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1.
Cell Rep ; 43(2): 113705, 2024 Feb 27.
Artículo en Inglés | MEDLINE | ID: mdl-38307025

RESUMEN

Nerve growth factor receptor (NGFR) is expressed by follicular dendritic cells (FDCs). However, the role of NGFR in the humoral response is not well defined. Here, we study the effect of Ngfr loss on lymph node organization and function, demonstrating that Ngfr depletion leads to spontaneous germinal center (GC) formation and an expansion of the GC B cell compartment. In accordance with this effect, stromal cells are altered in Ngfr-/- mice with a higher frequency of FDCs, characterized by CD21/35, MAdCAM-1, and VCAM-1 overexpression. GCs are located ectopically in Ngfr-/- mice, with lost polarization together with impaired high-affinity antibody production and an increase in circulating autoantibodies. We observe higher levels of autoantibodies in Bcl2 Tg/Ngfr-/- mice, concomitant with a higher incidence of autoimmunity and lower overall survival. Our work shows that NGFR is involved in maintaining GC structure and function, participating in GC activation, antibody production, and immune tolerance.


Asunto(s)
Receptor de Factor de Crecimiento Nervioso , Receptores de Factor de Crecimiento Nervioso , Animales , Ratones , Autoanticuerpos , Células Dendríticas Foliculares , Centro Germinal
2.
Eur J Immunol ; 54(1): e2350626, 2024 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-37837385

RESUMEN

To better understand the stoichiometry of CD95L required to trigger apoptotic and nonapoptotic signals, we generated several CD95L concatemers from dimer to hexamer conjugated via a flexible link (GGGGS)2 . These ligands reveal that although the hexameric structure is the best stoichiometry to trigger cell death, a dimer is sufficient to induce the apoptotic response in CD95-sensitive Jurkat cells. Interestingly, only trimeric and hexameric forms can implement a potent Ca2+ response, suggesting that while CD95 aggregation controls the implementation of the apoptotic signal, both aggregation and conformation are required to implement the Ca2+ pathway.


Asunto(s)
Apoptosis , Receptor fas , Humanos , Apoptosis/fisiología , Proteína Ligando Fas , Células Jurkat
3.
Front Immunol ; 14: 1155906, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37359540

RESUMEN

Introduction: In mature B cells, activation-induced deaminase reshapes Ig genes through somatic hypermutation and class switch recombination of the Ig heavy chain (IgH) locus under control of its 3' cis-regulatory region (3'RR). The 3'RR is itself transcribed and can undergo "locus suicide recombination" (LSR), then deleting the constant gene cluster and terminating IgH expression. The relative contribution of LSR to B cell negative selection remains to be determined. Methods: Here, we set up a knock-in mouse reporter model for LSR events with the aim to get clearer insights into the circumstances triggering LSR. In order to explore the consequences of LSR defects, we reciprocally explored the presence of autoantibodies in various mutant mouse lines in which LSR was perturbed by the lack of Sµ or of the 3'RR. Results: Evaluation of LSR events in a dedicated reporter mouse model showed their occurrence in various conditions of B cell activation, notably in antigen-experienced B cells Studies of mice with LSR defects evidenced increased amounts of self-reactive antibodies. Discussion: While the activation pathways associated with LSR are diverse, in vivo as well as in vitro, this study suggests that LSR may contribute to the elimination of self-reactive B cells.


Asunto(s)
Linfocitos B , Suicidio , Ratones , Animales , Cadenas Pesadas de Inmunoglobulina/genética , Cadenas Pesadas de Inmunoglobulina/metabolismo , Cambio de Clase de Inmunoglobulina/genética , Antígenos/metabolismo
4.
Cancers (Basel) ; 14(21)2022 Oct 29.
Artículo en Inglés | MEDLINE | ID: mdl-36358756

RESUMEN

Upregulated expression of the anti-apoptotic BCL2 oncogene is a common feature of various types of B-cell malignancies, from lymphoma to leukemia or myeloma. It is currently unclear how the various patterns of deregulation observed in pathology eventually impact the phenotype of malignant B cells and their microenvironment. Follicular lymphoma (FL) is the most common non-Hodgkin lymphoma arising from malignant germinal center (GC) B-cells, and its major hallmark is the t(14:18) translocation occurring in B cell progenitors and placing the BCL2 gene under the control of the immunoglobulin heavy chain locus regulatory region (IgH 3'RR), thus exposing it to constitutive expression and hypermutation. Translocation of BCL2 onto Ig light chain genes, BCL2 gene amplification, and other mechanisms yielding BCL2 over-expression are, in contrast, rare in FL and rather promote other types of B-cell lymphoma, leukemia, or multiple myeloma. In order to assess the impact of distinct BCL2 deregulation patterns on B-cell fate, two mouse models were designed that associated BCL2 and its full P1-P2 promoter region to either the IgH 3'RR, within a "3'RR-BCL2" transgene mimicking the situation seen in FL, or an Ig light chain locus context, through knock-in insertion at the Igκ locus ("Igκ-BCL2" model). While linkage to the IgH 3' RR mostly yielded expression in GC B-cells, the Igκ-driven up-regulation culminated in plasmablasts and plasma cells, boosting the plasma cell in-flow and the accumulation of long-lived plasma cells. These data demonstrate that the timing and level of BCL2 deregulation are crucial for the behavior of B cells inside GC, an observation that could strongly impact the lymphomagenesis process triggered by secondary genetic hits.

5.
Cell Death Dis ; 13(10): 895, 2022 10 23.
Artículo en Inglés | MEDLINE | ID: mdl-36274061

RESUMEN

CD95 is a death receptor that can promote oncogenesis through molecular mechanisms that are not fully elucidated. Although the mature CD95 membrane receptor is considered to start with the arginine at position 17 after elimination of the signal peptide, this receptor can also be cleaved by MMP7 upstream of its leucine at position 37. This post-translational modification occurs in cancer cells but also in normal cells such as peripheral blood leukocytes. The non-cleaved CD95 amino-terminal region consists in a disordered domain and its in silico reconstitution suggests that it might contribute to receptor aggregation and thereby, regulate the downstream death signaling pathways. In agreement with this molecular modeling analysis, the comparison of CD95-deficient cells reconstituted with full-length or N-terminally truncated CD95 reveals that the loss of the amino-terminal region of CD95 impairs the initial steps of the apoptotic signal while favoring the induction of pro-survival signals, including the PI3K and MAPK pathways.


Asunto(s)
Metaloproteinasa 7 de la Matriz , Receptor fas , Receptor fas/genética , Receptor fas/metabolismo , Metaloproteinasa 7 de la Matriz/metabolismo , Apoptosis/fisiología , Leucina , Fosfatidilinositol 3-Quinasas/metabolismo , Señales de Clasificación de Proteína , Arginina
6.
Leuk Lymphoma ; 63(9): 2114-2125, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35481805

RESUMEN

The 3' regulatory region (3'RR) located downstream from the Cα gene is the conductor of transcription, accessibility, and remodeling of the IgH locus at mature B-cell stages. Convincing demonstrations of the essential contributions of the 3'RR in B-cell lymphomagenesis have been provided by mouse models which bring the oncogene c-Myc under the 3'RR transcriptional control. In this study, we developed a mouse model of CD138+ plasma B-cell lymphomas. If the KI of c-myc directly into Cα just 5' to the 3'RR in iMycCα mice produced B-cell lymphomas with low kinetics, we enforced c-myc production in iMycCα mice by the generation of homozygous c-myc transgenic mice. Our results show that homozygous iMycCα mice lead to a mouse model of plasma CD138+ B-cell lymphomas with interesting and wide transcriptomic similarities to human multiple myeloma and appropriated emergence kinetics that can be used to test new experimental therapeutic approaches.


Asunto(s)
Cadenas Pesadas de Inmunoglobulina , Linfoma de Células B , Animales , Linfocitos B/patología , Modelos Animales de Enfermedad , Humanos , Cadenas Pesadas de Inmunoglobulina/genética , Linfoma de Células B/genética , Linfoma de Células B/patología , Ratones , Ratones Transgénicos , Secuencias Reguladoras de Ácidos Nucleicos
7.
J Allergy Clin Immunol ; 149(5): 1795-1801, 2022 05.
Artículo en Inglés | MEDLINE | ID: mdl-34740604

RESUMEN

BACKGROUND: Allergy regroups numerous complex and various diseases classified as IgE-dependent or non-IgE-dependent hypersensitivities. IgEs are expressed as membrane and secreted forms by B cells and plasma cells, respectively. In IgE-mediated hypersensitivity, IgE secretion and binding to the high-affinity IgE receptor FcεRI on effector cells are responsible for the onset of allergic symptoms; in contrast, surface IgE expression as a B-cell receptor is barely detectable. OBJECTIVE: Our aim was to test an innovative antisense approach to reducing IgE secretion. METHODS: We designed an antisense oligonucleotide (ASO) targeting the polyadenylation signal of human secreted IgE to redirect IgE transcript polyadenylation from the secreted form to the membrane form. ASO treatments were performed on B cells from transgenic mice expressing humanized IgE (InEps mice), as well as on human primary B cells and myeloma cells. In vivo ASO delivery was tested by using an InEps mouse model. RESULTS: We demonstrated that treatment with a morpholino ASO targeting the secreted IgE polyadenylation signal drastically decreased IgE secretion and inversely increased membrane IgE mRNA expression. In addition, ASO treatment induced apoptosis of IgE-expressing U266 myeloma cells, and RNA sequencing revealed attenuation of their plasma cell phenotype. Remarkably, systemic administration of an ASO coupled with Pip6a as an arginine-rich cell-penetrating peptide decreased IgE secretion in vivo. CONCLUSION: Altogether, this ASO strategy could be an effective way to decrease IgE secretion and allergic symptoms in patients with IgE-dependent allergies, and it could also promote allergen tolerance through apoptosis of IgE+ antibody-secreting cells.


Asunto(s)
Hipersensibilidad , Mieloma Múltiple , Animales , Supervivencia Celular , Humanos , Inmunoglobulina E/metabolismo , Ratones , Oligonucleótidos Antisentido/farmacología , Células Plasmáticas/metabolismo , Poliadenilación , Receptores de IgE/metabolismo
8.
Front Immunol ; 12: 641692, 2021.
Artículo en Inglés | MEDLINE | ID: mdl-34017329

RESUMEN

Activating mutations of MYD88 (MYD88L265P being the far most frequent) are found in most cases of Waldenström macroglobulinemia (WM) as well as in various aggressive B-cell lymphoma entities with features of plasma cell (PC) differentiation, such as activated B-cell type diffuse large B-cell lymphoma (DLBCL). To understand how MYD88 activation exerts its transformation potential, we developed a new mouse model in which the MYD88L252P protein, the murine ortholog of human MYD88L265P, is continuously expressed in CD19 positive B-cells together with the Yellow Fluorescent Protein (Myd88L252P mice). In bone marrow, IgM B and plasma cells were expanded with a CD138 expression continuum from IgMhigh CD138low to IgMlow CD138high cells and the progressive loss of the B220 marker. Serum protein electrophoresis (SPE) longitudinal analysis of 40 Myd88L252P mice (16 to 56 weeks old) demonstrated that ageing was first associated with serum polyclonal hyper gammaglobulinemia (hyper Ig) and followed by a monoclonal immunoglobulin (Ig) peak related to a progressive increase in IgM serum levels. All Myd88L252P mice exhibited spleen enlargement which was directly correlated with the SPE profile and was maximal for monoclonal Ig peaks. Myd88L252P mice exhibited very early increased IgM PC differentiation. Most likely due to an early increase in the Ki67 proliferation index, IgM lymphoplasmacytic (LP) and plasma cells continuously expanded with age being first associated with hyper Ig and then with monoclonal Ig peak. This peak was consistently associated with a spleen LP-like B-cell lymphoma. Clonal expression of both membrane and secreted µ chain isoforms was demonstrated at the mRNA level by high throughput sequencing. The Myd88L252P tumor transcriptomic signature identified both proliferation and canonical NF-κB p65/RelA activation. Comparison with MYD88L265P WM showed that Myd88L252P tumors also shared the typical lymphoplasmacytic transcriptomic signature of WM bone marrow purified tumor B-cells. Altogether these results demonstrate for the first time that continuous MYD88 activation is specifically associated with clonal transformation of differentiating IgM B-cells. Since MYD88L252P targets the IgM PC differentiation continuum, it provides an interesting preclinical model for development of new therapeutic approaches to both WM and aggressive MYD88 associated DLBCLs.


Asunto(s)
Diferenciación Celular/inmunología , Inmunoglobulina M/inmunología , Mutación Missense , Factor 88 de Diferenciación Mieloide/inmunología , Proteínas de Neoplasias/inmunología , Células Plasmáticas/inmunología , Sustitución de Aminoácidos , Animales , Diferenciación Celular/genética , Humanos , Inmunoglobulina M/genética , Linfoma de Células B Grandes Difuso/genética , Linfoma de Células B Grandes Difuso/inmunología , Linfoma de Células B Grandes Difuso/patología , Ratones , Ratones Transgénicos , Factor 88 de Diferenciación Mieloide/genética , Proteínas de Neoplasias/genética , Células Plasmáticas/patología
9.
Blood ; 136(14): 1645-1656, 2020 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-32559766

RESUMEN

Light chain (LC) deposition disease (LCDD) is a rare disorder characterized by glomerular and peritubular amorphous deposits of a monoclonal immunoglobulin LC, leading to nodular glomerulosclerosis and nephrotic syndrome. We developed a transgenic model using site-directed insertion of the variable domain of a pathogenic human LC gene into the mouse immunoglobulin κ locus, ensuring its production by all plasma cells (PCs). High free LC levels were achieved after backcrossing with mice presenting increased PC differentiation and no immunoglobulin heavy chain production. Our mouse model recapitulates the characteristic features of LCDD, including progressive glomerulosclerosis, nephrotic-range proteinuria, and finally kidney failure. The variable domain of the LC bears alone the structural properties involved in its pathogenicity. RNA sequencing conducted on PCs demonstrated that LCDD LC induces endoplasmic reticulum stress, likely accounting for the high efficiency of proteasome inhibitor-based therapy. Accordingly, reduction of circulating pathogenic LC was efficiently achieved and not only preserved renal function but also partially reversed kidney lesions. Finally, transcriptome analysis of presclerotic glomeruli revealed that proliferation and extracellular matrix remodeling represented the first steps of glomerulosclerosis, paving the way for future therapeutic strategies in LCDD and other kidney diseases featuring diffuse glomerulosclerosis, particularly diabetic nephropathy.


Asunto(s)
Cadenas Ligeras de Inmunoglobulina/metabolismo , Paraproteinemias/diagnóstico , Paraproteinemias/etiología , Animales , Biomarcadores , Ciclo Celular/genética , Modelos Animales de Enfermedad , Estrés del Retículo Endoplásmico , Matriz Extracelular , Citometría de Flujo , Perfilación de la Expresión Génica , Orden Génico , Marcación de Gen , Vectores Genéticos/genética , Cadenas Ligeras de Inmunoglobulina/genética , Cadenas kappa de Inmunoglobulina/genética , Cadenas kappa de Inmunoglobulina/metabolismo , Inmunohistoquímica , Riñón/metabolismo , Riñón/patología , Pruebas de Función Renal , Glomérulos Renales/metabolismo , Glomérulos Renales/patología , Glomérulos Renales/ultraestructura , Ratones , Ratones Transgénicos , Paraproteinemias/complicaciones , Paraproteinemias/mortalidad , Agregado de Proteínas , Agregación Patológica de Proteínas , Insuficiencia Renal/diagnóstico , Insuficiencia Renal/etiología , Insuficiencia Renal/metabolismo , Insuficiencia Renal/mortalidad
10.
J Am Soc Nephrol ; 30(7): 1238-1249, 2019 07.
Artículo en Inglés | MEDLINE | ID: mdl-31227634

RESUMEN

BACKGROUND: IgA nephropathy (IgAN) often follows infections and features IgA mesangial deposition. Polymeric IgA deposits in the mesangium seem to have varied pathogenic potential, but understanding their pathogenicity remains a challenge. Most mesangial IgA1 in human IgAN has a hypogalactosylated hinge region, but it is unclear whether this is required for IgA deposition. Another important question is the role of adaptive IgA responses and high-affinity mature IgA antibodies and whether low-affinity IgA produced by innate-like B cells might also yield mesangial deposits. METHODS: To explore the effects of specific qualitative variations in IgA and whether altered affinity maturation can influence IgA mesangial deposition and activate complement, we used several transgenic human IgA1-producing models with IgA deposition, including one lacking the DNA-editing enzyme activation-induced cytidine deaminase (AID), which is required in affinity maturation. Also, to explore the potential role of the IgA receptor CD89 in glomerular inflammation, we used a model that expresses CD89 in a pattern observed in humans. RESULTS: We found that human IgA induced glomerular damage independent of CD89. When comparing mice able to produce high-affinity IgA antibodies with mice lacking AID-enabled Ig affinity maturation, we found that IgA deposition and complement activation significantly increased and led to IgAN pathogenesis, although without significant proteinuria or hematuria. We also observed that hinge hypoglycosylation was not mandatory for IgA deposition. CONCLUSIONS: In a mouse model of IgAN, compared with high-affinity IgA, low-affinity innate-like IgA, formed in the absence of normal antigen-driven maturation, was more readily involved in IgA glomerular deposition with pathogenic effects.


Asunto(s)
Afinidad de Anticuerpos , Mesangio Glomerular/metabolismo , Glomerulonefritis por IGA/etiología , Inmunoglobulina A/metabolismo , Animales , Antígenos CD/fisiología , Activación de Complemento , Citidina Desaminasa/fisiología , Mesangio Glomerular/patología , Glomerulonefritis por IGA/inmunología , Glicosilación , Humanos , Inmunoglobulina A/toxicidad , Ratones , Receptores Fc/fisiología
11.
Blood Adv ; 2(3): 252-262, 2018 02 13.
Artículo en Inglés | MEDLINE | ID: mdl-29437640

RESUMEN

The immunoglobulin heavy chain (IgH) 3' regulatory region (3'RR) superenhancer controls B2 B-cell IgH transcription and cell fate at the mature stage but not early repertoire diversity. B1 B cells represent a small percentage of total B cells differing from B2 B cells by several points such as precursors, development, functions, and regulation. B1 B cells act at the steady state to maintain homeostasis in the organism and during the earliest phases of an immune response, setting them at the interface between innate and acquired immunity. We investigated the role of the 3'RR superenhancer on B1 B-cell fate. Similar to B2 B cells, the 3'RR controls µ transcription and cell fate in B1 B cells. In contrast to B2 B cells, 3'RR deletion affects B1 B-cell late repertoire diversity. Thus, differences exist for B1 and B2 B-cell 3'RR control during B-cell maturation. For the first time, these results highlight the contribution of the 3'RR superenhancer at this interface between innate and acquired immunity.


Asunto(s)
Linfocitos B/inmunología , Cadenas Pesadas de Inmunoglobulina/genética , Secuencias Reguladoras de Ácidos Nucleicos/fisiología , Inmunidad Adaptativa , Animales , Linfocitos B/citología , Línea Celular , Genes de las Cadenas Pesadas de las Inmunoglobulinas/genética , Inmunidad Innata , Ratones , Análisis de Secuencia de ADN , Transcripción Genética , Recombinación V(D)J
13.
J Am Soc Nephrol ; 27(9): 2748-61, 2016 09.
Artículo en Inglés | MEDLINE | ID: mdl-26825533

RESUMEN

IgA1 mesangial deposition is the hallmark of IgA nephropathy and Henoch-Schönlein purpura, the onset of which often follows infections. Deposited IgA has been reported as polymeric, J chain associated, and often, hypogalactosylated but with no information concerning the influence of the IgA repertoire or the link between immune stimuli and IgA structure. We explored these issues in the α1KI mouse model, which produces polyclonal human IgA1 prone to mesangial deposition. Compared with mice challenged by a conventional environment, mice in a specific pathogen-free environment had less IgA deposition. However, serum IgA of specific pathogen-free mice showed more galactosylation and much lower polymerization. Notably, wild-type, α1KI, and even J chain-deficient mice showed increased polymeric serum IgA on exposure to pathogens. Strict germfree conditions delayed but did not completely prevent deposition; mice housed in these conditions had very low serum IgA levels and produced essentially monomeric IgA. Finally, comparing monoclonal IgA1 that had different variable regions and mesangial deposition patterns indicated that, independently of glycosylation and polymerization, deposition might also depend on IgA carrying specific variable domains. Together with IgA quantities and constant region post-translational modifications, repertoire changes during immune responses might, thus, modulate IgA propensity to deposition. These IgA features are not associated with circulating immune complexes and C3 deposition and are more pertinent to an initial IgA deposition step preceding overt clinical symptoms in patients.


Asunto(s)
Mesangio Glomerular/metabolismo , Inmunoglobulina A/metabolismo , Animales , Formación de Anticuerpos , Femenino , Inmunoglobulina A/inmunología , Masculino , Ratones , Conformación Proteica
14.
Blood ; 126(6): 757-65, 2015 Aug 06.
Artículo en Inglés | MEDLINE | ID: mdl-26113545

RESUMEN

Randall-type heavy chain deposition disease (HCDD) is a rare disorder characterized by glomerular and peritubular amorphous deposits of a truncated monoclonal immunoglobulin heavy chain (HC) bearing a deletion of the first constant domain (CH1). We created a transgenic mouse model of HCDD using targeted insertion in the immunoglobulin κ locus of a human HC extracted from a HCDD patient. Our strategy allows the efficient expression of the human HC in mouse B and plasma cells, and conditional deletion of the CH1 domain reproduces the major event underlying HCDD. We show that the deletion of the CH1 domain dramatically reduced serum HC levels. Strikingly, even with very low serum level of truncated monoclonal HC, histologic studies revealed typical Randall-type renal lesions that were absent in mice expressing the complete human HC. Bortezomib-based treatment resulted in a strong decrease of renal deposits. We further demonstrated that this efficient response to proteasome inhibitors mostly relies on the presence of the isolated truncated HC that sensitizes plasma cells to bortezomib through an elevated unfolded protein response (UPR). This new transgenic model of HCDD efficiently recapitulates the pathophysiologic features of the disease and demonstrates that the renal damage in HCDD relies on the production of an isolated truncated HC, which, in the absence of a LC partner, displays a high propensity to aggregate even at very low concentration. It also brings new insights into the efficacy of proteasome inhibitor-based therapy in this pathology.


Asunto(s)
Antineoplásicos/farmacología , Ácidos Borónicos/farmacología , Enfermedad de las Cadenas Pesadas/tratamiento farmacológico , Cadenas Pesadas de Inmunoglobulina/química , Enfermedades Renales/tratamiento farmacológico , Inhibidores de Proteasoma/farmacología , Agregación Patológica de Proteínas/tratamiento farmacológico , Pirazinas/farmacología , Secuencia de Aminoácidos , Animales , Bortezomib , Modelos Animales de Enfermedad , Expresión Génica , Sitios Genéticos , Enfermedad de las Cadenas Pesadas/genética , Enfermedad de las Cadenas Pesadas/inmunología , Enfermedad de las Cadenas Pesadas/patología , Humanos , Cadenas Pesadas de Inmunoglobulina/genética , Cadenas Pesadas de Inmunoglobulina/inmunología , Enfermedades Renales/genética , Enfermedades Renales/inmunología , Enfermedades Renales/patología , Glomérulos Renales/efectos de los fármacos , Glomérulos Renales/inmunología , Glomérulos Renales/metabolismo , Glomérulos Renales/patología , Ratones , Ratones Transgénicos , Datos de Secuencia Molecular , Células Plasmáticas/efectos de los fármacos , Células Plasmáticas/inmunología , Células Plasmáticas/metabolismo , Células Plasmáticas/patología , Complejo de la Endopetidasa Proteasomal/efectos de los fármacos , Complejo de la Endopetidasa Proteasomal/metabolismo , Agregación Patológica de Proteínas/genética , Agregación Patológica de Proteínas/inmunología , Agregación Patológica de Proteínas/patología , Estructura Terciaria de Proteína , Eliminación de Secuencia , Respuesta de Proteína Desplegada/efectos de los fármacos , Respuesta de Proteína Desplegada/genética , Respuesta de Proteína Desplegada/inmunología
15.
Oncotarget ; 6(7): 4845-52, 2015 Mar 10.
Artículo en Inglés | MEDLINE | ID: mdl-25742787

RESUMEN

We report that the IgH 3' regulatory region (3'RR) has no role on µ chain transcription and pre-BCR expression in B cell progenitors. In contrast, analysis of heterozygous IgH aΔ3'RR/bwt mice indicated that the 3'RR controls µ chain transcripts in mature splenocytes and impacts membrane IgM density without obvious effect on BCR signals (colocalisation with lipid rafts and phosphorylation of Erk and Akt after BCR crosslinking). Deletion of the 3'RR modulates the B cell fate to less marginal zone B cells. In conclusion, the 3'RR is dispensable for pre-BCR expression and necessary for optimal commitments toward the marginal zone B cell fate. These results reinforce the concept of a dual regulation of the IgH locus transcription and accessibility by 5' elements at immature B cell stages, and by the 3'RR as early as the resting mature B cell stage and then along further activation and differentiation.


Asunto(s)
Linfocitos B/fisiología , Cadenas Pesadas de Inmunoglobulina/genética , Cadenas mu de Inmunoglobulina/genética , Animales , Linfocitos B/metabolismo , Humanos , Cadenas Pesadas de Inmunoglobulina/metabolismo , Cadenas mu de Inmunoglobulina/biosíntesis , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Secuencias Reguladoras de Ácidos Nucleicos
16.
Oncotarget ; 5(19): 8995-9006, 2014 Oct 15.
Artículo en Inglés | MEDLINE | ID: mdl-25229630

RESUMEN

Deregulation of c-myc by translocation onto immunoglobulin (Ig) loci can promote B cell malignant proliferations with phenotypes as diverse as acute lymphoid leukemia, Burkitt lymphoma, diffuse large B cell lymphoma, myeloma... The B cell receptor (BCR) normally providing tonic signals for cell survival and mitogenic responses to antigens, can also contribute to lymphomagenesis upon sustained ligand binding or activating mutations. BCR signaling varies among cell compartments and BCR classes. For unknown reasons, some malignancies associate with expression of either IgM or class-switched Ig. We explored whether an IgA BCR, with strong tonic signaling, would affect lymphomagenesis in c-myc IgH 3'RR transgenic mice prone to lymphoproliferations. Breeding c-myc transgenics in a background where IgM expression was replaced with IgA delayed lymphomagenesis. By comparison to single c-myc transgenics, lymphomas from double mutant animals were more differentiated and less aggressive, with an altered transcriptional program. Larger tumor cells more often expressed CD43 and CD138, which culminated in a plasma cell phenotype in 10% of cases. BCR class-specific signals thus appear to modulate lymphomagenesis and may partly explain the observed association of specific Ig classes with human B cell malignancies of differential phenotype, progression and prognosis.


Asunto(s)
Inmunoglobulina A/genética , Inmunoglobulina M/genética , Linfoma/genética , Proteínas Proto-Oncogénicas c-myc/genética , Receptores de Antígenos de Linfocitos B/genética , Animales , Linfocitos B/patología , Supervivencia Celular/genética , Transformación Celular Neoplásica/genética , Humanos , Inmunoglobulina A/inmunología , Cadenas Pesadas de Inmunoglobulina/genética , Inmunoglobulina M/inmunología , Leucosialina/biosíntesis , Linfoma/patología , Ratones , Ratones Endogámicos C57BL , Ratones Transgénicos , Receptores de Antígenos de Linfocitos B/biosíntesis , Receptores de Antígenos de Linfocitos B/inmunología , Sindecano-1/biosíntesis
17.
Nephrol Dial Transplant ; 27(12): 4368-77, 2012 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-23024220

RESUMEN

BACKGROUND: Fanconi syndrome (FS) is a rare renal disorder featuring proximal tubule dysfunction that may occur following tubular reabsorption of a monoclonal light chain (LC), in patients with multiple myeloma. FS may precede the recognition of multiple myeloma by several years. In most cases, crystalline inclusions of monoclonal κ LCs are observed within the lysosomes of proximal tubular cells (PTCs) and probably participate in their functional alteration. METHODS: To investigate the mechanism implicated in proximal tubule dysfunction, we compared the effects of κ LC-CHEB obtained from a patient with myeloma-associated FS to those of control κ LC-BON obtained from a patient without evidence of FS, on the viability and proliferation of two different PTC lines. RESULTS: Our data suggest that the tubular atrophy in myeloma-associated FS does not result from increased apoptosis of PTCs, but from their impaired capacity to proliferate and renew. Indeed, in vitro incubation of cultured PTCs with physiological amounts of the nephrotoxic κ LC-CHEB was sufficient to cause a depression in DNA synthesis and in cell proliferation. This effect was observed neither with control κ LC-BON nor in the absence of κ LC. CONCLUSIONS: The reduced turnover of PTCs may affect tubular repair and regeneration. In addition, the reduced proliferation of myeloma cells producing the same monoclonal κ LC might explain the frequent association of FS with smoldering multiple myeloma.


Asunto(s)
Proliferación Celular , Síndrome de Fanconi/fisiopatología , Cadenas kappa de Inmunoglobulina/fisiología , Túbulos Renales Proximales/patología , Atrofia , Células Cultivadas , Síndrome de Fanconi/etiología , Humanos
18.
Nephrol Dial Transplant ; 26(12): 3930-7, 2011 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21459784

RESUMEN

BACKGROUND: Henoch-Schönlein purpura (HSP) and IgA nephropathy (IgAN) are characterized by mesangial deposition of polyclonal IgA eventually showing aberrant glycosylation, affinity for mesangial cells and/or co-precipitation with antigen, bacterial peptides, autoantibodies or soluble receptors. IgA were also suggested to be negatively charged and predominantly of λ type but rarely in a monoclonal form. METHODS: A gammopathy case with HSP provided us with a unique molecularly defined nephritogenic IgA1λ. Immunological analysis, biological activities, glycosylation analysis and finally IgA sequence were determined. RESULTS: Compared to IgA1 from healthy subjects or IgAN patients, IgA1 CAT showed hyposialylation but no hypogalactosylation, in agreement with underexpression of sialyltransferase genes by the plasma cell clone. IgA variable domains had low pIs with negatively charged complementarity-determining regions. Weak reactivity appeared against the cationic autoantigen lactoferrin, which was, however, absent from kidney deposits. Deposition also occurred in mice upon injection of only the polymeric form of IgA1 CAT, despite whether or not co-injected with lactoferrin. CONCLUSIONS: This monoclonal model of IgA nephritogenicity strongly suggests that beside hinge region glycosylation, V domains play a role in IgA stability and pathogenicity and supports the hypothesis that responses against cationic epitopes from pathogens or autoantigens may select negatively charged complementarity-determining regions prone either to bind charged structures of the mesangium or to promote by themselves IgA aggregation and deposition.


Asunto(s)
Mesangio Glomerular , Glomerulonefritis por IGA/inmunología , Inmunoglobulina A , Paraproteinemias/inmunología , Anciano , Femenino , Glomerulonefritis por IGA/complicaciones , Humanos , Vasculitis por IgA/complicaciones
19.
J Immunol ; 178(1): 407-15, 2007 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-17182579

RESUMEN

Certain individuals are resistant to HIV-1 infection, despite repeated exposure to the virus. Although protection against HIV-1 infection in a small proportion of Caucasian individuals is associated with mutant alleles of the CCR5 HIV-1 coreceptor, the molecular mechanism underlying resistance in repeatedly HIV-1-exposed, uninfected individuals (EU) is unclear. In this study, we performed complementary transcriptome and proteome analyses on peripheral blood T cells, and plasma or serum from EU, their HIV-1-infected sexual partners, and healthy controls, all expressing wild-type CCR5. We report that activated T cells from EU overproduce several proteins involved in the innate immunity response, principally those including high levels of peroxiredoxin II, a NK-enhancing factor possessing strong anti-HIV activity, and IL-22, a cytokine involved in the production of acute-phase proteins such as the acute-phase serum amyloid A (A-SAA). Cell supernatants and serum levels of these proteins were up-regulated in EU. Moreover, a specific biomarker for EU detected in plasma was identified as an 8.6-kDa A-SAA cleavage product. Incubation of in vitro-generated myeloid immature dendritic cells with A-SAA resulted in CCR5 phosphorylation, down-regulation of CCR5 expression, and strongly decreased susceptibility of these cells to in vitro infection with a primary HIV-1 isolate. Taken together, these results suggest new correlates of EU protection and identify a cascade involving IL-22 and the acute phase protein pathway that is associated with innate host resistance to HIV infection.


Asunto(s)
Infecciones por VIH/inmunología , VIH-1 , Interleucinas/metabolismo , Proteína Amiloide A Sérica/metabolismo , Linfocitos T/inmunología , Proteínas de Fase Aguda/análisis , Proteínas de Fase Aguda/metabolismo , Biomarcadores/sangre , Células Dendríticas/inmunología , Femenino , Perfilación de la Expresión Génica , Infecciones por VIH/genética , Humanos , Inmunidad Innata/genética , Interleucinas/sangre , Interleucinas/genética , Activación de Linfocitos , Masculino , Células Mieloides/inmunología , Peroxidasas/sangre , Peroxidasas/genética , Peroxidasas/metabolismo , Peroxirredoxinas , Fosforilación , Proteoma/análisis , Proteoma/genética , Proteoma/metabolismo , Receptores CCR5/metabolismo , Proteína Amiloide A Sérica/análisis , Transcripción Genética , Regulación hacia Arriba , Interleucina-22
20.
AIDS ; 19(9): 897-905, 2005 Jun 10.
Artículo en Inglés | MEDLINE | ID: mdl-15905670

RESUMEN

OBJECTIVES: The soluble HIV-1 gp120 envelope glycoprotein, after being shed from infected cells, can cross-link its receptors on both HIV-1 infected and non-infected target cells, leading to their activation. We have assessed the impact of soluble gp120 on viral replication in CD4+/CXCR4+ T cells, via its effects on Tat-mediated transactivation of the HIV-1/LTR. MATERIALS AND METHODS: Primary cord blood-derived CD4+/CXCR4+ T cells were stimulated with soluble recombinant gp120 (rgp120) from the HIV-1/HXB2 clone. The level of gene or protein expression was assessed by serial analysis gene expression (SAGE), reverse transcriptase-polymerase chain reaction, western blotting or flow-cytometry analysis. Cellular division of rgp120-stimulated T cells was assessed by CFDA-SE labeling. Long terminal repeat (LTR) activity and HIV infection level were respectively measured by a chemiluminescent beta-gal Reporter Gene Assay and by p24 determination. RESULTS: We have demonstrated that rgp120 activates both PKCepsilon and its upstream effector PI3K/Akt, involved in the HIV-1 replication process. Moreover, rgp120 enhances the gene, as well as protein expression of the cellular Tat cofactors Tat-Sf1 and SPT5 in primary CD4+/CXCR4+ T cells. Finally, stimulation of HIV-1 infected T cells with rgp120 was found to result in both a higher LTR-activity and an increased production of viral particles. CONCLUSION: Taken together, these results show that soluble gp120 contributes to HIV-1 replication and dissemination, via the activation of multiple cell signaling pathways and the induction of Tat-cofactor expression, underscoring its potential as a therapeutic target in HIV-1-mediated pathogenesis.


Asunto(s)
Linfocitos T CD4-Positivos/virología , Productos del Gen tat/metabolismo , Proteína gp120 de Envoltorio del VIH/farmacología , VIH-1/fisiología , Replicación Viral/efectos de los fármacos , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/metabolismo , División Celular/efectos de los fármacos , Células Cultivadas , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/inmunología , Duplicado del Terminal Largo de VIH/inmunología , Humanos , Activación de Linfocitos/efectos de los fármacos , Receptores CXCR4/metabolismo , Proteínas Recombinantes/farmacología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Transducción de Señal/inmunología , Solubilidad , Activación Transcripcional/efectos de los fármacos , Replicación Viral/inmunología , Productos del Gen tat del Virus de la Inmunodeficiencia Humana
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