bluuebunny/crossref_metadata_2025_split huggingface dataset:
Dimethyl fumarate attenuates T helper type 2 (Th2) -mediated allergic airway inflammation by modulating dendritic cell function
Authors: Jaiswal Anil Kumar , Sandey Maninder , Suryawanshi Amol , Cattley Russell C , Mishra Amarjit 
Published: January 2023

Abstract
Dendritic cells (DCs) are primary antigen presenting cells in lungs that promote Th2 cell mediated immunity to house dust mite (HDM) allergen. DCs are attractive therapeutic targets to modulate allergen-induced airway inflammation and adaptive immune responses. Dimethyl fumarate (DMF) are methyl esters of fumaric acid and regulates immune cell differentiation and effector functions. However, the effect of DMF on lung DCs has not yet been addressed. Here, we investigated the effect of DMF treatment employing murine model of HDM-induced experimental allergic asthma and DC adoptive transfer experiment. We show that local DMF application attenuates airway inflammation, mucous cell hyperplasia, and airway resistance to increasing dosage of inhaled methacholine. Notably, DMF administration in the allergen challenge phase significantly inhibited the lung recruitment of CD11b+ conventional DCs (cDC2) and suppressed Th2 cytokine producing IL4, IL5, and IL13 CD4+ T cells. In lung draining mediastinal lymph node (mLNs), DMF treatment significantly induces CD4+T regulatory (Tregs) cells. Moreover, prior to administration of fluorescent-labeled HDM, single DMF application to naïve mice impeded the migration of CD11b+ cDC2 to the draining mLNs. Adoptive transfer of DMF treated CD11c+ HDM-pulsed DCs to naïve mice with subsequent intranasal HDM-challenge similarly attenuates the features of airway inflammation. Collectively, our findings suggest that local DMF application modulates lung DCs function, thereby attenuates airway inflammation and is a potent antiasthmatic therapeutic agent.
The role of tyrosine kinase Itk in T helper 17 and T regulatory cells in IL17 mediated hypersensitivity pneumonitis
Authors: Elmore Jessica , Carter Chavez , Huang Weishan , Koylas Nicholass , Bennett Amelia , August Avery 
Published: January 2023

Abstract
The balance of inflammatory and suppressive cytokines is critical in controlling inflammatory responses, and the pro- and anti-inflammatory cytokines IL17A and IL10 has been implicated in numerous pulmonary inflammatory diseases. The tyrosine kinase, Itk, plays a critical role in T cell activation. We have previously shown that Itk−/− mice do not develop allergic airway disease coupled with reduced production of IL17A in the lungs. In vitro, Itk is required for the development of Th17 cells and their production of the pro-inflammatory cytokine IL17A. Furthermore, Type I regulatory and Foxp3+ T regulatory (Tregs) cells, suppressive producers of IL10, are positively and negatively regulated by Itk respectively. Farmer’s lung is a subset of hypersensitivity pneumonitis caused by repeated exposure to the bacteria Saccharopolyspora rectivirgula (SR), and is dependent on IL17A and regulated by IL10. We find that surprisingly, exposure to SR drives robust IL17A response produced by CD4+ T cells even in the absence of Itk, accompanied by the development of pulmonary inflammation. SR also induced Itk independent development of a population of population of Foxp3+ Tregs cells that produce IL17A, accompanied by a significant decrease in Tr1 cells compared to WT mice. These studies suggest that TCR signaling through Itk differentially regulates the development of inflammatory Th17 cells and suppressive Tregs and Tr1 cells. Understanding how Itk modulates the development of Th17/Treg cytokine responses will allow us to better understand the precise role of Itk in the production of IL17A and IL10 in airway inflammation.
Augmented Co-Stimulatory Repertoire of γδ-T Cell Pool in Pediatric Peanut Allergic Patients
Authors: Watkin Levi B , Hassan Oluwatomi , Davis Carla M , Anvari Sara 
Published: January 2023

Abstract
The prevalence of Atopic disease has been on a steady increase over the last 20–30 years. Of these diseases, food allergy represents a major proportion of the anaphylaxis mediated deaths, particularly to peanut. While there are presently multiple theories to explain this increase in allergy, it is clear that in food allergic individuals there is a loss of peripheral tolerance to these innocuous antigens. One of the most effective means of generating peripheral tolerance is through the gut which is home to many specialized immune cells, including intra-epithelial lymphocytes (IELs). Of these IELs, γδ-T cells represent nearly 50% of the population. Due to the large representation of the IELs in the gut, we hypothesize that alteration in the γδ-T cell pool plays a role in the loss of tolerance to these innocuous antigens. Here we examined PBMCs from peanut allergic patients and compared them to healthy controls. We find that while there is no difference within the overall frequency of total γδ-T cells between healthy and peanut allergic patients, when separated based upon CD4 and CD8 expression, we find the CD8 population decreased in the peanut allergic population. Additionally, when investigating subsets within the CD8 γδ-T cells we find altered frequencies of cells expressing costimulatory molecules both classically thought of as being expressed on T-cells (CD7 and OX40), and on antigen presenting cells (CD86). Taken together, these data suggest that γδ-T cells may contribute to the pathogenesis of peanut allergy by way of augmenting the costimulatory dynamic taking place in the gut. Further investigation is required to determine whether this observation leads to the development of peanut allergy or is a result of the disease on the individual.
Gut oxidative responses by chronic exposure to pollutants enhance IgE production via PGE2-dependent mechanism
Authors: Kim Eunsoo , Bonnegarde-Bernard Astrid , Opiyo Stephen O. , Ahmer Brain M. , Comet-Boyaka Estelle , Boyaka Prosper N 
Published: January 2023

Abstract
Environmental pollutants are believed to contribute to the increased incidence of allergic diseases. However, mechanisms linking exposure to pollutants with allergy are poorly understood and researches about heavy metals are mostly focused on its toxicity. Groups of mice were chronically exposed to subtoxic doses of cadmium in the drinking water. We then analyzed the impact of this exposure on gut immune homeostasis, microbiota, allergic sensitization to ingested allergen and subsequent allergic responses after allergen challenge. Chronic ingestion of subtoxic doses of cadmium increased IgE responses of mice to ingested allergen and the severity of allergy symptoms upon subsequent allergen challenge. We also found that exposure to subtoxic doses of cadmium induces dysbiosis of the gut microbiota and luminal accumulation of oxidized vitamin D3 metabolites. More importantly, we found that this microenvironment selectively enhances the activation of BAFF and the production of IgE via PGE2-dependent mechanism by intestinal epithelial cells. These findings reveal a new mechanism for the induction of IgE responses via alteration of vitamin D3 metabolism by environmental pollutants which stimulate an oxidative and inflammatory environment in the gut.
Characterization of protein allergens in a mouse model of wheat-induced anaphylaxis
Authors: Gangur Venugopal , Jorgensen Rick , Gao Haoran , Jin Yining , Salloum Jillian , Jian Dan , Ng Perry KW 
Published: January 2023

Abstract
Wheat allergies are among the major types of food allergies that are potentially life-threatening because of anaphylaxis. Wheat contains 3 different types of protein allergens: water/saline-soluble albumins/globulins, alcohol-soluble gliadins and acid-soluble glutenins. Although gliadins have been linked to anaphylaxis, whether or not other proteins cause anaphylaxis is not well understood. Here, we sought to identify saline-soluble wheat proteins (SSWP) involved in anaphylaxis using a mouse model. Balb/cJ mice were sensitized to SSWP by four intraperitoneal (IP) injections along with alum as an adjuvant. Sensitization was confirmed by testing for SSWP-specific IgE using ELISA. Anaphylaxis was measured by hypothermia shock response. After multiple booster injections with SSWP, hyper-IgE plasma was collected and pooled to create a mini-plasma bank. Using hyper-IgE plasma, an IgE-Western blotting (WB) method was optimized. The IgE-binding proteins were identified by sequencing using LC/MS/MS method. The hyper-IgE plasma had an IgE titer of ~2560. The optimized IgE-WB included 3 overnight incubations followed by signal detection using an HRPO-based method. Sequencing of the IgE binding bands in WB identified the following proteins: globulin 3A (66 kDa), globulin 3B (57 kDa), serpin (43 kDa), glyceraldehyde-3-phosphate dehydrogenase (37 kDa), chitinase (28 kDa) and globulin 1 (25 kDa). This is the first study characterizing salt-soluble protein allergens in a mouse model of wheat-induced anaphylaxis. Funding: USDA, NIFA, Hatch project MICL02486 (Accession Number: 1012322), Agricultural and Food Research Initiative Competitive Program, grant number: 2018-67017-27876.
The bone marrow-derived Gr-1+CXCR2+ cells are required for T cell infiltration into the skin challenge site to elicit contact hypersensitivity responses are inflammatory monocytes expressing FasL and perforin
Authors: Kish Danielle , Fairchild Robert L 
Published: January 2023

Abstract
Contact hypersensitivity (CHS) is a CD8 T cell mediated response to skin sensitization with hapten and subsequent challenge. Hapten-primed CD8 T cell infiltration into the skin challenge site during elicitation of CHS requires prior infiltration and activation of Gr-1+CXCR2+cells expressing both FasL and perforin. This study tested factors generating these myeloid cells in the bone marrow (bm) of hapten sensitized mice. The myeloid cells were enriched from the bm of DNFB sensitized mice 2 hrs after hapten challenge by depleting CD3+, B220+, NK1+, F4/80+, class II MHC+and Ly6G+cells with &gt;90% of the remaining cells expressing Gr-1, CXCR2, CD11b, CD11c, Ly6C, CCR2, and CX3CR1, the phenotype of inflammatory monocytes (IM), plus FasL. The FasL+IM were at low numbers in the bm of naïve and hapten sensitized mice but increased to peak numbers 2–6 hrs. after hapten challenge of sensitized wild type (WT) mice. The FasL+IM were also generated in the bm of hapten sensitized and challenged Rag−/−mice and T-cell depleted WT mice, indicating that their generation can occur independently of T cells. The FasL+IM were not generated following hapten challenge of sensitized mice with targeted deletions in TLR4, IL-1β receptor or IFN-γ genes or in sensitized WT mice treated with anti-TNF-α mAb during hapten challenge. However, treatment with anti-GCSF antibody during hapten challenge increased their numbers in the bm and increased the magnitude of the CHS response elicited. Collectively, these results indicate that haptenreactive CD8 T cell infiltration into hapten challenged skin requires the TLR4-, TNF-α- and IFN-γ-dependentgeneration of bm-derived IM expressing FasL and perforin and their activation within the hapten challenged skin site.
Immune modulation caused by synthetic opioids during CD4+ T cell activation
Authors: Llorens Braulio Jose , Levine Alan D 
Published: January 2023

Abstract
Opioids are heavily prescribed medications used to treat chronic pain. Since their additional response includes euphoria, there is an exceedingly high addiction rate that has led to a country wide epidemic of opioid misuse. Opioid effects are not limited to the nervous system, since leukocytes express opioid receptors, which are G-protein coupled receptors known as μ, δ, and κ, suggesting that natural and synthetic opioids might regulate the immune system. Based on previously published reports, we hypothesized that opioids induce distinct regulatory pathways in CD4+ T cells that results in inhibition of the immune response. CD4+ T cells are isolated by negative selection from peripheral blood mononuclear cell and opioid receptor expression is assessed on resting and activated T cells using flow cytometry. Two models of opioid exposure in vitro were developed to study the effects of opioids on CD4+ T cell activation. Cells were treated with a δ opioid receptor specific agonist (DPDPE) for 24 h before or after activation and harvested 24 h after the second treatment and stained for expression of a well characterized early T cell activation marker – CD69. Cells treated with an opioid after activation showed no major shifts in CD69 intensity, suggesting opioids had no influence on a previously activated T cell. However, CD4+ T cells treated with an opioid before activation exhibited an increase in CD69 expression, suggesting an unexpected enhancement of T cell activation. In conclusion, opioids seem to play distinct biological activities depending on the activation state of the CD4+ T cell, and may function as both immunoinhibitory and immunostimulatory agents. Future studies will identify the mechanisms by which opioids regulate immunity.
Secondary stimulation of CD4 T cells generates an exhaustion phenotype
Authors: Wolf Gideon S , Singh Nevil J 
Published: January 2023

Abstract
Activated naïve T cells typically acquire effector functions over a 2–3 day period of differentiation in the secondary lymphoid organs, before migrating to the tissue sites to deliver effector functions. If the stimulating antigen is long lasting, as in the case of chronic infections, the T cells may undergo exhaustion, often through pathways involving co-inhibitory receptors such as PD1. Surprisingly, we find that a secondary and acute stimulation even short periods after the initial antigen encounter leads CD4+ T cells to exhibit properties of an exhausted phenotype. This is marked by the loss of proliferative potential and the upregulation of PD1. Naïve TCR-transgenic 5C.C7 T cells stimulated with their cognate peptide, proliferate rapidly between 2–4 days. Re-stimulation of these T cells on the 3rd day however, revealed a profound proliferative arrest accompanied by the rapid re-expression of PD1. The new levels of PD1 was proportional to the dose of re-stimulating antigen. This phenomenon was distinguished from Activation Induced Non-Responsiveness (AINR) as the 2nd stimulation arrested even the basal proliferation that was residual from the 1st antigen exposure. However, this is consistent with the property of effector T cell tuning that was previously described for effector T cells homing to tissue sites. We are currently examining the molecular machinery underlying this duality during T cell expansion to understand the paradoxical loss of T cell responsiveness to antigen during a primary effector response. These studies are expected to develop new approaches to not only improving primary T cell responses but also circumventing exhaustion during chronic microbial infections and tumors.
Runx1 is a key regulator of T cell anergy
Authors: Wilfahrt Drew Nicholas , Hsu Fan-Chi , McCue Shaylene , Rajcula Matthew , Shapiro Virginia Smith 
Published: January 2023

Abstract
The transcription factor Runx1 is critical for T cell development and maturation. CD4-Cre Runx1 conditional knockout mice have a block in T cell maturation, leading to complement deposition on recent thymic emigrants, and leaving very few CD4+T cells in peripheral lymphoid organs. Interestingly, thymic- and splenic-derived T cells from CD4-Cre Runx1 cKO mice exhibit increased expression of markers indicative of anergy including FR4 and CD73, which is not seen in other genetically modified mice with a block in T cell maturation. These data suggest Runx1 plays a role in regulating the development of T cell anergy in CD4+T cells. We have generated a 1:1 mixed bone marrow chimera (BMC) system with Estrogen Receptor-Cre (ER-Cre) Runx1 conditional knockout and wild type B6.SJL bone marrow to delete Runx1 in mature CD4+ T cells. Upon tamoxifen treatment and deletion of Runx1, the ER-Cre Runx1 cKO CD4+ T cells spontaneously develop an anergic phenotype as they fail to proliferate upon TCR/CD28 stimulation, and co-express CD73 and FR4. This cell surface phenotype is cell intrinsic since wild type B6.SJL CD4+ T cells in the mixed bone marrow chimera do not upregulate CD73 and FR4 upon tamoxifen treatment. Surprisingly, the mixed BMC system reveals both the ER-Cre Runx1 cKO and the wild type B6.SJL peripheral T cells fail to proliferate in response to TCR/CD28 stimulation, suggesting Runx1 also regulates a cell-extrinsic suppressive mechanism controlling T cell tolerance. This cell-extrinsic induction of anergy requires Runx1-deficient CD4+ T cells, but it is not dependent on Runx1-deficient Tregs. Thus, Runx1 is a key regulator of T cell anergy, and further analysis of this BMC model will serve to uncover the mechanism.
Negative feedback control of autoimmunity by a novel population of regulatory unconventional CD8+ T cells
Authors: Marrero Idania , Sheng Huiming , Maricic Igor , Fanchiang Shaohsuan S , Zhang Sai , Sant’Angelo Derek , Kumar Vipin 
Published: January 2023

Abstract
Immune tolerance mediated by regulatory T cells is important in the control of homeostasis. The characterization of regulatory CD8 T cells has been hampered by inability to distinguish them from conventional CD8 T cells. We have identified a novel population of unconventional cytotoxic TCRαβ+CD8+ T cells with regulatory properties, called uTr, in both naïve mice and humans. The uTr cells are innate-like, express the promyelocytic leukemia zinc finger (PLZF) transcription factor and display an activated-memory phenotype (CD122high, CD44high,CD62Llow, CD69+). They are CD8αα+, NK1.1+(CD161+ in human), express NK inhibitory receptors and are dependent upon IL-15/IL-2Rβ signaling. A significant decrease in the frequency of uTr cells in PLZF−/− mice suggests their dependency on PLZF expression. Similarly, a significant reduction of uTr cells in Qa-1b−/− mice indicates that a sizable proportion of them are restricted by Qa-1b MHC molecules. The deep-sequencing, RT-PCR and FACS analysis of sorted hepatic uTr cells indicate a polyclonal TCR repertoire in both mice and humans. Adoptive transfer of sorted hepatic uTr cells but not conventional CD8 T cells protects B6 mice from EAE as well as RAG1−/− mice from T cell-induced colitis. The uTr cells secrete IFNγ, IL-4 and IL-17 but not TGFβ and express perforin and granzyme B and the mechanism(s) of regulation is dependent upon perforin. The uTr cells are distinct from other unconventional T cells, including NKT cells, NKT-dependent innate-like CD8 T and MAIT cells. Collectively, we have identified a novel unconventional regulatory CD8 T cells that are involved in a negative feedback mechanism of regulation of autoimmunity and also have important implications for anti-cancer immunity.
LAG-3+ induced regulatory T cells confer infectious tolerance with suppression of IFN-γ response decoupled from reserved proliferation
Authors: Huang Ching-Tai , Dutta Avijit , Hung Chen-Yiu , Chen Tse-Ching , Lin Chun-Yen , Lin Yung-Chang , Chang Chia-Shiang , Chen Ting-An , Huang Yu-Lin 
Published: January 2023

Abstract
Induced regulatory T cells (iTreg) is an adaptive response in inflammation with disease processes such as infection, autoimmunity or cancer. A small number of iTreg may exert long-lasting regulation. Here we report the LAG-3+ iTreg transfer regulatory activity to the CD4+ T cells they suppress. This infectious propagation of regulatory activity exemplify a dominant immune regulation in vivo. Naïve hemagglutinin (HA) antigen-specific CD4+ T cells are induced to differentiate into IFN-γ competent effector T cells and then LAG-3+ Tregs in HA transgenic mice inherently expressing the cognate HA antigen. The LAG-3+ iTreg suppresses activation of effector function of cognate naïve CD4+ T cells in HA transgenic mice, with silenced IFN-γ machinery including STAT1 phosphorylation and T-bet induction. The LAG-3 iTreg suppressed, IFN-γ disarmed CD4+ T cells still proliferate and acquire LAG-3 expression on their surface in vivo. They become the next generation LAG-3+ iTregs with the ability of suppression of activation of effector function of cognate naïve CD4+ T cells in HA transgenic mice, with the same mechanism of silenced IFN-γ machinery. LAG-3+ iTreg exercises regulation in vivo by suppression of effector function decoupled from reserved proliferative response. The effector function suppressed CD4+ T cells still proliferate upon activation and acquire LAG-3+ iTreg phenotype, a reminiscence of the infectious tolerance proposed in 1970.
Construction of CAR-iTreg (Chimeric antigen receptor-inducible Treg) for inducing tolerance in transplantation
Authors: Won Hye-ji , Kim Yong-Hee , Park Chung-Gyu 
Published: January 2023

Abstract
Tolerance induction is ultimate goal in the field of transplantation. Cell-based therapy with regulatory T cells (Treg) has been effective and evolving, Chimeric antigen receptor (CAR) T cells to target specific antigen are new promising cell-based therapy. Engineering of CAR-Treg have many advantages that can be used in transplantation. Here, we designed development of CAR-iTreg (Chimeric antigen receptor-inducible Treg), which is engineered to become inducible Treg in vivo. DNA for single chain fragment variable (scFv) was obtained from MR1 hybridoma to target CD40L as the antigen. Cytoplasmic signaling part was constituted with CD3z and CD28 for T cell activation, and with TGFβR1 and IL-2Rβ for Treg induction. The CAR DNA construct was inserted into pLVX-IRES-ZsGreen1 expression plasmid vector. The expression vector and packaging vectors were used to produce lentiviral vector in HEK293FT cells. Harvested lentivirus has been tested in Jurkat and EL4 cells, and the expression of ZsGreen fluorescence and the CAR could be confirmed. This CAR-containing lentivirus can be used to transfect naïve CD4+ T cells and be transferred to syngeneic mice. These CAR-T cells target CD40L expressed on activated T cell and convert into inducible Treg cells in the graft environment. Thus, this uniquely engineered CAR construct allows tolerance induction in transplantation model. Furthermore, this CAR-iTreg strategy with other scFv has potential application in autoimmune diseases.
Helios deletion in CD4+T cells induces pTregs that inhibit memory recall responses
Authors: Penna Vinay , Skadow Mathias , Shevach Ethan M , Thornton Angela M 
Published: January 2023

Abstract
The transcription factor Helios is required for the maintenance of the suppressive phenotype of Foxp3+ regulatory T cells (Tregs) as evidenced by the spontaneous autoimmune activation that develops in mice with a Foxp3− specific Helios deficiency. Helios is also upregulated in a subset of activated CD4+Foxp3−T conventional (Tconv) cells, but its role in Tconv cells remains unknown. Mice with a CD4-Cre driven Helios deletion do not exhibit autoimmunity despite the defect in the suppressor function of their Treg population, pointing to a requisite function of Helios in the activation of Tconv cells in response to antigen. We examined the capacity of Helios-deficient, naïve Tconv cells to respond to their cognate antigen in vivo and found that Helios was not required for proliferation, differentiation into helper subsets, or the production of effector cytokines. Unexpectedly, we observed that Helios deficiency increased the susceptibility of naïve T cells to differentiate into pTregs under inflammatory conditions. These pTregs were then subsequently able to suppress the reactivation of memory T cells in an antigen-specific manner upon challenge with antigen. These findings suggest that increased pTreg differentiation in the periphery in mice with a CD4-Cre driven Helios deletion may prevent the development of autoimmunity normally associated with Helios-deficient Tregs.