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...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 cloning.In this study,we established an AID-based mutant accumulation and sorting system in adherent human cells.Mouse AID gene was first transfected into the human non-small cell lung carcinoma H1299 cells,and a stable cell clone(H1299-AID)was selected.Afterwards,anti-hTNF-αscFv(ATscFv)was transfected into H1299-AID cells and ATscFv was displayed on the surface of H1299-AID cells.By 4-round amplification/flow cytometric sorting for cells with the highest affinities to hTNF-alpha,two ATscFv mutant gene clones were isolated.Compared with the wild type ATscFv,the two mutants were much more efficient in neutralizing cytotoxicity of hTNF-alpha.The 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.展开更多
Antibody diversification is essential for an effective immune response,with somatic hypermutation(SHM)serving as a key molecular process in this adaptation.Activation-induced cytidine deaminase(AID)initiates SHM by in...Antibody diversification is essential for an effective immune response,with somatic hypermutation(SHM)serving as a key molecular process in this adaptation.Activation-induced cytidine deaminase(AID)initiates SHM by inducing DNA lesions,which are ultimately resolved into point mutations,as well as small insertions and deletions(indels).These mutational outcomes contribute to antibody affinity maturation.The mechanisms responsible for generating point mutations and indels involve the base excision repair(BER)and mismatch repair(MMR)pathways,which are well coordinated to maintain genomic integrity while allowing for beneficial mutations to occur.In this regard,translesion synthesis(TLS)polymerases contribute to the diversity of mutational outcomes in antibody genes by enabling the bypass of DNA lesions.This review summarizes our current understanding of the distinct molecular mechanisms that generate point mutations and indels during SHM.Understanding these mechanisms is critical for elucidating the development of broadly neutralizing antibodies(bnAbs)and autoantibodies,and has implications for vaccine design and therapeutics.展开更多
Mantle cell lymphoma （MCL） is an aggressive nonHodgkin＇s lymphoma, originating from naive B-cells. The blastoid MCL tumors often show complex cytogenetic aberrations. In this review, we summarized the data availabl...Mantle cell lymphoma （MCL） is an aggressive nonHodgkin＇s lymphoma, originating from naive B-cells. The blastoid MCL tumors often show complex cytogenetic aberrations. In this review, we summarized the data available on immunoglobulin heavy-chain （IgH） genes rearrangement for their importance in suggesting the MCL normal counterpart B-cell. Some new data suggesting an antigen selection process were also presented in this review.展开更多
A simple immune-based multi-objective optimizer(IBMO) is proposed, and a rigorous running time analysis of IBMO on three proposed bi-objective pseudo-Boolean functions(Bi-Trap, Bi-Plateau and Bi-Jump) is presented. Th...A simple immune-based multi-objective optimizer(IBMO) is proposed, and a rigorous running time analysis of IBMO on three proposed bi-objective pseudo-Boolean functions(Bi-Trap, Bi-Plateau and Bi-Jump) is presented. The running time of a global simple evolutionary multi-objective optimizer(GSEMO) using standard bit mutation operator with IBMO using somatic contiguous hypermutation(CHM) operator is compared with these three functions. The results show that the immune-based hypermutation can significantly beat standard bit mutation on some well-known multi-objective pseudo-Boolean functions. The proofs allow us to understand the relationship between the characteristics of the problems and the features of the algorithms more deeply. These analysis results also give us a good inspiration to analyze and design a bio-inspired search heuristics.展开更多
Pulmonary T and B cells are important for protection of this mucosal barrier site.While viral infections lead to the development of ectopic lymphoid structures highly similar to those in germinal centers in secondary ...Pulmonary T and B cells are important for protection of this mucosal barrier site.While viral infections lead to the development of ectopic lymphoid structures highly similar to those in germinal centers in secondary lymphoid organs,little is known about how T/B cooperation occurs in the unstructured,diffuse tissue infiltrates characteristic of autoimmune diseases and nonviral infections.Using a mouse model of interstitial lung inflammation,we found that naive B cells are directly activated in lung tissue.Despite the absence of any germinal center-like structures,the interaction of B cells with peripheral T helper cells results in efficient somatic hypermutation and class switching.As antigen-presenting cells,macrophages are critical for this process.Unique B-cell repertoires indicated that the lung response was autonomous from the lung-draining lymph node.Only lung GC-like B cells were switched to IgA and had a broader repertoire,making them ideal candidates for producing broadly neutralizing immunoglobulins against respiratory pathogens.展开更多
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 ...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.展开更多
SATB1(Special A-T rich Binding protein 1)is a cell type-specific factor that regulates the genetic network in developing T cells and neurons.In T cells,SATB1 is required for lineage commitment,VDJ recombination,develo...SATB1(Special A-T rich Binding protein 1)is a cell type-specific factor that regulates the genetic network in developing T cells and neurons.In T cells,SATB1 is required for lineage commitment,VDJ recombination,development and maturation.Considering that its expression varies during B-cell differentiation,the involvement of SATB1 needs to be clarified in this lineage.Using 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 subsets.Our 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 repertoire.Second,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,respectively.Third,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.展开更多
基金funded by grants from the Ministry of Science and Technology of People’s Republic of China(Nos.2011CBA00906 and 2011YQ03013404).
文摘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 cloning.In this study,we established an AID-based mutant accumulation and sorting system in adherent human cells.Mouse AID gene was first transfected into the human non-small cell lung carcinoma H1299 cells,and a stable cell clone(H1299-AID)was selected.Afterwards,anti-hTNF-αscFv(ATscFv)was transfected into H1299-AID cells and ATscFv was displayed on the surface of H1299-AID cells.By 4-round amplification/flow cytometric sorting for cells with the highest affinities to hTNF-alpha,two ATscFv mutant gene clones were isolated.Compared with the wild type ATscFv,the two mutants were much more efficient in neutralizing cytotoxicity of hTNF-alpha.The 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.
基金supported by the National Key Research and Development Program of China(2021YFA1301400)the National Natural Science Foundation of China(32370934)the Shanghai Jiao Tong University 2030 Initiative(2030-B23).
文摘Antibody diversification is essential for an effective immune response,with somatic hypermutation(SHM)serving as a key molecular process in this adaptation.Activation-induced cytidine deaminase(AID)initiates SHM by inducing DNA lesions,which are ultimately resolved into point mutations,as well as small insertions and deletions(indels).These mutational outcomes contribute to antibody affinity maturation.The mechanisms responsible for generating point mutations and indels involve the base excision repair(BER)and mismatch repair(MMR)pathways,which are well coordinated to maintain genomic integrity while allowing for beneficial mutations to occur.In this regard,translesion synthesis(TLS)polymerases contribute to the diversity of mutational outcomes in antibody genes by enabling the bypass of DNA lesions.This review summarizes our current understanding of the distinct molecular mechanisms that generate point mutations and indels during SHM.Understanding these mechanisms is critical for elucidating the development of broadly neutralizing antibodies(bnAbs)and autoantibodies,and has implications for vaccine design and therapeutics.
文摘Mantle cell lymphoma （MCL） is an aggressive nonHodgkin＇s lymphoma, originating from naive B-cells. The blastoid MCL tumors often show complex cytogenetic aberrations. In this review, we summarized the data available on immunoglobulin heavy-chain （IgH） genes rearrangement for their importance in suggesting the MCL normal counterpart B-cell. Some new data suggesting an antigen selection process were also presented in this review.
基金the National Natural Science Foundation of China(Nos.61703183,61773410,61375053)the Public Welfare Technology Research Plan of Zhejiang Province(No.LGG19F030010)
文摘A simple immune-based multi-objective optimizer(IBMO) is proposed, and a rigorous running time analysis of IBMO on three proposed bi-objective pseudo-Boolean functions(Bi-Trap, Bi-Plateau and Bi-Jump) is presented. The running time of a global simple evolutionary multi-objective optimizer(GSEMO) using standard bit mutation operator with IBMO using somatic contiguous hypermutation(CHM) operator is compared with these three functions. The results show that the immune-based hypermutation can significantly beat standard bit mutation on some well-known multi-objective pseudo-Boolean functions. The proofs allow us to understand the relationship between the characteristics of the problems and the features of the algorithms more deeply. These analysis results also give us a good inspiration to analyze and design a bio-inspired search heuristics.
基金supported by the German Research Foundation(DFG),project ID 509546995supported by DFG grants HU 1294/8-1,HU 1294/8-2,and SFB 1526-A08(project ID 454193335)+2 种基金funded by the Federal Ministry of Education and Research(BMBF)as part of the German Center for Child and Adolescent Health(DZKJ)under funding codes 01GL2401C,CONAN and TreATgrants from the Leibniz Association(Leibniz Collaborative Excellence,TargArt and ImpACt)supported by Germany’s Excellence Strategy grants EXC2151 and EXC2047.
文摘Pulmonary T and B cells are important for protection of this mucosal barrier site.While viral infections lead to the development of ectopic lymphoid structures highly similar to those in germinal centers in secondary lymphoid organs,little is known about how T/B cooperation occurs in the unstructured,diffuse tissue infiltrates characteristic of autoimmune diseases and nonviral infections.Using a mouse model of interstitial lung inflammation,we found that naive B cells are directly activated in lung tissue.Despite the absence of any germinal center-like structures,the interaction of B cells with peripheral T helper cells results in efficient somatic hypermutation and class switching.As antigen-presenting cells,macrophages are critical for this process.Unique B-cell repertoires indicated that the lung response was autonomous from the lung-draining lymph node.Only lung GC-like B cells were switched to IgA and had a broader repertoire,making them ideal candidates for producing broadly neutralizing immunoglobulins against respiratory pathogens.
文摘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.
文摘SATB1(Special A-T rich Binding protein 1)is a cell type-specific factor that regulates the genetic network in developing T cells and neurons.In T cells,SATB1 is required for lineage commitment,VDJ recombination,development and maturation.Considering that its expression varies during B-cell differentiation,the involvement of SATB1 needs to be clarified in this lineage.Using 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 subsets.Our 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 repertoire.Second,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,respectively.Third,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.
