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Antibody Repertoires in Humanized NOD-scid-IL2Rγ(null) Mice and Human B Cells Reveals Human-Like Diversification and Tolerance Checkpoints in the Mouse

Immunodeficient mice reconstituted with human hematopoietic stem cells enable the in vivo study of human hematopoiesis. In particular, NOD-scid-IL2Rγ(null) engrafted mice have been shown to have reasonable levels of T and B cell repopulation and can mount T-cell dependent responses; however, antigen...

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Autores principales: Ippolito, Gregory C., Hoi, Kam Hon, Reddy, Sai T., Carroll, Sean M., Ge, Xin, Rogosch, Tobias, Zemlin, Michael, Shultz, Leonard D., Ellington, Andrew D., VanDenBerg, Carla L., Georgiou, George
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3338711/
https://www.ncbi.nlm.nih.gov/pubmed/22558161
http://dx.doi.org/10.1371/journal.pone.0035497
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author Ippolito, Gregory C.
Hoi, Kam Hon
Reddy, Sai T.
Carroll, Sean M.
Ge, Xin
Rogosch, Tobias
Zemlin, Michael
Shultz, Leonard D.
Ellington, Andrew D.
VanDenBerg, Carla L.
Georgiou, George
author_facet Ippolito, Gregory C.
Hoi, Kam Hon
Reddy, Sai T.
Carroll, Sean M.
Ge, Xin
Rogosch, Tobias
Zemlin, Michael
Shultz, Leonard D.
Ellington, Andrew D.
VanDenBerg, Carla L.
Georgiou, George
author_sort Ippolito, Gregory C.
collection PubMed
description Immunodeficient mice reconstituted with human hematopoietic stem cells enable the in vivo study of human hematopoiesis. In particular, NOD-scid-IL2Rγ(null) engrafted mice have been shown to have reasonable levels of T and B cell repopulation and can mount T-cell dependent responses; however, antigen-specific B-cell responses in this model are generally poor. We explored whether developmental defects in the immunoglobulin gene repertoire might be partly responsible for the low level of antibody responses in this model. Roche 454 sequencing was used to obtain over 685,000 reads from cDNA encoding immunoglobulin heavy (IGH) and light (IGK and IGL) genes isolated from immature, naïve, or total splenic B cells in engrafted NOD-scid-IL2Rγ(null) mice, and compared with over 940,000 reads from peripheral B cells of two healthy volunteers. We find that while naïve B-cell repertoires in humanized mice are chiefly indistinguishable from those in human blood B cells, and display highly correlated patterns of immunoglobulin gene segment use, the complementarity-determining region H3 (CDR-H3) repertoires are nevertheless extremely diverse and are specific for each individual. Despite this diversity, preferential D(H)-J(H) pairings repeatedly occur within the CDR-H3 interval that are strikingly similar across all repertoires examined, implying a genetic constraint imposed on repertoire generation. Moreover, CDR-H3 length, charged amino-acid content, and hydropathy are indistinguishable between humans and humanized mice, with no evidence of global autoimmune signatures. Importantly, however, a statistically greater usage of the inherently autoreactive IGHV4-34 and IGKV4-1 genes was observed in the newly formed immature B cells relative to naïve B or total splenic B cells in the humanized mice, a finding consistent with the deletion of autoreactive B cells in humans. Overall, our results provide evidence that key features of the primary repertoire are shaped by genetic factors intrinsic to human B cells and are principally unaltered by differences between mouse and human stromal microenvironments.
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spelling pubmed-33387112012-05-03 Antibody Repertoires in Humanized NOD-scid-IL2Rγ(null) Mice and Human B Cells Reveals Human-Like Diversification and Tolerance Checkpoints in the Mouse Ippolito, Gregory C. Hoi, Kam Hon Reddy, Sai T. Carroll, Sean M. Ge, Xin Rogosch, Tobias Zemlin, Michael Shultz, Leonard D. Ellington, Andrew D. VanDenBerg, Carla L. Georgiou, George PLoS One Research Article Immunodeficient mice reconstituted with human hematopoietic stem cells enable the in vivo study of human hematopoiesis. In particular, NOD-scid-IL2Rγ(null) engrafted mice have been shown to have reasonable levels of T and B cell repopulation and can mount T-cell dependent responses; however, antigen-specific B-cell responses in this model are generally poor. We explored whether developmental defects in the immunoglobulin gene repertoire might be partly responsible for the low level of antibody responses in this model. Roche 454 sequencing was used to obtain over 685,000 reads from cDNA encoding immunoglobulin heavy (IGH) and light (IGK and IGL) genes isolated from immature, naïve, or total splenic B cells in engrafted NOD-scid-IL2Rγ(null) mice, and compared with over 940,000 reads from peripheral B cells of two healthy volunteers. We find that while naïve B-cell repertoires in humanized mice are chiefly indistinguishable from those in human blood B cells, and display highly correlated patterns of immunoglobulin gene segment use, the complementarity-determining region H3 (CDR-H3) repertoires are nevertheless extremely diverse and are specific for each individual. Despite this diversity, preferential D(H)-J(H) pairings repeatedly occur within the CDR-H3 interval that are strikingly similar across all repertoires examined, implying a genetic constraint imposed on repertoire generation. Moreover, CDR-H3 length, charged amino-acid content, and hydropathy are indistinguishable between humans and humanized mice, with no evidence of global autoimmune signatures. Importantly, however, a statistically greater usage of the inherently autoreactive IGHV4-34 and IGKV4-1 genes was observed in the newly formed immature B cells relative to naïve B or total splenic B cells in the humanized mice, a finding consistent with the deletion of autoreactive B cells in humans. Overall, our results provide evidence that key features of the primary repertoire are shaped by genetic factors intrinsic to human B cells and are principally unaltered by differences between mouse and human stromal microenvironments. Public Library of Science 2012-04-27 /pmc/articles/PMC3338711/ /pubmed/22558161 http://dx.doi.org/10.1371/journal.pone.0035497 Text en Ippolito et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Ippolito, Gregory C.
Hoi, Kam Hon
Reddy, Sai T.
Carroll, Sean M.
Ge, Xin
Rogosch, Tobias
Zemlin, Michael
Shultz, Leonard D.
Ellington, Andrew D.
VanDenBerg, Carla L.
Georgiou, George
Antibody Repertoires in Humanized NOD-scid-IL2Rγ(null) Mice and Human B Cells Reveals Human-Like Diversification and Tolerance Checkpoints in the Mouse
title Antibody Repertoires in Humanized NOD-scid-IL2Rγ(null) Mice and Human B Cells Reveals Human-Like Diversification and Tolerance Checkpoints in the Mouse
title_full Antibody Repertoires in Humanized NOD-scid-IL2Rγ(null) Mice and Human B Cells Reveals Human-Like Diversification and Tolerance Checkpoints in the Mouse
title_fullStr Antibody Repertoires in Humanized NOD-scid-IL2Rγ(null) Mice and Human B Cells Reveals Human-Like Diversification and Tolerance Checkpoints in the Mouse
title_full_unstemmed Antibody Repertoires in Humanized NOD-scid-IL2Rγ(null) Mice and Human B Cells Reveals Human-Like Diversification and Tolerance Checkpoints in the Mouse
title_short Antibody Repertoires in Humanized NOD-scid-IL2Rγ(null) Mice and Human B Cells Reveals Human-Like Diversification and Tolerance Checkpoints in the Mouse
title_sort antibody repertoires in humanized nod-scid-il2rγ(null) mice and human b cells reveals human-like diversification and tolerance checkpoints in the mouse
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3338711/
https://www.ncbi.nlm.nih.gov/pubmed/22558161
http://dx.doi.org/10.1371/journal.pone.0035497
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