Out of sight, but not out of season: Nitrifier distributions and population dynamics in a large oligotrophic lake.

Nitrification is an important control on the form and distribution of nitrogen in freshwater ecosystems. However, the seasonality of nitrogen pools and the diversity of organisms catalyzing this process have not been well documented in oligotrophic lakes. Here, we show that nitrogen pools and nitrifying organisms in Flathead Lake are temporally and vertically dynamic, with nitrifiers displaying specific preferences depending on the season. While the ammonia-oxidizing bacteria (AOB) Nitrosomonadaceae and nitrite-oxidizing bacteria (NOB) Nitrotoga dominate at depth in the summer, the ammonia-oxidizing archaea (AOA) Nitrososphaerota and NOB Nitrospirota become abundant in the winter. Given clear seasonality in ammonium, with higher concentrations during the summer, we hypothesize that the succession between these two nitrifying groups may be due to nitrogen affinity, with AOB more competitive when ammonia concentrations are higher and AOA when they are lower. Nitrifiers in Flathead Lake share more than 99% average nucleotide identity with those reported in other North American lakes but are distinct from those in Europe and Asia, indicating a role for geographic isolation as a factor controlling speciation among nitrifiers. Our study shows there are seasonal shifts in nitrogen pools and nitrifying populations, highlighting the dynamic spatial and temporal nature of nitrogen cycling in freshwater ecosystems.

Barriers to care and treatment experiences among individuals with postpartum psychosis.

PURPOSE: Postpartum psychosis (PP) is a severe psychiatric disorder affecting 1-2 per 1,000 deliveries. Prompt access to healthcare and timely initiation of treatment are crucial to minimizing harm and improving outcomes. This analysis seeks to fill gaps in knowledge surrounding barriers to care and treatment experiences among this population. METHODS: Participants were individuals with histories of PP who enrolled in the Massachusetts General Hospital Postpartum Psychosis Project (MGHP3). The MGHP3 Healthcare Access Survey, a cross-sectional questionnaire, assesses barriers to care, treatment-seeking behaviors, and experiences with treatment. Descriptive statistics were utilized to describe sample characteristics. RESULTS: 139 participants provided 146 episode-specific survey responses. Lack of available services was cited as the greatest barrier to care for PP. Among those who sought treatment, obstetric providers (34.5%) and emergency medical professionals (29.4%) were the most common initial points of contact. 82.2% of the respondents went to an emergency room or crisis center during their episode(s). Most (61.8%) reported being given insufficient information to manage their PP. Approximately half of participants were hospitalized (55.5%), the majority of whom had no access to their infant during hospitalization (70.4%). Of those breastfeeding or pumping at admission, 31.3% were not given access to a breast pump. 44.4% dealt with delivery-related medical issues during their hospitalization. CONCLUSION: This report is the first of its kind to assess key public health domains among individuals with PP. Findings point to several directions for future research and clinical practice to improve treatment timeliness and quality, potentially improving long-term outcomes related to this serious illness.

The lived experiences of individuals with postpartum psychosis: A qualitative analysis.

Postpartum psychosis (PP) is a severe psychiatric illness that occurs in about 1 to 2 per 1000 people in the perinatal period. To date, qualitative research investigating PP has focused on specific topics, such as treatment experiences or the impact of the illness on patients' lives and families. These studies have included small samples of women with histories of PP, often limited to certain geographical areas or treatment centers. Given the heterogeneity in presentations of PP and access to care, larger and geographically diverse samples are needed to broadly understand this complex illness. Initiated in 2018, the Massachusetts General Hospital Postpartum Psychosis Project (MGHP3) consists of a large, international sample of those who have experienced PP. In addition to the specific aims of MGHP3, which include to better understand the phenomenology and potential genetic underpinnings of PP, this investigation invites participants to qualitatively describe their narratives of postpartum psychosis. This analysis included 130 participants who reported on 133 episodes of PP. Participants' responses to the PP narrative prompt fell under several overarching categories: 1) broad psychosocial experiences surrounding postpartum psychosis, 2) impact on the mother-baby dyad, 3) treatment experiences, and 4) recovery experiences. Our findings shed light on a range of ways in which individuals' lives are impacted by this illness, and point to areas for future research and clinical directions to improve the support and care for individuals with PP and their families.

White matter hyperintensities modify relationships between corticospinal tract damage and motor outcomes after stroke.

Motor outcomes after stroke relate to corticospinal tract (CST) damage. Concurrent damage from white matter hyperintensities (WMHs) might impact neurological capacity for recovery after CST injury. Here, we evaluated if WMHs modulate the relationship between CST damage and post-stroke motor impairment outcome. We included 223 individuals from the ENIGMA Stroke Recovery Working Group. CST damage was indexed with weighted CST lesion load (CST-LL). Mixed effects beta-regression models were fit to test the impact of CST-LL, WMH volume, and their interaction on motor impairment. WMH volume related to motor impairment above and beyond CST-LL (ß = 0.178, p = 0.022). We tested if relationships varied by WMH severity (mild vs. moderate-severe). In individuals with mild WMHs, motor impairment related to CST-LL (ß = 0.888, p < 0.001) with a CST-LL x WMH interaction (ß = -0.211, 0.026). In individuals with moderate-severe WMHs, motor impairment related to WMH volume (ß = 0.299, p = 0.044), but did not significantly relate to CST-LL or a CST-LL x WMH interaction. WMH-related damage may be under-recognised in stroke research as a factor contributing to variability in motor outcomes. Our findings emphasize the importance of brain structural reserve in motor outcomes after brain injury.

Erratum: A library of cancer testis specific T cell receptors for T cell receptor gene therapy.

[This corrects the article DOI: 10.1016/j.omto.2022.11.007.].

Rising wildfire risk to houses in the United States, especially in grasslands and shrublands.

Wildfire risks to homes are increasing, especially in the wildland-urban interface (WUI), where wildland vegetation and houses are in close proximity. Notably, we found that more houses are exposed to and destroyed by grassland and shrubland fires than by forest fires in the United States. Destruction was more likely in forest fires, but they burned less WUI. The number of houses within wildfire perimeters has doubled since the 1990s because of both housing growth (47% of additionally exposed houses) and more burned area (53%). Most exposed houses were in the WUI, which grew substantially during the 2010s (2.6 million new WUI houses), albeit not as rapidly as before. Any WUI growth increases wildfire risk to houses though, and more fires increase the risk to existing WUI houses.

Using high-resolution land cover data to assess structure loss in the 2018 Woolsey Fire in Southern California.

There are growing concerns about increases in the size, frequency, and destructiveness of wildfire events. One commonly used mitigation strategy is the creation and maintenance of defensible space, a zone around buildings where vegetation is managed to increase potential for structures to survive during wildfires. Despite widespread acceptance and advocacy of defensible space, few studies provide empirical evidence documenting the efficacy of different fuel modification practices under real wildfire conditions. The 2018 Woolsey Fire in Los Angeles County, California, occurred a short time after high-resolution (0.07 m2) land cover data were created, providing a unique opportunity to quantify vegetation before the fire. We integrated measurements from this high-resolution land cover data with parcel data, building attributes, and environmental context. We then used Random Forests models to analyze the extent to which these factors predicted structure loss in the wildfire. Variable importance scores showed vegetation around buildings was not a strong predictor of building-level damage outcomes compared to building materials and landscape features such as paved land cover per parcel, elevation, building density, and distance to road networks. Among building materials, multi-paned windows and enclosed eaves were most highly associated with building survival. These results are consistent with other studies that conclude building materials and environmental context are more related to survivorship than defensible space.

Social vulnerability of the people exposed to wildfires in U.S. West Coast states.

Understanding of the vulnerability of populations exposed to wildfires is limited. We used an index from the U.S. Centers for Disease Control and Prevention to assess the social vulnerability of populations exposed to wildfire from 2000-2021 in California, Oregon, and Washington, which accounted for 90% of exposures in the western United States. The number of people exposed to fire from 2000-2010 to 2011-2021 increased substantially, with the largest increase, nearly 250%, for people with high social vulnerability. In Oregon and Washington, a higher percentage of exposed people were highly vulnerable (>40%) than in California (~8%). Increased social vulnerability of populations in burned areas was the primary contributor to increased exposure of the highly vulnerable in California, whereas encroachment of wildfires on vulnerable populations was the primary contributor in Oregon and Washington. Our results emphasize the importance of integrating the vulnerability of at-risk populations in wildfire mitigation and adaptation plans.

The global wildland-urban interface.

The wildland-urban interface (WUI) is where buildings and wildland vegetation meet or intermingle1,2. It is where human-environmental conflicts and risks can be concentrated, including the loss of houses and lives to wildfire, habitat loss and fragmentation and the spread of zoonotic diseases3. However, a global analysis of the WUI has been lacking. Here, we present a global map of the 2020 WUI at 10 m resolution using a globally consistent and validated approach based on remote sensing-derived datasets of building area4 and wildland vegetation5. We show that the WUI is a global phenomenon, identify many previously undocumented WUI hotspots and highlight the wide range of population density, land cover types and biomass levels in different parts of the global WUI. The WUI covers only 4.7% of the land surface but is home to nearly half its population (3.5 billion). The WUI is especially widespread in Europe (15% of the land area) and the temperate broadleaf and mixed forests biome (18%). Of all people living near 2003-2020 wildfires (0.4 billion), two thirds have their home in the WUI, most of them in Africa (150 million). Given that wildfire activity is predicted to increase because of climate change in many regions6, there is a need to understand housing growth and vegetation patterns as drivers of WUI change.

Oxygen-consumption based quantification of chemogenetic H2O2 production in live human cells.

Reactive Oxygen Species (ROS) in the form of H2O2 can act both as physiological signaling molecules as well as damaging agents, depending on their concentration and localization. The downstream biological effects of H2O2 were often studied making use of exogenously added H2O2, generally as a bolus and at supraphysiological levels. But this does not mimic the continuous, low levels of intracellular H2O2 production by for instance mitochondrial respiration. The enzyme d-Amino Acid Oxidase (DAAO) catalyzes H2O2 formation using d-amino acids, which are absent from culture media, as a substrate. Ectopic expression of DAAO has recently been used in several studies to produce inducible and titratable intracellular H2O2. However, a method to directly quantify the amount of H2O2 produced by DAAO has been lacking, making it difficult to assess whether observed phenotypes are the result of physiological or artificially high levels of H2O2. Here we describe a simple assay to directly quantify DAAO activity by measuring the oxygen consumed during H2O2 production. The oxygen consumption rate (OCR) of DAAO can directly be compared to the basal mitochondrial respiration in the same assay, to estimate whether the ensuing level of H2O2 production is within the range of physiological mitochondrial ROS production. In the tested monoclonal RPE1-hTERT cells, addition of 5 mM d-Ala to the culture media amounts to a DAAO-dependent OCR that surpasses ∼5% of the OCR that stems from basal mitochondrial respiration and hence produces supra-physiological levels of H2O2. We show that the assay can also be used to select clones that express differentially localized DAAO with the same absolute level of H2O2 production to be able to discriminate the effects of H2O2 production at different subcellular locations from differences in total oxidative burden. This method therefore greatly improves the interpretation and applicability of DAAO-based models, thereby moving the redox biology field forward.

PRAME and CTCFL-reactive TCRs for the treatment of ovarian cancer.

Recurrent disease emerges in the majority of patients with ovarian cancer (OVCA). Adoptive T-cell therapies with T-cell receptors (TCRs) targeting tumor-associated antigens (TAAs) are considered promising solutions for less-immunogenic 'cold' ovarian tumors. In order to treat a broader patient population, more TCRs targeting peptides derived from different TAAs binding in various HLA class I molecules are essential. By performing a differential gene expression analysis using mRNA-seq datasets, PRAME, CTCFL and CLDN6 were selected as strictly tumor-specific TAAs, with high expression in ovarian cancer and at least 20-fold lower expression in all healthy tissues of risk. In primary OVCA patient samples and cell lines we confirmed expression and identified naturally expressed TAA-derived peptides in the HLA class I ligandome. Subsequently, high-avidity T-cell clones recognizing these peptides were isolated from the allo-HLA T-cell repertoire of healthy individuals. Three PRAME TCRs and one CTCFL TCR of the most promising T-cell clones were sequenced, and transferred to CD8+ T cells. The PRAME TCR-T cells demonstrated potent and specific antitumor reactivity in vitro and in vivo. The CTCFL TCR-T cells efficiently recognized primary patient-derived OVCA cells, and OVCA cell lines treated with demethylating agent 5-aza-2'-deoxycytidine (DAC). The identified PRAME and CTCFL TCRs are promising candidates for the treatment of patients with ovarian cancer, and are an essential addition to the currently used HLA-A*02:01 restricted PRAME TCRs. Our selection of differentially expressed genes, naturally expressed TAA peptides and potent TCRs can improve and broaden the use of T-cell therapies for patients with ovarian cancer or other PRAME or CTCFL expressing cancers.

Broadly applicable TCR-based therapy for multiple myeloma targeting the immunoglobulin J chain.

BACKGROUND: The immunoglobulin J chain (Jchain) is highly expressed in the majority of multiple myeloma (MM), and Jchain-derived peptides presented in HLA molecules may be suitable antigens for T-cell therapy of MM. METHODS: Using immunopeptidomics, we identified Jchain-derived epitopes presented by MM cells, and pHLA tetramer technology was used to isolate Jchain-specific T-cell clones. RESULTS: We identified T cells specific for Jchain peptides presented in HLA-A1, -A24, -A3, and -A11 that recognized and lysed JCHAIN-positive MM cells. TCRs of the most promising T-cell clones were sequenced, cloned into retroviral vectors, and transferred to CD8 T cells. Jchain TCR T cells recognized target cells when JCHAIN and the appropriate HLA restriction alleles were expressed, while JCHAIN or HLA-negative cells, including healthy subsets, were not recognized. Patient-derived JCHAIN-positive MM samples were also lysed by Jchain TCR T cells. In a preclinical in vivo model for established MM, Jchain-A1, -A24, -A3, and -A11 TCR T cells strongly eradicated MM cells, which resulted in 100-fold lower tumor burden in Jchain TCR versus control-treated mice. CONCLUSIONS: We identified TCRs targeting Jchain-derived peptides presented in four common HLA alleles. All four TCRs demonstrated potent preclinical anti-myeloma activity, encouraging further preclinical testing and ultimately clinical development.

A library of cancer testis specific T cell receptors for T cell receptor gene therapy.

To increase the number of cancer patients that can be treated with T cell receptor (TCR) gene therapy, we aimed to identify a set of high-affinity cancer-specific TCRs targeting different melanoma-associated antigens (MAGEs). In this study, peptides derived from MAGE genes with tumor-specific expression pattern were identified by human leukocyte antigen (HLA) peptidomics. Next, peptide-HLA tetramers were generated, and used to sort MAGE-specific CD8+ T cell clones from the allogeneic (allo) HLA repertoire of healthy donors. To evaluate the clinical potential, most potent TCRs were sequenced, transferred into peripheral blood-derived CD8+ T cells, and tested for antitumor efficacy. In total we identified, seven MAGE-specific TCRs that effectively target MAGE-A1, MAGE-A3, MAGE-A6, and MAGE-A9 in the context of HLA-A∗01:01, -A∗02:01, -A∗03:01, -B∗07:02, -B∗35:01, or -C∗07:02. TCR gene transfer into CD8⁺ T cells resulted in efficient reactivity against a variety of different tumor types, while no cross-reactivity was detected. In addition, major in vivo antitumor effects of MAGE-A1 specific TCR engineered CD8⁺ T cells were observed in the orthotopic xenograft model for established multiple myeloma. The identification of seven MAGE-specific TCRs expands the pool of cancer patients eligible for TCR gene therapy and increases possibilities for personalized TCR gene therapy.

Analysis of supplemental wildlife feeding in Mississippi and environmental gastrointestinal parasite load.

Wildlife species are host to a variety of gastrointestinal parasites (GIPs). Artificially concentrating animals may increase the risk of disease spread due to increased GIP load and associated environmental load. Supplemental feeding of deer is common among hunters and known to concentrate animals, but there is limited knowledge of how it affects GIP environmental load. GIP load was compared between ecologically-equivalent pairs of sites in Mississippi with and without year-round supplemental feeding (average distance between pairs = 147 m). During May-August in 2019 and 2020, feces from white-tailed deer and raccoons were collected and examined for the presence of nematodes, coccidia, Giardia spp., Cryptosporidium spp., and Baylisascaris procyonis. On average, fed sites had 8 more deer (241% increase) and 2 more raccoon fecal piles (540% increase) than unfed sites. Average parasite loads for individual fecal samples did not differ between fed and unfed sites, but the greater number of deer and raccoon fecal piles at fed sites (p < 0.0001) produced 231% and 308% greater environmental loads of nematodes and coccidia, respectively. Spin feeders, the only feeder type that distributed feed on the ground, had 326% more coccidia in feces on average compared to other feeder types (p < 0.03). These results show that supplemental feeding of white-tailed deer, especially with spin feeders, increases environmental loads of GIP and the potential for transmission of parasitic diseases.

Cutting Edge: Unconventional CD8+ T Cell Recognition of a Naturally Occurring HLA-A*02:01-Restricted 20mer Epitope.

Unconventional HLA class I-restricted CD8+ T cell epitopes, longer than 10 aa, have been implicated to play a role in human immunity against viruses and cancer. T cell recognition of long peptides, centrally bulging from the HLA cleft, has been described previously. Alternatively, long peptides can contain a linear HLA-bound core peptide, with a N- or C-terminal peptide "tail" extending from the HLA peptide binding groove. The role of such a peptide "tail" in CD8+ T cell recognition remains unclear. In this study, we identified a 20mer peptide (FLPTPEELGLLGPPRPQVLA [FLP]) derived from the IL-27R subunit α gene restricted to HLA-A*02:01, for which we solved the crystal structure and demonstrated a long C-terminal "tail" extension. FLP-specific T cell clones demonstrated various recognition modes, some T cells recognized the FLP core peptide, while for other T cells the peptide tail was essential for recognition. These results demonstrate a crucial role for a C-terminal peptide tail in immunogenicity.

The wildland-urban interface in the United States based on 125 million building locations.

The wildland-urban interface (WUI) is the focus of many important land management issues, such as wildfire, habitat fragmentation, invasive species, and human-wildlife conflicts. Wildfire is an especially critical issue, because housing growth in the WUI increases wildfire ignitions and the number of homes at risk. Identifying the WUI is important for assessing and mitigating impacts of development on wildlands and for protecting homes from natural hazards, but data on housing development for large areas are often coarse. We created new WUI maps for the conterminous United States based on 125 million individual building locations, offering higher spatial precision compared to existing maps based on U.S. census housing data. Building point locations were based on a building footprint data set from Microsoft. We classified WUI across the conterminous United States at 30-m resolution using a circular neighborhood mapping algorithm with a variable radius to determine thresholds of housing density and vegetation cover. We used our maps to (1) determine the total area of the WUI and number of buildings included, (2) assess the sensitivity of WUI area included and spatial pattern of WUI maps to choice of neighborhood size, (3) assess regional differences between building-based WUI maps and census-based WUI maps, and (4) determine how building location accuracy affected WUI map accuracy. Our building-based WUI maps identified 5.6%-18.8% of the conterminous United States as being in the WUI, with larger neighborhoods increasing WUI area but excluding isolated building clusters. Building-based maps identified more WUI area relative to census-based maps for all but the smallest neighborhoods, particularly in the north-central states, and large differences were attributable to high numbers of non-housing structures in rural areas. Overall WUI classification accuracy was 98.0%. For wildfire risk mapping and for general purposes, WUI maps based on the 500-m neighborhood represent the original Federal Register definition of the WUI; these maps include clusters of buildings in and adjacent to wildlands and exclude remote, isolated buildings. Our approach for mapping the WUI offers flexibility and high spatial detail and can be widely applied to take advantage of the growing availability of high-resolution building footprint data sets and classification methods.

Enhanced antigen cross-presentation in human colorectal cancer-associated fibroblasts through upregulation of the lysosomal protease cathepsin S.

BACKGROUND: Cross-presentation of exogenous antigens in HLA-class I molecules by professional antigen presenting cells (APCs) is crucial for CD8+ T cell function. Recent murine studies show that several non-professional APCs, including cancer-associated fibroblasts (CAFs) also possess this capacity. Whether human CAFs are able to cross-present exogenous antigen, which molecular pathways are involved in this process and how this ultimately affects tumor-specific CD8+ T cell function is unknown. METHODS: In this study, we investigated the ability of human colorectal cancer (CRC)-derived CAFs to cross-present neoantigen-derived synthetic long peptides (SLPs), corresponding to tumor-derived mutant peptides, and how this affects tumor-specific T-cell function. Processing of the SLP was studied by targeting components of the cross-presentation machinery through CRISPR/Cas9 and siRNA-mediated genetic ablation to identify the key molecules involved in fibroblast-mediated cross-presentation. Multispectral flow cytometry and killing assays were performed to study the effect of fibroblast cross-presentation on T cell function. RESULTS: Here, we show that human CRC-derived CAFs display an enhanced capacity to cross-present neoantigen-derived SLPs when compared with normal colonic fibroblasts. Cross-presentation of antigens by fibroblasts involved the lysosomal protease cathepsin S. Cathepsin S expression by CAFs was detected in situ in human CRC tissue, was upregulated in ex vivo cultured CRC-derived CAFs and showed increased expression in normal fibroblasts after exposure to CRC-conditioned medium. Cognate interaction between CD8+ T cells and cross-presenting CAFs suppressed T cell function, reflected by decreased cytotoxicity, reduced activation (CD137) and increased exhaustion (TIM3, LAG3 and CD39) marker expression. CONCLUSION: These data indicate that CAFs may directly suppress tumor-specific T cell function in an antigen-dependent fashion in human CRC.

T cell receptor engineering of primary NK cells to therapeutically target tumors and tumor immune evasion.

BACKGROUND: T cell receptor (TCR)-engineered cells can be powerful tools in the treatment of malignancies. However, tumor resistance by Human Leukocyte antigen (HLA) class I downregulation can negatively impact the success of any TCR-mediated cell therapy. Allogeneic natural killer (NK) cells have demonstrated efficacy and safety against malignancies without inducing graft-versus-host-disease, highlighting the feasibility for an 'off the shelf' cellular therapeutic. Furthermore, primary NK cells can target tumors using a broad array of intrinsic activation mechanisms. In this study, we combined the antitumor effector functions of NK cells with TCR engineering (NK-TCR), creating a novel therapeutic strategy to avoid TCR-associated immune resistance. METHODS: BOB1, is a transcription factor highly expressed in all healthy and malignant B cell lineages, including multiple myeloma (MM). Expression of an HLA-B*07:02 restricted BOB1-specifc TCR in peripheral blood-derived NK cells was achieved following a two-step retroviral transduction protocol. NK-TCR was then compared with TCR-negative NK cells and CD8-T cells expressing the same TCR for effector function against HLA-B*07:02+ B-cell derived lymphoblastoid cell lines (B-LCL), B-cell acute lymphoblastic leukemia and MM cell lines in vitro and in vivo. RESULTS: Firstly, TCR could be reproducibly expressed in NK cells isolated from the peripheral blood of multiple healthy donors generating pure NK-TCR cell products. Secondly, NK-TCR demonstrated antigen-specific effector functions against malignancies which were previously resistant to NK-mediated lysis and enhanced NK efficacy in vivo using a preclinical xenograft model of MM. Moreover, antigen-specific cytotoxicity and cytokine production of NK-TCR was comparable to CD8 T cells expressing the same TCR. Finally, in a model of HLA-class I loss, tumor cells with B2M KO were lysed by NK-TCR in an NK-mediated manner but were resistant to T-cell based killing. CONCLUSION: NK-TCR cell therapy enhances NK cell efficacy against tumors through additional TCR-mediated lysis. Furthermore, the dual efficacy of NK-TCR permits the specific targeting of tumors and the associated TCR-associated immune resistance, making NK-TCR a unique cellular therapeutic.

Comparing CAR and TCR engineered T cell performance as a function of tumor cell exposure.

Chimeric antigen receptor (CAR) T cell therapies have resulted in profound clinical responses in the treatment of CD19-positive hematological malignancies, but a significant proportion of patients do not respond or relapse eventually. As an alternative to CAR T cells, T cells can be engineered to express a tumor-targeting T cell receptor (TCR). Due to HLA restriction of TCRs, CARs have emerged as a preferred treatment moiety when targeting surface antigens, despite the fact that functional differences between engineered TCR (eTCR) T and CAR T cells remain ill-defined. Here, we compared the activity of CAR T cells versus engineered TCR T cells in targeting the B cell malignancy-associated antigen CD20 as a function of antigen exposure. We found CAR T cells to be more potent effector cells, producing higher levels of cytokines and killing more efficiently than eTCR T cells in a short time frame. However, we revealed that the increase of antigen exposure significantly impaired CAR T cell expansion, a phenotype defined by high expression of coinhibitory molecules and effector differentiation. In contrast, eTCR T cells expanded better than CAR T cells under high antigenic pressure, with lower expression of coinhibitory molecules and maintenance of an early differentiation phenotype, and comparable clearance of tumor cells.