Novel in vitro booster vaccination to rapidly generate antigen-specific human monoclonal antibodies

Nandin, Irene Sanjuan; Fong, Carol Ho-Yan; Deantonio, Cecilia; Torreño-Pina, Juan A.; Pecetta, Simone; Maldonado, Paula; Gasparrini, Francesca; Ordovás-Montañés, José; Kazer, Samuel W.; Kjær, Svend; Borley, Daryl W.; Nair, Usha; Coleman, Julia; Lingwood, Daniel; Shalek, Alex K.; Meffre, Eric; Poignard, Pascal; Burton, Dennis R.; Batista, Facundo D.

doi:10.1084/jem.20170633

Cited by 19 publications

(11 citation statements)

References 60 publications

(81 reference statements)

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“…The R/S ratios for the CDRs, however, ranged from 1 (VK-CDR2) to 6.5 (Vλ-CDR2), indicating that all CDRs favored replacement mutations. These values trended toward being lower than published averages; however, the CDR R/S was higher than the FWR R/S, indicating a memory B-cell phenotype, and the ratio of CDR R/S: FWR R/S was similar to published values ( 49 , 52 – 55 ).…”

Section: Resultssupporting

confidence: 75%

Isolating Human Monoclonal Antibodies Against Adeno-Associated Virus From Donors With Pre-existing Immunity

et al. 2020

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With the advent of single B-cell cloning technology, we can isolate antibodies against virtually any antigen to study the interaction of a given pathogen with the immune system and develop novel therapeutic strategies. Antibodies directed against the capsid of adeno-associated viruses (AAV) are a significant obstacle to effectively leveraging AAV as a gene-delivery vector in seropositive individuals. In order to design next-generation vectors that can evade neutralization by these antibodies, studies have mapped the epitopes of mouse monoclonal antibodies generated by immunization with AAV. Although these studies provide critical information regarding capsid immunogenicity, they cannot address (1) differences in the antibody repertoire generated in humans following AAV natural infection; or (2) how reactions can vary when generated in response to vector administration. Here, we isolated and evaluated a panel of novel, fully human anti-AAV antibodies by cloning single memory B cells from a seropositive normal donor. We have validated the utility of this approach to study AAV immunology. Our goal is to leverage this knowledge to design novel AAV variants that can effectively transduce target tissues in individuals with AAV-neutralizing antibodies.

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Section: Resultssupporting

confidence: 75%

Isolating Human Monoclonal Antibodies Against Adeno-Associated Virus From Donors With Pre-existing Immunity

et al. 2020

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“…Recently, it has been described a novel in vitro platform to produce antigen-specific human antibodies based on the use of particles (0.11 lm) coated with anti-j together with CpG to costimulate at the same time BCR and TLR9 receptors, respectively. Authors claim that particle internalization allows CpG to reach TLR9 intracellular compartment, something that is not possible in a soluble manner [45]. This particle internalization results in the synergistic activation of both receptors inducing a more robust B-cell proliferation and differentiation into plasma cells A Confocal microscopy image of antigen aggregates generated with NIP-BSA-FITC complexed with alum.…”

Section: Alum-based Immunizations Induce a Potent Humoral Response Rementioning

confidence: 99%

Antigen phagocytosis by B cells is required for a potent humoral response

et al. 2018

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Successful vaccines rely on activating a functional humoral response that results from promoting a proper germinal center (GC) reaction. Key in this process is the activation of follicular B cells that need to acquire antigens and to present them to cognate CD4 T cells. Here, we report that follicular B cells can phagocytose large antigen‐coated particles, a process thought to be exclusive of specialized antigen‐presenting cells such as macrophages and dendritic cells. We show that antigen phagocytosis by B cells is BCR‐driven and mechanistically dependent on the GTPase RhoG. Using Rhog −/− mice, we show that phagocytosis of antigen by B cells is important for the development of a strong GC response and the generation of high‐affinity class‐switched antibodies. Importantly, we show that the potentiation effect of alum, a common vaccine adjuvant, requires direct phagocytosis of alum–antigen complexes by B cells. These data suggest a new avenue for vaccination approaches by aiming to deliver 1–3 μm size antigen particles to follicular B cells.

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“…63,[66][67][68][69] The antigenic reactivity of each cell can then be tested using the supernatant from each well through a number of methods including ELISA. Assuming that antibody secretion is constitutive in all PC, a property that remains to be formally established, the absence of autoreactive PC would represent the ultimate measure of effective tolerance.…”

Section: E Xperimental Approache S To a Ss E Ss H Uman B-cell Tolermentioning

confidence: 99%

“…To date, single-cell cultures and screening for antigenic reactivity have been successfully applied to transitional, naive, memory, and ASC. 63,[66][67][68][69] The antigenic reactivity of each cell can then be tested using the supernatant from each well through a number of methods including ELISA. In addition, the antigenic reactivity of single cells can also be determined directly using a variety of methods including microengraving and diverse microfluidics devices such as microdroplets, microfluidic chambers, and microcapillaries.…”

Section: Autoreactivity Measurementsmentioning

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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Novel in vitro booster vaccination to rapidly generate antigen-specific human monoclonal antibodies

Cited by 19 publications

References 60 publications

Isolating Human Monoclonal Antibodies Against Adeno-Associated Virus From Donors With Pre-existing Immunity

Isolating Human Monoclonal Antibodies Against Adeno-Associated Virus From Donors With Pre-existing Immunity

Antigen phagocytosis by B cells is required for a potent humoral response

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

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