The role of the light chain in the structure and binding activity of two cattle antibodies that neutralize bovine respiratory syncytial virus

Ren, Jingshan; Nettleship, Joanne E.; Harris, Gemma; Mwangi, William; Rhaman, N.; Grant, Clare; Kotecha, Abhay; Fry, Elizabeth E.; Charleston, Bryan; Stuart, D.I.; Hammond, John A.; Owens, Raymond J.

doi:10.1016/j.molimm.2019.04.026

Cited by 13 publications

(16 citation statements)

References 33 publications

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“…Given that the C10 LC swap (C10 H C27 L ) had lower PPS3 affinity and was less agglutinating than native C10, its superior binding and efficacy against B2 may depend on its V L structure. Notably, structure-function studies of viral antibodies have revealed that V L gene use and structure can dictate whether an antibody is neutralizing or nonneutralizing (46,47). Our data show that the C10 V L plays a critical role in its agglutinating activity, which was lost when we substituted its V L with the V L of C27.…”

Section: Discussionmentioning

confidence: 78%

Isolation and characterization of human monoclonal antibodies to pneumococcal capsular polysaccharide 3

Babb

Doyle²,

Pirofski

2021

Preprint

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The current pneumococcal capsular polysaccharide (PPS) conjugate vaccine (PCV13) is less effective against Streptococcus pneumoniae serotype 3 (ST3), which remains a major cause of pneumococcal disease and mortality. Therefore, dissecting structure-function relationships of human PPS3 antibodies may reveal characteristics of protective antibodies. Using flow cytometry, we isolated PPS3-binding memory B cells from pneumococcal vaccine recipients and generated seven human PPS3-specific monoclonal antibodies (humAbs). Five humAbs displayed ST3 opsonophagocytic activity, four induced ST3 agglutination in vitro, and four mediated both activities. For two humAbs, C10 and C27, that used the same variable heavy (VH) and light (VL) chain domains (VH3-9*01/VL2-14*03), C10 had fewer VL somatic mutations, higher PPS3 affinity, more ST3 opsonophagocytic and agglutinating activity, whilst both humAbs altered ST3 gene expression in vitro. After VL swaps, C10VH/C27VL exhibited reduced ST3 binding and agglutination, but C27VH/C10VL binding was unchanged. In C57Bl/6 mice, C10 and C27 reduced nasopharyngeal colonization with ST3 A66 and a clinical strain, B2, and prolonged survival following lethal A66 intraperitoneal infection, but only C10 protected against lethal intranasal infection with the clinical strain. Our findings, associate efficacy of PPS3-specific humAbs with ST3 agglutination and opsonophagocytic activity and reveal an unexpected role for the VL in functional activity in vitro and in vivo. These findings also provide insights that may inform antibody-based therapy and identification of surrogates of vaccine efficacy against ST3.IMPORTANCEDespite the global success of pneumococcal conjugate vaccination, serotype 3 (ST3) pneumococcus remains a leading cause of morbidity and mortality. In comparison to other vaccine-included serotypes, the ST3 pneumococcal capsular polysaccharide (PPS3) induces a weaker opsonophagocytic response, which is considered a correlate of vaccine efficacy. Previous studies of mouse PPS3 monoclonal antibodies identified ST3 agglutination as a correlate of reduced ST3 nasopharyngeal colonization in mice, however neither the agglutinating ability of human vaccine-elicited PPS3 antibodies nor their ability to prevent experimental murine nasopharyngeal colonization has been studied. We generated and analysed the functional and in vivo efficacy of human vaccine-elicited PPS3 monoclonal antibodies and found that ST3 agglutination associated with antibody affinity, protection in vivo, and limited somatic mutations in the light chain variable region. These findings provide new insights that may inform the development of antibody-based therapies and next generation vaccines for ST3.

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Section: Discussionmentioning

confidence: 78%

Isolation and characterization of human monoclonal antibodies to pneumococcal capsular polysaccharide 3

Babb

Doyle²,

Pirofski

2021

Preprint

View full text Add to dashboard Cite

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“…It was also shown that swapping the heavy and light chains of different antibodies can result in the abolishment of the antigen binding activity [34]. For a cattle IgG, a recent report showed that the light chain contributes to the neutralizing activity and spectrum [24]. However, the role of the light chain in binding activity has not been characterized in porcine antibodies.…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies demonstrated that pairing of a certain heavy chain with a range of light chains could change binding affinities of resultant antibodies to antigens in mouse, human and bovine repertoires [24,25]. To verify the characterization of porcine mAbs, we tested the reactivity of the reconstructed mAbs, which were produced by random pairing of the heavy and light chains of the five FMDV-specific mAbs.…”

Section: Role Of Light Chain In Binding Activity Of Fmdvspecific Porcine Mabsmentioning

confidence: 99%

Isolation and characterization of porcine monoclonal antibodies revealed two distinct serotype-independent epitopes on VP2 of foot-and-mouth disease virus

Zhu

Zhou

et al. 2021

Journal of General Virology

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Pigs are susceptible to foot-and-mouth disease virus (FMDV), and the humoral immune response plays an essential role in protection against FMDV infection. However, little information is available about FMDV-specific mAbs derived from single B cells of pigs. This study aimed to determine the antigenic features of FMDV that are recognized by antibodies from pigs. Therefore, a panel of pig-derived mAbs against FMDV were developed using fluorescence-based single B cell antibody technology. Western blotting revealed that three of the antibodies (1C6, P2-7E and P2-8G) recognized conserved antigen epitopes on capsid protein VP2, and exhibited broad reactivity against both FMDV serotypes A and O. An alanine-substitution scanning assay and sequence conservation analysis elucidated that these porcine mAbs recognized two conserved epitopes on VP2: a linear epitope (2KKTEETTLL10) in the N terminus and a conformational epitope involving residues K63, H65, L66, F67, D68 and L81 on two β-sheets (B-sheet and C-sheet) that depended on the integrity of VP2. Random parings of heavy and light chains of the IgGs confirmed that the heavy chain is predominantly involved in binding to antigen. The light chain of porcine IgG contributes to the binding affinity toward an antigen and may function as a support platform for antibody stability. In summary, this study is the first to reveal the conserved antigenic profile of FMDV recognized by porcine B cells and provides a novel method for analysing the antibody response against FMDV in its natural hosts (i.e. pigs) at the clonal level.

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“…For example, antibodies from λ chains may dominate the neutralizing effect against SIV gp-120 in blood 34,35 . A recombination experiment showed that exchanging the light chain could reduce the binding affinity of antibody 36 . A study of the structure of an H3-clade neutralizing human monoclonal antibody suggests that light chains modulate the neutralizing spectrum of antibodies by affecting the local conformation of heavy chains 37…”

Section: Discussionmentioning

confidence: 99%

Responding kinetic of B-cell receptor repertoire to the Toll-like receptor 7/8 stimulation in non-human primates

Wang¹,

Mandl²,

Feinberg³

et al. 2021

Preprint

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TLR7 and 8 regulate B cell immunity, but the precise details of the mechanism are still unclear. Here, we studied the kinetics of both heavy and light chains (IgKL) of B-cell receptor (BCR) repertoire responding to the TLR7/8 stimulation in two geniuses of non-human primates (NHPs), African green monkeys (AGMs) and rhesus macaques (RMs). We evaluated the activation of lymphocytes by flow cytometry and studied characteristics of BCR repertoire in terms of gene usage, repertoire diversity, and the number of lineages. Although AGMs had a weaker activation than RMs, and a different responding kinetic, both AGMs and RMs presented an increased IgKL repertoire diversity and lineages expansion. It suggested that the responding time rather than initiation of TLR7/8-induced IgKL repertoire response related to B cell activation. Expanded IgKL lineages with frequency from 0.001% to 1% had an elevated mutation rate and expanded IgH lineages used more IgA/G/E, suggesting that the TLR7/8 stimulation expanded low-frequent but high-mutated lineages. Besides, most of expanded IgKL lineages were lambda isotype. In conclusion, TLR7/8 selectively expands IgKL lineages with a high mutation rate, low frequency, and lambda isotype. The selective effect of TLR7/8 on BCR repertoire allows TLR7/8 agonists to be adjuvant for selectively accelerating antibody maturation.

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The role of the light chain in the structure and binding activity of two cattle antibodies that neutralize bovine respiratory syncytial virus

Cited by 13 publications

References 33 publications

Isolation and characterization of human monoclonal antibodies to pneumococcal capsular polysaccharide 3

Isolation and characterization of human monoclonal antibodies to pneumococcal capsular polysaccharide 3

Isolation and characterization of porcine monoclonal antibodies revealed two distinct serotype-independent epitopes on VP2 of foot-and-mouth disease virus

Responding kinetic of B-cell receptor repertoire to the Toll-like receptor 7/8 stimulation in non-human primates

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