Mechanisms That Shape Human Antibody Repertoire Development in Mice Transgenic for Human Ig H and L Chain Loci

Longo, Nancy S.; Rogosch, Tobias; Zemlin, Michael; Zouali, Moncef; Lipsky, Peter E.

doi:10.4049/jimmunol.1700133

Cited by 10 publications

(16 citation statements)

References 64 publications

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“…Interestingly, the mechanisms of this preference are due to decreased N additions, and a tendency to incorporate shorter DH gene segments. This result has also been seen in multiple other transgenic and wild-type rodents 25,26,28,29 . The specific reasons remain unclear, although the observation that wild-type rat germline D gene segment lengths are shorter suggests intrinsic species-specific mechanisms of selection as well as differences in TdT expression during bone marrow B cell development.…”

Section: Resultssupporting

confidence: 69%

“…Differences in gene segment usage between transgenic animal models and humans, as well as between tissues are expected. For example, multiple human Ig loci transgenic rodents are reported to have gene usage profiles that slightly vary from that of humans 25,26 . Furthermore, antibody repertoires from separate human tissues are known to deviate strongly enough to be clustered by hierarchical clustering 27 .…”

Section: Resultsmentioning

confidence: 99%

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Comparisons of the antibody repertoires of a humanized rodent and humans by high throughput sequencing

Joyce

Burton

Briney

2020

Sci Rep

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the humanization of animal model immune systems by genetic engineering has shown great promise for antibody discovery, tolerance studies and for the evaluation of vaccines. Assessment of the baseline antibody repertoires of unimmunized model animals will be useful as a benchmark for future immunization experiments. We characterized the heavy chain and kappa light chain antibody repertoires of a model animal, the OmniRat, by high throughput antibody sequencing and made use of two novel datasets for comparison to human repertoires. Intra-animal and inter-animal repertoire comparisons reveal a high level of conservation in antibody diversity between the lymph node and spleen and between members of the species. Multiple differences were found in both the heavy and kappa chain repertoires between OmniRats and humans including gene segment usage, CDR3 length distributions, class switch recombination, somatic hypermutation levels and in features of V(D)J recombination. The Inference and Generation of Repertoires (IGoR) software tool was used to model recombination in VH regions which allowed for the quantification of some of these differences. Diversity estimates of the OmniRat heavy chain repertoires almost reached that of humans, around two orders of magnitude less. Despite variation between the species repertoires, a high frequency of OmniRat clonotypes were also found in the human repertoire. These data give insights into the development and selection of humanized animal antibodies and provide actionable information for use in vaccine studies.A major challenge in human vaccine science is finding appropriate models for studying antibody responses. Animals such as mice, rabbits and monkeys have typically been used in the past and the small animals, in particular, have been favored for ease of immunization, cost reasons and the ability to extensively biopsy post-immunization. One limitation is their use of non-human immunoglobulin (V, D, J) genes in antibodies which can be restricted in their specificity 1 , and/or lack residues needed for priming by a germline targeting immunogen 2 . One approach to solving this problem of wild-type animal models is to use humanized immunoglobulin loci-transgenic rodents 3,4 . The first demonstration of a transgenic rodent with the ability to express human IgM was 30 years ago 5 . Since then, advances in genetic engineering technologies allowed for the first transgenic mice strains that express fully human antibodies 6,7 . Today, many new transgenic animal models have been developed including rodents, chickens, rabbits and cows 8 . These animal models have been used extensively for the discovery of monoclonal antibodies (mAbs) 9 , tolerance studies 10 and more recently for modelling human antibody responses to vaccine candidates 3,4 . Here we focus on one such animal model: a rat with expression of humanized chimeric antibodies.The generation of antibody diversity begins with the development of B cells in the bone marrow. Three unlinked loci contain the immunoglobulin gene segments ne...

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Section: Resultssupporting

confidence: 69%

Section: Resultsmentioning

confidence: 99%

Comparisons of the antibody repertoires of a humanized rodent and humans by high throughput sequencing

Joyce

Burton

Briney

2020

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Interestingly, the mechanisms of this preference are due to decreased N additions, and a tendency to incorporate shorter DH gene segments. This result has also been seen in multiple other transgenic and wild-type rodents 22,23,25,26 . The specific reasons remain unclear, although the observation that wild-type rat germline D gene segment lengths are shorter suggests intrinsic species-specific mechanisms of selection as well as differences in Tdt expression during bone marrow B cell development.…”

Section: Resultssupporting

confidence: 69%

“…Differences in gene segment usage between transgenic animal models and humans, as well as between tissues are expected. For example, multiple human Ig loci transgenic rodents are reported to have gene usage profiles that slightly vary from that of humans 22,23 . Furthermore, antibody repertoires from separate human tissues are known to deviate strongly enough to be clustered by hierarchical clustering 24 .…”

Section: Resultsmentioning

confidence: 99%

Comparisons of the antibody repertoires of a humanized rodent and humans by high throughput sequencing

Joyce

Burton

Briney

2019

Preprint

View full text Add to dashboard Cite

The humanization of animal model immune systems by genetic engineering has shown great promise for antibody discovery, tolerance studies and for the evaluation of vaccines. Assessment of the baseline antibody repertoire of unimmunized model animals will be useful as a benchmark for future immunization experiments. We characterized the heavy chain and kappa chain antibody repertoires of a model animal, the OmniRat, by high throughput antibody sequencing and made use of two novel datasets for comparison to human repertoires. Intra-animal and inter-animal repertoire comparisons reveal a high level of conservation in antibody diversity between the lymph node and spleen and between members of the species. Multiple differences were found in both the heavy and kappa chain repertoires between OmniRats and humans including gene segment usage, CDR3 length distributions, class switch recombination, somatic hypermutation levels and in features of V(D)J recombination. The Inference and Generation of Repertoires (IGoR) software tool was used to model recombination in VH regions which allowed for the quantification of some of these differences. Diversity estimates of the OmniRat heavy chain repertoires almost reached that of humans, around two orders of magnitude less. Despite variation between the species repertoires, a high frequency of OmniRat clonotypes were also found in the human repertoire. These data give insights into the development and selection of humanized animal antibodies and provide actionable information for use in vaccine studies.

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“…We performed single-cell PCR for amplification and further compared the plasmablast repertoires expressed in multiple chronically HIV-infected donors (HIVDs) and healthy donors (HDs). Productive rearrangements may be positively or negatively selected, as they are capable of interacting with Ags (8). Nonproductive rearrangements either do not encode an Ig receptor, code for an Ig receptor that cannot engage with Ag, or code for an Ig receptor that is not subjected to positive or negative selection.…”

Section: Introductionmentioning

confidence: 99%

Characteristics of Plasmablast Repertoire in Chronically HIV-Infected Individuals for Immunoglobulin H and L Chain Profiled by Single-Cell Analysis

Liao

et al. 2020

Front. Immunol.

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Characterization of the diversified immunoglobulin (Ig) repertoire may provide insight into pathways that shape an efficient antibody (Ab) repertoire for immune response against human immunodeficiency virus (HIV) infection. This study aimed to profile characteristics of the plasmablast repertoire during chronic HIV infection. Ig variable regions of plasmablasts from both chronically HIV-infected donors (HIVDs) previously treated with antiretroviral therapy (ART) and healthy donors (HDs) were amplified by single-cell PCR to establish the basis for further repertoire analysis. We compared the plasmablast repertoires expressed in multiple chronically HIVDs after ART treatment cessation and HDs. We also examined the non-productive repertoire to identify the indication of the immediate products of the rearrangement machinery without an impact of selection during HIV infection. We found multiple differences between the productive repertoires of HIVD and HD subjects, including biased usages of VH3-49, VH1-2, VH3-33, VH3-74, and VH5-51 in VH and D1-7, D1-14, D1-20, and D5-5/18 in D segments in the HIVD group, as well as shorter and preferential glycine usages in CDRH3 regions. Gene selections were also detected in light chains. Notably, differences between productive rearrangements of HIVDs and HDs outnumbered those between productive and non-productive rearrangements within HIVDs. HIV infection may exert a dominant impact on the development of the plasmablast repertoire. The impact of selection is of limited significance in shaping the plasmablast repertoire. Overall, the data indicate that the environment in which the plasmablasts live can affect the distribution of the VH and VL genes in the repertoire and the amino acid compositions of the expressed Abs.

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Mechanisms That Shape Human Antibody Repertoire Development in Mice Transgenic for Human Ig H and L Chain Loci

Cited by 10 publications

References 64 publications

Comparisons of the antibody repertoires of a humanized rodent and humans by high throughput sequencing

Comparisons of the antibody repertoires of a humanized rodent and humans by high throughput sequencing

Comparisons of the antibody repertoires of a humanized rodent and humans by high throughput sequencing

Characteristics of Plasmablast Repertoire in Chronically HIV-Infected Individuals for Immunoglobulin H and L Chain Profiled by Single-Cell Analysis

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