bluuebunny/crossref_metadata_2025_split huggingface dataset:
Constitutive expression of TIGIT defines a population of CD4+ regulatory T cells in Bcell non-Hodgkin lymphoma
Authors: Yang Zhi-Zhang , Kim Hyo Jin , Price-Troska Tammy , Jalali Shahrzad , Villasboas Jose C. , Novak Anne J. , Ansell Stephen M. 
Published: January 2023

Abstract
T cell Ig and ITIM domain (TIGIT) is an immune checkpoint molecule and its ligation delivers an inhibitory signal to T cells that negatively regulates anti-tumor responses. However, the expression and biological relevance of TIGIT in B-cell non-Hodgkin lymphoma (NHL) is completely unknown. To phenotypically characterize TIGIT+ T cell subsets, we profiled TIGIT-expressing CD4+ and CD8+ T cells from biopsy specimens of B-cell NHL. TIGIT expression was not detectable on T cells from peripheral blood, however, TIGIT is constitutively expressed on CD4+ T cells from biopsy specimens of B-cell NHL with a median of 44.8% (range: 25.9–62.2, n=8) of CD4+ T cells expressing TIGIT. By profiling the CD4+TIGIT+ T cells, we observed that CD25 was highly expressed on intratumoral CD4+TIGIT+ T cells. Intracellular staining revealed that Foxp3 was expressed by intratumoral CD4+TIGIT+ T cells. These intratumoral CD4+TIGIT+ T cells expressed high level of chemokine receptor CCR4 and were absent of IL-7 receptor-α (CD127). Functionally, CD4+TIGIT+ T cells displayed reduced cytokine production, as the number of IFN-γ- and TNF-α-producing cells was lower in the TIGIT+ population than in TIGIT− T cells. While TIGIT expression can be induced by TCR activation, treatment of T cells with IL-2 and TGF-β inhibited TCR-mediated TIGIT induction. Furthermore, lymphoma B cells were involved in TIGIT induction as in vitro depletion of lymphoma B cells altered upregulation of TIGIT on CD4+ T cells in B-cell NHL. Taken together, these results indicated that constitutive expression of TIGIT defines a CD4+ population with similar phenotype to regulatory T cells. Inhibition of TIGIT signaling may be an additional mechanism to prevent T-cell suppression in B-cell NHL
Targeting human acute myeloid leukemia with multi-epitope specific naïvederived anti-tumor CD4+ T cells
Authors: Chang Mark Jesse , Whitehill Gregory , Stegemann Scott , Scofield Eric , Wong Susan , Keyvanfar Keyvan , Jain Prachi , Ito Sawa , Chen Jichun , Barrett John , Muranski Pawel 
Published: January 2023

Abstract
Little is known about the anti-tumor potential of human CD4+ T cells. Hematological malignancies constitutively expressing MHC class II could be a target for tumor-specific CD4+ T cells. We studied antigen-specific human CD4+ T cells targeting leukemia associated antigens (LAA) WT-1, PRAME, MAGEA A3 and Ny-ESO1 in vitro and in a humanized NSG mouse acute myeloid leukemia (AML) model. We hypothesized that naïve CD4+ T cells are an important source of anti-tumor T cells. Bulk and naïve CD4+ T cells isolated from the blood of normal donors were stimulated with autologous dendritic cells pulsed with overlapping 15mer peptides spanning the entire protein. After culture, naïve-derived Th cells robustly recognized LAA in most donors, while bulk-derived CD4+ T cells had negligible activity, suggesting that pre-exiting memory Th cells have a competitive advantage over LAA-specific precursors. Addition of inflammatory cytokines (IL-1, 6, 7, 21, 23 and TGF-β) further enhanced reactivity. Naïve-derived donor LAA-specific Th cells (but not control CMV pp65-specific Th cells) specifically recognized HLA-matched sibling AML blasts. After adoptive transfer LAA-specific T cells significantly reduced leukemic burden in NSG mice bearing fully-HLA matched AML. Thus, it is feasible to generate naïve-derived multi-epitope specific anti-leukemia CD4+ T cells from normal donors. Removal of competing memory Th cells and culture in inflammatory cytokines unmasks strong LAA-specific reactivity. These Th cells demonstrate highly specific recognition of naturally processed LAA in HLA-matched leukemic blasts and can control AML in vivo, establishing a foundation for CD4+ T cell immunotherapy for hematological malignancies.
Naïve CD4+ T Cells, Recruited by CCL18 Secreted by Tumor-Associated Macrophages, Differentiate Within Breast Tumors into Tumor-Infiltrating Regulatory T Cells
Authors: Song Erwei , Su Shicheng , Liao Jianyou , Zhao Qiyi , Lieberman Judy , Liu Qiang 
Published: January 2023

Abstract
The origin of tumor-infiltrating Tregs, critical mediators of tumor immunosuppression, is unclear. Here we show that human breast tumor-infiltrating Tregs and naive CD4+ T cells have overlapping TCR repertoires, which barely overlap with circulating Treg sequences, suggesting that intratumoral Tregs develop from naive T cells in situ rather than from recruited Tregs. Naive CD4+ T cells, whose abundance in tumors is linked to poor prognosis, are recruited to human breast tumors when their PITPNM3 receptor recognizes the tumor-associated macrophage chemokine CCL18. In tumor-bearing humanized mice, blocking naive CD4+ T cell tumor recruitment by knocking down PITPNM3 reduces tumor-infiltrating Tregs and inhibits tumor progression. These findings suggest that breast tumor-infiltrating Tregs arise from chemotaxis of circulating naive CD4+ T cells that differentiate into Tregs in the tumor microenvironment. Inhibiting naïve CD4+ T cell recruitment into tumors by interfering with PITPNM3 recognition of CCL18 may be an attractive strategy for anticancer immunotherapy.
Loss of immune tolerance is controlled by ICOS in Sle1 mice
Authors: Mittereder Nanette , Kuta Ellen , Bhat Geetha , Dacosta Karma , Cheng Lily , Herbst Ronald , Carlesso Gianluca 
Published: January 2023

Abstract
ICOS, a member of the CD28 family, represents a key molecule that regulates adaptive responses to foreign Ags. ICOS is prominently expressed on T follicular helper (TFH) cells, a specialized CD4+ T cell subset that orchestrates B cell differentiation within the germinal centers and humoral response. However, the contribution of ICOS and TFH cells to autoAb profiles under pathological conditions has not been thoroughly investigated. We used the Sle1 lupus-prone mouse model to examine the role of ICOS in the expansion and function of pathogenic TFH cells. Genetic deletion of ICOS impacted the expansion of TFH cells in B6.Sle1 mice and inhibited the differentiation of B lymphocytes into plasma cells. The phenotypic changes observed in B6.Sle1-ICOS–KO mice were also associated with a significant reduction in class-switched IgG and anti-nucleosomal IgG-secreting B cells compared with B6.Sle1 animals. An upregulation in the IFN gene signature early in the life of B6.Sle1 mice modestly correlated with increased levels of autoAbs. The level of VCAM-1, a molecule that was shown to be elevated in patients with SLE and in lupus models, was also increased in an ICOS-dependent manner in Sle1 mice and correlated with autoAb levels. The elimination of ICOS-expressing CD4+ T cells in B6.Sle1 mice, using a glyco-engineered anti-ICOS–depleting Ab, resulted in a significant reduction in anti-nucleosomal autoantibodies. Our results indicate that ICOS regulates the ontogeny and homeostasis of B6.Sle1 TFH cells and influences the function of TFH cells during aberrant germinal center B cell responses. Therapies targeting the ICOS signaling pathway may offer new opportunities for the treatment of lupus and other autoimmune diseases.
CXCR3 blockade impedes the development of Sjögren’s syndrome-like xerostomia in non-obese diabetic mice
Authors: Zhou Jing , Yu Qing 
Published: January 2023

Abstract
The chemokine receptor CXCR3 plays an important role in T cell recruitment in various immune responses and autoimmune diseases. Expression of CXCR3 ligands, including CXCL9, CXCL10 and CXCL11, is elevated in the salivary glands of patients with Sjögren’s syndrome (SS). To elucidate whether interaction between CXCR3 and its ligands is required for the development of SS, we administrated an anti-CXCR3 blocking antibody to non-obese diabetic (NOD) mice, a well-defined model of SS, during the stage prior to disease onset. CXCR3 blockade significantly improved salivary secretion, indicating a remission of SS clinical manifestation. CXCR3 blockade did not affect the gross leukocyte infiltration of the submandibular glands (SMG) as assessed by H&E staining. However, flow cytometric analysis showed that anti-CXCR3 treatment markedly reduced the percentage of CXCR3+CD8 T cells and CD44+CD8 T cells, without affecting that of CXCR3+CD4 T cells and CD44+CD4 T cells in the SMG and submandibular lymph nodes, suggesting a selective effect of CXCR3 blockade on effector T cytotoxic 1 cells. Meanwhile, SMG expression of inflammatory factor TNF-α was markedly diminished by anti-CXCR3 treatment. Consistent with this, CXCR3 blockade significantly enhanced SMG expression of tight junction protein claudin-1 and water channel protein aquaporin-5, two factors that are crucially required for normal salivary secretion and can be down-regulated by TNF-α. Taken together, our findings demonstrated that the interaction between endogenous CXCR3 and its ligands plays a proinflammatory and pathogenic role in the development of SS-like xerostomia in NOD mouse model.
IL-17A inhibits expression of IL-17-lineage cytokines through a negative feedback loop involving IL-24 and controls autoimmune uveitis
Authors: Chong Wai Po , Raychaudhuri Kumarkrishna , Horai Reiko , Silver Phyllis , Jittayasothorn Yingyos , Chan Chi-Chao , Chen Jun , Caspi Rachel R 
Published: January 2023

Abstract
The Th17 response has been associated with autoimmune diseases in patients and in animal models. IL-17A is recognized as the Th17 signature cytokine and IL-17A-producing T cells are pathogenic effectors in models of autoimmunity, including experimental autoimmune uveitis (EAU). Paradoxically, however, injection of IL-17 ameliorates EAU (PMID: 19234216). Using a model of spontaneous uveitis in IRBP T cell receptor transgenic R161H mice, we investigated their susceptibility on the IL-17A−/− background. Surprisingly, IL-17A−/− R161H mice developed essentially undiminished uveitis and IL-17−/− R161H T cells, polarized to Th17 and infused into wild type recipients, induced similar disease to IL-17A sufficient R161H T cells. Interestingly, IL-17A−/− R161H T cells polarized under Th17 conditions produced elevated amounts of other Th17-related cytokines, i.e. IL-17F, GM-CSF and IL-22. Supplementing these cultures with recombinant IL-17A normalized the elevated production of these cytokines. RNAseq analysis revealed that IL-17A−/− T cells expressed lower levels of IL-24 compared to their IL-17A sufficient counterparts. Mechanistic studies indicated a negative feedback loop where IL-17A induces Th17 cells to produce IL-24, which subsequently suppresses production of Th17 lineage cytokines. Finally, injection of recombinant IL-24 ameliorated adoptive Th17-induced EAU, and conversely, silencing IL-24 expression in Th17 cells increased their pathogenicity and enhanced disease severity. Our data suggest that: (a) IL-17A exerts a negative feedback on uveitogenic Th17 cells via IL-24 production, and (b) IL-24 limits the expression of other Th17-related cytokines and controls their pathogenicity.
S1P1 Agonist SEW2871 Inhibits human Th17 cell generation and functions
Authors: Eken Ahmet , Okus Fatma Z , Yetkin Mehmet F , Cakir Mustafa , Ozdarendeli Aykut , Mirza Meral 
Published: January 2023

Abstract
S1P1 is a G-protein coupled receptor expressed by lymphocytes and is required for their egress out of secondary lymphoid organs. Fingolimod, an agonist of four S1P receptors including S1P1, is used in multiple sclerosis (MS) treatment in humans; however, less is known regarding the impact of S1P1 agonists on the egress-independent biology and functions of human Th17 and Treg cells. In this study, we explored the effects of S1P1 agonists SEW2871 and FTY720 on the ex vivo generation and functions of human Th17 and Treg cells. We report that a selective S1P1 agonist SEW2871 inhibited human Th17 development ex vivo, from naïve T cells, associated with reduced IL-17 and IL-22 production, demonstrated by ELISA and intracellular cytokine staining. On the other hand, Treg generation was augmented in SEW2871+ cultures. Additionally, SEW2871-exposed monocytes and dendritic cells produced less IL-23, a cytokine required for Th17 cell-expansion and pathogenicity. Th17 differentiation in the presence or absence of antigen presenting cells revealed that SEW2871 could inhibit Th17 generation either by blocking APC-derived IL-23 or directly affecting T cells. Our data provide novel information as to egress-independent effects of S1P1 agonists on Th17 cells.
Cellular misfolded proteins transported to the cell surface by aberrantly expressed MHC class II molecules are major autoantibody targets in autoimmune diseases
Authors: Arase Hisashi , Jin Hui , Tanimura Kenji , Suenaga Tadahiro , Arase Noriko , Lanier Lewis L 
Published: January 2023

Abstract
Cellular misfolded proteins are generally not transported outside the cells and thus may not be exposed to immune cells. This suggests that immune cells may not be tolerant to the cellular misfolded proteins. However, we recently found that such misfolded proteins in the endoplasmic reticulum are rescued from protein degradation and are transported to the cell surface by aberrantly expressed MHC class II molecules without being processed to peptides. Because structures of misfolded proteins associated with MHC class II molecules are different from those of normal proteins, their antigenicities are also different. On the other hand, it has been well known that MHC class II molecules are aberrantly expressed in many autoimmune diseased tissues. In this study, we extensively analyzed misfolded proteins transported to the cell surface by MHC class II molecules in autoimmune diseases and found that these misfolded proteins complexed with MHC class II molecules are specific targets for autoantibodies produced in autoimmune diseases such as rheumatoid arthritis and antiphospholipid syndrome. Furthermore, a strong correlation has been observed between autoantibody binding to misfolded protein/HLA-DR complex and the autoimmune disease susceptibility conferred by each HLA-DR allele, suggesting that misfolded protein/HLA-DR complexes are involved in the pathogenicity of autoimmune diseases. These findings imply that misfolded proteins rescued from protein degradation by aberrantly expressed MHC class II molecules function as a ‘neo-self’ antigens that might induce abnormal immune response in autoimmune diseases.
Visual function in retina-specific TCR-transgenic mice with spontaneous autoimmune uveitis
Authors: Jittayasothorn Yingyos , Horai Reiko , Caspi Rachel R 
Published: January 2023

Abstract
Autoimmune uveitis is a potentially blinding disease caused by circulating retina-specific T cells. To understand the mechanisms behind autoimmune uveitis, R161 transgenic mice expressing a TCR specific for IRBP, the retinal target protein for autoimmune uveitis in the mouse model, were generated. Three lines of R161 mice expressing the same TCR at different copy numbers have been established: R161H&gt;R161M&gt;R161L. The H and M lines developed spontaneous uveitis with 100% incidence by 2–3 months, whereas the L line developed disease only upon immune perturbation. The expression levels of the R161 TCR transgene correlated with frequency of IRBP-specific CD4 T cells in the peripheral repertoire, and with severity of disease. To study the association of visual function with pathology of uveitis, we examined visual function of R161 lines and compared with their parental strain B10.RIII (WT) by electroretinography (ERG), which measures electrical responses of retinal cells under dark- and light-adapted conditions. Uveitis was monitored and scored weekly by fundoscopy and ERG was performed in the next day following dark adaptation. Overall, as inflammatory scores increased, ERG responses decreased. R161L mice, which develop little or no disease, had visual function equivalent to WT mice. At 16 weeks old when disease in the H and M lines had reached its peak, R161H mice exhibited no and most of R161M mice exhibited poor visual function, whereas only minor reduction of ERG, which is normally observed with age, was detected in R161L and WT mice. The ERG data illustrate an association of visual function with uveitis scores among R161 lines, each of which may serve as a unique model of uveitis with different severity and visual loss.
Abacavir competes for HLA-B*57:01 restricted peptide presentation in HLA-transgenic mice.
Authors: Tilahun Mulualem E , Natarajan Kannan , Boyd Lisa F , Cardone Marco , Norcross Michael A , Margulies David H 
Published: January 2023

Abstract
A wide variety of genetically determined autoimmune phenomena and adverse drug reactions (ADR) are linked to specific HLA class I or class II genes. A particularly severe ADR is caused by the anti-HIV drug abacavir and is associated with HLA-B*57:01 expression. We have produced several transgenic mouse lines on the C57BL/6 background expressing HLA-B*57:01 as: 1) a full length HLA-B*57:01 protein; 2) a chimeric version in which the α3, transmembrane (TM) and cytoplasmic (CY) domains derived from H2-Dd; and 3) a single-chain molecule consisting of human β2-microglobulin linked to a chimeric version, HLA-B*57:01α1α2 and the H2-Dbα3, TM, and CY domains. Flow cytometry showed cell surface expression on all lymphoid cell populations tested. Expression of the transgenes on the H2-Kb−Db− background was significantly higher than in MHC-I sufficient mice. Immunization with an HLA-B*57:01-restricted peptide induced a CD8+ T-cell response, indicating the biological antigen presentation function of the transgenically expressed protein, and abacavir inhibited the peptide specific response. Furthermore, in vivo administration of abacavir followed by in vitro culture induced a CD8+ T-cell response and production of proinflammatory cytokines in the transgenics and not in controls. Thus, transgenic strains expressing HLA-B*57:01 present a specific HLA-B*57:01-restricted peptide, presentation is inhibited by abacavir, and abacavir alone can elicit CD8+ T cell responses. These observations support the view that HLA-B* 57:01 expression uniquely confers the abacavir sensitive phenotype. Such transgenic mouse strains provide useful models for exploring the mechanisms of HLA-B*57:01-linked adverse drug hypersensitivity reactions.
A novel flow cytometric assay for single-cell analysis of microRNA, messenger RNA and protein expression during cell differentiation and maturation.
Authors: Lai Chunfai , Stepniak Dariusz , Sias Leslie , Funatake Castle 
Published: January 2023

Abstract
We describe a novel assay that enables analysis of microRNA, mRNA, and protein expression in single cells by flow cytometry. This in situ hybridization-based method employs pairs of target-specific probes, branched DNA amplification technology, and fluorochrome-conjugated label probes, resulting in sensitive detection of different types of RNA targets including mRNA, microRNA, long non-coding RNA and viral RNA. To broaden our understanding of the applicability of this assay, we determined its limit of detection for mRNA and microRNA. Using in vitro bone marrow-derived macrophages as a model, we demonstrate how this assay can be used to study the process of cell differentiation and maturation. This novel assay enables the simultaneous detection of up to four RNA targets in the presence of fluorochrome-conjugated antibodies in single cells and is a powerful new tool that will enable new insights into cellular processes.
A method for determining antibody-dependent cellular phagocytosis
Authors: Kamen Lynn A , Kho Elviza , Ordonia Benjamin , Langsdorf Chris , Chung Shan 
Published: January 2023

Abstract
Antibody-based therapeutics are a powerful tool to treat diseases. While their mechanism of action (MOA) always involves binding to a specific target via the Fab region of the antibody, the induction of effector functions through the Fc region of the antibody is equally important for antibody therapeutics designed to deplete tumor cells. By binding of the Fc region to FcγRs on the surface of immune cells, antibody therapeutics exert effector functions such as antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), both of which induce target cell death and aid in the efficacy of the treatment. There are many in vitro assays designed to measure ADCC and CDC activity of therapeutics. However, another major Fc effector function is antibody-dependent cellular phagocytosis (ADCP). ADCP is the mechanism by which antibody-opsonized target cells activate the FcγRs on the surface of macrophages to induce phagocytosis, resulting in the internalization and degradation of the target cell through acidification of the phagosome. ADCP has been implicated as a major MOA of several biologics, but this activity is difficult to measure in an in vitro assay. Most assays measure the association between target cells and macrophages, however co-localization does not indicate phagocytosis. In this presentation, we describe the development of a robust method for measuring ADCP activity. By labeling target cells with a pH sensitive dye that only fluoresces in mature phagosomes, it is possible to quantitate the ADCP activity of different antibody therapeutics. This method was developed on parallel platforms, utilizing both flow cytometry as well as a novel method quantitating the ADCP activity on a cell imaging system.
Immunomagnetic separation technology with reversible labeling for direct isolation of leukocytes
Authors: Evaristo Cesar , Steinbrueck Philipp , Bissa Ursula , Yu Zhong , Brieden Jennifer , Dose Christian 
Published: January 2023

Abstract
Immunomagnetic enrichment of leukocytes plays an important role in both research and clinical applications. Accordingly, there is a continued need for improved and reliable cell separation methods. Current enrichment strategies with specific magnetic labeling of the target cells allow for efficient isolation but have the disadvantage of residual labeling on the cell surface after isolation. Therefore, the isolation of viable cells with a minimal potential of surface-initiated activation becomes increasingly more important.
To this end, we have combined magnetic cells separation, MACSTM, the proven state-of-the-art method for the isolation of viable cells from heterogeneous cell populations with a releasable-label technology developed at Miltenyi Biotec, which allows direct isolation of truly unmarked cells. This method provides an easy and fast solution for the highly specific separation of unlabeled leukocytes directly from PBMCs. In this poster we present our latest results from this novel technology applied to key markers in the immunology field.