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1.
Am J Kidney Dis ; 84(1): 120-125, 2024 07.
Artigo em Inglês | MEDLINE | ID: mdl-38266972

RESUMO

Monoclonal immunoglobulin (MIg) crystalline nephropathies are rare lesions resulting from precipitation of MIgs in the kidney as intracellular or extracellular crystals. We describe a patient with multiple myeloma (IgGλ) and diabetes who presented with nephrotic range proteinuria. Kidney biopsy revealed membranous nephropathy superimposed on diabetic glomerulosclerosis. Glomeruli were negative for PLA2R, THSD7A, and NELL-1. Ultrastructurally, the subepithelial deposits were composed of crystals (ranging from rhomboid to rod to needle shaped), which failed to stain for immunoglobulins by routine immunofluorescence but stained for IgG+λ by paraffin immunofluorescence after pronase digestion. RNA-based immunoglobulin repertoire sequencing performed on bone marrow aspirate identified an IgGλ (γ1) clone, which was highly atypical, combining an extensively mutated (23.6%) Ig heavy chain derived from the IGHV1-24 with low pI and unusual mutations and a light chain derived from an extremely rare germline gene (IGLV10-54). This report expands the pathologic spectrum of MIg crystalline nephropathies by describing a unique case of crystalline nephropathy with IgGλ deposits manifesting as membranous nephropathy.


Assuntos
Glomerulonefrite Membranosa , Humanos , Glomerulonefrite Membranosa/patologia , Glomerulonefrite Membranosa/genética , Glomerulonefrite Membranosa/imunologia , Masculino , Cristalização , Imunoglobulina G , Mieloma Múltiplo/complicações , Mieloma Múltiplo/diagnóstico , Pessoa de Meia-Idade , Anticorpos Monoclonais
2.
Kidney Int ; 103(3): 616-626, 2023 03.
Artigo em Inglês | MEDLINE | ID: mdl-36581019

RESUMO

Monoclonal immunoglobulin light chain (LC) crystalline inclusions within podocytes are rare, poorly characterized entities. To provide more insight, we now present the first clinicopathologic series of LC crystalline podocytopathy (LCCP) encompassing 25 patients (68% male, median age 56 years). Most (80%) patients presented with proteinuria and chronic kidney disease, with nephrotic syndrome in 28%. Crystalline keratopathy and Fanconi syndrome were present in 22% and 10%, respectively. The hematologic condition was monoclonal gammopathy of renal significance (MGRS) in 55% and multiple myeloma in 45%. The serum monoclonal immunoglobulin was IgG κappa in 86%. Histologically, 60% exhibited focal segmental glomerulosclerosis (FSGS), often collapsing. Ultrastructurally, podocyte LC crystals were numerous with variable effacement of foot processes. Crystals were also present in proximal tubular cells as light chain proximal tubulopathy (LCPT) in 80% and in interstitial histiocytes in 36%. Significantly, frozen-section immunofluorescence failed to reveal the LC composition of crystals in 88%, requiring paraffin-immunofluorescence or immunohistochemistry, with identification of kappa LC in 87%. The LC variable region gene segment, determined by mass spectrometry of glomeruli or bone marrow plasma cell sequencing, was IGKV1-33 in four and IGKV3-20 in one. Among 21 patients who received anti-plasma cell-directed chemotherapy, 50% achieved a kidney response, which depended on a deep hematologic response. After a median follow-up of 36 months, 26% progressed to kidney failure and 17% died. The mean kidney failure-free survival was 57.6 months and was worse in those with FSGS. In sum, LCCP is rare, mostly associates with IgG κappa MGRS, and frequently has concurrent LCPT, although Fanconi syndrome is uncommon. Paraffin-immunofluorescence and electron microscopy are essential to prevent misdiagnosis as primary FSGS since kidney survival depends on early diagnosis and subsequent clone-directed therapy.


Assuntos
Síndrome de Fanconi , Glomerulosclerose Segmentar e Focal , Nefropatias , Insuficiência Renal , Humanos , Masculino , Pessoa de Meia-Idade , Feminino , Glomerulosclerose Segmentar e Focal/patologia , Síndrome de Fanconi/patologia , Parafina , Rim/patologia , Nefropatias/patologia , Insuficiência Renal/patologia , Imunoglobulina G
3.
Kidney Int ; 101(2): 331-337, 2022 02.
Artigo em Inglês | MEDLINE | ID: mdl-34767830

RESUMO

The diagnostic approach of monoclonal gammopathy of renal significance is based on the detection of a monoclonal immunoglobulin in the blood and urine, and the identification of the underlying clone through bone marrow and/or peripheral blood cytologic and flow cytometry analysis. However, the monoclonal component and its corresponding clone may be undetectable using these routine techniques. Since clone identification is the cornerstone for guiding therapy and assessing disease response, more sensitive methods are required. We recently developed a high-throughput sequencing assay from bone marrow mRNA encoding immunoglobulins (RACE-RepSeq). This technique provides both full-length V(D)J region (variable, diversity and joining genes that generate unique receptors as antigen receptors) of the monoclonal immunoglobulin and the dominant immunoglobulin repertoire. This allows analysis of mutational patterns, immunoglobulin variable gene frequencies and diversity due to somatic hypermutation. Here, we evaluated the diagnostic performance of RACE-RepSeq in 16 patients with monoclonal-associated kidney lesions, and low serum monoclonal immunoglobulin and free light chain levels at diagnosis. Bone marrow immunohistochemical analysis was negative in all 11 patients so tested and 7 of 12 patients had no detectable clone matching the kidney deposits using flow cytometry analysis. By contrast, RACE-RepSeq detected a dominant clonal light chain sequence of matched isotype with respect to kidney deposits in all patients. Thus, high throughput mRNA sequencing appears highly sensitive to detect subtle clonal disorders in monoclonal gammopathy of renal significance and suggest this novel approach could help improve the management of this kidney disease.


Assuntos
Nefropatias , Paraproteinemias , Humanos , Cadeias Leves de Imunoglobulina , Rim/patologia , Nefropatias/diagnóstico , Nefropatias/genética , Nefropatias/terapia , Paraproteinemias/diagnóstico , Paraproteinemias/genética , Paraproteinemias/terapia , RNA
4.
Blood ; 135(20): 1750-1758, 2020 05 14.
Artigo em Inglês | MEDLINE | ID: mdl-32243509

RESUMO

Polyneuropathy, organomegaly, endocrinopathy, monoclonal gammopathy, and skin changes (POEMS) syndrome is a rare multisystem disease resulting from an underlying plasma cell (PC) dyscrasia. The pathophysiology of the disease remains unclear, but the role of the monoclonal immunoglobulin (Ig) light chain (LC) is strongly suspected because of the highly restrictive usage of 2 λ variable (V) domains (IGLV1-40 and IGLV1-44) and the general improvement of clinical manifestations after PC clone-targeted treatment. However, the diagnostic value of Ig LC sequencing, especially in the case of incomplete forms of the disease, remains to be determined. Using a sensitive high-throughput Ig repertoire sequencing on RNA (rapid amplification of cDNA ends-based repertoire sequencing [RACE-RepSeq]), we detected a λ LC monoclonal expansion in the bone marrow (BM) of 83% of patients with POEMS syndrome, including some in whom BM tests routinely performed to diagnose plasma cell dyscrasia failed to detect λ+ monoclonal PCs. Twenty-four (83%) of the 29 LC clonal sequences found were derived from the IGLV1-40 and IGLV1-44 germline genes, as well as 2 from the closely related IGLV1-36 gene, and all were associated with an IGLJ3*02 junction (J) gene, confirming the high restriction of VJ region usage in POEMS syndrome. RACE-RepSeq VJ full-length sequencing additionally revealed original mutational patterns, the strong specificity of which might crucially help establish or eliminate the diagnosis of POEMS syndrome in uncertain cases. Thus, RACE-RepSeq appears as a sensitive, rapid, and specific tool to detect low-abundance PC clones in BM and assign them to POEMS syndrome, with all the consequences for therapeutic options.


Assuntos
Análise Mutacional de DNA/métodos , Sequenciamento de Nucleotídeos em Larga Escala , Cadeias lambda de Imunoglobulina/genética , Síndrome POEMS/genética , Biópsia , Medula Óssea/metabolismo , Medula Óssea/patologia , Mutação em Linhagem Germinativa , Humanos , Cadeias Leves de Imunoglobulina/análise , Cadeias Leves de Imunoglobulina/genética , Cadeias lambda de Imunoglobulina/análise , Linfonodos/metabolismo , Linfonodos/patologia , Técnicas de Diagnóstico Molecular/métodos , Síndrome POEMS/patologia , Análise de Sequência de Proteína
5.
J Am Soc Nephrol ; 27(9): 2748-61, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-26825533

RESUMO

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.


Assuntos
Mesângio Glomerular/metabolismo , Imunoglobulina A/metabolismo , Animais , Formação de Anticorpos , Feminino , Imunoglobulina A/imunologia , Masculino , Camundongos , Conformação Proteica
6.
Proc Natl Acad Sci U S A ; 107(7): 3064-9, 2010 Feb 16.
Artigo em Inglês | MEDLINE | ID: mdl-20133609

RESUMO

Sequentially along B cell differentiation, the different classes of membrane Ig heavy chains associate with the Ig alpha/Ig beta heterodimer within the B cell receptor (BCR). Whether each Ig class conveys specific signals adapted to the corresponding differentiation stage remains debated. We investigated the impact of the forced expression of an IgA-class receptor throughout murine B cell differentiation by knocking in the human C alpha Ig gene in place of the S mu region. Despite expression of a functional BCR, homozygous mutant mice showed a partial developmental blockade at the pro-B/pre-BI and large pre-BII cell stages, with decreased numbers of small pre-BII cells. Beyond this stage, peripheral B cell compartments of reduced size developed and allowed specific antibody responses, whereas mature cells showed constitutive activation and a strong commitment to plasma cell differentiation. Secreted IgA correctly assembled into polymers, associated with the murine J chain, and was transported into secretions. In heterozygous mutants, cells expressing the IgA allele competed poorly with those expressing IgM from the wild-type allele and were almost undetectable among peripheral B lymphocytes, notably in gut-associated lymphoid tissues. Our data indicate that the IgM BCR is more efficient in driving early B cell education and in mucosal site targeting, whereas the IgA BCR appears particularly suited to promoting activation and differentiation of effector plasma cells.


Assuntos
Linfócitos B/citologia , Cadeias alfa de Imunoglobulina/metabolismo , Cadeias mu de Imunoglobulina/metabolismo , Linfopoese/imunologia , Plasmócitos/citologia , Receptores de Antígenos de Linfócitos B/metabolismo , Animais , Linfócitos B/imunologia , Bromodesoxiuridina , Citometria de Fluxo , Marcação de Genes , Humanos , Cadeias alfa de Imunoglobulina/imunologia , Cadeias mu de Imunoglobulina/imunologia , Imuno-Histoquímica , Camundongos , Camundongos Transgênicos , Plasmócitos/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa
7.
iScience ; 26(3): 106260, 2023 Mar 17.
Artigo em Inglês | MEDLINE | ID: mdl-36845033

RESUMO

To understand the fine differential elements that can lead to or prevent acute respiratory distress syndrome (ARDS) in COVID-19 patients, it is crucial to investigate the immune response architecture. We herein dissected the multiple layers of B cell responses by flow cytometry and Ig repertoire analysis from acute phase to recovery. Flow cytometry with FlowSOM analysis showed major changes associated with COVID-19 inflammation such as an increase of double-negative B-cells and ongoing plasma cell differentiation. This paralleled COVID-19-driven expansion of two disconnected B-cell repertoires. Demultiplexing successive DNA and RNA Ig repertoire patterns characterized an early expansion of IgG1 clonotypes with atypically long and uncharged CDR3, the abundance of this inflammatory repertoire being correlated with ARDS and likely pejorative. A superimposed convergent response included convergent anti-SARS-CoV-2 clonotypes. It featured progressively increasing somatic hypermutation together with normal-length or short CDR3 and it persisted until a quiescent memory B-cell stage after recovery.

8.
Eur J Immunol ; 41(12): 3632-44, 2011 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-22076801

RESUMO

B-cell fate and responses are modulated by soluble mediators and direct cellular interactions. Migration properties also vary during differentiation, commitment and activation. In many cells, modulation of responses to stimuli involves cell surface glycans, whose architecture depends on the simultaneous expression of multiple enzymes. By looking at the glycosylation-related gene expression patterns among B-cell populations, we determined in this study that the strongest variations were observed for CSGalNAcT-1 and EXTL1. These are enzymes involved in the biosynthesis of alternative forms of glycosaminoglycans (GAGs), namely chondroitin sulfate and heparan sulfate, respectively. These two enzymes showed inverse fluctuations in progenitors, resting B cells and activated B cells, suggesting a developmentally regulated switch between chondroitin and heparan sulfate synthesis. To explore whether these variations contributed to optimal B-cell differentiation, we overexpressed EXTL1 in the B-cell lineage of transgenic mice, yielding a partial differentiation blockade at the pro-B to pre-B transition. In the periphery, this defect was almost fully compensated for in vivo, with normal-size B-cell compartments and normal serum immunoglobulin levels in the transgenic EXTL1 mice. The peripheral B cells from EXTL1 transgenics were only affected with regard to their in vitro responses to polyclonal activation, showing reduced proliferation. Together the data suggest that despite their low amounts in lymphocytes, the heparan sulfate chains decorating the endogenous GAGs appear to be regulators of B-cell physiology.


Assuntos
Linfócitos B/citologia , Linfócitos B/metabolismo , Glicosaminoglicanos/biossíntese , Animais , Linfócitos B/imunologia , Diferenciação Celular/genética , Diferenciação Celular/imunologia , Linhagem Celular , Linhagem Celular Tumoral , Sulfatos de Condroitina/biossíntese , Sulfatos de Condroitina/imunologia , Variação Genética , Glicosaminoglicanos/genética , Heparitina Sulfato/biossíntese , Heparitina Sulfato/imunologia , Imunoglobulinas/biossíntese , Imunoglobulinas/imunologia , Ativação Linfocitária/genética , Ativação Linfocitária/imunologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Transgênicos , N-Acetilgalactosaminiltransferases/genética , N-Acetilgalactosaminiltransferases/imunologia , N-Acetilglucosaminiltransferases/genética , N-Acetilglucosaminiltransferases/imunologia , Proteínas Supressoras de Tumor/genética , Proteínas Supressoras de Tumor/imunologia
9.
Haematologica ; 97(11): 1686-94, 2012 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-22689689

RESUMO

BACKGROUND: While most antibody-based therapies use IgG because of their well-known biological properties, some functional limitations of these antibodies call for the development of derivatives with other therapeutic functions. Although less abundant than IgG in serum, IgA is the most abundantly produced Ig class in humans. Besides the specific targeting of its dimeric form to mucosal areas, IgA was shown to recruit polymorphonuclear neutrophils against certain targets more efficiently than does IgG1. DESIGN AND METHODS: In this study, we investigated the various pathways by which anti-tumor effects can be mediated by anti-CD20 IgA against lymphoma cells. RESULTS: We found that polymeric human IgA was significantly more effective than human IgG1 in mediating direct killing or growth inhibition of target cells in the absence of complement. We also demonstrated that this direct killing was able to indirectly induce the classical pathway of the complement cascade although to a lesser extent than direct recruitment of complement by IgG. Recruitment of the alternative complement pathway by specific IgA was also observed. In addition to activating complement for lysis of lymphoma cell lines or primary cells from patients with lymphoma, we showed that monomeric anti-CD20 IgA can effectively protect mice against tumor development in a passive immunization strategy and we demonstrated that this protective effect may be enhanced in mice expressing the human FcαRI receptor on their neutrophils. CONCLUSIONS: We show that anti-CD20 IgA antibodies have original therapeutic properties against lymphoma cells, with strong direct effects, ability to recruit neutrophils for cell cytotoxicity and even recruitment of complement, although largely through an indirect way.


Assuntos
Anticorpos Antineoplásicos/farmacologia , Antígenos CD20/imunologia , Antígenos CD/imunologia , Imunoglobulina A/farmacologia , Linfoma/imunologia , Receptores Fc/imunologia , Animais , Anticorpos Antineoplásicos/imunologia , Antígenos CD/genética , Humanos , Imunoglobulina A/imunologia , Imunoglobulina G/imunologia , Células Jurkat , Linfoma/tratamento farmacológico , Linfoma/genética , Linfoma/patologia , Camundongos , Camundongos Transgênicos , Infiltração de Neutrófilos/efeitos dos fármacos , Infiltração de Neutrófilos/genética , Infiltração de Neutrófilos/imunologia , Neutrófilos/imunologia , Neutrófilos/patologia , Receptores Fc/genética
10.
J Infect Public Health ; 15(12): 1455-1458, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36403401

RESUMO

The immune response is a key player in the course of SARS-CoV-2 infection, and is often seriously dysfunctional in severe Coronavirus Disease 2019. The hyperinflammatory status has been described to be accompanied by the appearance of autoantibodies. In a lethal COVID-19 infection, we observed the emergence of a de novo natural alloantibody which targeted the M antigen from the MNS blood group on red blood cells (RBC) without evidence of any cross-reaction with SARS-CoV-2 antigens. This IgM lambda alloantibody was unmutated and unswitched. Here, we describe for the first time the emergence of a bystander de novo natural alloantibody against RBCs in a severe COVID-19 patient, highlighting the extra-follicular humoral response reported in these cases.


Assuntos
Antígenos de Grupos Sanguíneos , COVID-19 , Humanos , SARS-CoV-2 , Eritrócitos
11.
Cytometry B Clin Cytom ; 102(1): 62-69, 2022 01.
Artigo em Inglês | MEDLINE | ID: mdl-33634586

RESUMO

BACKGROUND: Differential diagnosis of Waldenström macroglobulinemia (WM) with other indolent B-cell malignancies is still a challenge. Here, we propose an original and simple analysis of routine flow cytometry (FCM) unraveling the characteristic ongoing plasma cell (PC) differentiation of WM tumor B-cells. METHODS: FCM analysis of both B-cells and PC was performed on a series of 77 patients with IgM peak. MYD88 and CXCR4 mutations were studied using an allele-specific PCR and by high throughput sequencing. RESULTS: Twenty seven (35%), 46 (58%) and 4 (5%) patients were classified as WM, IgM monoclonal gammopathy of undetermined significance (MGUS) or other B-NHL respectively. MYD88 mutation was found in 25/27 WM (93%) and in 29/46 MGUS (63%). Using FCM, monotypic B-cells were found in 27/27 WM (100%) and 34/46 MGUS (74%). Monotypic CD138pos/CD38pos PCs were detected in 23/27 WM (85%) and 25/46 MGUS (54%). Highlighting the ongoing PC differentiation of WM tumor B-cells by FCM, we evidenced a CD138 expression continuum between monotypic B-cells and PCs. This pattern remained absent in control samples and was significantly associated with higher IgM peaks (p = 6.10-5 ) and MYD88 mutations (p = 10-3 ) in both WM and MGUS cases. CONCLUSIONS: FCM exploration of both B-cells and PC led to identify a CD138 expression continuum as an objective marker of ongoing PC differentiation of WM tumor cells and was strongly associated with increased IgM peak levels and MYD88 mutations. This approach could contribute to place FCM at the forefront of WM diagnosis.


Assuntos
Fator 88 de Diferenciação Mieloide , Sindecana-1/genética , Macroglobulinemia de Waldenstrom , Citometria de Fluxo , Humanos , Imunoglobulina M/genética , Mutação/genética , Fator 88 de Diferenciação Mieloide/genética , Plasmócitos/patologia , Macroglobulinemia de Waldenstrom/diagnóstico , Macroglobulinemia de Waldenstrom/genética
12.
J Clin Med ; 10(14)2021 Jul 16.
Artigo em Inglês | MEDLINE | ID: mdl-34300307

RESUMO

IgA nephropathy (IgAN) is the most common primary glomerulonephritis in the world. It was first described in 1968 by Jean Berger and Nicole Hinglais as the presence of intercapillary deposits of IgA. Despite this simple description, patients with IgAN may present very broad clinical features ranging from the isolated presence of IgA in the mesangium without clinical or biological manifestations to rapidly progressive kidney failure. These features are associated with a variety of histological lesions, from the discrete thickening of the mesangial matrix to diffuse cell proliferation. Immunofluorescence on IgAN kidney specimens shows the isolated presence of IgA or its inconsistent association with IgG and complement components. This clinical heterogeneity of IgAN clearly echoes its complex and multifactorial pathophysiology in humans, inviting further analyses of its various aspects through the use of experimental models. Small-animal models of IgAN provide the most pertinent strategies for studying the multifactorial aspects of IgAN pathogenesis and progression. Although only primates have the IgA1 subclass, several murine models have been developed in which various aspects of immune responses are deregulated and which are useful in the understanding of IgAN physiopathology as well as in the assessment of IgAN therapeutic approaches. In this manuscript, we review all murine IgAN models developed since 1968 and discuss their remarkable contribution to understanding the disease.

13.
Med Sci (Paris) ; 37(1): 35-40, 2021 Jan.
Artigo em Francês | MEDLINE | ID: mdl-33492216

RESUMO

Mucosal immunity has to deal with a patchy mix of commensal but also eventually pathogenic bugs. Immunoglobulins of the A class (IgA) are opposing to this duality a functional balance going from tolerance to protective response or even to hyper-inflammation. Recent reports have shown the binding of polyreactive natural IgA, but also of affinity maturated protective IgA to the commensal microbiota, to superantigens and also to vaccinal antigens. Diverse types of humoral responses thus altogether contribute to the homeostasis of mucosal immunity. Their knowledge has to be taken into consideration for defining strategies of immuno-intervention, for mucosal vaccination as much as for immunotherapy of chronic inflammatory bowel disease.


TITLE: Homéostasie de la réponse IgA et microbiote. ABSTRACT: L'immunité muqueuse s'établit en réponse à un ensemble de microorganismes qui sont surtout commensaux mais aussi, parfois, pathogènes. À cette dualité, les immunoglobulines de classe A (IgA) opposent une balance fonctionnelle allant de la tolérance à la protection, voire à une hyper-inflammation. Des travaux récents ont révélé la liaison d'IgA polyréactives naturelles ou, à l'inverse, d'IgA spécifiquement affines et protectrices, au microbiote commensal, mais aussi à des super-antigènes ou encore à des vaccins muqueux. Différents types de réponse humorale s'associent ainsi pour composer, ensemble, l'homéostasie de l'immunité muqueuse. Leur connaissance devrait ainsi influencer les stratégies de vaccination muqueuse et également les immunothérapies ciblant les maladies inflammatoires chroniques de l'intestin.


Assuntos
Imunidade/fisiologia , Imunoglobulina A/metabolismo , Microbiota/fisiologia , Animais , Microbioma Gastrointestinal/imunologia , Microbioma Gastrointestinal/fisiologia , Homeostase/fisiologia , Humanos , Tolerância Imunológica/fisiologia , Imunidade nas Mucosas/fisiologia , Doenças Inflamatórias Intestinais/imunologia , Doenças Inflamatórias Intestinais/microbiologia , Mucosa Intestinal/imunologia , Mucosa Intestinal/microbiologia , Microbiota/imunologia
14.
Med Sci (Paris) ; 25(12): 1149-54, 2009 Dec.
Artigo em Francês | MEDLINE | ID: mdl-20035696

RESUMO

The properties of monoclonal antibodies explain why they are such a successful class of therapeutic molecules. However, pionneered initial antibodies were of murine origin and triggered an immune response which limited the therapeutic potential of the antibody and generated deleterious effects. Consequently, tremendous efforts have been developped to engineer these murine Ig by introducing human sequences in vitro, or in vivo by humanization of murine antibodies, leading to chimeric immunoglobulins, and more recently generation of fully human antibodies in transgenic mice with a more or less diversified V repertoire. These approaches have led to the development of an increasing number of these chimeric or humanized monoclonal antibodies entering pharmaceutical pipelines.


Assuntos
Anticorpos Monoclonais/imunologia , Camundongos Transgênicos/imunologia , Animais , Anticorpos Anti-Idiotípicos/biossíntese , Anticorpos Monoclonais/biossíntese , Anticorpos Monoclonais/genética , Anticorpos Monoclonais/isolamento & purificação , Anticorpos Neutralizantes/biossíntese , Diversidade de Anticorpos , Previsões , Genes de Imunoglobulinas , Humanos , Hibridomas/imunologia , Região Variável de Imunoglobulina/genética , Região Variável de Imunoglobulina/imunologia , Camundongos , Engenharia de Proteínas , Proteínas Recombinantes de Fusão/biossíntese , Proteínas Recombinantes de Fusão/imunologia , Especificidade da Espécie , Transgenes
15.
Methods Mol Biol ; 901: 295-317, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22723109

RESUMO

Physiology usually combines polyclonal antibodies of multiple classes in a single humoral response. Beyond their common ability to bind antigens, these various classes of human immunoglobulins carry specific functions which can each serve specific goals. In many cases, the function of a monoclonal therapeutic antibody may thus be modulated according to the class of its constant domains. Depending on the immunoglobulin class, different functional assays will be used in order to evaluate the functional activity of a monoclonal antibody.


Assuntos
Anticorpos/uso terapêutico , Imunoglobulinas/imunologia , Animais , Anticorpos/efeitos adversos , Anticorpos/imunologia , Anticorpos/metabolismo , Anticorpos Monoclonais/efeitos adversos , Anticorpos Monoclonais/imunologia , Anticorpos Monoclonais/metabolismo , Anticorpos Monoclonais/uso terapêutico , Apoptose/efeitos dos fármacos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Humanos , Imunoglobulinas/efeitos adversos , Imunoglobulinas/metabolismo , Imunoglobulinas/uso terapêutico , Camundongos
16.
Methods Mol Biol ; 901: 149-59, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22723099

RESUMO

Mice are widely available laboratory animals that can easily be used for the production of antibodies against a broad range of antigens, using well-defined immunization protocols. Such an approach allows optimal in vivo affinity maturation of the humoral response. In addition, high-affinity antibodies arising in this context can readily be further characterized and produced as monoclonals after immortalizing and selecting specific antibody-producing cells through hybridoma derivation. Using such conventional strategies combined with mice that are either genetically engineered to carry humanized immunoglobulin (Ig) genes or engrafted with a human immune system, it is thus easy to obtain and immortalize clones that produce either fully human Ig or antibodies associating variable (V) domains with selected antigen specificities to customized human-like constant regions, with defined effector functions. In some instances, where there is a need for in vivo functional assays of a single antibody with a known specificity, it might be of interest to transiently express that gene in mice by in vivo gene transfer. This approach allows a rapid functional assay. More commonly, mice are used to obtain a diversified repertoire of antibody specificities after immunization by producing antibody molecules in the mouse B cell lineage from mouse strains with transgene Ig genes which are of human, humanized, or chimeric origin. After in vivo maturation of the immune response, this will lead to the secretion of antibodies with optimized antigen binding sites, associated to the desired human constant domains. This chapter focuses on two simple methods: (1) to obtain such humanized Ig mice and (2) to transiently express a human Ig gene in mice using hydrodynamics-based transfection.


Assuntos
Anticorpos Monoclonais Humanizados/metabolismo , Animais , Anticorpos Monoclonais Humanizados/genética , Genes de Imunoglobulinas/genética , Humanos , Camundongos , Camundongos Transgênicos
17.
Nat Med ; 17(11): 1456-65, 2011 Oct 23.
Artigo em Inglês | MEDLINE | ID: mdl-22019886

RESUMO

Anemia because of insufficient production of and/or response to erythropoietin (Epo) is a major complication of chronic kidney disease and cancer. The mechanisms modulating the sensitivity of erythroblasts to Epo remain poorly understood. We show that, when cultured with Epo at suboptimal concentrations, the growth and clonogenic potential of erythroblasts was rescued by transferrin receptor 1 (TfR1)-bound polymeric IgA1 (pIgA1). Under homeostatic conditions, erythroblast numbers were increased in mice expressing human IgA1 compared to control mice. Hypoxic stress of these mice led to increased amounts of pIgA1 and erythroblast expansion. Expression of human IgA1 or treatment of wild-type mice with the TfR1 ligands pIgA1 or iron-loaded transferrin (Fe-Tf) accelerated recovery from acute anemia. TfR1 engagement by either pIgA1 or Fe-Tf increased cell sensitivity to Epo by inducing activation of mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3-kinase (PI3K) signaling pathways. These cellular responses were mediated through the TfR1-internalization motif, YXXΦ. Our results show that pIgA1 and TfR1 are positive regulators of erythropoiesis in both physiological and pathological situations. Targeting this pathway may provide alternate approaches to the treatment of ineffective erythropoiesis and anemia.


Assuntos
Anemia/fisiopatologia , Proliferação de Células , Eritroblastos/fisiologia , Eritropoese/fisiologia , Imunoglobulina A/metabolismo , Animais , Células Cultivadas , Eritroblastos/citologia , Eritroblastos/efeitos dos fármacos , Eritropoetina/farmacologia , Humanos , Hipóxia/metabolismo , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Camundongos Transgênicos , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Fosfatidilinositol 3-Quinases/metabolismo , Receptores da Transferrina/metabolismo , Transdução de Sinais/fisiologia , Transferrina/farmacologia
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