scholarly journals Functional Enrichment and Analysis of Antigen-Specific Memory B Cell Antibody Repertoires in PBMCs

2019 ◽  
Author(s):  
Eric Waltari ◽  
Aaron McGeever ◽  
Peter S. Kim ◽  
Krista M. McCutcheon
Keyword(s):  
B Cell ◽  
Plasma Cells ◽  
Functional Enrichment ◽  
Cell Repertoire ◽  
Culture Methods ◽  
Memory B Cell ◽  
Low Frequencies ◽  
Specific Memory ◽  
B Cell Repertoire ◽  
And Function

Phenotypic screening of antigen-specific antibodies in human blood is a common diagnostic test for infectious agents and a correlate of protection after vaccination. In addition to long-lived antibody secreting plasma cells residing in the bone marrow, memory B cells are a latent source of antigen-experienced, long-term immunity that can be found at low frequencies in circulating PBMCs. Assessing the genotype, clonal frequency, quality, and function of antibodies resulting from an individual’s persistent memory B cell repertoire can help inform the success or failure of immune protection. We have applied ELISPOT cell culture methods to functionally expand the memory repertoire from PBMCs and clonally map monoclonal antibodies from this population. We show that combining deep sequencing of stimulated memory B cell repertoires with retrieving single antigen-specific cells is a promising approach in evaluating the latent, functional B cell memory in PBMCs.

scholarly journals Memory B cell repertoire for recognition of evolving SARS-CoV-2 spike

2021 ◽  
Author(s):  
Pei Tong ◽  
Avneesh Gautam ◽  
Ian Windsor ◽  
Meghan Travers ◽  
Yuezhou Chen ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
B Cell ◽  
Cell Receptor ◽  
B Cell Receptor ◽  
Cell Repertoire ◽  
Memory B Cell ◽  
Group Assignment ◽  
Specific Memory ◽  
B Cell Repertoire ◽  
Protective Antibodies

ABSTRACTMemory B cell reserves can generate protective antibodies against repeated SARS-CoV-2 infections, but with an unknown reach from original infection to antigenically drifted variants. We charted memory B cell receptor-encoded monoclonal antibodies (mAbs) from 19 COVID-19 convalescent subjects against SARS-CoV-2 spike (S) and found 7 major mAb competition groups against epitopes recurrently targeted across individuals. Inclusion of published and newly determined structures of mAb-S complexes identified corresponding epitopic regions. Group assignment correlated with cross-CoV-reactivity breadth, neutralization potency, and convergent antibody signatures. mAbs that competed for binding the original S isolate bound differentially to S variants, suggesting the protective importance of otherwise-redundant recognition. The results furnish a global atlas of the S-specific memory B cell repertoire and illustrate properties conferring robustness against emerging SARS-CoV-2 variants.

Development and Function of the Early B Cell Repertoire

1992 ◽  
Vol 8 (2-3) ◽  
pp. 247-257 ◽  
Author(s):  
John E. Kearney ◽  
Joachim Bartels ◽  
Ann Marie Hamilton ◽  
Agnès Lehuen ◽  
Nanette Solvason ◽  
...  
Keyword(s):  
B Cell ◽  
Cell Repertoire ◽  
B Cell Repertoire ◽  
And Function

scholarly journals Different B cell subpopulations show distinct patterns in their IgH repertoire metrics

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Marie Ghraichy ◽  
Valentin von Niederhäusern ◽  
Aleksandr Kovaltsuk ◽  
Jacob D Galson ◽  
Charlotte M Deane ◽  
...  
Keyword(s):  
B Cell ◽  
In Silico ◽  
Plasma Cells ◽  
Somatic Hypermutation ◽  
Bioinformatic Analysis ◽  
Cell Repertoire ◽  
Humoral Immune ◽  
B Cell Repertoire ◽  
Cell Subpopulations ◽  
The Individual

Several human B-cell subpopulations are recognized in the peripheral blood, which play distinct roles in the humoral immune response. These cells undergo developmental and maturational changes involving VDJ recombination, somatic hypermutation and class switch recombination, altogether shaping their immunoglobulin heavy chain (IgH) repertoire. Here, we sequenced the IgH repertoire of naïve, marginal zone, switched and plasma cells from 10 healthy adults along with matched unsorted and in silico separated CD19+ bulk B cells. Using advanced bioinformatic analysis and machine learning, we show that sorted B cell subpopulations are characterised by distinct repertoire characteristics on both the individual sequence and the repertoire level. Sorted subpopulations shared similar repertoire characteristics with their corresponding in silico separated subsets. Furthermore, certain IgH repertoire characteristics correlated with the position of the constant region on the IgH locus. Overall, this study provides unprecedented insight over mechanisms of B cell repertoire control in peripherally circulating B cell subpopulations.

scholarly journals Tissue memory B cell repertoire analysis after ALVAC/AIDSVAX B/E gp120 immunization of rhesus macaques

JCI Insight ◽  
2016 ◽  
Vol 1 (20) ◽  
Author(s):  
Kan Luo ◽  
Hua-Xin Liao ◽  
Ruijun Zhang ◽  
David Easterhoff ◽  
Kevin Wiehe ◽  
...  
Keyword(s):  
B Cell ◽  
Rhesus Macaques ◽  
Cell Repertoire ◽  
Memory B Cell ◽  
Repertoire Analysis ◽  
B Cell Repertoire

Bim establishes the B-cell repertoire from early to late in the immune response

2020 ◽  
Author(s):  
Akiko Sugimoto-Ishige ◽  
Michishige Harada ◽  
Miho Tanaka ◽  
Tommy Terooatea ◽  
Yu Adachi ◽  
...  
Keyword(s):  
Immune Response ◽  
T Cell ◽  
B Cells ◽  
B Cell ◽  
Plasma Cells ◽  
Affinity Maturation ◽  
Late Response ◽  
Cell Repertoire ◽  
High Affinity ◽  
B Cell Repertoire

Abstract In T cell-dependent antibody responses, some of the activated B cells differentiate along extrafollicular pathways into low-affinity memory and plasma cells, whereas others are involved in subsequent germinal center (GC) formation in follicular pathways, in which somatic hypermutation and affinity maturation occur. The present study demonstrated that Bim, a proapoptotic BH3-only member of the Bcl-2 family, contributes to the establishment of the B-cell repertoire from early to late stages of immune responses to T cell-dependent antigens. Extrafollicular plasma cells grew in the spleen during the early immune response, but their numbers rapidly declined with the appearance of GC-derived progeny in wild-type mice. By contrast, conditional Bim deficiency in B cells resulted in expansion of extrafollicular IgG1+ antibody-forming cells (AFCs) and this expansion was sustained during the late response, which hampered the formation of GC-derived high-affinity plasma cells in the spleen. Approximately 10% of AFCs in mutant mice contained mutated VH genes; thus, Bim deficiency appears not to impede the selection of high-affinity AFC precursor cells. These results suggest that Bim contributes to the replacement of low-affinity antibody by high-affinity antibody as the immune response progresses.

scholarly journals Clonal dissection of the human memory B-cell repertoire following infection and vaccination

2009 ◽  
Vol 39 (5) ◽  
pp. 1260-1270 ◽  
Author(s):  
Debora Pinna ◽  
Davide Corti ◽  
David Jarrossay ◽  
Federica Sallusto ◽  
Antonio Lanzavecchia
Keyword(s):  
B Cell ◽  
Human Memory ◽  
Cell Repertoire ◽  
Memory B Cell ◽  
B Cell Repertoire

scholarly journals Serotype-Specific and Age-Dependent Generation of Pneumococcal Polysaccharide-Specific Memory B-Cell and Antibody Responses to Immunization with a Pneumococcal Conjugate Vaccine

2007 ◽  
Vol 15 (2) ◽  
pp. 182-193 ◽  
Author(s):  
Elizabeth A. Clutterbuck ◽  
Sarah Oh ◽  
Mainga Hamaluba ◽  
Sharon Westcar ◽  
Peter C. L. Beverley ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
Conjugate Vaccine ◽  
Pneumococcal Conjugate Vaccine ◽  
Plasma Cells ◽  
Memory B Cells ◽  
Antibody Responses ◽  
Memory B Cell ◽  
Cell Responses ◽  
Specific Memory

ABSTRACT Glycoconjugate vaccines have dramatically reduced the incidence of encapsulated bacterial diseases in toddlers under 2 years of age, but vaccine-induced antibody levels in this age group wane rapidly. We immunized adults and 12-month-old toddlers with heptavalent pneumococcal conjugate vaccine to determine differences in B-cell and antibody responses. The adults and 12-month-old toddlers received a pneumococcal conjugate vaccine. The toddlers received a second dose at 14 months of age. The frequencies of diphtheria toxoid and serotype 4, 14, and 23F polysaccharide-specific plasma cells and memory B cells were determined by enzyme-linked immunospot assay. The toddlers had no preexisting polysaccharide-specific memory B cells or serum immunoglobulin G (IgG) antibody but had good diphtheria toxoid-specific memory responses. The frequencies of plasma cells and memory B cells increased by day 7 (P < 0.0001) in the adults and the toddlers following a single dose of conjugate, but the polysaccharide responses were significantly lower in the toddlers than in the adults (P = 0.009 to <0.001). IgM dominated the toddler antibody responses, and class switching to the IgG was serotype dependent. A second dose of vaccine enhanced the antibody and memory B-cell responses in the toddlers but not the ex vivo plasma cell responses. Two doses of pneumococcal conjugate vaccine are required in toddlers to generate memory B-cell frequencies and antibody class switching for each pneumococcal polysaccharide equivalent to that seen in adults.

scholarly journals Different B cell subpopulations show distinct patterns in their IgH repertoire metrics

2021 ◽  
Author(s):  
Marie Ghraichy ◽  
Valentin von Niederhäusern ◽  
Aleksandr Kovaltsuk ◽  
Jacob Daniel Galson ◽  
Charlotte M Deane ◽  
...  
Keyword(s):  
B Cell ◽  
In Silico ◽  
Plasma Cells ◽  
Somatic Hypermutation ◽  
Bioinformatic Analysis ◽  
Cell Repertoire ◽  
Humoral Immune ◽  
B Cell Repertoire ◽  
Cell Subpopulations ◽  
The Individual

Background: Several human B-cell subpopulations are recognized in the peripheral blood, which play distinct roles in the humoral immune response. These cells undergo developmental and maturational changes involving VDJ recombination, somatic hypermutation and class switch recombination, altogether shaping their immunoglobulin heavy chain (IgH) repertoire. Methods: Here, we sequenced the IgH repertoire of naïve, marginal zone, switched and plasma cells from 10 healthy adults along with matched unsorted and in silico separated CD19+ bulk B cells. We used advanced bioinformatic analysis and machine learning to thoroughly examine and compare these repertoires. Results: We show that sorted B cell subpopulations are characterised by distinct repertoire characteristics on both the individual sequence and the repertoire level. Sorted subpopulations shared similar repertoire characteristics with their corresponding in silico separated subsets. Furthermore, certain IgH repertoire characteristics correlated with the position of the constant region on the IgH locus. Conclusion: Overall, this study provides unprecedented insight over mechanisms of B cell repertoire control in peripherally circulating B cell subpopulations.

scholarly journals A role for gut-associated lymphoid tissue in shaping the human B cell repertoire

2013 ◽  
Vol 210 (9) ◽  
pp. 1665-1674 ◽  
Author(s):  
Anna Vossenkämper ◽  
Paul A. Blair ◽  
Niloufar Safinia ◽  
Louise D. Fraser ◽  
Lisa Das ◽  
...  
Keyword(s):  
B Cells ◽  
B Cell ◽  
Lupus Erythematosus ◽  
Lymphoid Tissue ◽  
Plasma Cells ◽  
Cell Subset ◽  
Cell Repertoire ◽  
Human B Cells ◽  
B Cell Repertoire ◽  
Immature B Cells

We have tracked the fate of immature human B cells at a critical stage in their development when the mature B cell repertoire is shaped. We show that a major subset of bone marrow emigrant immature human B cells, the transitional 2 (T2) B cells, homes to gut-associated lymphoid tissue (GALT) and that most T2 B cells isolated from human GALT are activated. Activation in GALT is a previously unknown potential fate for immature human B cells. The process of maturation from immature transitional B cell through to mature naive B cell includes the removal of autoreactive cells from the developing repertoire, a process which is known to fail in systemic lupus erythematosus (SLE). We observe that immature B cells in SLE are poorly equipped to access the gut and that gut immune compartments are depleted in SLE. Thus, activation of immature B cells in GALT may function as a checkpoint that protects against autoimmunity. In healthy individuals, this pathway may be involved in generating the vast population of IgA plasma cells and also the enigmatic marginal zone B cell subset that is poorly understood in humans.

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