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Affinity maturation of anti-TNF-alpha scFv with somatic hypermutation in non-B cells

Protein & Cell 2012年第6期3卷 460-469页

作者： Shaopeng Chen Junkang Qiu Chuan Chen Chunchun Liu Yuheng Liu Lili An Junying Jia Jie Tang Lijun Wu Haiying Hang Key Laboratory for Protein and Peptide Pharmaceuticals National Laboratory of BiomacromoleculesInstitute of BiophysicsChinese Academy of SciencesBeijing 100101China Key Laboratory of Ion Beam Bioengineering Institute of Technical Biology and Agriculture EngineeringChinese Academy of SciencesHefei 230031China Core Facility Center Institute of BiophysicsChinese Academy of SciencesBeijing 100101China􀀍

Activation-induced cytidine deaminase(AID)is required for the generation of antibody diversity through initiat-ing both somatic hypermutation(SHM)and class switch recombination.A few research groups have success-fully used the feature of AID for generating mutant li-braries in directed evolution of target proteins in B cells in vitro.B cells,cultured in suspension,are not con-venient for transfection and *** this study,we established an AID-based mutant accumulation and sorting system in adherent human *** AID gene was first transfected into the human non-small cell lung carcinoma H1299 cells,and a stable cell clone(H1299-AID)was ***,anti-hTNF-αscFv(ATscFv)was transfected into H1299-AID cells and ATscFv was displayed on the surface of H1299-AID *** 4-round amplification/flow cytometric sorting for cells with the highest affinities to hTNF-alpha,two ATscFv mutant gene clones were *** with the wild type ATscFv,the two mutants were much more efficient in neutralizing cytotoxicity of *** results indicate that directed evolution by somatic hypermutation can be carried out in adherent non-B cells,which makes directed evolution in mammalian cells easier and more efficient.

关键词： antibody activation-induced cytidine deaminase(AID) somatic hypermutation affinity maturation TNF-alpha

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Generation and repair of AID-initiated DNA lesions in B lymphocytes

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Frontiers of Medicine 2014年第2期8卷 201-216页

作者： Zhangguo Chen Jing H. Wang Integrated Department of Immunology University of Colorado Anschutz Medical Campus and National Jewish Health Denver CO 80206USA

Activation-induced deaminase （AID） initiates the secondary antibody diversification process in B lymphocytes. In mammalian B cells, this process includes somatic hypermutation （SHM） and class switch recombination （CSR）, both of which require AID. AID induces U：G mismatch lesions in DNA that are subsequently converted into point mutations or DNA double stranded breaks during SHM/CSR. In a physiological context, AID targets immunogiobulin （Ig） loci to mediate SHM/CSR. However, recent studies reveal genome-wide access of AID to numerous non-Ig loci. Thus, AID poses a threat to the genome of B cells if AID-initiated DNA lesions cannot be properly repaired. In this review, we focus on the molecular mechanisms that regulate the specificity of AID targeting and the repair pathways responsible for processing AID-initiated DNA lesions.

关键词： class switch recombination somatic hypermutation activation-induced deaminase DNA repair genomic instability

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A dual function for the chromatin organizer Special A-T rich Binding Protein 1 in B-lineage cells

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Cellular & Molecular Immunology 2023年第10期20卷 1114-1126页

作者： Morgane Thomas Charlotte Bruzeau Ophélie Alyssa Martin Justine Pollet Sébastien Bender Claire Carrion Sandrine Le Noir Eric Pinaud Laboratoire Contrôle de la Réponse Immune B et des Lymphoproliférations(CRIBL) Universitéde LimogesCNRS UnitéMixte de Recherche 7276Inserm Unité1262LimogesFrance Centre Hospitalier Universitaire Dupuytren Service d’ImmunopathologieLimogesFrance Centre Hospitalier Universitaire de Limoges Centre National de l’Amylose AL et Autres Maladies par Dépôt d’Immunoglobulines MonoclonalesLimogesFrance Laboratoire Suivi des Therapies Innovantes Institut de Genetique HumaineUMR 9002 CNRS-UMMontpellierFrance

SATB1(Special A-T rich Binding protein 1)is a cell type-specific factor that regulates the genetic network in developing T cells and *** T cells,SATB1 is required for lineage commitment,VDJ recombination,development and *** that its expression varies during B-cell differentiation,the involvement of SATB1 needs to be clarified in this *** a KO mouse model in which SATB1 was deleted from the pro-B-cell stage,we examined the consequences of SATB1 deletion in naive and activated B-cell *** model indicates first,unlike its essential function in T cells,that SATB1 is dispensable for B-cell development and the establishment of a broad IgH ***,we show that SATB1 exhibits an ambivalent function in mature B cells,acting sequentially as a positive and negative regulator of Ig gene transcription in naive and activated cells,***,our study indicates that the negative regulatory function of SATB1 in B cells extends to the germinal center response,in which this factor limits somatic hypermutation of Ig genes.

关键词： B cells Nuclear factor Ambivalent somatic hypermutation

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