bluuebunny/crossref_metadata_2025_split huggingface dataset:
Interleukin 4 increases the production of indduced regulatory T cells in C57BL/6 mice (120.12)
Authors: Haribhai Dipica , Schmitt Erica , Ziegelbauer Jennifer , Upchurch Kyle , Salzman Nita , Williams Calvin 
Published: January 2023

Abstract
Naïve peripheral T cells can express the transcription factor Foxp3 and develop regulatory function. These “induced” regulatory T (iTreg) cells are essential for tolerance and complement the function of other regulatory populations. In this study we used the lymphopenia-associated model of colitis to compare the kinetics of iTreg cell production in both C57BL/6 and BALB/c mice, the prototypical Th1- and Th2-type strains. Following the adoptive transfer of naïve CD4+ T cells into Rag-/- recipients, we tracked the development of iTreg cells over a period of 40 days. Whereas iTreg cell production in C57BL/6 was minimal, by 10 days there was a relative five-fold increase in the number of iTreg cells in BALB/c mice that grew to 35 fold by 40 days. This dramatic increase in iTreg cells correlated with a similar increase in IL-17A producing cells in the mesenteric lymph nodes and gut of BALB/c mice. In contrast, the proportion of CD4+ T cells that produced IFN-γ was comparable between the two strains. Pretreatment of C57BL/6 Rag-/- recipient mice with IL-4 resulted in a six-fold increase in the number of iTreg cells compared to untreated mice. Pretreatment with in vitro derived M2 macrophages also resulted in a five to fifteen fold increase in the number of iTreg cells compared to pretreated mice with M0 or M1 macrophages, respectively. These results indicate that a Th2 milieu supports iTreg cell induction.
The role of Ndfip1 in CD8+ T cell activation and effector function (121.25)
Authors: Kurzweil Vanessa , O'Leary Claire , Oliver Paula 
Published: January 2023

Abstract
Nedd4-family interacting protein 1 (Ndfip1) is an adapter protein for Nedd4-family E3 ubiquitin ligases. Mice lacking Ndfip1 in both CD4+ and CD8+ T cells develop a severe atopic inflammatory disease at sites of environmental exposure and die prematurely. In part, this is because Ndfip1 expression in CD4+ T cells is needed to inhibit IL-4 production and permit differentiation into non-Th2 lineages. However, Ndfip1-/- mice that lack MHC Class II-restricted CD4+ T cells still develop inflammation at a young age. This suggests that Ndfip1 might also be important in CD8+ T cells. We show that Ndfip1-/- CD8+ T cells are more sensitive to TCR stimulation than WT cells. After activation, Ndfip1-/- CD8+ T cells produce more IFN-γ than their WT counterparts. Consistent with this, Ndfip1-/- CD8+ T cells have higher levels of Eomesodermin than control cells. Future studies will attempt to identify E3 ligases and substrates that regulate these CD8+ T cell functions.
Mimicry epitope from Ehrlichia canis for interphotoreceptor retinoid-binding protein ameliorates autoimmune uveoretinitis by acting as altered peptide ligand in A/J mice (123.29)
Authors: Gangaplara Arunakumar , Massilamany Chandirasegaran , Madayiputhiya Nadakumar , Nandakumar Renu , Kuszynski Charles , Steffen David , Reddy Jay 
Published: January 2023

Abstract
Uveoretinitis is one of the sight-threatening inflammatory diseases of eyes, and autoimmune response to retinal antigens is suspected in the causation of disease. Although environmental triggers are suspected, their association remains tenuous and clinically not proven. We report here identification of novel mimicry epitopes for interphotoreceptor retinoid-binding protein (IRBP) 201-216 and they represent Ehrlichia canis (EHC), Pseudomonas putida, Trypanosoma cruzi, Chlamydia trachomatis, Clostridium novyi, Hepatitis C virus and Paracoccidoides brasiliensis. We demonstrate that one mimic from E. canis (EHC 44-59) although can generate cross-reactive T cells for IRBP 201-216, capable of producing T helper (Th)1 and Th17 cytokines, fail to induce autoimmune uveoretinitis by active immunization in A/J mice. Likewise, the animals previously immunized with IRBP 201-216 and subsequently challenged with EHC 44-59 fail to show the disease. The data suggest that exposure to microbial products containing the mimicry sequences for retinal antigens does not necessarily leads to the development of eye autoimmunity rather they can behave as natural altered peptide ligands.
Regulation of selection and autoimmunity by miR-181 family miRNAs (123.36)
Authors: Schaffert Steven , Wang Song , Axtell Robert , Newell Evan , Steinman Lawrence , Davis Mark , Chen Chang-Zheng 
Published: January 2023

Abstract
T cell sensitivity to antigens is regulated during development to ensure proper immunity and tolerance. We previously showed that miR-181a regulates the strength and threshold of T cell receptor signaling by targeting negative feedback molecules. Here we used miR-181 knockout mice to examine the role of miR-181 miRNAs in selection and T cell reactivity to antigens. We found that loss of mir-181ab1 caused an increase in the percent of positively selected CD8 single-positive (SP) cells in the thymi of female HY TCR transgenic mice. However, loss of miR-181 had no effect on the deletion CD8 SP cells in the thymi of male HY mice. Moreover, we showed that loss of mir-181ab1 resulted in two-fold more self-reactive T cells in the periphery after immunization with a self-peptide derived from myelin oligodendrocyte glycoprotein (MOG). Paradoxically, knockout mice exhibited a delayed disease onset in a MOG-dependent mouse model of multiple sclerosis, experimental autoimmune encephalomyelitis (EAE). In particular, the central nervous systems of knockout mice during disease had similar percentages of MOG-specific T cells, but an underrepresentation of TH17 and T H1-polarized CD4 T cells compared to the periphery. These findings demonstrate that mir-181ab1 also plays a role in modulating the differentiation and function of self-reactive T cells. Together, data from our study illustrate the diverse roles of mir-181ab1 in selection and autoimmunity.
Inhibition of p300 impairs Foxp3+ T-regulatory (Treg) function and promotes anti-tumor immunity (127.40)
Authors: Liu Yujie , Wang Liqing , Han Rongxiang , Predian Jarrod , Beier Ulf , Cole Philip , Albelda Steven , Wayne Wayne 
Published: January 2023

Abstract
Foxp3+ Treg cells limit autoimmunity and maintain immune homeostasis but also restrict immune responses to cancer. Thus, Tregs are a promising target for modulation of host anti-tumor immune responses. We have shown that targeting of Foxp3+ Tregs using histone/protein deacetylase inhibitors (HDACi) promotes Foxp3 acetylation and increases Treg production and suppression function. By contrast, there are no studies concerning the role of histone/protein acetyltransferases (HAT) targeting in Tregs. HATs are classified into 3 main families; GNAT, MYST and p300/CBP. Here, we found that conditional deletion of p300 in Tregs caused lymphoproliferation, weight loss, arthritis and dermatitis in mice. Likewise, deletion of p300 in Tregs, or its pharmacologic inhibition, decreased Foxp3+ Treg production and impaired Treg-mediated suppression of homeostatic proliferation, and blocked Treg-dependent donor-specific allograft tolerance. In contrast with these data, p300 targeting decreased TC1 lung tumor volume in immunocompetent C57BL/6 mice, but had no direct effect on TC1 tumor cell proliferation in vitro, or on the growth of TC1 implanted tumors in RAG1-/- immunodeficient mice. We conclude that in real world tumor models using immunocompetent mice, p300 targeting impaired Treg function without overt impairing effector T cell responses or inducing autoimmunity. We thereby identify p300 as a critical target for modulation of Foxp3+ Treg function in the context of anticancer therapy.
MARCKS protein plays a critical role in proinflammatory cytokine production (160.7)
Authors: Nordone Shila , Li Jingjing , D’Annibale Melissa , Birkenheuer Adam , Levy Mike , Yoder Jeff , Jones Sam , Adler Ken 
Published: January 2023

Abstract
Myristoylated alanine-rich C kinase substrate (MARCKS) is a ubiquitously expressed Protein Kinase C substrate that has emerged as a potential therapeutic target for amelioration of mucin secretion during chronic obstructive pulmonary disease. MARCKS also plays a key role in regulation of neutrophil adhesion, migration and degranulation. Given its biological role in immune cells, we hypothesized that MARCKS may play an integral role in cytokine secretion. MARCKS protein is highly conserved between the dog and human, and importantly, the N-terminal 24 amino acids are virtually identical between species, allowing us to use the well-characterized MANS peptide to inhibit canine MARCKS function through the N-terminus. Using isolated neutrophils, we evaluated the effect of MARCKS inhibition on LPS-induced cytokine production. We found that pre-treatment of canine neutrophils with MANS peptide significantly reduced the secretion of a broad range of LPS-induced cytokines, including IL-8, CXCL1 and TNFα, in comparison to untreated cells or those treated with a scrambled control peptide. This reduction in IL-8 was maintained when MANS was administered 2 hours after LPS-treatment of cells. Reduction in IL-8 secretion was found not to be due to protein retention or cell death. Our observations identify MARCKS protein as a promising therapeutic target in treatment of inflammatory disease, particularly those syndromes attributed to neutrophil influx and inflammatory cytokine production.
Anti-IL-20 monoclonal antibody suppresses breast cancer progression and bone osteolysis in murine models (161.10)
Authors: Hsu Yu-Hsiang , Chang Ming-Shi 
Published: January 2023

Abstract
IL-20 is a proinflammatory cytokine involved in rheumatoid arthritis and osteoporosis. However, little is known about the role of IL-20 in breast cancer. We explored the function of IL-20 in tumor growth and metastasis, and its clinical outcome. Tumor expression of IL-20 was assessed by IHC staining among 198 breast invasive ductal carcinoma (IDC) patients with available clinical and survival data. IL-20 expression was associated with advanced tumor stage, high tumor metastasis, and worse survival. RTQ-PCR showed that clinical breast tumor tissue expressed higher IL-20 and its receptors than did non-tumorous breast tissue. IL-20 was also highly expressed in breast cancer bone-metastasis tissue. In vitro, IL-20 upregulated matrix metalloproteinase-9, matrix metalloproteinase-12, cathepsin K, and cathepsin G, and enhanced proliferation and migration of breast cancer cells, which were inhibited by anti-IL-20 monoclonal antibody (mAb) 7E. In vivo, 7E reduced tumor growth, suppressed bone colonization, reduced tumor-mediated osteolysis, and lessened bone density decrement in mice injected with breast cancer cells. In conclusion, our results suggest that IL-20 plays pivotal roles in the tumor progression of breast cancer. IL-20 expression in breast cancer tissue is associated with a poor clinical outcome. Anti-IL-20 mAb 7E suppressed bone colonization, and decreased osteolytic bone lesions. Therefore, IL-20 may be a novel target in treating breast tumor-induced osteolysis.
Immune-mediated cure of established B16F10 melanoma with a nonvirulent strain of Toxoplasma gondii (162.12)
Authors: Fiering Steven , Baird Jay , Byrne Katelyn , Lizotte Patrick , Mullins David , Turk Mary Jo 
Published: January 2023

Abstract
Immune recognition and elimination of tumors protects us from cancer. Tumors frequently manifest immunosuppressive mechanisms to avoid anti-tumor immunity and thereby become clinical challenges. The immune system is quite sensitive to microbial pathogens and microbes or microbial extracts are very effective adjuvants to stimulate an immune response. Here we present the results of treatment of established cutaneous B16F10 murine melanoma by intratumoral injection of a fully attenuated uracil auxotroph strain of Toxoplasma gondii (ΔCPS) that cannot replicate in vivo. ΔCPS exposure breaks immunosuppression and stimulates a strong Th1 antitumor immune response in vivo that cures established primary melanoma and develops immune memory that confers protection against subsequent rechallenges. This treatment modifies the tumor microenvironment, increases MHC Class I expression on tumor cells and reverses immunosuppression, permitting a tumor-antigen specific CD8 T cell response to eliminate the tumor. The approach has efficacy against multiple tumor types and has potential for clinical utility.
Myeloid derived suppressor cells(MDSCs) emergence from distinct splenic precursors (162.28)
Authors: Park Young-Jun , Song Boyeong , Kim Yun-Sun , Kim Eun-Kyung , Lee Jung-Mi , Lee Ga-Eun , Kim Jae-Ouk , Kim Yeon-Jeong , Chang Woo-Sung , Kang Chang-Yuil 
Published: January 2023

Abstract
Myeloid derived suppressor cells(MDSCs) are a heterogenous population of immature myeloid cells, which accumulate in various pathological conditions, especially in tumor. In mice, MDSCs are characterized by the co-expression of Gr1 and CD11b molecules, and these cells could be classified into two subsets, CD11b+Ly6G+Ly6Clow polymorphonuclear neutrophil-like(PMN) MDSCs and CD11b+Ly6G-Ly6Chigh monocytic(Mo) MDSCs, by their expression of Ly6C and Ly6G molecules. Functional characteristics and fate of MDSCs are defined quite well. On the other hands, accumulation mechanism and origins of MDSC need to be more elucidated, and have not been fully understood whether MDSC could be generated in the periphery from distinct precursors in tumor microenvironment. For the first time, we identified the presence of splenic MDSC precursors in tumor bearing mice, characterized by CD11b+Ly6G-Ly6Cneg/low. We demonstrated that cytokines related to MDSC accumulation convert CD11b+Ly6G-Ly6Cneg/low cells into CD11b+Ly6G+Ly6Clow PMN-MDSC and/or CD11b+Ly6G-Ly6Chigh Mo-MDSC. These converted cells exerted strong suppressive activity on DO11.10 Tg splenocytes proliferation. Thus, these data suggest that there are peripheral distinct precursors, CD11b+Ly6Cneg/low cells, from which immunosuppressive MDSC could be generated.
Avidity maturation of γ9δ2T-cell receptor engineered T-cells by CDR3 modulation (162.37)
Authors: Kuball Jürgen , Gründer Cordula , van Dorp Suzanne , Drent Esther , Hol Samantha , Scholten Kirsten , Heijhuurs Sabine , Strong Roland 
Published: January 2023

Abstract
Immunotherapy with innate immune cells has recently evoked a broad interest as a novel treatment option for cancer patients. γ9δ2T-cells are an emerging innate cell population with strong anti-tumor-reactivity and a promising candidate for immune interventions. Especially γ9δ2T-cell receptors (TCR) which recognize a broad panel of tumor cells with high avidity are clinically attractive since αβT-cells can be efficiently redirected against a variety of tumor cells by introducing a γ9δ2TCR. Here we demonstrate that distinct γ9δ2TCRs mediate different functional avidities and present the concept of combinatorial-γδTCR-chain exchange (CTE) as an efficient method to create γ9δ2TCRs that mediate strong anti-tumor-responses. Thereby γ9- and δ2-chains derived from individual γ9δ2T-cell clones are newly combined allowing the design of γ9δ2TCRs that mediate a higher functional avidity against a broad tumor cell panel in vitro and in vivo when compared to a reference γ9δ2TCR. We furthermore demonstrate that this phenomenon is selectively caused by differences in the CDR3-domains of γ9- and δ2-chain. Accordingly, alanine-scanning-mutations were performed and elucidated important residues within the CDR3 sequence and stress the impact of the CDR3 length for optimal γ9δ2TCR function. This knowledge allowed improving tumor-control by engineered T-cells not only in vitro but also in vivo in a humanized mouse model.
Novel functions of cFLIP in lymphocyte death signaling (163.22)
Authors: Zhang Haibing , Zhang Jianke 
Published: January 2023

Abstract
Fas/Apo-1 signals through the FADD (Fas-associated death domain) adaptor protein, which recruits and activates the apical caspase 8 and leads to apoptosis. cFLIP is a homologue of caspase 8 but lacks caspase activities, and presumably inhibits apoptosis induced by the death receptors by competing with caspase 8 for binding to FADD. FADD, Caspase 8, and cFLIP form a signaling complex during death receptor-induced apoptosis. In addition to the cell death function, deletion of FADD, Caspase 8, and cFLIP results in embryonic lethality and lymphocytes proliferation defects upon TCR-induced or Toll-like receptor stimulation. Previous studies have shown that death receptors also induce a particular type of necrotic death in certain cell types, termed necroptosis, which is dependent on RIP1 and RIP3. Recently, our group and other groups showed that RIP1 or RIP3 deficiency could rescue the embryonic lethality and T cell proliferation defects caused by FADD or Caspase 8 deficiency. These studies identified a critical function of FADD and caspase-8 in vivo in suppressing RIP1/RIP3-dependent necrosis that is essential for embryonic development and T cell proliferation. Using a conditional cFLIP knockout mouse model and various in vitro systems, our recent studies reveal novel functions for cFLIP in regulating apoptosis and necrosis in lymphocytes.
Multiparameter flow cytometric analysis of RORγt expression in mouse models of T cell development and differentiation (163.25)
Authors: Wu Xiaowei , Le Hai , Nobida Naomi , Boyce Christopher , Sepulveda Homero , Rohrer Jurg , Ernst David 
Published: January 2023

Abstract
Retinoic acid-related Orphan Receptor gamma t (RORγt) is a DNA-binding transcription factor that is selectively expressed by lymphoid cells including CD4+CD8+ thymocytes, CD4+ Th17 and CD8+ Tc17 cells, NKT cells and lymphoid tissue inducer (LTi) cells. RORγt plays essential roles in thymopoiesis, T cell homeostasis, effector T cell differentiation and the development of secondary lymphoid tissues, eg, lymph nodes and Peyer’s patches. A new mouse mAb, Q31-378, that recognizes mouse RORγt was developed to provide a sensitive probe for analyzing the cellular and molecular mechanisms that underlie the development and differentiation of T cells and lymphoid tissues. With a newly developed transcription factor buffer, Q31-378 was successfully used in multicolor flow cytometric analyses to analyze RORγt expression in thymocyte subsets defined by CD4 and CD8 expression. Among all thymocytes, CD4+CD8+ cells comprised the main cell subset that expressed RORγt. Analyses of splenocytes cultured under Th17-inducing conditions revealed that most activated CD4+IL-17A+IFN-γ± T cells expressed RORγt when compared with other T cell subsets. These new reagents, along with additional fluorescent antibodies to other cytokines, transcription factors and phenotypic markers are being applied to analyze RORγt expression patterns in the development and differentiation of T cells and LTi cells.
Signals via TCR and common-γ-chain receptors concurrently coordinate the generation of antigen-specific human Th17 (163.3)
Authors: Ni Ling , Joo HyeMee , Duluc Dorothee , Kluca Peter , Xue Yaming , Zurawski Sandy , Zurawski Gerard , Banchereau Jacques , Oh SangKon 
Published: January 2023

Abstract
IL-17-producing CD4+ T cells (Th17) can confer protective and therapeutic immunity against microbial infections and cancers. However, mechanisms for the induction of antigen-derived peptide-specific human Th17 remain unknown. To study this, we employed an in vitro system in which influenza viral antigen HA1 was delivered to dendritic cells (DCs) via Dectin-1. Notably, MHC II/peptide complexes presented by DCs expanded memory Th17, but could not induce HA1-specific Th17 even with Th17-promoting cytokines. We found that signals via TCR and common-γ-chain receptors, which utilize Stat1/3, first induce naïve CD4+ T cells to express IL-1R1 followed by programming naïve CD4+ T cells to express Rorc in response to IL-1β in a MyD88 dependent manner. IL-6-mediated Stat3 activation simply amplifies IL-1β-MyD88-mediated Rorc expression. A failure in any of those signaling components, Stat1, Stat3, or MyD88, significantly diminished HA1-specific Th17 induction. Thus, the differentiation of naïve CD4+ T cells into pathogen-specific human Th17 cells requires interplays between innate and adaptive immune signals.
Secondary CD4+ T cell effectors retain heterogeneity during recall responses but are functionally superior to primary effectors (163.8)
Authors: Strutt Tara , McKinstry Karl , Kuang Yi , Bradley Linda , Swain Susan 
Published: January 2023

Abstract
It is not known whether or how distinctions between secondary (2°) CD4+ T cell effectors, generated from memory precursors, and primary (1°) effectors, arising from naïve cells, contribute to protective recall responses. Here, we compare these two populations in vivo following influenza A virus infection (IAV). We show that 2° effectors are functionally superior to 1° effectors, and that their generation is critical to memory CD4+ T cell mediated protection against IAV. 2° effectors develop and migrate to infected lungs with similar kinetics, but 2° effectors accumulate in greater number and demonstrate enhanced multifunctional capabilities. The 2° effector response in the lung is a critical component of protection against influenza mediated by memory CD4+ T cells and upon transfer to unprimed hosts, 2° effectors are superior to 1° effectors at mediating protection. Transcriptome analysis indicates that 1° and 2° effectors responding in different organs share key genetic pattterns, however, 2° effectors possess unique gene signatures. Finally, by targeting differentially expressed surface molecules, we demonstrate specific regulation of 1° and 2° effector functions in vivo. Our approach provides novel insight into the regulation of effector CD4+ T cells during pathogen challenge and may reveal ideal candidates for manipulation with the goal of directing the generation of larger populations of multifunctional effectors in a variety of settings apart from IAV infection.
Anaplasma marginale infection results in an impaired pathogen-specific CD4 T-cell response (164.16)
Authors: Brown Wendy , Turse Joshua 
Published: January 2023

Abstract
Anaplasma marginale infection of cattle previously immunized with a non-protective outer membrane (OM) protein resulted in complete loss of functional CD4 T cell responses to the specific protein and rapid deletion of tetramer+ CD4 T cells in peripheral blood. Furthermore, lack of A. marginale-specific responses during persistent infection suggested a failure of infection to prime for normal effector and memory T-cell responses. Dysfunctional memory T-cell responses were confirmed following infection of naïve cattle, as only transient and sporadic A. marginale-specific responses were detected in blood and spleen. We hypothesize that high antigen load during acute and persistent infection results in apoptosis and/or suppression of antigen-specific T cells. The objective of this study was to determine if immunization with a protective whole OM vaccine, which prevents disease, and in some cases, infection, would similarly result in loss of memory T cells primed by OM immunization. Upon infection of OM vaccinees with A. marginale, OM-specific T cell responses disappeared in all cattle, even though infection was blunted compared with a naïve control. To determine if Anaplasma-specific T cell responses recover once pathogen load is removed, infection was cleared with long-acting oxytetracycline and oral Aureomycin. The response did recover in all animals by 11 weeks. The induction of suppressive Treg cells by A. marginale infection is currently under investigation.