bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL-27 promotes pathogenic Tfh-like CD4 effectors and exhausted-like CD8 T cells in a mouse model of Sjögren’s
Authors: Debreceni Ivy L , Lieberman Scott 
Published: August 2023

Abstract
Sjögren’s is an autoimmune disease that targets the exocrine glands leading to profound dryness, extra glandular manifestations, and increased risk of developing lymphoma with no therapy to treat the underlying immune dysregulation. Nonobese diabetic (NOD) mice spontaneously develop T cell mediated inflammation of the lacrimal gland, similar to human disease. IL-27 is elevated in the salvia and serum of people with Sjögren’s and is required for lacrimal gland autoimmunity in NOD mice. Utilizing IL-27Rα KO NOD mice, I have previously found both CD4 and CD8 effector T cells require IL-27 to transfer lacrimal gland autoimmunity to NOD-SCID mice. Here, we utilized an adoptive co-transfer system that promotes infiltration of lacrimal glands by both WT and IL-27Ra KO T cells to define the T cell intrinsic effects of IL-27 signaling in lacrimal gland infiltrating T cells through 28 color spectral flow cytometry and performed unsupervised dimensionality reduction analyses to identify subsets of T cells reliant on intrinsic IL-27 signaling. Among the total concatenated T cell population including both WT and KO T cells, 12 total subsets were identified. Strikingly, when visualizing WT and KO separately, we identified populations of CD4 effector T cells and CD8 T cells that were either heavily skewed or only present in WT. Through phenotyping these populations, I found IL-27 signaling induced Tfh-like PD-1+ICOS+ CD4 effector T cells and PD-1+ICOS+CD39+ exhausted-like CD8 T cells in the glands of recipient mice. Taken together, these data demonstrate a role for IL-27 in promoting pathogenic T cells in the immune dysregulation of Sjögren’s.
Supported by grants from NIH(T32 AI007485, R01 EY027731)
Cutaneous T cells promote distinct outcomes in human skin structural cells associated with inflammatory disease
Authors: Morawski Peter A , DeBerg Hannah , Pribitzer Stephan , Fahning Mitch , Stefani Caroline , Lacy-Hulbert Adam , Gratz Iris , Campbell Daniel J 
Published: August 2023

Abstract
The skin is an immunologically active barrier tissue specialized to deal with a litany of external stresses. Immune cells coordinate with structural cells in the skin to promote appropriate inflammatory responses and subsequent restoration of barrier integrity following insult. Recent single cell analyses of human skin have defined previously unappreciated transcriptional heterogeneity in structural cells, including disease-associated changes in keratinocytes and fibroblasts. Cutaneous T cell activity is implicated in the development of inflammatory skin disease, but the mechanisms by which T cells promote disease-associated changes in the skin remain unclear. We now show that distinct subsets of circulating and resident CD4+ cutaneous T cells promote a diverse array of cytokine-dependent transcriptional outcomes in human keratinocytes and fibroblasts. We used these in vitro generated transcriptional signatures to identify T cell-dependent gene modules associated with inflammatory skin disease in vivo. For example, we find that Thelper17 cells regulate the expression of epigenetic modifiers in keratinocytes, genes that are enriched in the skin of patients with psoriasis and normalized in response to anti-IL-17 therapy. Thus, we have identified T cell-dependent cytokine-driven transcriptional changes associated with inflammatory skin disease that can be used to dissect immune mechanisms of anti-cytokine therapeutic activity.
Targeting Pim2 improves effector function and longevity of T cells in cancer immunotherapy
Authors: Wu Yongxia , Tian Linlu , Hitchingham Brianna , Choi Heejin , Yang Kaipo , Yu Xue-Zhong 
Published: August 2023

Abstract
The Provirus Integration sites for Moloney murine leukemia virus (Pim) kinase family has been studied extensively in tumorigenesis. We previously reported a key role of Pim2 in negatively regulating T-cell responses to alloantigen. We further investigated how Pim2 regulates T-cell mediated anti-tumor responses using murine models of adoptive cell therapy (ACT) and autologous hematopoietic cell transplantation (HCT). We found that Pim2−/− Pmel CD8 T cells showed enhanced ability for controlling B16 melanoma growth following ACT. Pim2−/− T cells were more potent to control C1498 leukemia growth after syngeneic HCT. Mechanistically, Pim2−/− CD8 T cells exhibited increased effector cytokine production, metabolic activities, and unexpectedly CD62L expression than WT controls following activation. Activated Pim2−/− CD8 T cells had less apoptosis and exhaustion during expansion in IL-2. In vivo, Pim2−/− CD8 T cells differentiated more into effector, but less into exhausted or suppressive subsets in tumors. On the other hand, Pim2−/− CD8 T cells exhibited increased longevity in spleen and tumor draining lymph nodes. Our data indicates that Pim2 negatively regulates CD8 T-cell anti-tumor activity by controlling their persistence in secondary lymphoid organs and effector function in tumors. Blocking Pim2 with a Pim2-specific inhibitor improved graft-versus-leukemia activity after autologous HCT and enhanced anti-melanoma effects after ACT. Pim2 inhibition also increased anti-tumor activity of human melanoma-specific T cells and CD19 CAR-T cells. Taken together, targeting Pim2 may serve as a novel strategy for improving cancer immunotherapy through enhancing differentiation and persistence of effector T cells.
NIH R01 CA258440 and R21 CA263140 to Xue-Zhong Yu, MCW Center for Immunology Pilot Award to Yongxia Wu
BRD4 and BET proteins regulate CD8 T cell exhaustion in mouse models of cancer
Authors: Pratt Brandon M , Green William D , Mullins Genevieve , Green Jarred , Milner Justin 
Published: August 2023

Abstract
Harnessing the cytotoxic capacity of CD8 T cells is central to the efficacy of immunotherapy and has revolutionized the fight against cancer. However, despite encouraging success for select indications, immunotherapies still fail to achieve durable and effective outcomes for certain cancer types and in many patients. Failure of immunotherapies often results from CD8 T cells differentiating into an epigenetically controlled and reinforced state known as T cell exhaustion. Therefore, resolving the molecular programs governing T cell differentiation within the tumor microenvironment (TME) is critical to enhancing treatment. Using mouse models of breast cancer and pancreatic cancer, we investigated the role of the bromodomain and extraterminal (BET) family of proteins in mediating T cell differentiation and exhaustion. Depletion of BRD2, BRD3, and BRD4 in CD8 T cells differentially affected CD8 T cell accumulation and the exhausted phenotype within the tumor microenvironment. Further, small molecule inhibition of the BET family similarly impacted CD8 T cell differentiation and exhaustion in vivo. Our findings may inform new strategies to circumvent CD8 T cell exhaustion during infection.
supported by a training grant from the UNC Pharmacology Department (5T32GM135095-02)
Novel mechanisms of androgen receptor-centered transcriptional regulatory network in regulating CD8+ T cell exhaustion and sex bias in cancer
Authors: Xiao Tong , Schafer Johanna , Song No-joon , Wang Cankun , Weltge Payton , Li Xue , Ma Qin , Li Zihai 
Published: August 2023

Abstract
Sex bias in cancers arising from nonreproductive organs is known but poorly understood. We recently reported on a T cell-intrinsic role of androgen receptor (AR) in driving CD8+ T cell exhaustion which underlies the poorer tumor control in males (Kwon et al., Sci Immunol, 2022). This study aims to understand how AR regulates CD8+ tumor-infiltrating lymphocytes (TILs) by identifying its genome-wide targets. We created a CD8+ T cell-specific AR knockout (KO) mouse model by crossing E8I-cre mice with Ar-floxed mice, which we challenged with syngeneic bladder tumor MB49. We then monitored tumor growth and performed spectral flow cytometry using a T cell exhaustion panel. Further, we used Cleavage Under Targets and Tagmentation – sequencing (CUT&Tag-seq) to map the entire AR targets in CD8+ TILs. Loss of AR in CD8+ T cells significantly slowed the growth of MB49 in male but not female mice. CD8+ TILs from male CD8 AR KO mice showed reduced TOX+ TCF1− terminally exhausted subset (69%; P≤ 0.05) and TOX expression (72%; P = 0.0583) compared to controls. Similarly, TOX expression decreased by 43% in AR-deleted CD8+ T cells following chronic TCR stimulation in vitro (P ≤ 0.0001). Finally, using CUT&Tag-seq, we found that AR binds directly to promoters of multiple key transcriptional regulators of T cell exhaustion, including Tcf7 and Tox.
Through identifying the genome-wide targets of AR in CD8+ TILs, we characterized the cell-intrinsic mechanism by which AR regulates CD8+ T cell exhaustion. Our work suggests AR is a master transcriptional regulator that negatively impacts CD8+ T cell immunity, which may contribute to poorer survival in males and supports combining androgen deprivation therapy with immune checkpoint blockage for cancer treatment.
Special Institutional Funding
A new slow-releasing formulation of butyrate prolongs allograft survival
Authors: Sepulveda Martin N , Chen Luqiu , Cao Shijie , Wang Ruyi , Hubbell Jeffrey , Nagler Cathryn , Alegre Maria-Luisa 
Published: August 2023

Abstract
The microbiota consists of diverse microbes producing metabolites that can influence the immune system locally as well as systemically. These bioactive metabolites (postbiotics) have therapeutic potential and may prove useful to facilitate graft acceptance. Short-chain fatty acids (SCFA) promote intestinal homeostasis and have been linked to health and disease. The Hubbell-Nagler laboratories at the University of Chicago have developed a micelle-based platform that can deliver the SCFA butyrate systemically, with long-term retention and induction of tolerogenic responses. We hypothesized that the administration of this butyrate formulation will improve graft survival by suppressing the alloimmune response. Our preliminary data showed that only oral delivery of the butyrate formulation prolonged survival of a C57Bl/6 male to female minor-mismatched skin graft significantly, whereas sub-cutaneous and intra-peritoneal administration were ineffective. Improved graft survival was associated with diminished production of TNFα by splenic and skin-draining lymph node myeloid cells upon LPS stimulation, suggesting that butyrate modulates the myeloid compartment to reduce levels of pro-inflammatory cytokines. The postbiotic also enhanced survival of a BALB/c to C57Bl/6 major-mismatched skin graft, without the use of immunosuppressive drugs, and resulted in lower levels of donor-specific alloantibodies. Prolonging the administration of butyrate post-transplantation ameliorated graft survival even further, confirming the micelle-based platform has therapeutic potential. Current efforts focus on determining if the butyrate formulation drives myeloid cell polarization toward an anti-inflammatory phenotype.
This project was supported by grant NIH/NIAID 2R01AI115716
Prohibitin is Upregulated on Donor T cells in Murine Acute Graft-versus-Host Disease and Promotes Th17 Response
Authors: Snyder Katiri Jenna , Gao Yandi , Neidemire-Colley Lotus M , Sell Natalie , Ranganathan Parvathi 
Published: August 2023

Abstract

Introduction
Acute graft-versus-host disease (GVHD) is a T cell-mediated disorder and the main cause of non-relapse mortality in allogeneic hematopoietic cell transplantation recipients. Here, we investigate PHB in Th17-mediated inflammation and as a therapeutic target for acute GVHD.

Methods
Irradiated mice received T cell depleted bone marrow cells and allogeneic or syngeneic splenocytes. At day 25 post-transplant, blood, spleen, liver, and colon were harvested and analyzed for surface PHB, IFNγ, and IL-17 on donor T cells by flow cytometry. Splenocytes were cultured in the presence of PHB pharmacological inhibitor rocaglamide (RocA) and analyzed for PHB and IL-17 expression in donor T cells. Naïve murine CD4 T cells were cultured in Th17 polarizing conditions with RocA. CRISPR/Cas9-mediated knockout of PHB was performed followed by evaluation of Il17a, Il17f, and Il22 gene expression by quantitative PCR.

Results
Surface expression of PHB is increased in murine allogeneic donor T cells in liver, spleen, colon, and peripheral blood in acute GVHD. PHB-expressing splenic T cells secrete IL-17, which is reduced with treatment with a PHB inhibitor in ex vivo experiments. PHB inhibition of murine CD4 T cells in vitro impairs differentiation into the Th17 subtype resulting in reduced IL-17 secretion and downregulation of Il17a, Il17f, and Il22 genes. Knockout of PHB reduces transcription of Il17 and Il22 genes.

Conclusions
These findings show relevancy of PHB in Th17-mediated inflammation and suggest a potential role for PHB in acute GVHD. To our knowledge, this study is the first to investigate PHB in acute GVHD, and our findings will offer greater support for clinical use of PHB inhibition to treat this fatal disease.

Supported by grants from AAI (KS, PR), ACS (PR; RSG-22-053-01-IBCD), and NIH NCI (PR; R01 CA252469).
Gene Expression Profiling and Signaling Pathway Analysis of Tolerant T cells Circulating in Adult Patients after PTCy Haplotransplantation
Authors: Chen Xiaohua , Nee Taylor , Rajasundaram Dhivyaa , Farah Rafic , Szabolcs Paul 
Published: August 2023

Abstract
The mechanisms for the signaling pathways of T cell-dependent tolerance after haploidentical hematopoietic stem cell transplantation (haplo-HSCT), are incompletely understood. We tested 8 patients (Pt) with leukemia or myelodysplastic syndromes (median age:51-yrs old) following IRB approved consenting. All received Flu+TBI (+/− Cy) pre-conditioning and haplo-HSCT with post-transplant Cytoxan (PTCy) and FK506 as GvHD prophylaxis.
The circulating T cells were hyporeactive in mixed lymphocyte reaction (MLR) assay to recipient DC in Pts at 1yr (or 2yrs in 1 Pt) post-haplo-HSCT (~6m post-ISD withdrawal), while these T cells responded vigorously to 3rd party APC. Neither depletion of Tregs, nor blockade of IL10R or PD1 could reverse T-cell hyporeactivity. Low-dose IL2 triggered mild anti-host responses. Cytokine profiling further confirmed the MLR responses. ImmunoSEQ™ revealed the disappearance of alloreactive T-cell clones from peripheral pool with time, even though a few alloreactive T clones remained. Low dose IL2 had minimal impact to reverse the diminishment.
Unlike graft T-cell responses against host DC, tolerant T cells in circulation against hDC showed relative downregulation of allograft rejection, T-helper response, co-stimulation, and cell proliferation pathway, that was paired with upregulation of co-inhibition, PD1-PDL1, Ferroptosis and CREB pathways.
In summary, hyporeactivity of donor-derived circulating T cells to recipient DC were evident. We found Treg, Tr1 activity to be insignificant in maintaining long-term tolerance. T-cell clonal deletion appears to be the dominant mechanism, along with systemically altered signaling pathways in tolerant T-cells.
Characterization of ZG16b protein exocrine function and biomarker potential for salivary gland damage in chronic graft vs. host disease
Authors: Mody Drashty Paresh , da Silva Ana Costa , Sharma Rubina , Mays Jacqueline 
Published: August 2023

Abstract
The oral cavity is a frequent target of chronic Graft versus Host Disease (cGVHD) following allogenic hematopoietic stem cell transplant (allo-HSCT). Detection of early cGVHD-induced damage to exocrine tissues, such as salivary glands, is challenging to detect prior to irreversible tissue fibrosis. Recent work in our laboratory used shotgun proteomics to identify putative salivary biomarkers at salivary gland cGVHD onset. Saliva samples from patients affected with oral cGVHD post allo-HSCT had a drop in protein content of zymogen granule 16b (ZG16b), recapitulated in immunohistochemical staining of human minor salivary gland (MSG) biopsy sections. ZG16b loss correlated with CD45+ cells infiltration. This protein is poorly characterized, and immunofluorescence tissue staining suggested ZG16b protein localization to acinar cells. ZG16b cellular expression localized to serous and serous mucous acinar cell populations in single cell RNAseq (scRNA seq) data from healthy human labial MSG. Little is known about the function of this ZG16b in other exocrine tissues such as pancreas. scRNA seq analysis of repository data suggests ZG16b colonization in the pancreatic ductal cells. Similar to MSG, the pancreatic Zg16b gene was expressed in mucus secretion pathways and additional interactions with cell migration and homeostasis markers. Ongoing work is being done to further probe for binding partners for ZG16b (endogenous proteins, components of the oral microbiome and immune cell receptors) to distinguish the protein from its homologs. In parallel, the utility of salivary ZG16b as a cGVHD and general salivary gland damage biomarker is being tested. This work will bring clarity Zg16b’s role in exocrine tissues and alloimmune disease.
NIH, NIDCR Intramural program
Elevated exhausted effector tissue resident CD8+ T-cells in Chronic Graft versus Host Disease patient Oral mucosa unveiled by spatial transcriptomics and scRNA seq analysis
Authors: Sharma Rubina , da Silva Ana Costa , Martin Daniel , Pavletic Steve , Mays Jacqueline 
Published: August 2023

Abstract
Chronic graft-versus-host disease (cGVHD), an autoimmune-like disease following allogeneic hematopoietic stem cell transplantation where T cells can attack host tissue. Oral mucosa (OM) is a most common cGVHD effector site in the oral cavity. We studied T cell populations and tissue-specific signals that regulate their differentiation and functionality. Patient OM biopsies from ongoing clinical trials (NCT03602599, NCT00331968) were taken at 6 months post-transplant. We found enrichment of immune cells in the OM mainly in the CD8 T-cell cluster and identified two unique populations in the OM of cGVHD patients: one characterized by expression of proliferative markers and the other more prominent, characterized by exhaustion makers (TIM3, LAG3, TIGIT). The latter had low expression of CD69 but expressed markers of tissue residency; CD103, BLIMP1, PD1, FABP5. Interestingly, this population showed high expression of BATF, IRF4, and RUNX3, transcription factors that induce sustained effector CD8 T-cell programming. These exhausted cells displayed higher expression of effector molecules IFNG, GZMB, PRF1 inducing sustained CD8 T-cell effector function. Ligand-receptor analysis revealed strong interactions between CXCL9-CXCR3 in CD8 T-cells and myeloid cells. Spatial transcriptomic analysis confirmed the CD8 T-cell infiltration in the submucosal region of OM in cGVHD colocalizing with the elevated markers of exhaustion, surrounded by myeloid cells, thus confirming the striking interaction of CXCL9 and CXCR3. These exhausted CD8 T-cells may play an important role in cGVHD pathogenesis causing by driving sustained tissue damage and could serve as a might be used as a novel target for cGVHD therapy.
This work was supported by the intramural programs of the NIDCR and NCI.
Amelioration of graft versus host disease in humanized mice by exopolysaccharide from a commensal bacterium
Authors: Kalinina Olga , Knight Katherine L 
Published: August 2023

Abstract
Graft versus host disease (GVHD) is the major and often lethal complication of allogenic bone marrow transplantation. Prophylactic regimens for GVHD are given as standard pre-transplantation therapy; however, up to 60% of these patients develop GVHD, and require additional immunosuppressive intervention. We identified a probiotic exopolysaccharide (EPS) that induces inhibitory macrophages and tolerogenic dendritic cells (DCs) that inhibit T cell proliferation. We tested if EPS-treated human DCs, derived in vitro from CD34+ cord blood cells, would inhibit allogeneic T cell proliferation in a mixed lymphocyte reaction (MLR) and found that they did, demonstrating that EPS can induce tolerogenic DCs in vitro. We translated these studies in vivo using a humanized mouse model of aGvHD and demonstrated that EPS-treated human DCs administered at the time of transfer of human peripheral blood mononuclear cells (hPBMCs) significantly increased survival of these mice compared to mice given PBS-treated DCs, or no DCs. Several subsets of DCs have been described, e.g., cDCs, moDCs and plasmacytoid DCs, each with different phenotypes and functions. Using labeled EPS, we showed that the tolerogenic CD34-derived DCs generated by EPS treatment are conventional DCs (cDCs). We found that EPS binds to two cDC populations, conventional DC1 (cDC1) and conventional DC2 (cDC2), but only cDC1 were inhibitory in our MLR cultures. Our data indicate that EPS-DCs, potentially cDC1, can be used as a cell-based therapy to mitigate aGvHD in humans.
NIH 1R41AI155281-01
Deleting Runx3 in CD4+ T Cells Disrupts Effector Functions and Facilitates Tolerance Induction in Solid Organ Transplantation
Authors: Xie Stephen Zhiyuan , Allocco Jennifer B , Cassano Alexandra , Chen Luqiu , McKeague Michelle L , Chong Anita S , Alegre Maria-Luisa 
Published: August 2023

Abstract
Long-term transplantation tolerance depends on the control of graft-reactive T cells. In particular, the alloreactivity of CD4+ T cells needs to be suppressed as these cells are both necessary and sufficient for cardiac allograft rejection. Although experimental protocols such as anti-CD154-mediated co-stimulation blockade can stably induce donor-specific tolerance in MHC-mismatched cardiac allografts, other more immunogenic organs such as skin are less susceptible to tolerogenic treatments, in part due to additional reactivity of the host against donor commensals that accompany the graft. The transcriptional mechanisms that enable CD4+ T cells to drive rejection and whose targeting can promote tolerance are not well understood. RNAseq comparison of the transcriptome in graft-reactive T cells from rejecting versus tolerant heart allograft recipients revealed several transcription factors differentially expressed between rejected and tolerant mice in both spleen and graft. Here, we report that the transcription factor Runx3 in alloreactive CD4+ T cells promotes Th1 and cytotoxic response against the allograft and resistance to tolerance induction. Conversely, ablating Runx3 in alloreactive CD4+ T cells augments the efficacy of anti-CD154 blockade treatment enabling skin graft acceptance.