Immunity

Memory B cells (MBCs) are a critical cellular reservoir for long-term humoral immunity. MBCs display heterogeneous isotypes and surface markers and can arise through both germinal center (GC)-dependent and GC-independent extrafollicular (EF) pathways. Both the mechanisms controlling EF MBC differentiation and the identities of the B cell populations from which EF MBCs derive remain poorly understood. To capture MBC diversity, we applied a broad selection strategy followed by transcriptional profiling, identifying a subset of MBCs characterized by minimal class switching, limited somatic hypermutation, and an innate-like gene signature. Using genetic models, we demonstrated that this subset arises independently of GC responses and derives from innate B1 cells. These innate-like MBCs differentiate into antigen-specific antibody-secreting cells and confer protection against lethal viral infection. Together, our findings define a previously unrecognized arm of MBC responses, demonstrating that innate B1 cells contribute a non-redundant antibody repertoire to protective immunological memory.

Induction of broadly neutralizing antibody (bnAb) responses remains a central challenge to designing efficacious HIV vaccines. Lineage design strategies, in which bnAb precursors are guided via iterative immunizations to their mature forms, depend on high levels of somatic mutation and the recall of antigen-specific B cells. Recent studies have identified spatial context as an important determinant of boosting efficacy, but the application of this to HIV vaccines and the underlying mechanisms are incompletely understood. Here, using mice harboring a V3-glycan bnAb lineage precursor knock-in combined with lineage-tracing and single-cell analyses, we show that persistent germinal centers (GCs) support continued affinity maturation of founder clones and ipsilateral boosting preferentially engages these lineages in secondary GCs. In contrast, contralateral boosting predominantly recruits naive B cells and memory B cells not directed towards the immunizing antigen. The few memory cells recruited at this site were biased towards a plasma cell fate. Finally, we identify disfavored mutational trajectories within the V3-glycan bnAb lineage, revealing intrinsic constraints on bnAb lineage evolution.

While aging is the primary risk factor for cancer development and a critical driver of metastasis, yet how aging selectively remodels specific distant organs to favor tumor colonization remains unclear. Here, we demonstrate that host aging selectively enhances susceptibility to lung metastasis in an inflammation-dependent manner, without uniformly affecting other organs. Single-cell RNA sequencing reveals that the aged lung accumulates PD-L1CTLA-4IgMIgD- regulatory B (Breg)-like cells, representing the immune population most amplified by the cooperative effects of aging and tumor burden. Trajectory analysis delineates their differentiation from naive B cells, driven by Il10, Ctla4, Cd274, and Egr1 upregulation. This program is evolutionarily conserved, increasing progressively with human chronological lung age. Mechanistically, aged CD140a adventitial fibroblasts drive CXCL13-dependent B-cell reprogramming into a senescent state. These Breg-like cells directly impair natural killer cell cytotoxicity and CD4 T-cell responses, accelerating pulmonary colonization. Our findings reveal a targetable, lung-specific age-associated stromal-immune axis driving metastatic organotropism.

Mouse models have been instrumental in defining immune mechanisms but often fail to capture the complexity of human immunity, limiting clinical translation. A major limitation is the immunological immaturity of specific pathogen-free (SPF) mice relative to pathogen-experienced adult humans. Here, we use a sequential infection (SI) model that recapitulates cumulative pathogen exposure and define its impact on immune composition and function. Beyond the previously reported expansion of memory T cells, SI induced durable, system-wide remodeling across lymphoid and non-lymphoid tissues, reshaping innate and adaptive immune populations, tissue-resident immunity, and hematopoietic output. Single-cell transcriptomic analyses revealed inflammatory imprinting of naive CD4 and CD8 T cells, whereas memory T cells acquired enhanced effector programs coupled with reduced biosynthetic activity, transcriptional states that more closely resemble those of pathogen-experienced adult humans. Functionally, SI mice recapitulated the human response to anti-CD28 super-agonist and exhibited altered magnitude and differentiation of acute and chronic antiviral T cell responses, demonstrating that cumulative pathogen exposure reshapes both existing immunity and the generation of future immune responses. Thus, cumulative pathogen exposure coordinately remodels hematopoiesis and naive and memory lymphocyte states, establishing a durable inflammation-experienced immune landscape that reshapes both immune memory and future immune responses, with broad implications for the translational fidelity of preclinical mouse models.

Staphylococcus aureus remains a major global pathogen with no licensed vaccine and high recurrent infection burden, yet correlates of protection remain undefined. In a prospective pediatric cohort, we profiled 319 children spanning non-carriers, asymptomatic carriers, those with skin and soft tissue infection (SSTI), or invasive disease. We interrogated 182,149 antibody features, generating the most comprehensive S. aureus immune profiling dataset to date. Antibody responses increased with age, marked by expansion of IgG subclasses and Fc-receptor engagement. Asymptomatic carriage was associated with functional antibody profiles targeting conserved surface antigens and select toxins. Multivariate modeling robustly distinguished clinical phenotypes and identified high-value antigens associated with disease resilience. Protection from recurrent disease converged on enhanced Fc{gamma}R binding and antibody effector function. These findings nominate key antigen targets, and highlight anti-Hla neutralizing antibodies and functional antibodies to additional surface antigens that can be recapitulated through Fc engineering, informing next-generation vaccine and monoclonal antibody strategies.

Food allergies are associated with progressive gastrointestinal symptoms driven by exacerbated mucosal type-2 immunity. Here, we investigated whether cellular interactions between the gut epithelium and innate immune cells regulate the severity of allergic symptoms in mice. Using the BALB/c OVA-alum food allergy model, we observed that repeated oral allergen challenges remodel the gut epithelium, expanding tuft cells, while shifting the intestinal stem cell niche toward a fetal-like repair state. Using uLIPSTIC, we systematically characterized in vivo immune-epithelial interactions and found that eosinophils and mast cells directly interact with intestinal epithelial cells (iECs) in an allergen challenge-dependent manner. Epithelial subset-specific uLIPSTIC provided further resolution and revealed that eosinophils and mast cells contact enteroendocrine cells and Paneth cells in a regionally compartmentalized manner. Allergic challenge was associated with rapid eosinophil migration towards the crypts and modulation of iEC differentiation. Depletion of eosinophils using two independent approaches reversed key markers of food allergy-associated epithelial remodeling, while exacerbating allergic diarrhea and mortality from anaphylactic shock. These findings establish eosinophils as orchestrators of protective epithelial remodeling in food allergies.

Personalized neoantigen (neoAg) vaccines have shown clinical promise in solid tumors1-8, yet their efficacy and mechanism of action in hematopoietic malignancies remain poorly defined9-11. Herein, we establish an immunocompetent syngeneic A20 B-cell lymphoma platform to test the efficacy of neoAg vaccines used either as mono- or combinatorial therapies with other immunotherapies12-17. Whereas subcutaneous A20 tumors were refractory to single-agent PD-1 or CTLA4 therapy, they were eradicated in a T cell-dependent manner in 90% of syngeneic hosts treated with dual immune checkpoint therapy (dual ICT, i.e., PD-1 + CTLA4). By mapping antigen specificity of dual-ICT-elicited T cells, we identified and validated dominant endogenous A20 MHC-I and MHC-II neoantigens and designed therapeutic synthetic long peptide (SLP) vaccines containing these neoepitopes. This vaccine (A20 neoVAX) promoted robust neoAg-specific CD4{square} and CD8{square} T cell responses in naive syngeneic BALB/c mice and induced tumor rejection in [~]70% of subcutaneous tumor-bearing mice. In addition, nearly all mice rejected their subcutaneous A20 tumors when A20 neoVAX was combined with PD-1. To render the results of this study more physiologic, we developed a systemic A20 lymphoma model and found that dual ICT failed to control tumor progression and A20 neoVAX delayed tumor progression and prolonged animal survival but did not induce tumor rejection. In contrast, A20 neoVAX plus dual ICT achieved durable systemic tumor elimination. Mechanistically, the combination of A20 neoVAX plus dual ICT amplified priming of A20 neoAg-specific T cells, prevented T cell dysfunction, sustained the cytotoxic capacity of tumor-specific CD8+ T cells, and induced Th1-skewing of CD4+ T cells in tumor and peripheral compartments. To increase the clinical relevance of these findings and to minimize potential adverse events in tumor-bearing, therapeutically treated individuals, we substituted CD8-targeted cytokine muteins (CD8-IL2 or CD8-IL21) for CTLA4. These agents represent genetically modified forms of IL-2 or IL-21 that selectively stimulate CD8+ T cells but have significantly reduced capacity to activate chronic inflammation and immunosuppressive functions of other immune cells. Whereas mice bearing systemic A20 lymphoma treated with either nothing, A20 neoVAX, or A20 neoVAX + CD8-IL2 failed to control tumor outgrowth, 66.7% of tumor-bearing mice treated with A20 neoVAX + CD8-IL2 + PD-1 rejected their tumors. In similar experiments in which CD8-IL21 was substituted for CD8-IL2, tumor clearance was also observed in two-thirds of A20-bearing mice but now rejection occurred in the absence of PD1. Together, these data define a framework for optimal personalized neoAg vaccination in B-lymphoma and demonstrate that neoAg vaccines can safely synergize with CD8+ T cell-selective immunotherapies to prevent T-cell dysfunction and generate durable systemic anti-tumor immunity.

Tissue-resident memory B cells (BRM) provide powerful localized protection against microbial infection in barrier tissues. It is unknown if analogous BRM populations survey solid tumors and contribute to anti-cancer immunity. We profiled B cells from patients with colorectal cancer and cutaneous basal cell carcinoma and identified a CD69+ memory B cell population consistent with a tissue-resident phenotype. Integrative analysis of transcriptomic datasets identified an optimized signature enriched across cancer types. Tumor infiltrating BRM-like cells preferentially exhibited autoreactivity and their signature correlated with patient outcomes and response to immunotherapy. Skin and lung targeted vaccination established localized BRM that provided IgA dependent organ specific protection upon tumor challenge in murine models. These findings establish BRM as an active component of anti-cancer immunity via preferential reactivity to tumor associated self-antigens.

Glioblastoma is a lethal brain tumor that is unresponsive to current cancer immunotherapeutic approaches, including immune checkpoint blockade (ICB). This suggests that initial priming of T cells, rather than their expansion and licensing as effectors, is a restricting feature in this tumor setting. To overcome the limited initiation of CD8+ T cell responses, we employed a strong heterologous prime-boost vaccination with the simian adenovirus ChAdOx1 and poxvirus modified vaccinia Ankara (MVA). Vaccination conferred therapeutic efficacy against orthotopic, immune checkpoint-blockade (ICB)-refractory SB28 murine glioblastoma. Vaccination was effective against both the murine tumor antigen, P1A, and a newly identified glioblastoma-associated antigen, Gpr149. Additional treatment with ICB provided no additional benefit. Systemic ChAdOx1/MVA vaccination induced robust infiltration of antigen-specific T cells in tumor-challenged brains, the majority of which exhibited a CD103+CD69+CD8+ tissue-resident memory (TRM)-like phenotype. These cells were polyfunctional, durable in brains with sustained tumor control, and mediated tissue-specific immunological memory. Moreover, intracranial adoptive transfer of glioblastoma-derived antigen-specific TRM-like cells was sufficient to protect naive recipients from subsequent orthotopic tumor challenge. Together, these findings establish that viral vector vaccination can generate tumor-specific TRM-like cells that mediate effective anti-glioblastoma immunity, providing a rationale for clinical evaluation of ChAdOx1/MVA-based strategies in glioblastoma.

Segmented filamentous bacteria (SFB) are a canonical model of microbiota-driven Th17 immunity, but how distinct gut-associated lymphoid tissues shape the quality and effector potential of commensal-specific T-cell responses remains unclear. Here we show that Peyer's patches (PPs) and mesenteric lymph nodes (MLNs) generate transcriptionally, clonally, and functionally distinct SFB-reactive CD4 T-cell programs. In PPs, CCR2-dependent monocyte-derived LysoDCs capture luminal SFB and locally prime antigen-specific CD4 T cells. PP priming drives robust T cell activation, Th17 differentiation with type 1 regulatory (Tr1)-like features, preferential clonal expansion within Th17-Tfh17 lineages, and tissue-retention programs. In contrast, CCR2-independent MLN priming induces a less differentiated, recirculating profile dominated by non-expanded clonotypes. Notably, these distinct programs carry functional consequences. Upon transfer into Citrobacter rodentium-infected lymphopenic mice, PP-primed T cells preserve barrier integrity and limit pathology, whereas MLN-primed cells from the same donors fail to provide equivalent protection. Together, these findings establish PP LysoDCs as specialized orchestrators of compartmentalized microbiota-specific immunity and identify the anatomical site of commensal priming as a key determinant of T-cell functional diversification and mucosal immune outcome.

Antibody (Ab) feedback impacts germinal center (GC) responses. Mice given IgG1 or IgG2c recombinant Ab (rAb) specific for the receptor binding site (RBS) of H3 hemagglutinin (HA) and later immunized with the H3 HA trimer exhibited altered GC repertoires. Passive RBS rAb had no effect on the magnitude of ensuing primary GC responses but reduced the numbers of RBS-specific GC B cells. These losses were matched by increases in "unspecific" B cells which did not bind the HA immunogen, with no changes in B cells specific for distal epitopes. These effects were independent of IgG subclass. Higher doses of passive rAb resulted in reduced epitope-specific affinity maturation and clonal proliferation in GCs. Passive rAb generated Ab:HA complexes and favored recruitment of rare HA-specific GC B cells with enhanced avidity for the rAb:HA immune complex (IC). We show that some no- and low-affinity GC B cells represent responses to local ICs.

Type 1 diabetes (T1D) is caused by T cell-mediated autoimmune destruction of insulin-producing islet beta-cells. Treatment with T-cell depleting therapies delays the progression of stage 2 and 3 T1D, but these agents exert broad immunosuppressive effects on T cell populations, including T regulatory cells (Tregs), which are key in promoting immune tolerance. We evaluated non-obese diabetic (NOD) mice and recently diagnosed T1D patients and identified CD38 as a marker for pathogenic T cell populations. Using adoptive T-cell transfer in Recombination Activating Gene 1 knockout NOD mice and in a humanized mouse model of autoimmune diabetes, we demonstrated that CD38-expressing autoreactive T cells drive diabetes pathogenesis. Furthermore, we found that selective depletion of CD38+ cells, using an anti-CD38 monoclonal antibody (mAb), prevents insulitis and diabetes onset without depleting CD4+CD25+ Tregs. Administration of anti-CD38 mAb did not adversely affect islet function and may selectively eliminate immunogenic senescent islet beta-cells. These results support the strategy of selectively depleting diabetogenic T cells using an anti-CD38 mAb to treat T1D and restore immune tolerance. Therefore, transient depletion of autoreactive T cells using anti-CD38 mAb may provide a novel strategy to prevent or abrogate autoimmunity in T1D.

The theoretical diversity of T cell receptors (TCRs), generated through V(D)J recombination, is enormous, yet the diversity of TCRs capable of recognizing the same epitope remains unknown. Defining this TCR solution space is essential for uncovering basic principles that govern TCR specificity. Using single-cell RNA and TCR sequencing, we generated ultra-deep (more than 4000 unique TCRs per epitope) epitope-specific TCR libraries derived from 560 immunized C57BL/6 mice, identifying over 27,000 unique epitope-reactive TCRs across three distinct CD8+ T cell epitopes presented by two major histocompatibility complex (MHC) class I alleles. Saturation analyses indicated that the solution space for all studied epitopes comprises many tens of thousands of unique TCRs. Despite highly skewed and peptide-dependent VJ-usage patterns, nearly the entire set of functional germline V/ and J/ segments was detected at least once within each epitope-specific repertoire. Therefore, diversity of epitope-specific TCRs is not limited by distinct germline combinations but rather can emerge from a near-to-complete combinatorial space of - and -chain, V and J segments paired with compatible CDR3 sequences.

Age-associated thymic involution is a major driver of immunosenescence, yet the cellular and spatial mechanisms coordinating age-related thymic remodeling remain incompletely understood. Combining single-cell transcriptomics, chromatin accessibility profiling, and spatial transcriptomics, we generated a spatially resolved multi-omic atlas of the aging mouse thymus. We show that thymic aging is not simply a process of epithelial loss, but a spatial reorganization of the stroma into new microenvironments, including age-associated epithelial states, a fibroblast-supported epithelial progenitor niche, and tertiary lymphoid structures. This remodeling displaces niches supporting positive and negative thymocyte selection and coincides with an intrinsic decline in cortical thymic epithelial cell function. Ligand-receptor mapping identifies medullary fibroblasts as a signaling hub sustaining epithelial progenitors and promoting tertiary lymphoid structure neogenesis, linking these hallmarks of thymic aging. Together, our findings reframe thymic involution as spatial stromal reorganization that links stromal remodeling to impaired thymopoiesis, central tolerance, and immune aging.

Conventional dendritic cells navigate complex tissues and sample peripheral antigens, balancing immune suppression and activation to achieve tissue homeostasis. However, it remains unclear how an individual dendritic cells reconciles co-incident signals from immunological opposing antigens within the tissue or tumor microenvironment, where tolerogenic self-antigen and immunogenic tumor antigen or microbes coexist. Here, using complementary in vivo fluorescent reporter systems, high-resolution imaging and endosomal profiling, we simultaneously tracked uptake, intracellular processing and cross-presentation of cutaneous self, tumor and microbial antigens in murine skin tissue, tumors and draining lymph nodes. We find that a substantial fraction of dendritic cells acquire antigen from multiple sources and that localization within endosomal compartments is dictated by antigen source. Notably, type 1 conventional dendritic cells that co-process self and tumor antigen represent a significant proportion of tumor antigen-bearing dendritic cells in the tumor and tumor draining lymph node. These dual-antigen loaded dendritic cells display a diminished capacity to prime tumor-specific CD8+ T cells and a marked reduction in tumor derived peptide presented on surface MHCI, while cross-priming of self-antigen specific T cells is significantly increased. These changes occur despite equivalent or greater tumor antigen uptake relative to self-antigen and high expression of surface MHCI and costimulatory molecules. Together, these data support a model in which multiantigen processing within dendritic cells can bias peptide loading away from tumor-derived epitopes, thereby limiting tumor-specific cross-priming. Modulating the antigenic context of the tumor microenvironment or endosomal routing after antigen uptake may therefore represent a strategy to restore effective dendritic cell-mediated antitumor immunity.

Shingles (herpes zoster), caused by reactivation of varicella zoster virus (VZV), affects approximately one third of the global population. Besides environmental factors, host genetics play a role in determining susceptibility to shingles. Here, we performed a large-scale genome-wide association study (GWAS) meta-analysis of shingles across five cohorts comprising 72,935 cases and 1,644,597 controls of European ancestry. We identified seven genome-wide significant loci, including novel associations at IGHG1, IFNAR2, MPV17L2 (IL12RB1), BACH2, and RHOBTB1, implicating MHC class I antigen presentation, type I interferon signaling, humoral immunity, and T-cell memory maintenance as key genetic determinants of shingles susceptibility. HLA fine-mapping identified eight independently associated HLA alleles, mapping predominantly to HLA-B (HLA-B*44:02), with additional associations at HLA-C (HLA-C*02:02) and an independent association at HLA-DQB1 (HLA-DQB1*05:02). Gene set analysis and stratified LD score regression identified significant enrichment of shingles heritability in immune tissues and pathways. Phenome-wide association study, genetic correlation analysis, and bidirectional two-sample Mendelian randomization identified causal effects of shingles on stroke, herpes simplex infection, and systemic lupus erythematosus, and suggested pain conditions and arthrosis as risk factors for shingles. These findings advance understanding of the genetic architecture of VZV reactivation and its causal relationships with other diseases.

Natural SARS-CoV-2 infections or vaccinations induce neutralizing antibodies (nAbs) offer protection from severe disease. The shared use of IGHV3-53/3-66 genes makes this class of monoclonal antibodies (mAbs) a public clonotype and is well established, but the evolution and structural basis of how these public antibodies maintain broad binding and acquire potent neutralizing activity is not completely understood. To understand how these features are facilitated by the IGHV3-53/3-66 germline segments and enhanced by somatic mutations, we investigated the biology of a panel of 242 human mAbs isolated from an individual infected with SARS-CoV-2 BA.1 strain and recovered. Interestingly, a mAb designated COV2-3731 encoded by IGHV3-53/IGKV1-33 retained potent neutralizing activity against SARS-CoV-2 variants BA.2.86, JN.1, KP.2, BA.3.2, and, to some extent, KP.3. Studies using deep mutational scanning with a BA.2 lentiviral library and determination of the structural complex of the BA.2 S protein and COV2-3731 Fab fragments using cryo-EM revealed key contact residues. Further, germline revertant analysis of the COV2-3731 mAb provided additional insights into how this COV2-3731 and other IGHV3-53/3-66-encoded public antibodies evolve to gain breadth against antigenically distinct SARS-CoV-2 variants such as BA.2.86, JN.1, KP.2, and BA.3.2.

Upon skin colonization, the prevalent human skin commensal S. epidermidis can elicit a CD8+ T cell response that protects against pathogens or clears tumors. The microbial features that drive this response are undefined, limiting our ability to understand and predict commensal-immune crosstalk and to engineer potent commensal-derived immunotherapies. To uncover these microbial features, we harnessed both the natural variation in CD8+ T cell induction across primary human isolates of Staphylococcus and our ability to genetically manipulate these strains. Stimulatory strains exhibit increased quorum sensing activation, which turns on a unique commensal-associated gene family, called phenol-soluble modulin {varepsilon} (PSM{varepsilon}), that is required for CD8+ T cell activation. PSM{varepsilon} not only acts as the immunodominant CD8+ T cell antigen but also enhances cross-presentation in an antigen-agnostic manner. Co-delivering PSM{varepsilon} promotes CD8+ T cell priming to an exogenous antigen via a mechanism that is independent of formyl peptide receptor and co-stimulatory receptor upregulation. Thus, we demonstrate that specific aspects of microbiome-immune crosstalk can be distilled to molecular components, which engage in previously undescribed mechanisms and can be harnessed for immunotherapy without requiring live bacterial colonization.

Interactions between Human Leukocyte Antigen (HLA) molecules and their cognate immunoreceptors are essential for regulating innate and adaptive immune cell functions. Leukocyte Immunoglobulin-like Receptors (LILRs) are key regulators of HLA-mediated immune responses, owing to their broad expression across immune cell populations and their ability to modulate both immune activation and tolerance. Among these, LILRB1-HLA interactions are increasingly recognised as important in transplantation, chronic infection and cancer therapies. Unlike other HLA-binding receptors, which recognise epitopes specific to HLA subsets, LILRB1 primarily engages the relatively conserved 3 and {beta}2-microglobulin components of HLA molecules, supporting its role as a broad regulator of pan-HLA class I-mediated functions. Nonetheless, there have been conflicting findings regarding the breadth of LILRB1-HLA-I interactions. While direct affinity studies on a limited subset of HLA-I molecules have revealed no significant differences in LILRB1 binding, broader analyses using single-antigen bead arrays suggest underlying variability. Here, we show through a broad binding assay that, while LILRB1 is a broad HLA-I-binding receptor, it exhibits differential preferences across HLA-I allotypes. Molecular dynamics analyses of the HLA-I-LILRB1 interface suggest that HLA-3 domain dynamism underlies these binding differences. We further determined the crystal structure of LILRB1 and used it to highlight intrinsic structural flexibility within its domains. Finally, these structural insights were leveraged to refine our understanding of the binding modalities of therapeutic monoclonal antibodies currently described. Together, our findings establish structural and mechanistic bases for differential HLA-I recognition by LILRB1 and provide insights into immunotherapeutic targeting of LILRB1.

HER2 mutations are oncogenic drivers in 1-6% of non-small cell lung cancers (NSCLC), but therapeutic resistance limits the durability of current HER2-targeted treatments. Here, we identify T-cell receptors (TCRs) targeting recurrent HER2 hotspot mutations as a potential immunotherapeutic strategy for HER2-mutant NSCLC. Using neoepitope prediction and antigen-specific T-cell enrichment, we isolated HLA-A*02:01restricted TCRs recognizing HER2 A775insYVMA, S310F, and G776delinsVC mutations, collectively covering approximately 60% of HER2-mutant NSCLC. These TCRs selectively recognized mutant HER2 epitopes without detectable wild-type reactivity and some displayed cross-recognition of related hotspot variants, expanding the spectrum of targetable tumors. The G776delinsVC-specific TCR also exhibited co-receptorindependent activity showcased by its ability to activate CD4+ T cells. Importantly, timelapse single-cell flow cytometry analyses demonstrated that TCR-engineered T cells repeatedly reacquired activated polyfunctional states following serial antigen stimulation, while serial tumor rechallenge assays confirmed sustained cytotoxic activity across multiple rounds of tumor killing. These findings identify recurrent HER2 mutations as shared immunotherapeutic targets and provide a foundation for the development of TCR-based therapies for HER2-mutant NSCLC.