H3N2 Influenza Infection Elicits More Cross-Reactive and Less Clonally Expanded Anti-Hemagglutinin Antibodies Than Influenza Vaccination

Moody, M. Anthony; Zhang, Ruijun; Walter, Emmanuel B.; Woods, Christopher W.; Ginsburg, Geoffrey S.; McClain, Micah T.; Denny, Thomas N.; Chen, Xi; Munshaw, Supriya; Marshall, Dawn J.; Whitesides, John F.; Drinker, Mark; Amos, Joshua D.; Gurley, Thaddeus C.; Eudailey, Joshua; Foulger, Andrew; Derosa, K; Parks, Robert; Meyerhoff, Robert; Yu, Jia; Kozink, Daniel M.; Barefoot, Brice E.; Ramsburg, Elizabeth; Khurana, Surender; Golding, Hana; Vandergrift, Nathan; Alam, S. Munir; Tomaras, Georgia D.; Kepler, Thomas B.; Kelsoe, Garnett; Liao, Hua‐Xin; Haynes, Barton F.

doi:10.1371/journal.pone.0025797

Cited by 159 publications

(189 citation statements)

References 56 publications

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“…In total, our analysis of the CCH rabbit revealed that the antigen-specific polyclonal response is composed of ∼34 IgG antibodies that are classified into 30 different clonotypes [same V and J, same CDRH3 length, 80% aa homology (14,15)]; 4 of the 34 antibodies that constitute the majority of the antigen-specific IgG repertoire in this animal differed by only 1-3 aa and therefore corresponded to clonally related antibodies ( Fig. 3B; also SI Appendix, Fig.…”

Section: Resultsmentioning

confidence: 97%

“…Composed of the V(D)J join with its inherent junctional diversity, the CDRH3 specifies the V H clonotype. The V H clonotype is defined as the group of V H sequences that share germ-line V and J segments, have identical CDRH3 lengths, and exhibit greater than 80% amino acid identity in the CDRH3 sequences (14,15). The V H clonotype is an important immunological concept because it accounts for antibodies that likely originate from a single B-cell lineage and may provide insight on the evolution of the antigenspecific response of that lineage.…”

Section: Resultsmentioning

confidence: 99%

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Molecular deconvolution of the monoclonal antibodies that comprise the polyclonal serum response

Wine

Boutz

Lavinder

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

126

156

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We have developed and validated a methodology for determining the antibody composition of the polyclonal serum response after immunization. Pepsin-digested serum IgGs were subjected to standard antigen-affinity chromatography, and resulting elution, wash, and flow-through fractions were analyzed by bottom-up, liquid chromatography-high-resolution tandem mass spectrometry. Identification of individual monoclonal antibodies required the generation of a database of IgG variable gene (V-gene) sequences constructed by NextGen sequencing of mature B cells. Antibody V-gene sequences are characterized by short complementarity determining regions (CDRs) of high diversity adjacent to framework regions shared across thousands of IgGs, greatly complicating the identification of antigen-specific IgGs from proteomically observed peptides. By mapping peptides marking unique V H CDRH3 sequences, we identified a set of V-genes heavily enriched in the affinity chromatography elution, constituting the serum polyclonal response. After booster immunization in a rabbit, we find that the antigenspecific serum immune response is oligoclonal, comprising antibodies encoding 34 different CDRH3s that group into 30 distinct antibody V H clonotypes. Of these 34 CDRH3s, 12 account for ∼60% of the antigen-specific CDRH3 peptide mass spectral counts. For comparison, antibodies with 18 different CDRH3s (12 clonotypes) were represented in the antigen-specific IgG fraction from an unimmunized rabbit that fortuitously displayed a moderate titer for BSA. Proteomically identified antibodies were synthesized and shown to display subnanomolar affinities. The ability to deconvolute the polyclonal serum response is likely to be of key importance for analyzing antibody responses after vaccination and for more completely understanding adaptive immune responses in health and disease.antibody proteomics | antibody repertoire | serum immunoprofiling | B-cell response | humoral response T he first Nobel Prize in Medicine was awarded to Emil von Behring, who in collaboration with Kitasato Shibasaburo and Paul Ehrlich discovered serum antitoxins (1, 2). Remarkably, after more than 100 y of intense research in immunology, little is known about the clonality, relative concentrations, and binding properties of the monoclonal antibodies that constitute the antigen-specific Ig pool in serum.At steady state, circulating antibodies are produced by terminally differentiated B lymphocytes (plasma cells) within the bone marrow, and thus cannot be accessed in living individuals (3). Although recent single B-cell cloning methods (4, 5) have led to the identification of peripheral antigen-specific B memory and/or antibody-secreting cells (plasmablasts), it is generally unknown whether the Igs encoded by peripheral blood B cells correspond to the antibodies present in circulation and especially whether they are present at physiologically relevant levels (i.e., at serum concentrations above K D corresponding to >1 μg/mL for an average affinity of individual antibodies of 5 nM)...

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Section: Resultsmentioning

confidence: 97%

Section: Resultsmentioning

confidence: 99%

Molecular deconvolution of the monoclonal antibodies that comprise the polyclonal serum response

Wine

Boutz

Lavinder

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

126

156

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“…The V H repertoires for each donor, encoded by day 7 plasmablasts and by IgD − mBCs collected on both day 7 and 3 mo postboost, were determined by 454 (Roche Diagnostics GmbH) sequencing (70,326 and 157,089 high-quality V H reads for HD1 and HD2, respectively; Table S1) and indexed by their V H clonotype. The V H clonotype, which represents a cluster of antibodies that likely originate from a single B-cell lineage (27,28), is defined here as the group of V H sequences that share germ-line V and J segments and also exhibit greater than 90% amino acid identity in the complementarity-determining region (CDR)-H3 (threshold for CDR-H3 amino acid identity determined by analysis of test sets from clustered deep-sequencing data; Fig. S2).…”

Section: Resultsmentioning

confidence: 99%

Identification and characterization of the constituent human serum antibodies elicited by vaccination

Lavinder

Wine

Giesecke

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

227

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Most vaccines confer protection via the elicitation of serum antibodies, yet more than 100 y after the discovery of antibodies, the molecular composition of the human serum antibody repertoire to an antigen remains unknown. Using high-resolution liquid chromatography tandem MS proteomic analyses of serum antibodies coupled with next-generation sequencing of the V gene repertoire in peripheral B cells, we have delineated the human serum IgG and B-cell receptor repertoires following tetanus toxoid (TT) booster vaccination. We show that the TT + serum IgG repertoire comprises ∼100 antibody clonotypes, with three clonotypes accounting for >40% of the response. All 13 recombinant IgGs examined bound to vaccine antigen with K d ∼ 10 −8 -10 −10 M. Five of 13 IgGs recognized the same linear epitope on TT, occluding the binding site used by the toxin for cell entry, suggesting a possible explanation for the mechanism of protection conferred by the vaccine. Importantly, only a small fraction (<5%) of peripheral blood plasmablast clonotypes (CD3 − CD14 − CD19 + CD27 ++ CD38 ++ CD20 − TT + ) at the peak of the response (day 7), and an even smaller fraction of memory B cells, were found to encode antibodies that could be detected in the serological memory response 9 mo postvaccination. This suggests that only a small fraction of responding peripheral B cells give rise to the bone marrow long-lived plasma cells responsible for the production of biologically relevant amounts of vaccine-specific antibodies (near or above the K d ). Collectively, our results reveal the nature and dynamics of the serological response to vaccination with direct implications for vaccine design and evaluation.B-cell repertoire | proteomics M ost approved vaccines confer protection against infectious diseases by the induction of long-lived plasma cells (LLPCs), which secrete antibodies that serve to neutralize and opsonize the pathogen for many years or decades (1-3). Additionally, the generation of memory B cells (mBCs) provides both a mechanism for the rapid synthesis of affinity matured, antigenspecific antibodies following rechallenge and a means to diversify the humoral immune response to confer protection against rapidly evolving viruses or bacteria (4). Although some vaccines elicit antibody titers that remain virtually constant for many decades, for others, including the tetanus toxoid (TT) vaccine, antibody titers wane monotonically over time (5). Booster immunization triggers the rapid expansion and differentiation of cognate B cells, generating antigen-specific plasmablasts that peak in concentration in peripheral blood after 6-7 d and subsequently rapidly decline to nearly undetectable levels (6, 7). Some, but not all, of these peak-wave plasmablasts migrate to specialized niches overwhelmingly located in the bone marrow (BM) and survive as LLPCs (8), which constitute the major source of all classes of Ig in the serum (9).The establishment of serological memory following either primary or booster vaccination is not understood well (10-14)...

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“…The mean heavy chain mutation frequency was 4.00 % ± 1.61 % (range 1.72-6.16 %), a mutation frequency typical of antibodies isolated from vaccine recipients [46,47]. None of the recovered mAbs were clonally related to each other, and a range of VH genes was used (Supplementary Table 1).…”

Section: Discussionmentioning

confidence: 99%

Interrogation of individual intratumoral B lymphocytes from lung cancer patients for molecular target discovery

Campa

Moody

Zhang

et al. 2016

Cancer Immunol Immunother

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H3N2 Influenza Infection Elicits More Cross-Reactive and Less Clonally Expanded Anti-Hemagglutinin Antibodies Than Influenza Vaccination

Cited by 159 publications

References 56 publications

Molecular deconvolution of the monoclonal antibodies that comprise the polyclonal serum response

Molecular deconvolution of the monoclonal antibodies that comprise the polyclonal serum response

Identification and characterization of the constituent human serum antibodies elicited by vaccination

Interrogation of individual intratumoral B lymphocytes from lung cancer patients for molecular target discovery

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