Rapid generation of fully human monoclonal antibodies specific to a vaccinating antigen

Smith, Kenneth; Garman, Lori; Wrammert, Jens; Zheng, Na; Capra, J D; Ahmed, Rafi; Wilson, Patrick C.

doi:10.1038/nprot.2009.3

Cited by 433 publications

(478 citation statements)

References 20 publications

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“…Of the 16 identified anti-LASV nAbs, 13 require the assembled GPC for binding, whereas 3 require only the GP1 (36). Thus, emerging techniques in mAb generation, such as the isolation of antigen-specific B cells (37) against an intact prefusion GPC spike antigen, may reveal antibodies capable of targeting alternative non-RBS epitopes on the JUNV glycoprotein surface.…”

Section: Resultsmentioning

confidence: 99%

Convergent immunological solutions to Argentine hemorrhagic fever virus neutralization

Zeltina

Krumm

Mehmet

et al. 2017

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

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Transmission of hemorrhagic fever New World arenaviruses from their rodent reservoirs to human populations poses substantial public health and economic dangers. These zoonotic events are enabled by the specific interaction between the New World arenaviral attachment glycoprotein, GP1, and cell surface human transferrin receptor (hTfR1). Here, we present the structural basis for how a mouse-derived neutralizing antibody (nAb), OD01, disrupts this interaction by targeting the receptor-binding surface of the GP1 glycoprotein from Junín virus (JUNV), a hemorrhagic fever arenavirus endemic in central Argentina. Comparison of our structure with that of a previously reported nAb complex (JUNV GP1-GD01) reveals largely overlapping epitopes but highly distinct antibody-binding modes. Despite differences in GP1 recognition, we find that both antibodies present a key tyrosine residue, albeit on different chains, that inserts into a central pocket on JUNV GP1 and effectively mimics the contacts made by the host TfR1. These data provide a molecular-level description of how antibodies derived from different germline origins arrive at equivalent immunological solutions to virus neutralization. . These viruses are endemic to rodent populations in rural areas of Argentina, Bolivia, and Venezuela, respectively, and viral spillover into human populations can result in severe hemorrhagic fever (HF) (3). Novel pathogenic NW arenaviruses continue to be identified (4-6), underscoring a wide-scale need for effective vaccines and therapeutics.JUNV, the etiological agent of Argentine hemorrhagic fever (AHF), constitutes one of the most dangerous NW arenaviruses, putting an estimated 5 million people at risk (7,8). JUNV infection typically exhibits a rapid onset of disease (7-14 d) and high mortality rates (15-30%) (7, 9, 10). There are no internationally approved drugs for preventing or treating NW arenavirus HF. However, the successful development of a live, attenuated JUNV vaccine, Candid#1, has proven that AHF can be controlled (11). Similarly, virus-neutralizing immune plasma from convalescent individuals has been successfully used for the treatment of AHF (10, 12).The JUNV envelope surface is decorated by trimeric multifunctional glycoprotein complex (GPC) spikes. Each protomer in the trimer consists of: a myristoylated (13) stable signal peptide, an attachment subunit (GP1), and a transmembrane fusion subunit (GP2) (14-19). Host-cell entry of JUNV and other clade B arenaviruses is initiated by the interaction between the arenaviral GP1 and the host transferrin receptor (TfR1) (20). The GP1 subunit interacts with the apical domain of TfR1, distal from natural transferrin and hereditary hemochromatosis protein recognition sites (21, 22). A primary determinant of zoonotic spread of clade B arenaviruses is the ability of the GP1 to recognize the human TfR1 ortholog (20, 23).The JUNV GPC spike comprises the primary target for neutralizing immune responses (24) and three GPC-specific mousederived nAbs-GD01, OD01, and GB03-have shown prom...

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Section: Resultsmentioning

confidence: 99%

Convergent immunological solutions to Argentine hemorrhagic fever virus neutralization

Zeltina

Krumm

Mehmet

et al. 2017

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

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“…Ab V region gene repertoires were characterized essentially as described previously (23)(24)(25). Single CD19 + IgM + CD27 + CD21 2/low B cells were sorted into 96-well PCR plates containing RNase inhibitor (Promega).…”

Section: Sequencing and Repertoire Analysismentioning

confidence: 99%

“…To provide a more precise definition of the phenotype of this expanded population, CD21 Previous reports found large expansions of V H 1-69 and V H 4-34 gene segments that encode RF of the WA idiotype (9-13). To characterize more precisely the repertoire of the CD21 2/low MZ B cell population characterized herein, we used single-cell PCR and sequencing (24,25). In all three HCV-MC patients, the CD21 2/low MZ B cell repertoires were dominated by a V H 1-69-utilizing B cell clone or, for patient 3, a V H 1-69-and a V H 4-34-expressing clone (Fig.…”

Section: Hcvmentioning

confidence: 99%

Expansion of Functionally Anergic CD21−/low Marginal Zone-like B Cell Clones in Hepatitis C Virus Infection-Related Autoimmunity

Terrier

Joly²,

Vazquez

et al. 2011

The Journal of Immunology

108

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Homeostasis of peripheral B cell subsets is disturbed during chronic hepatitis C virus (HCV) infection, leading to the occurrence of autoimmunity and B cell lymphoproliferation. However, mechanisms by which HCV causes lymphoproliferation remain controversial. We report in this article on the elevated number of clonal CD21−/lowIgM+CD27+ marginal zone (MZ)-like B cells, which correlates with autoimmunity and lymphoproliferation in HCV patients. We found an increase in autoreactive BCRs using VH1–69 and VH4–34 genes in CD21−/low MZ B cells. CD21−/low MZ B cells showed impaired calcium-mediated signaling, did not upregulate activation markers, and did not proliferate in response to BCR triggering. CD21−/low MZ B cells also were prone to dying faster than their CD21+ counterparts, suggesting that these B cells were anergic. CD21−/low MZ B cells, in contrast, remained responsive to TLR9 stimulation. Gene array analyses revealed the critical role of Early growth response 2 and Cbl-b in the induction of anergy. Therefore, HCV patients who display high frequencies of unresponsive CD21−/low MZ B cells are more susceptible to developing autoimmunity and/or lymphoproliferation. These cells remain in peripheral blood controlled by functional anergy instead of being eliminated, and chronic antigenic stimulation through TLR stimulation may create a favorable environment for breaking tolerance and activating these cells.

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“…The cloning method was performed based on an established protocol (24), with several modifications. Details are provided in SI Materials and Methods.…”

Section: Methodsmentioning

confidence: 99%

Genetic engineering of hematopoietic stem cells to generate invariant natural killer T cells

Smith

Liu

et al. 2015

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

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Invariant natural killer T (iNKT) cells comprise a small population of αβ T lymphocytes. They bridge the innate and adaptive immune systems and mediate strong and rapid responses to many diseases, including cancer, infections, allergies, and autoimmunity. However, the study of iNKT cell biology and the therapeutic applications of these cells are greatly limited by their small numbers in vivo (∼0.01-1% in mouse and human blood). Here, we report a new method to generate large numbers of iNKT cells in mice through T-cell receptor (TCR) gene engineering of hematopoietic stem cells (HSCs). We showed that iNKT TCR-engineered HSCs could generate a clonal population of iNKT cells. These HSC-engineered iNKT cells displayed the typical iNKT cell phenotype and functionality. They followed a two-stage developmental path, first in thymus and then in the periphery, resembling that of endogenous iNKT cells. When tested in a mouse melanoma lung metastasis model, the HSC-engineered iNKT cells effectively protected mice from tumor metastasis. This method provides a powerful and high-throughput tool to investigate the in vivo development and functionality of clonal iNKT cells in mice. More importantly, this method takes advantage of the self-renewal and longevity of HSCs to generate a long-term supply of engineered iNKT cells, thus opening up a new avenue for iNKT cell-based immunotherapy. αβ T lymphocytes highly conserved from mice to humans. Like conventional αβ T cells, iNKT cells are derived from hematopoietic stem cells (HSCs) and develop in the thymus. However, they differ from conventional T cells in several important aspects, including their display of NK cell markers, their recognition of glycolipid antigens presented by the nonclassical monomorphic major histocompatibility complex (MHC) molecule CD1d, and their expression of semi-invariant T-cell receptors (identical α chains paired with a limited selection of β chains) (1, 2). Despite their small numbers in vivo (∼0.1-1% in mouse blood and ∼0.01-1% in human blood), iNKT cells have been suggested to play important roles in regulating many diseases, including cancer, infections, allergies, and autoimmunity (3). When stimulated, iNKT cells rapidly release a large amount of effector cytokines like IFN-γ and IL-4, both as a cell population and at the single-cell level. These cytokines then activate various immune effector cells, such as natural killer (NK) cells and dendritic cells (DCs) of the innate immune system, as well as CD4 helper and CD8 cytotoxic conventional αβ T cells of the adaptive immune system via activated DCs (3, 4). Because of their unique activation mechanism, iNKT cells can attack multiple diseases independent of antigen and MHC restrictions, making them attractive universal therapeutic agents (3, 4). Notably, because of the capacity of effector NK cells and conventional αβ T cells to specifically recognize diseased tissue cells, iNKT cell-induced immune reactions result in limited off-target side effects (3, 4).Restricted by their extremely low numbers, ...

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Rapid generation of fully human monoclonal antibodies specific to a vaccinating antigen

Cited by 433 publications

References 20 publications

Convergent immunological solutions to Argentine hemorrhagic fever virus neutralization

Convergent immunological solutions to Argentine hemorrhagic fever virus neutralization

Expansion of Functionally Anergic CD21−/low Marginal Zone-like B Cell Clones in Hepatitis C Virus Infection-Related Autoimmunity

Genetic engineering of hematopoietic stem cells to generate invariant natural killer T cells

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