The engineering of antibodies and antibody fragments for affinity maturation, stability, and other biophysical characteristics is a common aspect of therapeutic development. Maturation of antibodies in B cells during the adaptive immune response is the result of a process called somatic hypermutation (SHM), in which the activation-induced cytidine deaminase (AID) acts to introduce mutations into immunoglobulin (Ig) genes. Iterative selection and clonal expansion of B cells containing affinity-enhancing mutations drive an increase in the overall affinity of antibodies. Here we describe the use of SHM coupled with mammalian cell surface display for the maturation of antibodies in vitro and the complementarity of these methods with the mining of immune lineages using next-generation sequencing (NGS).
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http://dx.doi.org/10.1007/978-1-4939-8648-4_24 | DOI Listing |
Int J Biol Macromol
January 2025
Henan Province Engineering Research Center of Innovation for Synthetic Biology, School of Life Sciences and Technology, Xinxiang Medical University, Xinxiang, 453003, Henan, China; School of Basic Medical Sciences, Xinxiang Medical University, Xinxiang, 453003, Henan, China. Electronic address:
The mechanisms underlying antigen receptor germline gene diversification have always been a topic of intensive study. Here, we discovered that the frequency of stem-loop sequences in the antigen receptor germline gene region is remarkably higher than the genomic background. By analyzing these stem-loop sequences' similarity and distribution patterns, we found that clustered regularly interspaced homologous stem-loop pairs (CRIHSP) are widely present on the germline genes of antigen receptors in different species.
View Article and Find Full Text PDFImmunity
January 2025
Garvan Institute of Medical Research, Darlinghurst, NSW, Australia; St Vincent's Clinical School, UNSW Sydney, Sydney, NSW, Australia. Electronic address:
The unexplained association between infection and autoimmune disease is strongest for hepatitis C virus-induced cryoglobulinemic vasculitis (HCV-cryovas). To analyze its origins, we traced the evolution of pathogenic rheumatoid factor (RF) autoantibodies in four HCV-cryovas patients by deep single-cell multi-omic analysis, revealing three sources of B cell somatic mutation converged to drive the accumulation of a large disease-causing clone. A method for quantifying low-affinity binding revealed recurring antibody variable domain combinations created by V(D)J recombination that bound self-immunoglobulin G (IgG) but not viral E2 antigen.
View Article and Find Full Text PDFCell Rep
January 2025
Immunology Program, Memorial Sloan Kettering Cancer Center, New York, NY 10065, USA; Immunology and Microbial Pathogenesis Program, Weill Cornell Graduate School of Medical Sciences, New York, NY 10065, USA; Gerstner Sloan Kettering Graduate School of Biomedical Sciences, New York, NY 10065, USA. Electronic address:
The T-cell-derived cytokine IL-21 is crucial for germinal center (GC) responses, but its precise role in B cell function has remained elusive. Using IL-21 receptor (Il21r) conditional knockout mice and ex vivo culture systems, we demonstrate that IL-21 has dual effects on B cells. While IL-21 induced apoptosis in a STAT3-dependent manner in naive B cells, it promoted the robust proliferation of pre-activated B cells, particularly IgG1 B cells.
View Article and Find Full Text PDFFront Immunol
January 2025
Public Health Sciences Division, Fred Hutchinson Cancer Center, Seattle, WA, United States.
Background: Microsatellite instability-high (MSI-high) tumors comprise ~15% of sporadic colorectal cancers (CRC) and are associated with elevated T cell infiltration. However, the universality of this response across T cell subtypes with distinct functions is unknown.
Methods: Including 1,236 CRC tumors from three observational studies, we conducted T cell profiling using a customized 9-plex (CD3, CD4, CD8, CD45RA, CD45RO, FOXP3, KRT, MKI67, and DAPI) multispectral immunofluorescence assay.
Int Cancer Conf J
January 2025
Department of Urology, The Second Affiliated Hospital of Xi'an Jiaotong University, No. 157 Xiwu Road, Xi'an, 710004 China.
Mismatch repair deficiency (MMRd) or microsatellite instability high (MSI-H) is rare in prostate cancer and more frequently observed in cases with ductal histology. MLH1 copy number loss is extremely rare in MMRd tumors. Herein, we describe a case of prostate ductal adenocarcinoma with MLH1 copy number loss, microsatellite instability high and BRCA2 mutation could derive benefit from immunotherapy plus ADT.
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