Affinity Maturation Drives Epitope Spreading and Generation of Proinflammatory Anti–Citrullinated Protein Antibodies in Rheumatoid Arthritis

Elliott, Serra E.; Kongpachith, Sarah; Lingampalli, Nithya; Adamska, Julia Z.; Cannon, Bryan J.; Mao, Rong; Blum, Lisa K.; Robinson, William H.

doi:10.1002/art.40587

Cited by 66 publications

(59 citation statements)

References 51 publications

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“…Cit‐tets were generated by incubating phycoerythrin (PE)–conjugated streptavidin with biotinylated citrullinated peptides (a list of the peptides used is provided in Supplementary Table 2, available on the Arthritis & Rheumatology web site at http://onlinelibrary.wiley.com/doi/10.1002/art.40760/abstract). These peptides were previously shown to be targeted by ACPAs in RA sera (for more details, see Supplementary Methods, http://onlinelibrary.wiley.com/doi/10.1002/art.40760/abstract).…”

Section: Methodsmentioning

confidence: 99%

“…Cell sorting of CD19+CD20−CD3−CD14−IgD−CD27+CD38 high plasmablasts, which were either IgG+ or IgA+, was performed in the same manner as used previously in our prior studies . A mix of the PE‐conjugated cit‐tets was included to identify ACPA‐producing plasmablasts (details in Supplementary Methods, http://onlinelibrary.wiley.com/doi/10.1002/art.40760/abstract).…”

Section: Methodsmentioning

confidence: 99%

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Affinity Maturation of the Anti–Citrullinated Protein Antibody Paratope Drives Epitope Spreading and Polyreactivity in Rheumatoid Arthritis

Kongpachith

Lingampalli

et al. 2019

Arthritis & Rheumatology

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Objective. Anti-citrullinated protein antibodies (ACPAs) are a hallmark of rheumatoid arthritis (RA). While epitope spreading of the serum ACPA response is believed to contribute to RA pathogenesis, little is understood regarding how this phenomenon occurs. This study was undertaken to analyze the antibody repertoires of individuals with RA to gain insight into the mechanisms leading to epitope spreading of the serum ACPA response in RA.Methods. Plasmablasts from the blood of 6 RA patients were stained with citrullinated peptide tetramers to identify ACPA-producing B cells by flow cytometry. Plasmablasts were single-cell sorted and sequenced to obtain antibody repertoires. Sixty-nine antibodies were recombinantly expressed, and their anticitrulline reactivities were characterized using a cyclic citrullinated peptide enzyme-linked immuosorbent assay and synovial antigen arrays. Thirty-six mutated antibodies designed either to represent ancestral antibodies or to test paratope residues critical for binding, as determined from molecular modeling studies, were also tested for anticitrulline reactivities.Results. Clonally related monoclonal ACPAs and their shared ancestral antibodies each exhibited differential reactivity against citrullinated antigens. Molecular modeling identified residues within the complementarity-determining region loops and framework regions predicted to be important for citrullinated antigen binding. Affinity maturation resulted in mutations of these key residues, which conferred binding to different citrullinated epitopes and/or increased polyreactivity to citrullinated epitopes.Conclusion. These results demonstrate that the different somatic hypermutations accumulated by clonally related B cells during affinity maturation alter the antibody paratope to mediate epitope spreading and polyreactivity of the ACPA response in RA, suggesting that these may be key properties that likely contribute to the pathogenicity of ACPAs.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Affinity Maturation of the Anti–Citrullinated Protein Antibody Paratope Drives Epitope Spreading and Polyreactivity in Rheumatoid Arthritis

Kongpachith

Lingampalli

et al. 2019

Arthritis & Rheumatology

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“…73,79 Both rheumatoid factor and anti-cyclic citrullinated peptide (CCP) reactivity of B-cells and PC have been studied by flow cytometry in RA through surface and intracellular staining, respectively. 71,78,82…”

Section: Flow Cytometrymentioning

confidence: 99%

Understanding and measuring human B‐cell tolerance and its breakdown in autoimmune disease

Cashman

Jenks

Woodruff

et al. 2019

Immunological Reviews

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It is well established that normal immune systems are endowed with a significant degree of autoreactivity. Accordingly, effective tolerogenic mechanisms are required to censor autoreactive cells and to ultimately prevent their differentiation into pathogenic effector cells. In this review, we will discuss how immunological tolerance is enforced in the human B-cell compartment. We shall also discuss the breakdown of B-cell tolerance in human autoimmune diseases.Finally, we will review different experimental approaches to measure B-cell tolerance in human disease. | B-CELL TOLER AN CE . CHECK P OINTS AND MECHANIS MS . LE SSONS LE ARNED THROUG H MOUS E S TUD IE SMouse models have been instrumental in defining multiple checkpoints and distinct mechanisms underpinning the enforcement of B-cell tolerance. Tolerance checkpoints are classified into central and peripheral mechanisms based on anatomical location and stage AbstractThe maintenance of immunological tolerance of B lymphocytes is a complex and critical process that must be implemented as to avoid the detrimental development of autoreactivity and possible autoimmunity. Murine models have been invaluable to elucidate many of the key components in B-cell tolerance; however, translation to human homeostatic and pathogenic immune states can be difficult to assess.Functional autoreactive, flow cytometric, and single-cell cloning assays have proven to be critical in deciphering breaks in B-cell tolerance within autoimmunity; however, newer approaches to assess human B-cell tolerance may prove to be vital in the further exploration of underlying tolerance defects. In this review, we supply a comprehensive overview of human immune tolerance checkpoints with associated mechanisms of enforcement, and highlight current and future methodologies which are likely to benefit future studies into the mechanisms that become defective in human autoimmune conditions.

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“…In fact, the RA field has been especially active in making use of mAb derived from the immunoglobulin sequences from patient‐derived B cells and plasma cells. Thus, several recently published studies , including our own, extend the first generation of RA patient B cell–derived mAb .…”

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confidence: 99%

“…Although our study delved deeply into the characteristics of plasma cell–derived ACPAs from synovial fluid, the sample studied was from a single individual, and our investigation was therefore not powered to address the matter of frequencies. We instead acknowledge that valid enumeration and phenotyping of antigen‐specific B cells between patients, time points, and anatomic sites represent an interesting area for future investigation, especially given the use of tetramers and high‐throughput approaches.…”

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confidence: 99%

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Stéen

Klareskog

2019

Arthritis & Rheumatology

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Affinity Maturation Drives Epitope Spreading and Generation of Proinflammatory Anti–Citrullinated Protein Antibodies in Rheumatoid Arthritis

Cited by 66 publications

References 51 publications

Affinity Maturation of the Anti–Citrullinated Protein Antibody Paratope Drives Epitope Spreading and Polyreactivity in Rheumatoid Arthritis

Affinity Maturation of the Anti–Citrullinated Protein Antibody Paratope Drives Epitope Spreading and Polyreactivity in Rheumatoid Arthritis

Understanding and measuring human B‐cell tolerance and its breakdown in autoimmune disease

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