bluuebunny/crossref_metadata_2025_split huggingface dataset:
Characterization of lymphocytes emerging early after nonmyeloablative conditioning and hematopoietic stem cell transplant supported with sirolimus (P2186)
Authors: Weitzel R. , Biancotto Angelique , Fitzhugh Courtney , McCoy Philip , Hsieh Matthew , Tisdale John 
Published: January 2023

Abstract
Hematopoietic stem cell transplant is a curative option for patients with severe congenital anemias such as Sickle Cell Disease. Due to the higher transplant-related mortality associated with myeloablative conditioning and the reversal of the SCD phenotype achieved with as low as 11% stable donor chimerism, we developed a successful non-myeloablative regimen for patients with matched-sibling donors consisting of pre-transplant alemtuzumab, 300cGy TBI, and post-transplant sirolimus. However, mechanisms that promote tolerance in these patients remain unclear. We hypothesized that lymphocytes emerging early post-transplant would display a toleragenic phenotype. Samples were thawed from time points near day 60 or day 100 post-transplant. 17-parameter flow cytometry was performed, revealing dramatically high proportion of Tregs (6.3%-32.8%) and T-helper-17 (Th17) (8.2-21.2%) cells, elevated markers of proliferation, activation, and B cell maturity in engrafting patients. Using multiplex cytokine immunoassay, 67 analytes were measured, revealing high expression of IL-17A in the engrafting patients, MCP-1 in the rejecting patients, but little difference in Treg/Th1/Th2-associated cytokines. These unexpected observations have provided insight into reconstitution following nonmyeloablative conditioning and sirolimus-based immunosupporession and suggest targets for further investigation into both the long-term, stable mixed chimerism and the lack of GvHD observed in our patients.
Varying frequencies of T cells exhibiting CMV peptide-specific reactivity to allo-MHC are present in cord blood transplant units (P2228)
Authors: White Jason , Kurche Jonathan , Gutman Jonathan , Kedl Ross 
Published: January 2023

Abstract
Cord blood transplants are a desirable treatment option for a number of hematopoietic diseases due partly to the increased tolerance of HLA mismatches between transplant donor and recipient, manifest in the decreased occurrence of GVHD. Unfortunately, these patients also have an increased risk of reactivating latent infections (e.g. CMV) post-transplant which will ultimately be fatal; likely due to the same decreased T cell functioning that results in HLA mismatch tolerance. Viral reactivation is by no means universal after transplant, however, and while certain factors are known to increase a patient’s risk (e.g. GVHD or in-vivo t cell depletion), there remains no good method for predicting susceptibility. Using MHC tetramers, our lab has found a population of T cells specific to the HLA-A2 CMV peptide in not only A2, but also non-A2 cord blood samples. The frequency of these cells varies widely between cord blood units, which could be a potential signifier of a unit’s efficacy in fighting viral reactivation. Further, preliminary studies suggest that these cells have a specificity and functionality that would make them therapeutically useful.
Retention of autoreactive CD4+ recent thymic emigrants in the liver (BA8P.127)
Authors: Xu Xi , Ge Qing 
Published: January 2023

Abstract
T cells escape from negative selection in the thymus require peripheral tolerance enforcement to prevent them from triggering T cell-mediated autoimmunity. Using adoptive transfer model and RAG2p-GFP transgenic mouse system, we demonstrated that autoreactive CD4+ recent thymic emigrants (RTEs) were retained and some were activated in the liver directly after their egress from the thymus. Educations from secondary lymphoid organs were not required for this hepatic retention but TCR-MHC interaction in the liver was important for the activation of CD4+ liver RTEs. Compared to RTEs in the lymph nodes, liver RTEs exhibited higher TCR reactivity. CD4+ liver RTEs preferentially repopulated the liver and peyer’s patches when adoptively transferred into lymphoreplete mice. When transferred into RAG1-/- mice, liver but not lymph node RTEs caused T cell infiltration and severe inflammation in the lung. These data suggest that the liver may be the first checkpoint in the periphery to filter and retain autoreactive CD4+ thymic emigrants that escape from negative selection. Once unchecked in the absence of regulatory T cells in lymphopenic hosts, these cells could induce Th1 type autoimmunity primarily in the lung.
Cytokines secreted by Th2 cells negatively regulate lymphangiogenesis (CAM3P.202)
Authors: Kim Dong-Eon , Shin Kihyuk , Lee Seung-Hyo 
Published: January 2023

Abstract
Abstract Lymphatic vessels (LVs) consist of a unidirectional network that absorbs protein-rich fluid from peripheral tissues and drains it into the venous circulation through a conduit system of collecting vessels, lymph nodes (LNs), and ducts. In addition to fluid homeostasis, LVs also play an important role in immune surveillance by regulating immune cell migration and antigen delivery to draining LNs. Previous study showed IFN-g, a major Th1 cell-associated cytokine, negatively regulates lymphangiogenesis. However, the effects of the Th2 cell cytokines, IL-4 and IL-13, are still unclear. To reveal this, by using mouse lymphatic endothelial cells (mLECs), human LECs (hLECs), and Prox-1-GFP transgenic mice, we investigated the effects of IL-4 and IL-13 on lymphangiogenesis. Here we show that treatment of mLECs and hLECs with IL-4 and Il-13 down-regulated expression of Prox-1, essential transcription factor expressed by LECs. IL-4 and IL-13 also inhibited tube formation by LECs in vitro. Similarly, LECs co-cultured with Th2 cells showed decreased tube formation, and this effect was fully restored by cytokine neutralization with anti-IL-4 and anti-IL-13 antibodies. Moreover, in vivo blockade of IL-4 and IL-13 increased not only the density of LVs, but also the function of LVs in the airways. Therefore, our findings indicate a novel anti-lymphangiogenic function for Th2 cells via production of IL-4 and IL-13.
LFA-1 and CD62L dependent transient surface CCR5 expression in naïve CD8+ T cells enhances T cell immune memory (CCR3P.214)
Authors: Huang Alex , Askew David , Su Charles , Liou Rachel , Nthale Joseph , Barkauskas Deborah 
Published: January 2023

Abstract
The initiation of an effective adaptive immune response requires choreographed interaction among rare antigen-bearing dendritic cells (DCs) with antigen-specific CD4+ and CD8+ T cells in the lymph node (LN). This interaction is especially critical for the magnitude of memory T cell generation. We and others have previously demonstrated that CCL3/CCL4 secretion from productive interaction between antigen-specific T cell and antigen-bearing DCs promote local recruitment of naïve CD8+ T cells in the inflamed LNs, implying that naïve CD8+ T cells can express CCR5 in the inflamed LN microenvironment. Here, we examined the kinetics of CCR5 expression on naïve T cells upon entering an inflamed LN. Maximum surface CCR5 expression occurs in 10-30% of newly arrived naive T cells within the first 6 hours via mobilization of pre-existing CCR5 protein found in intracellular vesicles, and this transient up-regulation of surface CCR5 expression in CD8+ T cells was dependent on direct physical contact with LFA and CD62L, as ligation of CD8+ T cells with anti-CD11a and/or anti-CD62L antibodies enhances surface CCR5 expression within 1 hour in the absence of TCR stimulation. More importantly, CCR5+ fraction of naïve CD8+ T cells exhibit enhanced ability to become memory T cells. Our study implicates an important functional role for CCR5 expression by naïve lymphocytes prior to cognate antigen recognition for enhanced memory T cell development.
IL-36 contributes to robust effector function of CD8+ T cells co-stimulated with 4-1BB (CD137) and OX40 (CD134) through a TCR independent mechanism (CCR3P.216)
Authors: Tsurutani Naomi , St. Rose Marie-Clare , Adler Adam , Vella Anthony 
Published: January 2023

Abstract
IL-36 cytokines are members of the IL-1 family of cytokines. It has been reported that murine bone barrow-derived dendritic cells and CD4+ T lymphocytes constitutively express IL-36R and respond to IL-36. Also, during CD4+ T cell stimulation, inclusion of IL-36 is known to alter cytokine potential. However, the role of IL-36 in CD8+ T cells is not well understood. In this study, we investigated whether IL-36R is expressed on CD8+ T cells and whether IL-36 exerts effects on CD8+ effector T cell function and phenotype. We found that by quantitative real-time RT-PCR IL-36R is highly expressed on CD8+ T cells after 4-1BB (CD137) and OX40 (CD134) dual co-stimulation. Importantly, these effector CD8+ T cells responded to IL-36β and IL-36γ. We found enhanced IFN-γ production when IL-36 was combined with IL-12, or IL-2. Surprisingly, IFN-γ production was induced in resting effector CD8+ T cells even in the absence of any known TCR trigger. Secondly, the synergy of IL-36 with IL-2 was time dependent and evident at IFN-γ mRNA and protein levels. Together, these data suggest that IL-36 can enhance the effector function of CD8+ T cells and may be a promising immunotherapy candidate.
TL1A level (low versus high) differentially affects expansion and function of regulatory and effector T cells. (CCR3P.218)
Authors: Sidhu Maninder , Shih David , Targan Stephan 
Published: January 2023

Abstract
Patients with Crohn’s disease have increased mucosal expression of TL1A. Mice generated with sustained and elevated T cell expression of Tl1a (L-Tg) have increased Foxp3+ Treg cells yet develop worsened ileitis, proximal migration of colitis and intestinal fibrosis. The aim of this study was to investigate the role of Tl1a in the differentiation and function of Tregs. In the L-Tg mice, the CD4+ T cell population was comprised of GFPhigh and GFPlow cells that correlated with the level of Tl1a expression. We found that Foxp3 expression was significantly higher in the GFPlow population of CD4+ T cells compared to the GFPhigh cells. The GFPlowFoxp3+ T cells expressed high Nrp-1, GARP, and LAG-3 surface markers and were more suppressive in function whereas the GFPhigh Foxp3+ T cells had an activated effector memory phenotype and were highly proliferative. Induced Tregs differentiated in-vitro from naïve GFPlow CD4+ T cells expanded into TGF-β producing Foxp3 T cells whereas naïve T cells from the GFPhigh fraction expanded into cells with a Th2 phenotype, producing IL-4, IL-13 and IL-10. Preliminary data from a T cell co-transfer study suggested that mice cotransferred with the GFPlowFoxp3+ T cells were better protected from weight loss compared to mice cotransferred with GFPhighFoxp3+ T cells. These data suggest that level of Tl1a (high v. low) differentially influenced the expansion of effector and Treg cells leading to either a pro-inflammatory or protective mucosal environment.
Expression of secreted osteoclastogenic factor of activated T cells (SOFAT) by T- and B-lineage cells may contribute significantly to bone loss in periodontitis. (CCR3P.221)
Authors: Napimoga Marcelo , Martinez Elizabeth , Peruzzo Daiane , Carregaro Vanessa , de Araújo Vera , Weitzmann M. 
Published: January 2023

Abstract
A novel T cell-secreted cytokine, termed secreted osteoclastogenic factor of activated T cells (SOFAT), that induces osteoclastic bone resorption in a RANKL-independent manner, has recently been described. We have previously reported that SOFAT is highly expressed in gingival tissues of patients with chronic periodontitis (CP) suggesting a putative role in the bone loss associated with CP. Here we characterized the cell types producing SOFAT in CP. Gingival tissues were biopsied from subjects without periodontal disease (n=5) and patients with CP (n=5) and SOFAT quantified by immunohistochemistry and immunofluorescence staining. Our data reveal marked SOFAT staining in CP that was predominantly associated with the lymphocytic infiltration of gingival tissues. Interestingly, besides CD3+ T cells, B-lineage cells including plasma cells also exhibited strong staining for SOFAT. Because SOFAT has not previously been reported in B-linage cells, we further purified splenic T cells and B cells from Balb/c mice and activated them using LPS or CD3/CD28 antibodies respectively. SOFAT was quantified by RT-qPCR and revealed significant induction of SOFAT expression in both activated T cells and B cells in comparison to unstimulated cells. Our data support a putative role of SOFAT in the bone loss associated with chronic periodontitis and further demonstrates for the first time that in addition to T cells, B-linage cells may also be a significant source of SOFAT in inflammatory states.
The biological function of membrane versus soluble TL1A (CCR3P.205)
Authors: Ferdinand John , Meylan Francoise , Siegel Richard , Al-Shamkhani Aymen 
Published: January 2023

Abstract
The Tumour Necrosis Factor Superfamily (TNFSF) are an important source of costimulatory signals required for activation of T cells. The TNFSF is composed of 29 receptors and 18 ligands; one such pair is Death receptor 3 (DR3) and TNF-like ligand 1A (TL1A). Signalling via DR3 costimulates lymphocytes and polymorphysims in TL1A and DR3 have been associated with Crohn's Disease. In mice, signalling via DR3 has been shown to be required for maximal pathology in models of asthma and lupus and transgenic constitutive expression of TL1A results in an IL-13 and IL-17a driven intestinal pathology. Several TNFSF ligands, including TL1A, can be expressed on the membrane and cleaved to yield a soluble product. Some cytokines, such as TNF are active in soluble form whereas others, such as FasL, are not. The biological roles of these distinct forms of TL1A in vivo are not known. To investigate the biological function of membrane vs. soluble TL1A we used retrotransgenic technology to express cleavable vs. membrane bound TL1A throughout the murine hematopoietic system. Membrane restricted TL1A promoted a strong inflammatory pathology in both the ileum and lung. Full length TL1A expression yielded little membrane TL1A. These mice failed to develop pathology despite a greater than physiological level of TL1A in the serum. This work demonstrated that membrane restricted TL1A is a more potent ligand for DR3. We have generated membrane-restricted TL1A transgenic mice to investigate this further.
Inhibiting TWEAK (TNF-like weak inducer of apoptosis) signaling ameliorates blood brain barrier integrity and neuronal damage in neuropsychiatric lupus prone MRL/lpr mice (CCR3P.212)
Authors: Wen Jing , Doerner Jessica , Stock Ariel , Sellers Rani , Burkly Linda , Gulinello Maria , Putterman Chaim 
Published: January 2023

Abstract
Neuropsychiatric disease (NPSLE) is one of the most common manifestations of human lupus, but remains poorly understood. TWEAK is a cytokine member of the TNF superfamily; its sole receptor, Fn14, is expressed in brain endothelial cells, astrocytes, microglia and neurons. In vitro, TWEAK signaling promotes expression of MCP-1, IL-6 and IL-8, decreases ZO-1 expression, and increases the permeability of human brain microvascular endothelial cells. Our previous study showed that Fn14 knockout (KO) in the MRL/lpr lupus strain led to markedly attenuated neuropsychiatric disease, as revealed by significant reduction in depressive-like behavior and improved cognitive function. The purpose of the present study was to determine the mechanisms by which TWEAK signaling is instrumental in the pathogenesis of NPSLE. We found that Fn14KO mice had improved BBB integrity as shown by decreased fibronectin and IgG deposition in the brain. Further, there were fewer cellular infiltrates in Fn14KO mice. Additionally, Fn14KO mice displayed reduced neuron apoptosis and hippocampal gliosis. Interestingly, there were no differences in neurogenesis and microglia activation, or in circulating autoantibody titers, between Fn14KO and Fn14WT mice. Our studies indicate that TWEAK/Fn14 interactions play a central role in the pathogenesis of NPSLE by improving BBB integrity and reducing neuronal damage, suggesting a novel target for therapeutic intervention for this major disease manifestation.
Recruitment of adult thymic progenitors is regulated by lymphotoxin beta receptor (HEM4P.240)
Authors: Shi Yaoyao , Wu Weiwei , Fu Yang-Xin , Zhu Mingzhao 
Published: January 2023

Abstract
Continuous homing of hematopoietic progenitors from peripheral blood to thymus is essential for the functional maintenance of thymus, especially upon thymic injury. Here we identified the signaling of lymphotoxin beta receptor (LTβR), is essential for the thymic progenitor settling. Dramatically reduced frequency and number of early T-lineage progenitors (ETP) were found in the steady thymus of LTβR knockout (KO) mice. This is probably not due to impaired development of hematopoietic progenitors, since normal frequencies of T-cell precursor were found in the bone marrow of LTβR KO mice. Further experiment by tracing adoptively transferred bone marrow cells directly confirmed a thymic homing defect in LTβR KO mice. Progenitor thymic settling is mediated by a cascade of adhesion and signal events, which involves P-selectin, CCL25, VCAM-1/ICAM-1, etc. We identified P-selectin is down-regulated in the thymi of LTβR KO mice while the expression levels of CCL25, VCAM-1, VCAM-1 are normal. Data from bone marrow chimeric mice suggest that LTβR on radio-resistant, likely stromal cells, regulates thymic progenitor receptivity. Finally, LTβR KO mice demonstrated severely impaired thymus regeneration upon thymus-targeted injury. Taken together, our data reveal a novel role of LTβR on the regulation of thymic progenitor homing. Manipulation of this signaling pathway may boost thymic regeneration and immune reconstitution during chemotherapy, radiotherapy or AIDS HAART therapy.
Thymic aging in the absence of klotho (HEM4P.243)
Authors: Smith Michelle , Goetz Christine , Min Dullei , Stefanski Heather , Hollander Georg , Weinberg Kenneth , Blazar Bruce 
Published: January 2023

Abstract
Age-related thymic involution, characterized by a decrease in thymic epithelial cell (TEC) proliferation, absolute number, and organization, results in overall defective support of T cell development and proliferation and is a major cause of thymic dysfunction. Using a genetic model of advanced aging caused by deficiency of Klotho, an anti-aging gene and modifier of FGF signaling, we investigated TEC-intrinsic aging in the absence of a globally aging environment. By one month of age, Klotho deficient (Kl-/-) mice display a disrupted thymic architecture, a 4-6 fold decrease in TEC numbers, as well as significant decreases in all thymocyte populations and also a 3-8 fold decrease in peripheral T cell numbers. To separate thymus-intrinsic aging from the effects of organismal aging, we implanted fetal thymi from Kl-/- mice under the kidney capsule of mice lacking thymi. Kl-/- and WT fetal thymi supported comparable levels of thymopoiesis as measured by the numbers of T cells in peripheral blood and lymphoid organs up to 4 months post-implantation, with both cohorts of implants not detectable past 5 months. In this aging model, isolation from the systemic effects of Klotho deficiency attenuates age-related thymic dysfunction, suggesting that extrathymic factors affect thymic aging.
Human T cell development in HSC-engrafted NSG mice is dependent on age of the recipient (HEM4P.247)
Authors: Hasgur Suheyla , Shultz Leonard , Brehm Michael 
Published: January 2023

Abstract
Humanized mouse models are an exciting alternative for the investigation of human hematopoiesis, immunobiology, autoimmunity, infectious diseases, cancer biology, transplantation biology and regenerative medicine. However the efficient generation of humanized mice is complicated by the availability of numerous mouse strains and the large diversity of engraftment protocols. NOD-scid IL2rγnull (NSG) mice injected with human HSC support high levels of engraftment and develop a functional human immune system including both innate and adaptive immune cells. Previous studies have shown that injection of human HSC into newborn NSG mice results in more efficient and rapid T cell development as compared to adult NSG (&gt;6 wk) mice. Here we compared engraftment of human HSC and immune system development following intracardiac injection into irradiated newborn NSG mice or intravenous injection into irradiated 3-wk old NSG mice. Total human cell chimerism was monitored in the blood between 6 and 18 wk after transplantation. We also evaluated the development of human T cells, B cells, monocytes, NK cells, NKT cells, conventional and plasmacytoid DC. Our results show that newborn NSG mice have higher levels of T cell reconstitution at earlier time points as compared to 3-wk old NSG recipients but that the older mice achieved comparable T cell levels at later times. Together these findings show that T cell development in humanized mice occurs with slower kinetics in older NSG mice.
Alteration of TCR Dβ sequence content alters Treg development (HEM4P.248)
Authors: Levinson Michael , Sanchez-Silva Aaron , Zhuang Yingxin , Sleckman Barry , Schroeder Harry 
Published: January 2023

Abstract
Diversity of the T cell receptor is mainly developed through V(D)J rearrangement and N addition during TCR development. The product of V(D)J rearrangement is the CDRβ3, a region of high variability that recognizes antigen and includes all of the D gene. The Dβ sequence is highly conserved across various species, from trout to mouse to humans. This suggests that there are some natural constraints on the TCR; these constraints are thought to limit deleterious T cells from reaching the periphery. We hypothesize that altering the D region will have an effect on the development of thymocytes. To do this we have created Dβ altered mice. These mutants are a Dβ2 KO (Dβ1); a replacement of the Dβ locus with a charged Dβ (DβDKRQ); and a replacement of the Dβ locus with a hydrophobic DH (DβYTL). When compared to WT mice, the mutant Dβ mice have an altered T cell repertoire in both CDRβ3 amino acid composition and length and these differences can be attributed to the changes in the germline sequence. Changing the Dβ also changes the total T cell number in both developing and mature T cells, with an altered Dβ being selected against. However, Treg numbers from the DβDKRQ mice are increased in the mucosa, suggesting a link between charge, self-reactivity and nTreg induction. Ongoing experiments will elucidate the role of the Dβ sequence on the development of T cells and the role it plays in infection and inflammation.
Ikaros and aiolos undergo different post-transcriptional regulation in mice and humans (HEM4P.249)
Authors: Mitchell Julie , Szarejko John , Yankee Thomas 
Published: January 2023

Abstract
The Ikaros family of transcription factors is important for T cell development and leukemogenesis. In mice, homozygous expression of an Ikaros isoform that is unable to bind DNA leads to a complete block in T cell development, while heterozygous expression of the same Ikaros isoform results in leukemogenesis. To better understand the roles of Ikaros family members in human and murine T cell development, we used quantitative RT-PCR and Western blot analysis to determine the mRNA and protein levels of Ikaros family members in FACS-purified murine and human thymocytes. As thymocytes mature from the double negative (DN) to double positive (DP) stages, Ikaros mRNA levels increase 3.0±1.8 fold (p&lt;0.01, n=6) in mice and 4.4±1.7 fold (p&lt;0.001, n=5) in humans. Similarly, Aiolos mRNA levels increase 27±2.3 fold (p&lt;0.001, n=6) in mice and 18±2.8 fold (p&lt;0.001, n=5) in humans. However, during the same DN to DP transition, protein levels only increase significantly in mice for both Ikaros (2.5±0.79 fold, p&lt;0.001, n=6) and Aiolos (11±5.4 fold, p&lt;0.01, n=5). These results suggest that Ikaros and Aiolos undergo different post-transcriptional control in murine and human thymocytes. Further studies are needed to understand the method of this post-transcriptional control, and how it regulates human T cell development and leukemogenesis.