High diversity of the immune repertoire in humanized NOD.SCID.γc<sup>−/−</sup> mice

Marodon, Gilles; Desjardins, Delphine; Mercey, Laetitia; Baillou, Claude; Parent, Pierric; Manuel, Manuarii; Caux, Christophe; Bellier, Bertrand; Pasqual, Nicolas; Klatzmann, David

doi:10.1002/eji.200939480

Cited by 53 publications

(58 citation statements)

References 38 publications

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“…The detailed protocol has been published previously [5]. Briefly, NOD.Cg-Prkdc scid Il2rg tm1Wjl/SzJ (NSG) (stock 005557, The Jackson Laboratory) mice were bred in our own animal facilities in Specific Pathogen-Free conditions (accreditation number from the Veterinary services: A75-13-10).…”

Section: Mice and Reconstitutionmentioning

confidence: 99%

“…Thymus sample from a child undergoing corrective cardiovascular surgery has been previously described [5]. Multiplex PCR was performed as described [5]. Quantile normalized data were used for repertoire perturbation analysis with the ISEApeaks software platform as described by Collette et al [13].…”

Section: Combinatorial Diversity Analysismentioning

confidence: 99%

“…Despite studies using high-throughput sequencing of TCR b chains [2,[7][8][9], a global view on the distribution of V-J rearrangement combinations of the TCR b chain in humans was lacking. Here, we have reanalyzed T-cell repertoire data that we generated in humanized NOD.SCID.gc À/À (NSG) mice [5] with the aim to determine the extent of the overlap between the T-cell repertoire of humans and humanized mice at the level of V-J rearrangements. In order to reveal the underlying structure of this complex data set, we applied an original combination of statistical and bioinformatics tools, including multivariate ranking and bootstrap analysis for credibility and confidence assessment.…”

Section: Introductionmentioning

confidence: 99%

“…Briefly, genomic DNA was extracted from human CD45 1 CD3 1 cells sorted on an FACSAria (Becton Dickinson, le Pont-de-Claix, France) with purity greater than 90% from mouse spleen or from mononuclear peripheral blood cells in humans after informed consent. Thymus sample from a child undergoing corrective cardiovascular surgery has been previously described [5]. Multiplex PCR was performed as described [5].…”

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confidence: 99%

“…The extent of the human T-cell repertoire diversity generated in mice may represent an important functional indicator [3], useful for immunological studies in the models. However, although some studies have reported on the TCR repertoire diversity of humanized mice [4,5], a comprehensive analysis of T-cell repertoire combinatorial diversity in humanized mice is lacking. We believe that this analysis is relevant since it may provide new information about the rules governing V-J combinations distribution in humans.…”

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Half of the T‐cell repertoire combinatorial diversity is genetically determined in humans and humanized mice

Pham

Manuel²,

Petit

et al. 2011

Eur J Immunol

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In humanized mice, the T-cell repertoire is derived from genetically identical human progenitors in distinct animals. Thus, careful comparison of the T-cell repertoires of humanized mice with those of humans may reveal the contribution of genetic determinism on T-cell repertoire formation. Here, we performed a comprehensive assessment of the distribution of V-J combinations of the human b chain of the T-cell receptor (hTRBV) in NOD.SCID.cc À/À (NSG) humanized mice. We observed that numerous V-J combinations were equally distributed in the thymus and in the periphery of humanized mice compared with human references. A global analysis of the data, comparing repertoire perturbation indices in humanized NSG mice and unrelated human PBMCs, reveals that 50% of the hTRBV families significantly overlapped. Using multivariate ranking and bootstrap analyses, we found that 18% of all possible V-J combinations contributed close to 50% of the expressed diversity, with significant over-representation of BV5-J1.111.2 and BV6-J1.111.2 rearrangements. Finally, comparison of CD3 À and CD3 1 thymocyte repertoires indicated that the observed V-J combination overlap was already present before TCR-MHC selection in the thymus. Altogether, our results show that half of the T-cell repertoire combinatorial diversity in humans is genetically determined.Key words: Humanized mice . ISEApeaks . Multivariate score . T-cell repertoire . V-J rearrangements Supporting Information available online Introduction T-cell repertoire diversity is based upon tightly ordered genetic rearrangements of the T-cell receptor V, D and J genes. The available numbers of these genes in the genome constitutes a first level of diversity: to date, 33 and 65 genes encoding variable elements of the human T-cell receptor b chain (hTRBV) from 30 hTRBV families have been characterized in mice and humans respectively (according to the ImMunoGeneTics (IMGT) database (http://www.imgt.org)). An additional level of diversity comes from the association of single V, (D) and J elements together. The major determinant of repertoire diversity is then random addition of nucleotides at the junction of these V(D)J gene segments, constituting the CDR3 of the TCR. Finally, a fourth level of complexity originates from the association of the rearranged a and b chains of the TCR. In theory, these sequential processes occurring during T-cell differentiation in the thymus could Eur. J. Immunol. 2012. 42: 760-770 760 generate up to 10 15 different TCRs, a number far in excess of the number of T-cells in the body. The 'real' size of the repertoire is probably much lower though, with recent estimations of unique TCR b chains ranging from 10 6 to 4.10 6 in humans [1,2]. Mice reconstituted with a human immune system generated from hematopoietic precursors represent a new animal model for human immunology studies. The extent of the human T-cell repertoire diversity generated in mice may represent an important functional indicator [3], useful for immunological studies in the models. However, a...

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Section: Mice and Reconstitutionmentioning

confidence: 99%

Section: Combinatorial Diversity Analysismentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

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Half of the T‐cell repertoire combinatorial diversity is genetically determined in humans and humanized mice

Pham

Manuel²,

Petit

et al. 2011

Eur J Immunol

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Infectious diseases in humanized mice

et al. 2013

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Despite many theoretical incompatibilities between mouse and human cells, mice with reconstituted human immune system components contain nearly all human leukocyte populations. Accordingly, several human-tropic pathogens have been investigated in these in vivo models of the human immune system, including viruses such as human immunodeficiency virus (HIV) and Epstein-Barr virus (EBV), as well as bacteria such as Mycobacterium tuberculosis and Salmonella enterica Typhi. While these studies initially aimed to establish similarities in the pathogenesis of infections between these models and the pathobiology in patients, recent investigations have provided new and interesting functional insights into the protective value of certain immune compartments and altered pathology upon mutant pathogen infections. As more tools and methodologies are developed to make these models more versatile to study human immune responses in vivo, such improvements build toward small animal models with human immune components, which could predict immune responses to therapies and vaccination in human patients. Keywords:Dengue virus r EBV r HIV r Mycobacterium tuberculosis r Salmonella enterica typhi IntroductionThe complexity of infections and the corresponding elicited immune responses are best investigated in animal models that allow the manipulation of the timing and dose of infection, as well as of the responding immune compartments. Small animal models, such as the mouse, are preferred for these types of investigations due to low costs and ease of handling. However, divergent evolution between these small mammals and humans in the past 65 million years has rendered the immune system the third most different organ system between the two species, after olfaction and reproduction [1]. Many of these differences are found in the innate immune system, which ensures the initial survival of the infected host and also recognizes pathogens by their molecular patterns [2]. This divergence probably results from the different Correspondence: Prof. Christian Münz e-mail: christian.muenz@uzh.ch infectious disease challenges associated with the respective ecological niches that these two species inhabit. Unfortunately, these differences between the mouse and human immune systems also result in dissimilar inflammatory responses to burns, trauma, and endotoxemia at the gene expression level, such as integrin, ICOS-ICOSL, CD28, and PKC signaling [3]. Therefore, alternatives to classical mouse models, which more closely model human immune system behavior during infection in vivo, would be of significant benefit for the development of immunomodulatory treatments.The category of new models, which comes closest to achieving this goal, is mice with reconstituted human immune system components. These mice are mainly generated by neonatal injection of human hematopoietic progenitor cells in mice that lack murine innate and adaptive lymphocytes, namely NOD-scid γ c −/− (NSG), (Fig. 1). For some studies, a fetal organoid of liver and thymic tissue is implanted ...

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