenitor cells (EPCs; CD31+ CD34(bright)CD133+ CD45(dim) cells) are novel markers of endothelial dysfunction and related to inflammatory processes such as acute graft-versus-host disease (aGvHD). PATIENTS AND METHODS: 47 patients with acute myeloid leukaemia (AML) who were in complete remission as they underwent allogeneic hematopoietic stem cell transplantation with myeloablative conditioning with PBSC as stem cell source were enrolled in the study. Blood samples for the quantitative analysis of circulating EPC levels were drawn at different time points in patients with and without aGvHD. CD34+ VEGFR2/KDR+ CD133+ triple-positive cells identified among CD34+ cells by FACS. EPC were quantified and data are presented as cells/ml whole blood. RESULTS: Circulating EPC levels were not significantly different in patients with and without aGvHD prior to conditioning (baseline) and at the time of engraftment. However, at diagnosis of aGvHD≥grade 2, EPC levels increased whereas in patients without aGvHD the EPC levels remained significantly lower (3021±278 versus 2322±195 cells/ml; p<0.001). Patients with steroid-refractory aGvHD had high levels of EPC throughout. EPC levels fell in responding patients. CONCLUSION: Our results demonstrate that the number of circulating EPCs is increased in patients with aGvHD compared to patients without aGvHD.
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Células Progenitoras Endoteliales/patología , Enfermedad Injerto contra Huésped/sangre , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Leucemia Mieloide Aguda/terapia , Enfermedad Aguda , Adulto , Anciano , Biomarcadores/sangre , Recuento de Células , Femenino , Enfermedad Injerto contra Huésped/patología , Humanos , Leucemia Mieloide Aguda/complicaciones , Masculino , Persona de Mediana Edad , Inducción de Remisión , Trasplante HomólogoRESUMEN
Allogeneic haematopoietic stem cell transplantation (HSCT) is used to treat an increasing number of congenital and acquired disorders of the haematopoietic system. Even though cytoreductive conditioning regimens vary in intensity, all clinically used protocols invariably cause side effects that compromise transiently or long-term the response of the natural and the adaptive immune systems. However, in the context of the reconstruction of immunity, the generation of naïve T cells constitutes a slow process, and requires a functionally competent thymus. Unfortunately, regular thymic function is frequently suppressed by transplant-related toxicities. Most notably, graft-versus-host disease (GVHD) causes a state of posttransplantation immune deficiency. Here we discuss preclinical allogeneic HSCT models and clinical observations that have contributed to a detailed understanding of the cellular and molecular mechanisms responsible for the thymic dysfunction caused by acute GVHD. An in-depth knowledge of the mechanisms that control regular thymopoiesis and, conversely, affect thymus function is expected to provide the factual basis for the design of innovative therapies to recover T-cell numbers and function following allogeneic HSCT.
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Enfermedad Injerto contra Huésped/inmunología , Trasplante de Células Madre Hematopoyéticas , Timo/inmunología , Inmunidad Adaptativa/efectos de los fármacos , Inmunidad Adaptativa/inmunología , Animales , Modelos Animales de Enfermedad , Células Epiteliales/efectos de los fármacos , Células Epiteliales/inmunología , Factor 7 de Crecimiento de Fibroblastos/farmacología , Efecto Injerto vs Leucemia/inmunología , Humanos , Inmunidad Innata/efectos de los fármacos , Inmunidad Innata/inmunología , Síndromes de Inmunodeficiencia/inmunología , Recuento de Linfocitos , Regeneración/efectos de los fármacos , Regeneración/inmunología , Linfocitos T/efectos de los fármacos , Linfocitos T/inmunología , Timo/efectos de los fármacos , Acondicionamiento PretrasplanteRESUMEN
The thymus constitutes the primary lymphoid organ for the generation of T cells. Its function is particularly susceptible to various negative influences ranging from age-related involution to atrophy as a consequence of malnutrition, infection or harmful iatrogenic influences such as chemotherapy and radiation. The loss of regular thymus function significantly increases the risk for infections and cancer because of a restricted capacity for immune surveillance. In recent years, thymus-stimulatory, thymus-regenerative, and thymus-protective strategies have been developed to enhance and repair thymus function in the elderly and in individuals undergoing hematopoietic stem cell transplantation. These strategies include the use of sex steroid ablation, the administration of growth and differentiation factors, the inhibition of p53, and the transfer of T cell progenitors to alleviate the effects of thymus dysfunction and consequent T cell deficiency.
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Envejecimiento/fisiología , Linfocitos T/metabolismo , Timo/fisiopatología , Antagonistas de Andrógenos/farmacología , Animales , Antineoplásicos/efectos adversos , Atrofia/etiología , Enfermedad Injerto contra Huésped/complicaciones , Enfermedad Injerto contra Huésped/fisiopatología , Hormona del Crecimiento/metabolismo , Humanos , Infecciones/complicaciones , Infecciones/fisiopatología , Desnutrición/complicaciones , Desnutrición/fisiopatología , Radioterapia/efectos adversos , Somatomedinas/metabolismo , Timo/efectos de los fármacos , Timo/patologíaRESUMEN
The clinical success of allogeneic hematopoietic stem cell transplantation (HSCT) depends on the appropriate reconstitution of the host's immune system. While recovery of T-cell immunity may occur in transplant recipients via both thymus-dependent and thymus-independent pathways, the regeneration of a population of phenotypically naive T cells with a broad receptor repertoire relies entirely on the de novo generation of T-cells in the thymus. Preclinical models and clinical studies of allogeneic HSCT have identified the thymus as a target of graft-versus-host disease (GVHD), thus limiting T-cell regeneration. The present review focuses on recent insight into how GVHD affects thymic structure and function and how this knowledge may aid in the design of new strategies to improve T-cell reconstitution following allogeneic HSCT.
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Enfermedad Injerto contra Huésped/inmunología , Enfermedad Injerto contra Huésped/patología , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Timo/inmunología , Timo/patología , Animales , HumanosRESUMEN
Acute graft-versus-host disease (aGVHD) impairs thymus-dependent T-cell regeneration in recipients of allogeneic bone marrow transplants through yet to be defined mechanisms. Here, we demonstrate in mice that MHC-mismatched donor T cells home into the thymus of unconditioned recipients. There, activated donor T cells secrete IFN-gamma, which in turn stimulates the programmed cell death of thymic epithelial cells (TECs). Because TECs themselves are competent and sufficient to prime naive allospecific T cells and to elicit their effector function, the elimination of host-type professional antigen-presenting cells (APCs) does not prevent donor T-cell activation and TEC apoptosis, thus precluding normal thymopoiesis in transplant recipients. Hence, strategies that protect TECs may be necessary to improve immune reconstitution following allogeneic bone marrow transplantation.
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Apoptosis/inmunología , Trasplante de Médula Ósea/inmunología , Endotelio/inmunología , Enfermedad Injerto contra Huésped/inmunología , Linfocitos T/inmunología , Timo/inmunología , Enfermedad Aguda , Animales , Células Presentadoras de Antígenos/inmunología , Células Presentadoras de Antígenos/patología , Endotelio/patología , Interferón gamma/deficiencia , Interferón gamma/inmunología , Activación de Linfocitos/inmunología , Ratones , Ratones Noqueados , Recuperación de la Función/inmunología , Linfocitos T/patología , Timo/patología , Quimera por Trasplante/inmunología , Trasplante HomólogoRESUMEN
The tumor suppressor Smad4 mediates signaling by the transforming growth factor beta (TGF-beta) superfamily of ligands. Previous studies showed that several TGF-beta family members exert important functions in hematopoiesis. Here, we studied the role of Smad4 in adult murine hematopoiesis using the inducible Mx-Cre/loxP system. Mice with homozygous Smad4 deletion (Smad4(Delta/Delta)) developed severe anemia 6 to 8 weeks after induction (mean hemoglobin level 70 g/L). The anemia was not transplantable, as wild-type mice reconstituted with Smad4(Delta/Delta) bone marrow cells had normal peripheral blood counts. These mice did not develop an inflammatory disease typical for mice deficient in TGF-beta receptors I and II, suggesting that the suppression of inflammation by TGF-beta is Smad4 independent. The same results were obtained when Smad4 alleles were deleted selectively in hematopoietic cells using the VavCre transgenic mice. In contrast, lethally irradiated Smad4(Delta/Delta) mice that received wild-type bone marrow cells developed anemia similar to Smad4(Delta/Delta) mice that did not receive a transplant. Liver iron stores were decreased and blood was present in stool, indicating that the anemia was due to blood loss. Multiple polyps in stomach and colon represent a likely source of the bleeding. We conclude that Smad4 is not required for adult erythropoiesis and that anemia is solely the consequence of blood loss.