RESUMEN
B-cell maturation antigen (BCMA) is a promising target for the treatment of multiple myeloma (MM) because the expression of this protein is largely limited to B-cell sets, plasma cells, MM, and other B-cell malignancies. Early studies assessing BCMA protein expression and localization have used insufficiently qualified immunohistochemistry assays, which have reported broad ranges of BCMA expression. As a result, our understanding of BCMA tissue expression derived from these data is limited, specifically the prevalence of BCMA expression on the cell surface/membrane, which has mechanistic relevance to the antimyeloma activity of several novel biotherapeutics. Here, we report on the qualification and application of a novel anti-BCMA immunohistochemistry antibody, 805G12. This antibody shows robust detection of BCMA in formalin-fixed, decalcified bone marrow tissue and provides key insights into membrane BCMA expression. The clone 805G12, which was raised against an intracellular C-terminal domain peptide of membrane BCMA, exhibited increased sensitivity and superior specificity across healthy and diseased tissue compared with the frequently referenced commercial reagent AF193. The new clone also demonstrated a broad range of expression of BCMA in MM and diffuse large B-cell lymphoma specimens. Additionally, cross-reactivity with closely related tumor necrosis factor receptor family members was observed with AF193 but not with 805G12. Furthermore, via established 805G12 and other independent BCMA assays, it was concluded that proteolytic processing by γ-secretase contributes to the levels of BCMA localized to the plasma membrane. As BCMA-directed therapeutics emerge to address the need for more effective treatment in the relapsed or refractory MM disease setting, the implementation of a qualified assay would ensure that reliable and consistent data on BCMA surface expression are used to inform clinical trial decisions and patient responses.
Asunto(s)
Mieloma Múltiple , Humanos , Mieloma Múltiple/patología , Inmunohistoquímica , Inmunoterapia Adoptiva , Antígeno de Maduración de Linfocitos B/metabolismo , Células Plasmáticas/patologíaRESUMEN
AA amyloidosis is characterized by amyloid deposition in systemic organs, but amyloid deposition in the central nervous system (CNS) or peripheral nervous system (PNS) is rare. In this study, AA amyloidosis was observed in 31 of 48 flamingos that died at a Japanese zoo. Almost all cases developed AA amyloidosis secondary to inflammatory diseases such as enteritis. Affected flamingos had AA amyloid deposition around blood vessels in periventricular white matter of the brain and in peripheral nerves. In addition, cerebral Aß amyloidosis was observed in one of the 31 cases with AA amyloidosis. In conclusion, flamingos in the zoo commonly developed systemic amyloidosis with frequent amyloid deposition in the CNS and PNS, which seems to be a unique distribution in this avian species. Comparative pathological analyses in flamingos may help elucidate the pathogenesis of amyloid neuropathy.
Asunto(s)
Amiloide/metabolismo , Amiloidosis/veterinaria , Enfermedades de las Aves/patología , Amiloidosis/patología , Animales , Aves , Sistema Nervioso Central/patología , Femenino , Masculino , Nervios Periféricos/patología , Sistema Nervioso Periférico/patologíaRESUMEN
BAFF is a B cell survival and maturation factor implicated in the pathogenesis of systemic lupus erythematosus (SLE). In this in vitro study, we describe that soluble BAFF in combination with IL-2 and IL-21 is a T cell contact-independent inducer of human B cell proliferation, plasmablast differentiation, and IgG secretion from circulating CD27+ memory and memory-like CD27-IgD- double-negative (DN) B cells, but not CD27-IgD+ naive B cells. In contrast, soluble CD40L in combination with IL-2 and IL-21 induces these activities in both memory and naive B cells. Blood from healthy donors and SLE patients have similar circulating levels of IL-2, whereas SLE patients exhibit elevated BAFF and DN B cells and reduced IL-21. B cell differentiation transcription factors in memory, DN, and naive B cells in SLE show elevated levels of Aiolos, whereas Ikaros levels are unchanged. Treatment with CC-220, a modulator of the cullin ring ligase 4-cereblon E3 ubiquitin ligase complex, reduces Aiolos and Ikaros protein levels and BAFF- and CD40L-induced proliferation, plasmablast differentiation, and IgG secretion. The observation that the soluble factors BAFF, IL-2, and IL-21 induce memory and DN B cell activation and differentiation has implications for extrafollicular plasmablast development within inflamed tissue. Inhibition of B cell plasmablast differentiation by reduction of Aiolos and Ikaros may have utility in the treatment of SLE, where elevated levels of BAFF and Aiolos may prime CD27+ memory and DN memory-like B cells to become Ab-producing plasmablasts in the presence of BAFF and proinflammatory cytokines.
Asunto(s)
Factor Activador de Células B/sangre , Factor Activador de Células B/inmunología , Subgrupos de Linfocitos B/inmunología , Factor de Transcripción Ikaros/genética , Memoria Inmunológica , Lupus Eritematoso Sistémico/inmunología , Péptido Hidrolasas/metabolismo , Proteínas Adaptadoras Transductoras de Señales , Formación de Anticuerpos/efectos de los fármacos , Factor Activador de Células B/metabolismo , Subgrupos de Linfocitos B/efectos de los fármacos , Ligando de CD40/farmacología , Diferenciación Celular , Compuestos Heterocíclicos de 4 o más Anillos/farmacología , Humanos , Factor de Transcripción Ikaros/sangre , Memoria Inmunológica/efectos de los fármacos , Interleucina-2/sangre , Interleucina-2/farmacología , Interleucinas/farmacología , Morfolinas , Ftalimidas , Piperidonas , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/deficiencia , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/genética , Miembro 7 de la Superfamilia de Receptores de Factores de Necrosis Tumoral/inmunología , Ubiquitina-Proteína LigasasRESUMEN
The authors describe a spontaneous case of amyloid A (AA) amyloidosis in an adult female Japanese quail ( Coturnix japonica). The bird developed AA amyloidosis secondary to chronic peritonitis caused by a Gram-negative bacillus infection. Mild amyloid deposition was also identified in the intestinal tract of apparently healthy adult individuals, suggesting that quail may develop intestinal amyloidosis with age. Based on these observations, it was hypothesized that quail can develop AA amyloidosis following inflammatory stimulation with lipopolysaccharide (LPS). Therefore, adult quail were repeatedly injected with LPS and the development of AA amyloidosis was confirmed. The amyloid deposition in this model increased when quail amyloid was intravenously injected as an amyloid-enhancing factor. The experiments were repeated with young quail, but amyloid deposits were not observed following LPS injections. However, AA amyloidosis did develop when quail amyloid was injected in addition to LPS. These results indicated that adult quail develop AA amyloidosis after inflammatory stimulation with LPS. Furthermore, quail AA amyloidosis was shown to have transmissibility regardless of age. Interestingly, the authors found that administration of chicken amyloid fibrils also induced AA amyloidosis in young quail. This is the first report of cross-species transmission of avian AA amyloidosis.
Asunto(s)
Amiloide/administración & dosificación , Amiloidosis/veterinaria , Enfermedades de las Aves/transmisión , Coturnix , Transmisión de Enfermedad Infecciosa/veterinaria , Glicoproteínas/administración & dosificación , Administración Intravenosa , Secuencia de Aminoácidos , Amiloide/metabolismo , Amiloidosis/inducido químicamente , Amiloidosis/patología , Animales , Enfermedades de las Aves/inducido químicamente , Enfermedades de las Aves/patología , Pollos , Modelos Animales de Enfermedad , Inflamación/inducido químicamente , Inflamación/patología , Inflamación/veterinaria , Lipopolisacáridos/administración & dosificación , Datos de Secuencia Molecular , Alineación de Secuencia/veterinaria , Proteína Amiloide A Sérica/metabolismoRESUMEN
Regulatory T cells (Treg) are important in maintaining immune homeostasis and in regulating a variety of immune responses, making them attractive targets for modulating immune-related diseases. Success in using induction or transfer of Treg in mice to mediate transplant tolerance suggests Treg-based therapies as mechanisms of long-term drug-free transplant tolerance in human patients. Although more work is needed, critical analyses suggest that key factors in Treg induction, migration, and function are important areas to concentrate investigative efforts and therapeutic development. Elucidation of basic biology will aid in translating data gleaned from mice to humans so that Treg therapies become a reality for patients.
Asunto(s)
Movimiento Celular/inmunología , Linfocitos T Reguladores/inmunología , Inmunología del Trasplante , Tolerancia al Trasplante , Traslado Adoptivo/métodos , Animales , Humanos , RatonesRESUMEN
Immunochemotherapy has been the mainstay of treatment for newly diagnosed diffuse large B-cell lymphoma (ndDLBCL) yet is inadequate for many patients. In this work, we perform unsupervised clustering on transcriptomic features from a large cohort of ndDLBCL patients and identify seven clusters, one called A7 with poor prognosis, and develop a classifier to identify these clusters in independent ndDLBCL cohorts. This high-risk cluster is enriched for activated B-cell cell-of-origin, low immune infiltration, high MYC expression, and copy number aberrations. We compare and contrast our methodology with recent DLBCL classifiers to contextualize our clusters and show improved prognostic utility. Finally, using pre-clinical models, we demonstrate a mechanistic rationale for IKZF1/3 degraders such as lenalidomide to overcome the low immune infiltration phenotype of A7 by inducing T-cell trafficking into tumors and upregulating MHC I and II on tumor cells, and demonstrate that TCF4 is an important regulator of MYC-related biology in A7.
Asunto(s)
Regulación Neoplásica de la Expresión Génica , Factor de Transcripción Ikaros , Lenalidomida , Linfoma de Células B Grandes Difuso , Proteínas Proto-Oncogénicas c-myc , Factor de Transcripción 4 , Transcriptoma , Linfoma de Células B Grandes Difuso/genética , Linfoma de Células B Grandes Difuso/inmunología , Linfoma de Células B Grandes Difuso/patología , Humanos , Proteínas Proto-Oncogénicas c-myc/genética , Proteínas Proto-Oncogénicas c-myc/metabolismo , Lenalidomida/uso terapéutico , Lenalidomida/farmacología , Factor de Transcripción Ikaros/genética , Factor de Transcripción Ikaros/metabolismo , Factor de Transcripción 4/genética , Factor de Transcripción 4/metabolismo , Linfocitos B/metabolismo , Linfocitos B/inmunología , Pronóstico , Animales , Línea Celular Tumoral , Perfilación de la Expresión Génica/métodos , Ratones , Linfocitos T/inmunología , Linfocitos T/metabolismo , Variaciones en el Número de Copia de ADNRESUMEN
PURPOSE OF REVIEW: The mechanisms of tolerance induction and maintenance remain incompletely understood and have yet to be translated to clinical practice. Advances in imaging techniques have allowed precise examination of cell interactions in the lymph node, often in real time. Herein we review evidence that lymph node structure is dynamic and controls the character of the immune response in a multistep, multiplayer dance. T-cell responses in particular can be initiated or influenced in regions beyond the canonical T-cell zone. We propose that the cortical ridge is one such region required for induction and maintenance of tolerance. RECENT FINDINGS: Lymph node domains are more complex than T-cell and B-cell zones. Different domains are important for different types of immune responses. These domains are in part defined by dynamic, malleable physical structures that guide cell interactions and influence immune outcomes. SUMMARY: Further probing as to how lymph node stromal cells and fibers interact with and determine the character of immune responses should yield fundamental insights into tolerance and immunity. Manipulation of lymph node structure and associated unique cell types and molecules may allow therapeutic interventions in the tolerogenic process.
Asunto(s)
Linfocitos B/inmunología , Tolerancia Inmunológica/fisiología , Ganglios Linfáticos/inmunología , Linfocitos T/inmunología , Animales , Comunicación Celular , HumanosRESUMEN
Type I interferons (IFNs) are known to mediate viral control, and also promote survival and expansion of virus-specific CD8+ T cells. However, it is unclear whether signaling cascades involved in eliciting these diverse cellular effects are also distinct. One of the best-characterized anti-viral signaling mechanisms of Type I IFNs is mediated by the IFN-inducible dsRNA activated protein kinase, PKR. Here, we have investigated the role of PKR and Type I IFNs in regulating viral clearance and CD8+ T cell response during primary and secondary viral infections. Our studies demonstrate differential requirement for PKR, in viral control versus elicitation of CD8+ T cell responses during primary infection of mice with lymphocytic choriomeningitis virus (LCMV). PKR-deficient mice mounted potent CD8+ T cell responses, but failed to effectively control LCMV. The compromised LCMV control in the absence of PKR was multifactorial, and linked to less effective CD8+ T cell-mediated viral suppression, enhanced viral replication in cells, and lower steady state expression levels of IFN-responsive genes. Moreover, we show that despite normal expansion of memory CD8+ T cells and differentiation into effectors during a secondary response, effective clearance of LCMV but not vaccinia virus required PKR activity in infected cells. In the absence of Type I IFN signaling, secondary effector CD8+ T cells were ineffective in controlling both LCMV and vaccinia virus replication in vivo. These findings provide insight into cellular pathways of Type I IFN actions, and highlight the under-appreciated importance of innate immune mechanisms of viral control during secondary infections, despite the accelerated responses of memory CD8+ T cells. Additionally, the results presented here have furthered our understanding of the immune correlates of anti-viral protective immunity, which have implications in the rational design of vaccines.
Asunto(s)
Interferón Tipo I/fisiología , Listeriosis/virología , Coriomeningitis Linfocítica/virología , Vaccinia/virología , eIF-2 Quinasa/fisiología , Animales , Western Blotting , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/patología , Linfocitos T CD8-positivos/virología , Ensayo de Inmunoadsorción Enzimática , Femenino , Citometría de Flujo , Técnica del Anticuerpo Fluorescente , Proteínas de Homeodominio/fisiología , Memoria Inmunológica , Listeria monocytogenes/fisiología , Listeriosis/inmunología , Listeriosis/patología , Coriomeningitis Linfocítica/inmunología , Coriomeningitis Linfocítica/patología , Virus de la Coriomeningitis Linfocítica/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Linfocitos T Citotóxicos/inmunología , Linfocitos T Citotóxicos/patología , Linfocitos T Citotóxicos/virología , Vaccinia/inmunología , Vaccinia/patología , Virus Vaccinia/fisiología , Replicación ViralRESUMEN
It is known that C3 is required for optimal expansion of T cells during acute viral infections. However, it is not yet determined whether T cell responses to intracellular bacterial infections require C3. Therefore, we have investigated the requirement for C3 to elicit potent T cell responses to Listeria monocytogenes (LM). We show that expansion of Ag-specific CD8 and CD4 T cells during a primary response to LM was markedly reduced in the absence of C3 activity. Further studies indicated that, unlike in an influenza virus infection, the regulation of LM-specific T cell responses by C3 might not involve the downstream effector C5a. Moreover, reduced T cell responses to LM was not linked to defective maturation of dendritic cells or developmental anomalies in the peripheral T cell compartment of C3-deficient mice. Experiments involving adoptive transfer of C3-deficient CD8 T cells into the C3-sufficient environment of wild-type mice showed that these T cells do not have intrinsic proliferative defects, and a paracrine source of C3 will suffice for clonal expansion of CD8 T cells in vivo. However, stimulation of purified C3-deficient CD8 T cells by plastic-immobilized anti-CD3 showed that C3 promotes T cell proliferation directly, independent of its effects on APC. On the basis of these findings, we propose that diminished T cell responses to LM in C3-deficient mice might be at least in part due to lack of direct effects of C3 on T cells. These studies have furthered our understanding of C3-mediated regulation of T cell immunity to intracellular pathogens.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/microbiología , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/microbiología , Proliferación Celular , Complemento C3/fisiología , Listeriosis/inmunología , Listeriosis/patología , Animales , Linfocitos T CD4-Positivos/citología , Linfocitos T CD8-positivos/citología , Complemento C3/deficiencia , Complemento C3/genética , Listeria monocytogenes/inmunología , Listeriosis/metabolismo , Activación de Linfocitos/genética , Activación de Linfocitos/inmunología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Ratones Transgénicos , Receptor de Anafilatoxina C5a/deficiencia , Receptor de Anafilatoxina C5a/genética , Receptor de Anafilatoxina C5a/fisiologíaRESUMEN
BACKGROUND: Pallister-Killian syndrome (PKS) is a rare disorder caused by the mosaic tetrasomy of chromosome 12p, and is characterized by facial dysmorphism, developmental delay, hypotonia and seizures. RESULTS: We report a patient with PKS showing unique polymicrogyria with calcification. He had delayed development and dysmorphic facial features including frontal bossing, hypertelorism, and high arched palate at 6 months of age. Neuroimaging revealed unilateral polymicrogyria with spot calcifications, which predominantly affected the right perisylvian region. Chromosome G-banding showed the karyotype 46,XY, however, array-based comparative genomic hybridization analysis showed mosaic duplication of chromosome 12p, in which CCND2, which encodes cyclin D2 and is a downstream mediator of PI3K-AKT pathway, is located. Supernumerary chromosome of 12p was detected in 58% of buccal mucosa cells by the interphase fluorescence in situ hybridization analysis using chromosome 12 centromere-specific D12Z3 probe. The diagnosis of PKS was made based on distinctive clinical features of our patient and the results of cytogenetic analyses. CONCLUSION: This report is, to our knowledge, the first case of a patient with PKS who clearly demonstrates polymicrogyria colocalized with calcifications, as shown by CT scans and MRI, and suggests that a patient with PKS could show structural brain anomalies with calcification. We assume that somatic mosaicism of tetrasomy could cause asymmetrical polymicrogyria in our patient, and speculate that increased dosages of CCND2 at chromosome 12p might be involved in the abnormal neuronal migration in PKS.
Asunto(s)
Calcinosis/genética , Trastornos de los Cromosomas/genética , Trastornos de los Cromosomas/patología , Polimicrogiria/genética , Encefalopatías/genética , Encefalopatías/patología , Cromosomas Humanos Par 12/genética , Hibridación Genómica Comparativa , Humanos , Lactante , Masculino , Análisis por MicromatricesRESUMEN
B cells are known to control CD4T cell differentiation in secondary lymphoid tissues. We hypothesized that IL-10 expression by marginal zone precursor (MZP) regulatory B cells controls the differentiation and positioning of effector and regulatory T cells during tolerization. Costimulatory blockade with donor-specific transfusion (DST) and anti-CD40L mAb in C57BL/6 mice induced tolerance to allogeneic cardiac allograft. B cell depletion or IL-10 deficiency in B cells prevented tolerance, resulting in decreased follicular regulatory CD4(+) T cells (Tfr) and increased IL-21 expression by T follicular helper (Tfh) cells in the B cell and T cell zones. IL-21 acted with IL-6 to induce CCR6(+) Th17 that caused rejection. Deficiency or blockade of IL-6, IL-21, IL-21R, or CCR6 prevented B cell depletion-induced acute cellular rejection; while agonistic mCCL20-Ig induced rejection. Adoptive transfer of IL-10(+/+) MZP in tolerogen treated CD19-Cre(+/-):IL-10(fl/fl) mice rescued the localization of Tfh and Tfr cells in the B cell follicle and prevented allograft rejection. MZP B cell IL-10 is necessary for tolerance and controls the differentiation and position of Th17, Tfh and Tfr cells in secondary lymphoid tissues. This has implications for understanding tolerance induction and how B cell depletion may prevent tolerance.
Asunto(s)
Diferenciación Celular , Centro Germinal/inmunología , Interleucina-10/metabolismo , Células Precursoras de Linfocitos B/inmunología , Subgrupos de Linfocitos T/inmunología , Tolerancia al Trasplante , Traslado Adoptivo , Aloinjertos , Animales , Biomarcadores , Centro Germinal/metabolismo , Rechazo de Injerto/inmunología , Inmunofenotipificación , Interleucina-10/deficiencia , Depleción Linfocítica , Ratones , Ratones Noqueados , Ratones Transgénicos , Fenotipo , Células Precursoras de Linfocitos B/metabolismo , Subgrupos de Linfocitos T/citología , Subgrupos de Linfocitos T/metabolismo , Linfocitos T Colaboradores-Inductores/citología , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Colaboradores-Inductores/metabolismo , Células Th17/citología , Células Th17/inmunología , Células Th17/metabolismoRESUMEN
BACKGROUND: Costimulatory blockade with anti-CD40L monoclonal antibody (mAb) plus donor-specific splenocyte transfusion (DST) induces alloantigen-specific tolerance. We previously showed that lymphotoxin signaling in the fibroblastic reticular cell (FRC) stromal subset was required for proper lymph node structure and function during tolerization in murine cardiac transplantation. Here we focused on FRC functions and hypothesized that DST and anti-CD40L mAb-modulated FRC interactions with CD4(+) T cells in mice. METHODS: Mice were immunized or tolerized by DST or DST plus anti-CD40L mAb. Fibroblastic reticular cells were flow-sorted at different timepoints for characterization and in vitro proliferation and activation assays. RESULTS: Fibroblastic reticular cells responded rapidly to DST by transcribing inflammatory cytokine and chemokine messenger RNAs, such as CXCL2, CXCL9, CXCL10, and CCL21. Conversely, anti-CD40L mAb inhibited FRC inflammatory responses. CD40 was expressed on FRC and agonistic anti-CD40 mAb activated FRC, which supported CD4(+) T-cell proliferation, whereas unstimulated FRC did not. Anti-CD3 mAb-activated CD4(+) T cells induced inflammatory cytokine and chemokine expressions by FRC, which were inhibited by anti-CD40L mAb. Thus, FRC phenotype was altered by interaction with CD4(+) T cells through CD40-CD40L, and activated FRC interacted directly with CD4(+) T cells to support T cell activation and proliferation in vitro. CONCLUSIONS: Taken together, these results demonstrated that CD40 on FRC facilitated bidirectional communication between FRC and CD4(+) T cells via CD40-CD40L, thereby altering FRC gene expression of immune regulatory molecules. Because blockade of CD40-CD40L interactions results in tolerance in mice, identification of FRC-T cell interactions provides a new research target for tolerance induction.
Asunto(s)
Linfocitos T CD4-Positivos/metabolismo , Antígenos CD40/metabolismo , Ligando de CD40/metabolismo , Comunicación Celular , Fibroblastos/metabolismo , Ganglios Linfáticos/metabolismo , Traslado Adoptivo , Animales , Anticuerpos Monoclonales/farmacología , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/trasplante , Antígenos CD40/inmunología , Ligando de CD40/deficiencia , Ligando de CD40/genética , Ligando de CD40/inmunología , Comunicación Celular/efectos de los fármacos , Proliferación Celular , Células Cultivadas , Técnicas de Cocultivo , Citocinas/genética , Citocinas/metabolismo , Fibroblastos/efectos de los fármacos , Fibroblastos/inmunología , Regulación de la Expresión Génica , Tolerancia Inmunológica , Mediadores de Inflamación/metabolismo , Ganglios Linfáticos/citología , Ganglios Linfáticos/efectos de los fármacos , Ganglios Linfáticos/inmunología , Activación de Linfocitos , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Fenotipo , Transducción de Señal , Factores de TiempoRESUMEN
BACKGROUND: Trafficking and differentiation of naive CD4+ and regulatory T cells (Treg) within the lymph node (LN) are integral for tolerance induction. The LN is comprised of stromal fibers that dictate lymphocyte migration and LN structure, organization, and microanatomic domains. Distribution of the stromal fiber ER-TR7 changes within the LN after antigenic challenge, but the contributions of ER-TR7 to the resulting immune response remain undefined. We hypothesized that these stromal fiber structural changes affect T cell fate and subsequently allograft survival. METHODS: C57BL/6 mice were left naive (untreated) or made immune or tolerant (donor-specific BALB/c splenocyte transfusion -/+ anti-CD40L monoclonal antibody), or made tolerant and received anti-ER-TR7 monoclonal antibody. Donor-specific T-cell migration was visualized by adoptive transfer of carboxyfluorescein diacetate, succinimidyl ester-labeled TEa T cell receptor transgenic CD4+ cells. Immunohistochemistry was performed on LNs to detect stromal fiber distribution, structure, CCL21 presence, and Treg and donor-specific cell location relative to high endothelial venules (HEV). Naive, tolerant, and tolerant + anti-ER-TR7 mice received BALB/c heterotopic cardiac allografts and graft survival was monitored. RESULTS: The ER-TR7 distribution changed after the induction of tolerance vs. immunity. Treating tolerant mice with anti-ER-TR7 altered HEV basement membrane structure and the distribution of CCL21 within the LN. These differences were mirrored by changes in the migration of naive and Treg cells within and surrounding the HEV. Anti-ER-TR7 prevented tolerance induction and resulted in allograft inflammation and rejection. CONCLUSIONS: These results identify ER-TR7 as an important component of LN structure in tolerance and a direct target for immune modulation.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Ganglios Linfáticos/inmunología , Tolerancia al Trasplante , Traslado Adoptivo , Aloinjertos , Animales , Quimiocina CCL21/metabolismo , Trasplante de Corazón , Ganglios Linfáticos/citología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Transgénicos , Linfocitos T Reguladores/inmunología , Donantes de TejidosRESUMEN
The late health effects of low-dose rate radiation exposure are still a serious public concern in the Fukushima area even four years after the accident at Fukushima Daiichi Nuclear Power Plant (FNPP). To clarify the factors associated with residents' risk perception of radiation exposure and consequent health effects, we conducted a survey among residents of Kawauchi village in May and June 2014, which is located within 30 km of FNPP. 85 of 285 residents (29.8%) answered that acute radiation syndrome might develop in residents after the accident, 154 (54.0%) residents responded that they had anxieties about the health effects of radiation on children, and 140 (49.1%) residents indicated that they had anxieties about the health effects of radiation on offspring. Furthermore, 107 (37.5%) residents answered that they had concerns about health effects that would appear in the general population simply by living in an environment with a 0.23 µSv per hour ambient dose for one year, 149 (52.2%) residents reported that they were reluctant to eat locally produced foods, and 164 (57.5%) residents believed that adverse health effects would occur in the general population by eating 100 Bq per kg of mushrooms every day for one year. The present study shows that a marked bipolarization of the risk perception about the health effects of radiation among residents could have a major impact on social well-being after the accident at FNPP.
Asunto(s)
Accidente Nuclear de Fukushima , Exposición a la Radiación/análisis , Monitoreo de Radiación , Agaricales , Anciano , Niño , Ingestión de Alimentos , Femenino , Humanos , Masculino , Persona de Mediana Edad , Plantas de Energía Nuclear , Radiación , RiesgoRESUMEN
BACKGROUND: Blocking CD40-CD40L costimulatory signals induces transplantation tolerance. Although B-cell depletion prevents alloantibody formation, nonhumoral functions of B cells in tolerance have not been well characterized. We investigated whether specific subsets of B cell or B cell-derived interleukin (IL)-10 contribute to tolerance. METHODS: Wild type C57BL/6, or B cell-specific interleukin (IL)-10 (CD19-Cre::IL-10) mice, received vascularized BALB/c cardiac allografts. BALB/c donor-specific splenocyte transfusion and anti-CD40L monoclonal antibody were used as tolerogen. B cells were depleted with antimouse CD20 monoclonal antibody. Various B-cell subsets were purified and characterized by flow cytometry, reverse transcription polymerase chain reaction, and adoptive transfer. RESULTS: B-cell depletion prevented costimulatory blockade-induced allogeneic tolerance. Costimulatory blockade increased IL-10 in marginal zone precursor (MZP) B cells, but not other subsets. In particular, costimulatory blockade did not change other previously defined regulatory B-cell subsets (Breg), including CD5CD1d Breg or expression of TIM1 or TIM4 on these Breg or other Breg cell subsets. Costimulatory blockade also induced IL-21R expression in MZP B cells, and IL-21R MZP B cells expressed even more IL-10. B-cell depletion or IL-10 deficiency in B cells prevented tolerance in a cardiac allograft model, resulting in rapid acute cardiac allograft rejection. Adoptive transfer of wild type MZP B cells but not other subsets to B cell-specific IL-10 deficient mice prevented graft rejection. CONCLUSIONS: CD40 costimulatory blockade induces MZP B cell IL-10 which is necessary for tolerance. These observations have implications for understanding tolerance induction and how B cell depletion may prevent tolerance.
Asunto(s)
Rechazo de Injerto/prevención & control , Supervivencia de Injerto , Trasplante de Corazón , Interleucina-10/metabolismo , Miocardio/metabolismo , Células Precursoras de Linfocitos B/metabolismo , Tolerancia al Trasplante , Traslado Adoptivo , Animales , Anticuerpos Monoclonales/farmacología , Antígenos CD40/inmunología , Antígenos CD40/metabolismo , Ligando de CD40/inmunología , Ligando de CD40/metabolismo , Células Cultivadas , Rechazo de Injerto/inmunología , Rechazo de Injerto/metabolismo , Rechazo de Injerto/patología , Trasplante de Corazón/efectos adversos , Humanos , Interleucina-10/deficiencia , Interleucina-10/genética , Interleucina-10/inmunología , Subunidad alfa del Receptor de Interleucina-21/inmunología , Subunidad alfa del Receptor de Interleucina-21/metabolismo , Depleción Linfocítica , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Ratones Noqueados , Miocardio/inmunología , Miocardio/patología , Fenotipo , Células Precursoras de Linfocitos B/inmunología , Células Precursoras de Linfocitos B/trasplante , Transducción de Señal , Factores de TiempoRESUMEN
An isocitrate dehydrogenase (ICDH) with an unique coenzyme specificity from Acidithiobacillus thiooxidans was purified and characterized, and its gene was cloned. The native enzyme was homodimeric with a subunit of M(r) 45000 and showed a 78-fold preference for NAD(+) over NADP(+). The cloned ICDH gene (icd) was expressed in an icd-deficient strain of Escherichia coli EB106; the activity was found in the cell extract. The gene encodes a 429-amino acid polypeptide and is located between open reading frames encoding a putative aconitase gene (upstream of icd) and a putative succinyl-CoA synthase beta-subunit gene (downstream of icd). A. thiooxidans ICDH showed high sequence similarity to bacterial NADP(+)-dependent ICDH rather than eukaryotic NAD(+)-dependent ICDH, but the NAD(+)-preference of the enzyme was suggested due to residues conserved in the coenzyme binding site of the NAD(+)-dependent decarboxylating dehydrogenase.
Asunto(s)
Gammaproteobacteria/enzimología , Isocitrato Deshidrogenasa/genética , Isocitrato Deshidrogenasa/metabolismo , NAD/metabolismo , Secuencia de Aminoácidos , Secuencia de Bases , Clonación Molecular , Gammaproteobacteria/genética , Isocitrato Deshidrogenasa/aislamiento & purificación , Datos de Secuencia Molecular , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
DBA/2 CrSlc mice infected with the D variant of encephalomyocarditis virus (EMC-D) (10 PFU/head) developed biphasic hind limb paralysis due to spinal cord lesion. The early phase lesion was characterized by demyelination with infiltration of macrophages in the funiculus lateraris and the late phase lesion by degeneration of motor neurons with infiltration of CD4(+) T cells in the cornu ventrale. In the present study, treatment with anti-Mac1 monoclonal antibody (MAb) or anti-CD4 MAb prior to virus infection (-3 to -1 days) reduced the early phase lesion and the incidence of the first paralysis. Signals of viral RNAs were observed only in a few oligodendrocytes in the funiculus lateraris. Treatment with anti-CD4 MAb from 31 to 33 days post infection when mice showed recovery from the first paralysis reduced the late phase lesion and prevented the second paralysis. Signals of viral RNAs were still detected in a few degenerated neurons in the cornu ventrale. These results indicate that while macrophages and CD4(+) T cells participate in the early phase lesion and paralysis and only CD4(+) T cells in the late phase lesion and paralysis.
Asunto(s)
Linfocitos T CD4-Positivos/inmunología , Infecciones por Cardiovirus/inmunología , Virus de la Encefalomiocarditis , Paraplejía/inmunología , Enfermedades de la Médula Espinal/inmunología , Animales , Anticuerpos Monoclonales/uso terapéutico , Ratones , Ratones Endogámicos DBA , Paraplejía/virología , ARN Mensajero/metabolismo , Enfermedades de la Médula Espinal/virologíaRESUMEN
Pregnant mice of the BALB/c CrSlc strain were experimentally infected with the D variant of encephalomyocarditis virus (EMC-D, 5 x 10(2) PFU/head) on three different gestational days (GD). Mice were intraperitoneally inoculated with EMC-D on 11, 13 and 15 GD and sacrificed 3 days post inoculation. There was no significant difference in the fetal mortality among all inoculation groups. Placenta showed higher virus titer than fetus and dam's serum in all inoculation groups, and the virus titer of the fetus was lowest in the 15GD group. Histopathological changes and signals of viral RNAs detected by in situ hybridization were observed almost restricted to the spongiotrophoblast layer of the placenta in all inoculation groups, and the signals were strongest in the 11GD group. In the fetus of the11GD group, signals of viral RNAs were also seen in myocardium and hepatocytes. Ultrastructurally, intracytoplasmic aggregations of virus-like particles in crystalline array were observed in trophoblast cells and giant cells in the spongiotrophoblast layer in all inoculation groups.
Asunto(s)
Infecciones por Cardiovirus/virología , Virus de la Encefalomiocarditis/genética , Ratones Endogámicos BALB C/virología , Preñez , Animales , Infecciones por Cardiovirus/sangre , Infecciones por Cardiovirus/patología , Modelos Animales de Enfermedad , Susceptibilidad a Enfermedades/virología , Femenino , Feto/virología , Edad Gestacional , Técnicas Histológicas , Hibridación in Situ , Ratones , Placenta/ultraestructura , Placenta/virología , Embarazo , ARN Viral/genéticaRESUMEN
A 12-year-old male Shiba dog showed anemia and the swelling of systemic lymph nodes. X-ray and post mortal examinations revealed a anterior mediastinal mass. Histologically, the tumor mass consisted of four different elements; cord-like proliferation of cuboidal epithelial cells, tubular or cystic structures lined with ciliated epithelial cells, proliferation of large round-shaped epithelial cells with PAS-slightly positive granular cytoplasm, and diffuse proliferation of neoplastic lymphocytes. Epithelial cells in cord-like or cystic structures were strongly positive for cytokeratin. Granular or foamy cells were negative for all markers examined and had myelin-like bodies in the cytoplasm by electron microscopy. The neoplastic lymphocytes in the tumor mass were considered being derived from concurrent multicentric lymphoma. Based on these findings, the present case was diagnosed as thymoma with a part of granular cell proliferation and concurrent lymphoma cells.