Humanization of T cell–mediated immunity in mice

Moore, Michael J.; Zhong, Maggie; Hansen, Johanna K.; Gärtner, Hans; Grant, Craig; Huang, Mingliang; Harris, Faith M.; Tu, Naxin; Bowerman, Natalie A.; Edelmann, Kurt H.; Barry, Thomas; Herbin, Olivier; Tay, Chin-Siean; DiLillo, David J.; Decker, Corinne E.; Levenkova, Natasha; Shevchuk, James; Dhanik, Ankur; Meagher, Karoline A; Karr, Amanda; Roos, Jan C.; Lee, Wen‐Yi; Suh, David; Eckersdorff, Mark; Meagher, T. Craig; Koss, Matthew; Esau, Lakeisha; Sleeman, Matthew A.; Babb, Robert; Chen, Gang; Kyratsous, Christos A.; Poueymirou, William; McWhirter, John R.; Voronina, Vera A.; Guo, Chunguang; Gurer, Cagan; Yancopouloš, George D.; Murphy, Andrew; Macdonald, Lynn E.

doi:10.1126/sciimmunol.abj4026

Cited by 11 publications

(11 citation statements)

References 34 publications

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“…Taken together with previous cryoEM studies that focused on a different a/b TCR pair 28,34 , our PN45545 TCR-CD3 complex structure supports the notion that the overall structure and assembly of TCR-CD3 is unaffected by differences in TCR variable region sequence. Furthermore, the PN45545 TCR-CD3 structure confirms that antigen-specific TCRs discovered in humanized mice 29 have expected structural features.…”

Section: Cryoem Structure Of Full-length Pn45545 Tcr-cd3supporting

confidence: 63%

“…Two α/β TCRs, PN45545 and PN45428, were isolated from humanized VelociT mice 29 immunized with the MAGEA4 (230-239) peptide. We first focused our studies on the PN45545 TCR.…”

Section: Resultsmentioning

confidence: 99%

“…Here, we used cryoEM to investigate the structural basis for the recognition of the cancer antigen HLA-A2/MAGEA4 by two TCRs (PN45545 and PN45428) isolated from mice with humanized T-cell immunity 29 . Our structures of two full-length, glycosylated TCR-CD3 complexes bound to antigen showed overall canonical antigen docking modes, though they both displayed a distinct shift toward the N-terminal side of the peptide.…”

Section: Discussionmentioning

confidence: 99%

“…TCRs were expressed in primary human T cells by targeting AAV-encoded TCR constructs to the TRAC locus as previous described 29 . Three days after activation, beads were removed, and cells were nucleofected with Crispr RNP consisting of Cas9 protein (Life Tech A36499) complexed with a mixture of modified synthetic guide RNAs (sgRNAs, IDT) targeting the TRAC and TRBC1/2 genes in their first exons.…”

Section: Flow Cytometrymentioning

confidence: 99%

“…Here, we use cryoEM to investigate TCR-pMHC recognition, focusing on two a/b TCRs derived from humanized mice 29 that target a peptide epitope containing residues 230-239 from the cancer-testis antigen MAGEA4. This peptide is presented by the MHC molecule HLA-A2 in numerous solid tumors, but not healthy adult tissues, making it an attractive target for TCR-based therapies and cancer vaccines [30][31][32] .…”

Section: Introductionmentioning

confidence: 99%

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Structural analysis of cancer-relevant TCR-CD3 and peptide-MHC complexes by cryoEM

Saotome

Dudgeon

Colotti

et al. 2022

Preprint

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The recognition of antigenic peptide-MHC (pMHC) molecules by T-cell receptors (TCR) initiates the T-cell mediated immune response. Structural characterization is key for understanding the specificity of TCR-pMHC interactions and informing the development of therapeutics. Despite the rapid rise of single particle cryoelectron microscopy (cryoEM), x-ray crystallography has remained the preferred method for structure determination of TCR-pMHC complexes. Here, we report cryoEM structures of two distinct full-length a/b TCR-CD3 complexes bound to their pMHC ligand, the cancer-testis antigen HLA-A2/MAGEA4 (230-239). We also determined cryoEM structures of pMHCs containing MAGEA4 (230-239) peptide and the closely related MAGEA8 (232-241) peptide in the absence of TCR, which provided a structural explanation for the MAGEA4 preference displayed by the TCRs. These findings provide insights into the TCR recognition of a clinically relevant cancer antigen and demonstrate the utility of cryoEM for high-resolution structural analysis of TCR-pMHC interactions.

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Section: Cryoem Structure Of Full-length Pn45545 Tcr-cd3supporting

confidence: 63%

“…Two α/β TCRs, PN45545 and PN45428, were isolated from humanized VelociT mice 29 immunized with the MAGEA4 (230-239) peptide. We first focused our studies on the PN45545 TCR.…”

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Flow Cytometrymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Structural analysis of cancer-relevant TCR-CD3 and peptide-MHC complexes by cryoEM

Saotome

Dudgeon

Colotti

et al. 2022

Preprint

Self Cite

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Human MHC Class II and Invariant Chain Knock‐in Mice Mimic Rheumatoid Arthritis with Allele Restriction in Immune Response and Arthritis Association

Romero‐Castillo,

Li,

et al. 2024

Advanced Science

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Transgenic mice expressing human major histocompatibility complex class II (MHCII) risk alleles are widely used in autoimmune disease research, but limitations arise due to non‐physiologic expression. To address this, physiologically relevant mouse models are established via knock‐in technology to explore the role of MHCII in diseases like rheumatoid arthritis. The gene sequences encoding the ectodomains are replaced with the human DRB1*04:01 and 04:02 alleles, DRA, and CD74 (invariant chain) in C57BL/6N mice. The collagen type II (Col2a1) gene is modified to mimic human COL2. Importantly, DRB1*04:01 knock‐in mice display physiologic expression of human MHCII also on thymic epithelial cells, in contrast to DRB1*04:01 transgenic mice. Humanization of the invariant chain enhances MHCII expression on thymic epithelial cells, increases mature B cell numbers in spleen, and improves antigen presentation. To validate its functionality, the collagen‐induced arthritis (CIA) model is used, where DRB1*04:01 expression led to a higher susceptibility to arthritis, as compared with mice expressing DRB1*04:02. In addition, the humanized T cell epitope on COL2 allows autoreactive T cell‐mediated arthritis development. In conclusion, the humanized knock‐in mouse faithfully expresses MHCII, confirming the DRB1*04:01 alleles role in rheumatoid arthritis and being also useful for studying MHCII‐associated diseases.

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What the Omicron wave is revealing about human immunity

Willyard¹

2022

Nature

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Humanization of T cell–mediated immunity in mice

Abstract: Knock-in mice with humanized TCR, MHC, and co-receptor genes enable therapeutic human TCR discovery and functional assessment in mice.

Cited by 11 publications

References 34 publications

Structural analysis of cancer-relevant TCR-CD3 and peptide-MHC complexes by cryoEM

Structural analysis of cancer-relevant TCR-CD3 and peptide-MHC complexes by cryoEM

Human MHC Class II and Invariant Chain Knock‐in Mice Mimic Rheumatoid Arthritis with Allele Restriction in Immune Response and Arthritis Association

What the Omicron wave is revealing about human immunity

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