Scalable TCR synthesis and screening enables antigen reactivity mapping in vitiligo

Stephanie A. Gaglione; Rachit S Mukkamala; Chirag Krishna; Blake E. Smith; Marc H. Wadsworth; Scott A. Jelinsky; Caleb R. Perez; Laura Schmidt-Hong; Erica L. Katz; Kyle J. Gellatly; Lestat R. Ali; Jie Shen; Patrick V. Holec; Qingyang Henry Zhao; Amanda O. Chan; Ellen J.K. Xu; Kellie M. Kravarik; Julia A. Guzova; Connor S. Dobson; Harshabad Singh; Manuel G. Garber; Michael Lawrence Dougan; Stephanie K. Dougan; John E. Harris; Aaron R. Winkler; Michael E. Birnbaum

doi:10.1016/j.immuni.2026.01.001

ImmunologyImmunityJan 28, 2026

Peer-reviewed journal article

Scalable TCR synthesis and screening enables antigen reactivity mapping in vitiligo

Stephanie A. Gaglione iD, Rachit S Mukkamala, Chirag Krishna iD, Blake E. Smith iD, Marc H. Wadsworth iD, Scott A. Jelinsky iD, Caleb R. Perez iD, Laura Schmidt-Hong iD, Erica L. Katz iD, Kyle J. Gellatly iD, Lestat R. Ali iD, Jie Shen, Patrick V. Holec iD, Qingyang Henry Zhao iD, Amanda O. Chan iD, Ellen J.K. Xu iD, Kellie M. Kravarik iD, Julia A. Guzova, Connor S. Dobson iD, Harshabad Singh iD, Manuel G. Garber iD, Michael Lawrence Dougan iD, Stephanie K. Dougan iD, John E. Harris iD, Aaron R. Winkler iD, Michael E. Birnbaum iD

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Abstract

T cell receptors (TCRs) mediate antigen recognition in adaptive immunity, yet large-scale mapping of TCR-antigen interactions remains a major challenge. Current approaches to synthesize and functionally screen TCRs remain technically complex and limited in throughput. We introduce a modular strategy, TCRAFT, to rapidly construct tens of thousands of TCRs for 99% accuracy. We integrate this approach with a high-throughput antigen discovery platform to enable library-on-library TCR-antigen screening. We reconstruct and screen 3,808 TCRs from vitiligo lesions, linking TCR specificity to transcriptional phenotypes for antigen-reactive T cells. To demonstrate scalability, we synthesize and screen 30,810 TCRs from donors with pancreatic ductal adenocarcinoma to capture antigen-specific TCRs. This workflow reduces the cost and complexity of large-scale TCR screening, enabling the expansion of the known landscape of antigen-specific TCRs in vitiligo with a method that can be readily extended to other immunological applications.