bluuebunny/crossref_metadata_2025_split huggingface dataset:
IFN-gamma mediated inflammatory monocyte recruitment neutralizes iNOS-dependent parasite killing by expanding the permissive host cell reservoir during early Leishmania amazonensis infection
Authors: Carneiro Matheus Batista , de Moura Lopes Mateus Eustaquio , Romano Audrey , Campos Alexandre Camargo , Sacks David , Vieira Leda Q , Peters Nathan Colin 
Published: January 2023

Abstract
IFN-γ is a key factor in the elimination of intracellular parasites. However, during early L. amazonensis infection IFN-γ production does not correlate with a reduction in parasite growth and IFN-γ−/− mice do not have enhanced parasite loads. The reason why IFN-γ production does not reduce parasite loads during early L.a. infection is controversial. In order to investigate the role of IFN-γ during early L.a. infection we infected C57BL/6 Wt and IFN-γ−/− mice with 104 RFP+ parasites in the ear and followed disease for 6 wks. IFN-γ−/− mice showed equivalent lesion sizes and parasite numbers compared to Wt mice for the first 4 wks p.i., despite higher expression of the putative disease promoting factors IL-4 and arginase I. IFN-γ−/− mice had no CD11b+iNOS+ phagocytes while iNOS production was detected as early as 2 wks p.i. in Wt mice. Interestingly, Wt mice had higher numbers of inflammatory monocytes per ear during early infection. Inflammatory monocytes represented the majority of total infected cells and parasites within inflammatory monocytes cells were viable. Inflammatory monocytes were a small proportion of CD11b+iNOS+ cells in Wt mice and had significantly reduced levels of IFN-γR versus dendritic cells and macrophages. The enhanced host cell infiltrate observed in Wt mice may counteract the protective role of IFN-γ mediated iNOS expression. This could explain, in part, why IFN-γ production does not lead to a reduction in parasite load during early L. amazonesis infection. We propose IFN-γ plays a dual role, inducing both the expression of iNOS and parasite killing and the recruitment of inflammatory monocytes that support parasite replication during early infection.
Local antigen in non-lymphoid tissue promotes resident memory CD8+ T cell formation during viral infection.
Authors: Nolz Jeffrey C , Khan Tahsin N , Mooster Jana L , Kilgore Augustus M , Osborn Jossef F 
Published: January 2023

Abstract
Tissue-resident memory (TRM) CD8+ T cells are functionally distinct from their circulating counterparts and are potent mediators of host protection against re-infection. One largely accepted paradigm is that activated CD8+ T cells are able to freely migrate to non-lymphoid tissues and differentiate into TRM independent of local antigen recognition. In contrast, using skin infections with Vaccinia virus (VacV) expressing model antigens, we found that local antigen recognition had a profound impact on TRM formation. Activated CD8+ T cells trafficked to VacV-infected skin in an inflammation-dependent, but antigen-independent manner. However, following viral clearance, there was a subsequent ~50-fold increase in CD103+/CD69+ TRM formation when antigen was present in the tissue microenvironment. Secondary antigen stimulation in non-lymphoid tissue caused CD8+ T cells to rapidly express CD69 and be retained at the site of infection. Finally, TRM CD8+ T cells that formed during VacV infection in an antigen-dependent manner became potent stimulators of localized antigen-specific inflammatory responses in the skin. Thus, our studies indicate that the presence of antigen in the non-lymphoid tissue microenvironment plays a critical role in the formation of functional TRM CD8+ T cell populations, a finding with relevance for both vaccine design and prevention of inflammatory disorders.
Route of immunization governs the development of distinct resident memory CD8 T cells and protective immunity in the intestinal mucosa
Authors: Qiu Zhijuan , Sheridan Brian S 
Published: January 2023

Abstract
We previously demonstrated that oral immunization with a murinized Listeria monocytogenes induced rapid formation of intestinal resident memory CD8 T (TRM) cells that were capable of providing secondary protection. It is well appreciated that systemic vaccination also induces a population of protective TRM cells in the intestine. We sought to understand the impact of immunization route on the development of intestinal TRM cells and protective immunity. Oral immunization resulted in significantly more antigen-specific effector CD8 T cells in the intestinal compartments than intravenous (i.v.) immunization, likely due to differential imprinting and migration of T cells. While both oral and i.v. immunization induced a4b7 expression on CD8 T cells, only oral immunization resulted in CCR9 expression. Moreover, compared to i.v.-induced CD8 T cells, oral-induced cells rapidly adopted a CD127+ KLRG-1− memory precursor phenotype and a CD103+ CD69+ residency phenotype. Furthermore, oral immunization led to a substantially larger TRM population in the intestine compared to i.v. immunization. While the majority of oral-induced TRM cells were CD103+, a substantial proportion of i.v.-induced TRM cells did not express CD103. Collectively, these data suggest that oral immunization would provide an optimal sentinel population of memory cells that are more efficient in providing protection against secondary intestinal exposure. Indeed, oral-immunized mice had reduced bacterial burden after challenge infection compared to i.v.-immunized mice. In conclusion, our studies demonstrate that route of immunization greatly affects the induction and maintenance of TRM cells and thus the outcome of immunization.
Failure of gut-associated pDCs to express membrane bound APRIL and BAFF prevents their ability to promote low-affinity IgA expression in ME/CFS
Authors: Lombardi Vincent C , Khaiboullina Svetlana F , De Meirleir Kenny L , Mijatovic Tanja , Hulstaert Jan 
Published: January 2023

Abstract
Myalgic encephalomyelitis/chronic fatigue syndrome (ME/CFS) is a heterogeneous illness characterized by a number of comorbid conditions; gastrointestinal (GI) dysregulation make up one subgroup of this disease. IgA is the most abundant antibody isotype found in mucosal secretions including the gut. In a process of class switch recombination (CSR), that relies on the interaction of plasmacytoid dendritic cells (pDCs) with B cells, in a T cell independent (TI) manner, low-affinity IgA are produced that limit the adhesion of commensal bacteria to intestinal epithelia without neutralizing them. These low-affinity antibodies also limit bacterial overgrowth and potential bacterial translocation thus maintaining gut homeostasis. This process is known as “immune exclusion”. Two ligands on the surface of pDCs that are obligatory for the process; the membrane bound form of APRIL and BAFF. The upregulation of APRIL and BAFF on the surface of pDCs is dependent on low-level expression of type I interferon (IFN) which is produced by intestinal stromal cells in response to Toll-like receptor (TLR) engagement. Previous studies suggest that peripheral pDCs are significantly lower in subjects with ME/CFS when compared to controls and studies conducted by us further suggest these cells likely redistribute from the periphery to the gut. We have observed that, in contrast to controls, gut-associated pDCs in subjects with ME/CFS lack APRIL and BAFF expression. These data support a model of gut pathology in ME/CFS whereby dysregulated pDCs fail to promote the production of low-affinity IgA through the process of TI activation of B cells, thereby leading to bacterial overgrowth, dysbiosis, bacterial translocation and systemic immune activation.
Investigating distinct immunization routes in zebrafish on mucosal immunity
Authors: Young Matt , Li Ellen S. , Branham Erica , Browne Zachary , Koufteros Christiana A , Pham Sydney E L , Kim Joshua C , Stebbins Elyssa A , Wenck Fallon E , Zhang Annie L , Kommein Swathi , Russo Maximillian A X , Criscitiello Michael F 
Published: January 2023

Abstract
Model organisms such as Danio rerio are used to explore and model physiological reactions to stress, illness, and disease. Immune response is measured by quantifying lymphocyte presence and the production of mRNA encoding different immunoglobulin proteins. Because the method through which D. rerio is exposed to an antigen affects its immune response and the degree to which immunity is developed, we will expand on experiments comparing the immune response after visceral or mucosal introduction of DNP-KLH in D. rerio. In our visceral treatment, we will combine DNP-KLH with Freund’s incomplete adjuvant and inject the mixture into the fish (100 uL at 1 mg/mL). For our mucosal exposure, we will combine DNP-KLH with cholera toxin as an adjuvant and introduce the mixture to their water (35 mg/L). Two control groups will account for intravenous immune response by injections solely with sterile saline solution as well as a control for the anesthetic used, MS-222. Viscera tissue samples will be collected from the immunized fish and two control groups of fish at weekly points throughout the experiment and will be analyzed for lymphocyte versus granulocyte ratios with flow cytometry and expression of IgM, IgZ-1, and IgZ-2 levels via qPCR. We expect to see an increase in lymphocyte percentages for both immunized groups compared to the control groups, with a higher lymphocyte ratio in the mucosal group than the visceral group. Furthermore, we expect more mucosal IgZ isotypes in the mucosal group and higher levels of the IgM, IgZ-1, and IgZ-2 antibodies in the visceral group than in the mucosal group. For future experiments, we would like to determine specific tissues of interest that provide data for analysis of mucosal and visceral immunological responses.
3H-1,2-Dithiole-3-thione as a novel therapeutic agent for the treatment of experimental autoimmune encephalomyelitis
Authors: Kuo Ping-Chang , Brown Dennis A. , Scofield Barbara A. , Yu I-Chen , Chang Fen-Lei , Yen Jui-Hung 
Published: January 2023

Abstract
3H-1,2-Dithiole-3-thione (D3T), the simplest member of the sulfur-containing dithiolethiones, is found in cruciferous vegetables, and has been previously reported to be a potent inducer of antioxidant genes and glutathione biosynthesis by activation of the transcription factor Nrf2. D3T is a cancer chemopreventive agent and possesses anti-inflammatory properties. Although D3T has been shown to protect against neoplasia, the effect of D3T in the autoimmune inflammatory disease Multiple Sclerosis/Experimental Autoimmune Encephalomyelitis (EAE) is unknown. The present study is the first report of the therapeutic effect of D3T in EAE. Our results show D3T, administered post immunization, not only delays disease onset but also dramatically reduces disease severity in EAE. Strikingly, D3T, administered post disease onset of EAE, effectively prevents disease progression and exacerbation. Mechanistic studies revealed that D3T suppresses the expression of co-stimulatory molecules and production of pro-inflammatory cytokines by dendritic cells, inhibits pathogenic Th1 and Th17 differentiation, reduces the production of inflammatory mediators by microglia, and induces the expression of antioxidant phase II enzymes in microglia. In summary, these results indicate that D3T affects both innate and adaptive immune cells, and the protective effect of D3T in EAE might be attributed to its effects on modulating dendritic cell and microglia activation and pathogenic T cell differentiation.
Inhibition of MIF as a novel treatment for autoimmune myocarditis and dilated cardiomyopathy.
Authors: Nalawade Saisha Abhay , Forsthuber Thomas Gunter 
Published: January 2023

Abstract
Myocarditis is an inflammatory disease of the myocardium and a major cause of sudden death in young adults. It is characterized by the presence of immune infiltrates and necrotic myocytes in the myocardium. Importantly, patients often progress to a more severe form of the disease termed dilated cardiomyopathy (DCM), which leads to ventricular dilation and impaired cardiac function. Despite the inflammatory and autoimmune nature of this disease condition, immunosuppressive treatments such as corticosteroids (CSs) have not been very effective in preventing myocarditis and its progression to DCM. We hypothesized that macrophage migration inhibitory factor (MIF) may play a role in resistance to CSs, as it is the only known pro-inflammatory cytokine to be induced by CSs. Importantly, MIF counter-regulates CS-mediated immunosuppression. Using the experimental autoimmune myocarditis (EAM) animal model, we observed that MIF−/− mice treated with Dexamethasone (Dex) were highly resistant to EAM and progression to DCM. Furthermore, using small molecule inhibitors of MIF combined with Dex treatment recapitulated this phenotype in wild type mice. Investigating the mechanism we found that treatment with MIF inhibitors and Dex decreased the expression of key chemokines and adhesion molecules and implicated these molecules in the progression of EAM to DCM. Our results suggest that MIF promotes the recruitment of inflammatory cells to the myocardium to promote DCM. Moreover, the results suggest therapeutic inhibition of MIF in combination with CSs as a novel treatment approach to prevent DCM. Last, our studies may provide new insights into the mechanisms driving DCM.
Th17 and Th1 play distinct roles in obliterative bronchiolitis pathogenesis after transplantation.
Authors: Xu Jiangnan , Zhang Yamei , Wang Lifeng , Chen Qirui , Li Hui , Bromberg Jonathan S , Ding Yaozhong 
Published: January 2023

Abstract
Development of obliterative bronchiolitis (OB) is the biggest obstacle that limits long-term allograft survival and clinical application of lung transplantation. Mouse orthotopic trachea or lung transplantations were utilized to examine roles of Th17 and Th1 during OB pathogenesis after transplantation. Allografts of wild type recipients displayed elevated levels of Th1 and Th17 cytokines IFNγ, IL-6 and IL-17 expression. Recipients deficient in Th1 key transcriptional factor T-bet resulted in decreased cell infiltration and improved airway integrity in orthotopic transplanted trachea grafts; Lack of Th2 transcriptional factor Stat6 had little effect; While absence of Th17 master transcriptional factor RORγt had striking effects in abolishing airway epithelial injury, a leading factor of OB pathogenesis. Administering anti-IL-6 or anti-IL-17 but not anti-IFNγ mAbs restored normal airway epithelial morphology. Furthermore, we found that anti-CD40L treatment resulted in increased FoxP3 and regulatory T cells, and decreased IFNγ expression; However it had very limited effect on IL-17 suppression and airway epithelial protection. While CTLA4-Ig treatment significantly inhibit IL-17 expression and reduced epithelial injury, and the combination of CTLA4-Ig and anti-CD40L markedly restored airway morphology of grafts. Together, our findings provide new insights for understanding Th17 and Th1 responses during OB pathogenesis and demonstrate the critical importance of suppressing Th17 for OB development after transplantation.
LRBA is essential for allogeneic responses in bone marrow transplantation
Authors: Sudan Raki , Park Mi Young , Srivastava Neetu , Neelam Sudha , Youngs Christie , Wang Jia-Wang , Engelman Robert W , Kerr William 
Published: January 2023

Abstract
Members of the PH-BEACH-WD40 repeat gene family (PBW) play a role in coordinating receptor signaling and intracellular vesicle trafficking. LPS-Responsive Beige-like Anchor (LRBA) is a PBW protein whose immune function has not been defined. Here we show that LRBA−/− mice are viable with normal and healthful life span, but exhibit compromised rejection of allogeneic, xenogeneic and missing self bone marrow grafts. Further we demonstrate that LRBA−/− Natural Killer (NK) cells exhibit impaired signaling by the key NK activating receptors, NKp46 and NKG2D. However, induction of γ-IFN by co-culture with IL12 and IL18 remains intact, indicating LRBA selectively facilitates signals by receptors for ligands expressed on the surface of NK targets. Surprisingly, LRBA limits immunoregulatory cell numbers in tissues where Graft versus Host Disease (GvHD) is primed or initiated, and consistent with this LRBA−/− mice demonstrate resistance to lethal GvHD. These findings demonstrate that LRBA is redundant for host longevity while being essential for both host and donor-mediated immune responses in allogeneic transplantation and thus represent a unique and novel molecular target in transplant immunology.
Thymus is required for T cell reconstitution following lymphoablation
Authors: Ayasoufi Katayoun , Fan Ran , Valujskikh Anna 
Published: January 2023

Abstract
Lymphoablation is used as induction therapy in transplantation. Following lymphoablation, peripheral lymphopenia triggers homeostatic T cell proliferation and thymopoiesis. However, the relative contribution of each pathway to T cell recovery is unknown. The goal of this study was to test the role of thymus during T cell reconstitution following lymphoablation. We use a mouse model of heterotopic heart transplantation (BALB/c to B6) and a murine analog of Thymoglobulin (mATG). We previously reported that the recovery of CD8 T cells after mATG depletion requires residual memory CD4 T cells and CD40 expressing B cells. CD4 T cell depletion or CD154 blockade significantly reduced the numbers of single positive CD8 thymocytes generated in mATG treated mice. To test if thymus is required for T cell reconstitution, thymectomized and euthymic heart allograft recipients were treated with mATG on days 0 and 4. Thymectomy impaired recovery of CD4 and CD8 T cells compared to euthymic mice (10% vs. 1.7% CD4 and 3% vs 0.06% CD8 T cells on d 12). The transplantation of thymi from B6 WT or RAG1−/− mice under the kidney capsule of thymectomized mice partially restored T cell reconstitution following depletion. Adoptive transfer of CD4 T cells (10×106 i.v. d −1) or treatment with agonistic anti-CD40 mAb (0.1mg i.v. d 0, 2) also restored T cell recovery in thymectomized mice. These results indicate that depletion-resistant memory CD4 T cells and thymus are both required for effective CD8 T cell recovery. Our findings may lead to therapies that specifically target reconstitution of naïve vs memory T cells following lymphoablation.
Generation of alloantigen-specific T regulatory cells via a chimeric antigen receptor targeting HLA-A2
Authors: MacDonald Katherine Grace , Hoeppli Romy , Huang Qing , Gillies Jana , Luciani Dan , Orban Paul , Broady Raewyn , Levings Megan K 
Published: January 2023

Abstract
CD4+ T regulatory cells (Tregs) are of clinical interest due to their potent immunosuppressive activity in both transplantation and autoimmunity models. Clinical efficacy in humans has been limited by aspects of Treg biology, including the difficulty of isolating sufficient numbers for potency without sacrificing purity and thus function. Enriching for antigen-specific Tregs greatly increases potency, however, antigen-specific Tregs are extremely rare. The success of chimeric antigen receptor (CAR) technology in generating antigen-specific effector T cells suggested that a similar approach could be used to generate alloantigen-specific Tregs for use in transplantation. We generated a new CAR specific for HLA-A2 (A2-CAR) and tested its application in the generation of alloantigen-specific human Tregs. Naïve Tregs were sorted from human peripheral blood, activated, and transduced with A2-CAR lentivirus. Stimulation through the A2-CAR upregulated Treg activation markers and functional molecules, and resulted in proliferation. Importantly, in vitro studies showed that A2-CAR-expressing Tregs maintained their expected phenotype and suppressive function, before, during and after HLA-A2-mediated stimulation. In vivo, A2-CAR Tregs were superior to Tregs expressing an irrelevant CAR at preventing xenogeneic graft-versus-host disease caused by HLA-A2+ T cells. Together, these data suggest that alloantigen-specific Tregs generated by the introduction of an A2-CAR may be a potent and specific immunosuppressive treatment in transplantation.
MicroRNA let 7i regulates DCs maturation targeting IL-10 via JAK1-STAT3 signal pathway subsequently induces prolonged cardiac allograft survival in rats
Authors: Jin Xiangyuan , Sun Yong , Liu Xianglan , Wang Junfeng , Zhang Maomao , Yu Bo , Wu Jian 
Published: January 2023

Abstract

Objective
In this study, we investigated whether microRNA (MiR) let 7i regulates DCs maturation targeting IL-10 via JAK1-STAT3 signal pathway subsequently prolongs rat cardiac allograft survival.

Methods
QRT-PCR, ELISA, dual luciferase assay were performed to verify whether IL-10 was the target of let 7i. Tregs were assessed by flow cytometry and immunohistochemical study, and JAK1, STAT3 and pSTAT3 expressions were detected by western bolt. Lewis recipients of DA heart were transfused with PBS, LPS-mDCs, or let 7i inhibitor-mDCs. Allograft survival times were recorded, and histologic study were performed.

Results
Expression of IL-10 mRNA level and production of IL-10 were increased in let 7i inhibitor-mDCs as compared to LPS-mDCs. Luciferase activity showed that the translational level of the IL-10 luciferase reporter was decreased by let 7i mimic, but increased by let 7i inhibitor. MiR let 7i inhibitor suppressed DC maturation by depressing CD80 and CD86, however， pretreatment of IL-10 SiRNA attenuated the suppression. Expressions of JAK1, STAT3, and pSTAT3 in mDCs were suppressed by let 7i mimic and pretreatment of IL-10 SiRNA, however up regulated by let 7i inhibitor. Lewis recipients transfused with let 7i inhibitor-mDCs significantly prolonged DA cardiac allograft survival. The allografts transfused with let 7i inhibitor-mDCs showed slight cell infiltration and significantly preserved graft structure. Inhibition of let 7i increased Tregs and modulated cytokine profiles both in vivo and in vitro.

Conclusion
MiR let 7i regulated DCs maturation and function targeting the IL-10 through JAK1-STAT3 pathway. Moreover, transfusion of let 7i inhibitor-mDCs induced prolonged cardiac allograft survival and generated Tregs.
The NK killer receptor Ncr1 shapes tumor architecture to prevent metastasis
Authors: Glasner Ariella , Mandelboim Ofer 
Published: January 2023

Abstract
Natural killer (NK) cells were so-named because of their excellent ability to kill tumors. NKp46 (Ncr1 in mice) is a prominent NK killer receptor, critically involved in controlling various pathogens as well as tumors and tumor metastases. We and others and have demonstrated the ability of the NK killer receptor Ncr1 to contain tumor growth, however, the mechanisms underlying the ability of Ncr1 to control tumor growth and metastases in vivo are yet undetermined
Metastatic melanoma is an extremely aggressive disease and the leading cause of death from skin cancers. Here, we used the spontaneous metastases model B16F10.9 (B16), the Ncr1 knockout mice (Ncr1gfp/gfp), Ifnγ−/− and Tnfα−/− knockout mice to visualize the tumors’ architecture in-vivo, using Reflectance Confocal Microscopy (RCM).
We demonstrate that Ncr1, through Ifnγ prevents tumor metastasis by a novel mechanism, modulating the structural properties of the developing tumors, at the level of the tissue organization.
Using RNAseq we have analyzed the transcriptome of Ifnγ treated B16 cells and reveled a 24 fold increase in the integrin binding ECM protein Fibronictin1 (FN1), the absence of which was reported to correlate with increased metastases.
The finding that Ncr1, through Ifnγ, shapes the tumors’ structures and thus controls metastasis may lead to generation of new drugs aimed at targeting specifically the tumors’ architecture and new treatment options for melanoma patients.