bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lactoferrin and TGF-β1 synergize to enhance differentiation of OVA-specific Foxp3+ iTregs
Authors: Jo Hyeon-Ju , Park Sun-Hee , Jang Young-Saeng , Park Hui-Won , Kim Tae-Gyu , Kim Pyeung-Hyeun 
Published: January 2023

Abstract
We recently found that lactoferrin (LF), like TGF-β1, enhanced Ag non-specific Foxp3+ iTreg differentiation and the two molecules synergized to express Foxp3. In the present study, we investigated effect of LF and TGF-β1 on Ag-specific Foxp3+ iTreg differentiation. Naïve CD4+ T cells isolated from OT-II mice were co-cultured with OVA323–339-loaded T cell-depleted splenocytes as APCs. OVA-specific Foxp3+ Treg population was increased by LF and TGF-β1 alone, and synergistically increased by combination of the two molecules. However, these increases were diminished by additional CD28 stimulation using anti-CD28 Ab. In parallel, blockade of B7 molecules using CTLA4-Ig markedly increased the Foxp3 expression. These results imply that lactoferrin alone or particularly along with TGF-β1 can cause naïve T cells to differentiate into Foxp3+ Tregs, greater under weak co-stimulatory signals.
Temporal, TCR signal strength, and Notch signaling requirements for γδ T-cell functional programming and execution in vivo
Authors: Chen Edward , Lee Christina , Wiest David , Zúñiga-Pflücker Juan Carlos 
Published: January 2023

Abstract
γδ T-cells are critical mediators of both protective and destructive immunity. In this study, we aimed to understand how ontogeny, T-cell receptor (TCR) signal strength, and Notch signaling dictate γδ T-cell functional programming and execution. We employed RBPJ-inducible (RBPJind) mice, which enabled control of the initiation and duration of Notch responsiveness in hematopoietic cells. Our results revealed that generation of γδ T-cell IL-17 producers occurred more readily during fetal life. However, while generation of lymph node IL-17 producers occurred exclusively during fetal life, generation of lung IL-17 producers still occurred postnatally. Using KN6-transgenic (KN6tg) and RBPJind mice, we observed that strong TCR signals programmed γδ T-cells towards the IL-4 fate. Conversely, weak TCR signals programmed γδ T-cells towards the IL-17 fate, and was required for the generation of innate-like IL-17 producers. Notch signaling played a role in promoting the IL-4 fate but, contrary to previous reports, did not play a role in dictating the IL-17 fate. To investigate the role of ontogeny and Notch signaling in the ability of lung γδ T-cells to execute their IL-17 function during infection, we employed an M. tuberculosis model using trehalose dimycolate (TDM). We show that fetal-derived γδ T-cells produced IL-17 in response to TDM while adult-derived γδ T-cells did not. Notch was dispensable for the fetal-derived lung γδ T-cells to produce IL-17, contrary to previous reports suggesting that Notch was required in the periphery for γδ T-cell IL-17 production. Taken together, this study revealed the precise temporal, TCR signal strength, and Notch signaling requirements for γδ T-cell functional programming and execution in vivo.
ACLY regulates CD4 T cell activation and differentiation
Authors: Dahunsi Debolanle , Wolf Melissa , Wellen Kathryn , Rathmell Jeffrey C 
Published: January 2023

Abstract
ATP citrate lyase (ACLY) is a transferase that catalyzes the cytosolic conversion of citrate and coenzyme A (CoA) to acetyl-CoA and oxaloacetate. ACLY has been shown to link glucose/mitochondrial (TCA) metabolism to cytosolic acetylation of proteins, nuclear acetylation of histones and fatty acid synthesis. T cells dramatically increase their metabolic activity upon activation and activated effector T cell subsets utilize distinct metabolic programs. It has been shown that small molecule inhibition of ACLY decreases histone acetylation and effector differentiation in T cells. Here, we examine the effects of T cell-specific genetic knockout of ACLY. ACLY deficient CD4 T cells were found to have lower proliferation and activation marker expression compared to WT. In vitro differentiation of CD4 T cells into Th1 and Th17 subsets was also reduced in the absence of ACLY. Future directions include comparing ACLY KO to small molecule inhibition and examining the metabolic and epigenetic consequences of ACLY deficiency in CD4 T cells in inflammatory disease models.
Apparent sterilizing immunity in BCG-immunized mice challenged with an ultra-low dose of Mycobacterium tuberculosis
Authors: Plumlee Courtney R , Duffy Fergal J , Gern Benjamin H , Delahaye Jared L , Cohen Sara B , Rustad Tige R , Aitchison John D , Sherman David R , Zak Daniel E , Gerner Michael Y , Urdahl Kevin B 
Published: January 2023

Abstract
Tuberculosis (TB) is a highly heterogeneous disease that develops in a subset of individuals infected with aerosolized Mycobacterium tuberculosis (Mtb). To improve upon the standard experimental mouse model, we infected mice with an ultra-low Mtb dose (ULD), consisting of 1-3 founding bacteria, which reflects the physiologic inoculum of humans. These mice exhibited a broad range of outcomes, with bacterial burdens ranging from &lt;10 to ~106 CFUs in individual lungs. Furthermore, they exhibited well-circumscribed granulomas that share features with human granulomas. To monitor outcomes in live mice, we identified a blood RNA signature that correlated with lung bacterial burdens. Remarkably, this mouse-derived signature predicted Mtb infection outcomes across species, including risk of progression to active TB in humans. Given these improvements in the mouse model, we wondered whether ULD infection would provide a better platform for assessing vaccine-induced immunity. We now report that BCG-immunized mice, compared to unimmunized controls, exhibited a lower percentage of infected mice at late timepoints after ULD Mtb challenge. Interestingly, this difference was not observed at earlier timepoints. Taken together, these results suggest that some BCG-immunized mice, upon subsequent Mtb aerosol challenge, are capable of eradicating their initial infection, a finding that has not previously been observed in the TB mouse model. We are currently developing methods to verify initial infection prior to potential clearance and are also investigating immune correlates of protection.
VAMP3 as a negative regulator of Leishmania amazonensis infection and antigen cross-presentation
Authors: Seguin Olivier , Descoteaux Albert 
Published: January 2023

Abstract
To colonize host immune cells, the protozoan parasite Leishmania creates a parasitophorous vacuole (PV) that can be either individual and tight-fitting or communal, large and spacious depending on the parasite species. We aimed to investigate the role of SNAREs in both PV formation and parasite survival. Using bone marrow-derived macrophages (BMM) and confocal microscopy, we found that L. amazonensis communal PV recruits SNAREs and membrane from the plasma membrane (SNAP23), the recycling endosome (VAMP3), the endoplasmic reticulum (STX18) and the trans-golgi (Vti1a) with fusion with the lysosomes (LAMP1) while only the late endosomes (VAMP8) are accumulated to L. major individual PV without fusion with the lysosome. In the absence of VAMP3 we found that replication of L. amazonensis and PV volume roughly doubled. The V0 subunit of the V-ATPase has been proposed as a fusion promoter and as anti-inflammatory. More recently, the V0 subunit was shown to controls Leishmania PV biogenesis via cholesterol homeostasis. We then linked the PV volume augmentation to an augmented V-ATPase V0 subunit TCIRG1. We verified the inflammatory capacity of VAMP3 KO BMMs and found that NO production reduced. IFN-ɣ and LPS stimulated VAMP3 KO BMMs killing capacity was also halved during L. amazonensis infection. Furthermore, cholesterol homeostasis was also shown to play a role in antigen presentation and we found that VAMP3 KO BMDCs had augmented antigen cross-presentation capacities. In conclusion, we report that VAMP3 acts as a negative regulator of L. amazonensis replication by limiting cholesterol availability and through its role in limiting the availability of the V0 subunit of the V-ATPase.
CD4+ T cells modulates efficacy of microbiome transplantation in treatment of Clostridiodes difficile infection
Authors: Abt Michael C , Littmann Eric R , Denny Joshua E , Lee Jung-Jin , Bittenger Kyle 
Published: January 2023

Abstract
Clostridiodes difficile is an opportunistic pathogen that infects the large intestine following perturbation of the intestinal microbiome causing epithelial damage and debilitating, potentially fatal, colitis. Fecal microbiome transplantation (FMT) is a clinically proven therapy to treat recurrent C. difficile infection. Despite remarkable efficacy, implementation of FMT therapy is limited and the mechanism of action remains poorly understood. Here, we demonstrate a critical role for CD4+ T cells in supporting FMT success using a murine C. difficile infection system. Following FMT, chronically infected wild-type mice resolve C. difficile while cohoused, littermate Rag1−/− mice fail to clear the infection. FMT in C. difficile infected mice that lack B cells, CD8+ T cells or CD4+ T cells reveals a necessary role for CD4+ T cells, but not B cells or CD8+ T cells, in resolution of C. difficile following FMT. Analysis of intestinal bacterial communities demonstrate the microbiota of FMT-responsive mice assimilates toward the composition of the FMT donor, while the non-responsive Rag1−/− and CD4+ T cell deficient mice exhibit impaired FMT engraftment. Further, FMT does not restore secondary bile acid pools in the cecum of non-responsive mice, an important metabolite associated with FMT efficacy. Combined, these data support a role for host immunity in determining efficacy of microbiota-based therapeutics.
Vitamin C insufficiency aggravates dextran sulfate sodium (DSS)-induced colitis and colitis-associated colon cancer by regulating IL-6 and IL-22 production in Gulo(−/−) mice
Authors: Kim Yejin , Lee Dahae , Jo Hyejung , Kang Jae Seung 
Published: January 2023

Abstract
Mucosal damage in inflammatory bowel diseases (IBDs) involves the dysfunctional immunoregulation of the gut. Among immunoregulatory factors, oxidative stress is abnormally high level in IBDs, and their destructive effects may contribute to the initiation or propagation of the disease. Vitamin C has anti-oxidant, anti-inflammatory and immunomodulatory effects. So, we investigated the effect of vitamin C on dextran sulfate sodium (DSS)-induced colitis in Gulo(−/−) mice which cannot synthesize vitamin C. Vitamin C-insufficient Gulo(−/−) mice showed the increased oxidative stress and inflammation in the colon. The production of interleukin (IL)-22, the recruitment of NKp46(+) cells, and the activation of p38 MAPK were decreased in the vitamin C-insufficient Gulo(−/−) mice accompanied by decreased mucin-1 expression in compared with vitamin C-sufficient Gulo(−/−) mice and wild-type mice. Moreover, the production of IL-6 was increased, and STAT3 and Akt were activated, which involved the decreased expression of E-cadherin and granyhead like-2 (GRHL2). Consequently, vitamin C-insufficient Gulo(−/−) mice eventually go with severe colitis and cancer development. Therefore, our results suggest that vitamin C has a protective effect on DSS-induced colitis, and has a progressive role in the colon cancer by regulating the production of cytokines and the induction of inflammation.
Formation of mesenteric tertiary lymphoid organs shapes the immune cell and lymph trafficking from the intestine in a Crohn’s disease-like mouse model
Authors: Czepielewski Rafael Sanguinetti , Onufer Emily J , Erlich Emma , Kim Ki-Wook , Saunders Brian , Young Shannon , Kumar Shashi , Zinselmeyer Bernd , Hsieh Chyi-Song , Davis Michael J , Randolph Gwendalyn J 
Published: January 2023

Abstract
Current understanding of Crohn’s disease (CD) is that hereditary factors render the mucosal surface susceptible by either deregulated interactions between the bacterial flora and surface epithelial cells or an overwhelming host immune response. Given that lymph flow is essential for leukocyte trafficking and lipid absorption, understanding the effects of chronic inflammation in the intestinal-draining lymphatics is crucial for IBD treatment. Proper immune responses in the gut require immune cell migration towards secondary lymphoid organs via mesenteric collecting vessels. Our group described newly developed tertiary lymphoid organs (TLO) in mesenteric fat of CD biopsies, that are formed in association with lymphatic collecting vessels. However, mouse models that resemble lymphatic impairment and TLO formation in IBD are missing. Using a unique IBD model of Crohn’s disease-like ileitis (TNFΔARE mice) we demonstrate that these mice develop mesenteric TLO. Under TNFΔARE background, we generated a lymphatic reporter and a transgenic photoconvertible lines. Our results show that mesenteric TLO are integrated with collecting lymphatic vessels, obstructing leukocyte and lymph traffic from the intestine. Using adoptive transfer of microbiota-dependent TCR Tregs we found that TLO have the capacity to differentiate naïve T cells, yet they harbor tissue-resident T cells, demonstrating their duality. Strikingly, although anti-TNF therapy reduce ileum inflammation, TLO are resistant to the treatment, contributing to TNF therapy relapse or inefficacy. Our results demonstrate that chronic intestinal inflammation extend to the mesentery, impairing immune surveillance of the gut mucosal barrier, altering homeostasis.
Epithelial cell intrinsic IL-1R signaling regulates intestinal homeostasis and inflammation
Authors: Overcast Garrett , Eshleman Emily , Alenghat Theresa , Pasare Chandrashekhar 
Published: January 2023

Abstract
Intestinal epithelial cells (IECs) provide a critical first line of defense against microbes through barrier function, production of antimicrobial peptides (AMPs), secretion of cytokines, and recruitment of inflammatory cells. Whether single or multiple upstream cues regulate this diverse set of responses from IECs is not clear. Here we discover an unexpected role for Interleukin-1 Receptor (IL-1R) signaling in IECs in regulating intestinal homeostasis and inflammation. Specifically, absence of IL-1R in epithelial cells abrogates a homeostatic anti-microbial program including production of AMPs. Although IL-1R signaling alone is not sufficient to drive the AMP gene induction in IECs, the IL-22-driven anti-microbial program requires synergistic IL-1R signaling. Mechanistically, IL-1R signaling results in enhancement and temporal stabilization of STAT3 phosphorylation downstream of IL-22R. Thus, we reveal an unexpected cooperation between IL-1R and IL-22R in directly regulating an intestinal anti-microbial program. In contrast, absence of IL-1R in IECs protects mice from chemically-induced acute intestinal inflammation suggesting that IEC specific IL-1R signaling contributes to inflammation and pathology. We find that IL-1R signaling directly induces expression of chemokines as well as genes involved in the production of reactive oxygen species in IECs, a process that likely evolved to protect against enteric pathogens. Our findings establish a critical role for IEC specific IL-1R signaling in both steady state anti-microbial gene induction as well as intestinal inflammation. Further studies are likely to reveal potential targets for developing therapies against Crohn’s Disease and Ulcerative Colitis.
Human T cell immunity in barrier sites is phenotypically distinct but clonally connected to other sites
Authors: Poon Maya ML , Meng Wenzhao , Dogra Pranay , Lam Nora , Szabo Peter A , Kubota Masaru , Matsumoto Rei , Rahman Adeeb , Luning Prak Eline T , Farber Donna L 
Published: January 2023

Abstract
The gut, lung, and skin are physical and immunological barriers that serve as the first line of defense against pathogens and foreign substances. Due to sampling limitations, previous studies have been limited to studying human barrier sites in isolation and often in the context of specific diseases. Our donor tissue resource established in collaboration with LiveOnNY offers a unique opportunity to examine human barrier T cell immunity as a network and elucidate how barrier sites are connected to each other and to lymphoid sites (lymph nodes, bone marrow, spleen). Here, using cytometry by time-of-flight (CyTOF) and T cell receptor (TCR) CDR3b sequencing, we investigate the unique defining characteristics and connectivity of T cells in barrier tissue – gut, lung, and skin – to lymphoid sites and blood. Through high-dimensional marker analysis by CyTOF, our results show that T cell subset composition and phenotype across 8 different sites comprise 33 clusters that are highly conserved between donors of different ages (22–70 yrs). Skin and gut T cells are distinct from each other and from lung, lymphoid sites, and blood, while lung T cells share features with lymphoid sites and blood. TCR repertoire analysis shows that, though majority of gut and skin T cell clones are unique to the tissue, expanded clones within these barrier sites share clonal overlap with other barrier and lymphoid sites. Moreover, T cells in the lung exhibit increased sharing with lymphoid sites and blood. Together, these results reveal how human T cells are compartmentalized in the skin and gut through site-specific adaptations while demonstrating that gut, lung, and skin barrier T cells are connected through shared specificities in a network of immune protection across the body.
Lupus pathogenesis and autoimmunity are exacerbated by high fat diet-induced obesity in mice
Authors: Zhang Xin , Meng Juan , Shi Xuhua , Ali Hiba , Hellmers Linh , Mathew Neeha , Washington Talon , Quinet Robert , Davis William , Zakem Jerald , Keshavamurthy Chandana , You Zongbing 
Published: January 2023

Abstract

Objectives
System lupus erythematosus (SLE)patients have increased prevalence of metabolic syndrome. Our previous data showed that circulating Tfh (Tfh) cells were increased in SLE patients and correlated to their disease activities. Here, we investigated the pathophysiologic link between fat-diet induced obesity and SLE and underlying mechanism using MRL/lpr lupus prone mice.

Methods
Twenty MRL/lpr mice were fed with a regular diet (RD) or high fat diet (HFD, 60% calories comprised of fat). Body weight and skin lesion were recorded weekly. Urine protein using Bradford assay was assessed. Blood was collected for IgG, anti-dsDNA and anti-nuclear antibody (ANA) detection. At the end of experiment of week 14, kidney and skin biopsy were embedded in paraffin for H&E, PAS, and Masson’s staining to detect lupus histopathological lesions and quantified as kidney index based on fibrinoid necrosis, periglomerular infiltrate, and inflammatory dermatitis. Spleen paraffin specimens were stained for Tfh cells markers.

Results
Obesity was achieved with a significant difference of mouse body weight in HFD groups by week 3 (p&lt;0.01). The aggravated SLE pathogenesis in HFD mice was characterized by skin rash, proteinuria, IgG glomerular deposition with increased histopathology index in kidney, and splenomegaly (p&lt;0.05). Interestingly, increased of Tfh cells were found in the spleen of HFD group.

Conclusion
Our results demonstrated that HFD-induced obesity with increase Tfh cells contributes to the acceleration of SLE progression and metabolic abnormalities. Changing eating habit or interventions reducing obesity could improve both SLE symptoms and metabolic inflammation, especially in genetically predisposed individuals.
Therapeutic Benefit of Regulating Iron Metabolism in Spontaneous Lupus Nephritis
Authors: Scindia Yogesh , Wlazlo Ewa , Ghias Elizabeth , Voi Valentina , Cechova Sylvia , Leeds Joseph , Swaminathan Sundararaman 
Published: January 2023

Abstract

Background
Lupus nephritis (LN) is an end-organ complication of Systemic lupus erythematosus and is more common in premenopausal women. Hepcidin, the master regulator of iron homeostasis, modulates inflammation and is negatively regulated by estrogen. Therefore, we hypothesized that exogenous hepcidin may reduce the severity and delay the onset of LN.

Methods
Pre-nephritic 8-week-old or nephritic 16-week-old MRL/lpr female mice were injected Hepcidin (50ug, i.p) or vehicle twice a week and markers of renal injury and inflammation were examined at 18 and 20 weeks of age. The direct effect of hepcidin on macrophages was studied in-vitro.

Results
Hepcidin reduced intrarenal iron accumulation, and increased H-ferritin. This was associated with reduced renal inflammation and immune cell infiltration, which collectively mitigated microalbuminuria and tubular injury, independent of immune complex deposits and autoantibodies. The increase in H-ferritin, was associated with a reduced number of renal Ki-67+-F4/80+ macrophages. In-vitro, hepcidin induced H-ferritin in macrophages and reduced labile (Fe2+) iron. H-ferritinhi macrophages proliferated less upon Mcsf-1 stimulation and secreted less IL-1b, and IL-6 upon TLR-3 activation. Hepcidin reduced microalbuminuria when administered to nephritic, 16-week-old mice without worsening lupus-associated anemia.

Conclusions
We have identified that hepcidin targets iron homeostasis and reduces cardinal pathogenic features of LN. Importantly, Hepcidin treatment ameliorates kidney disease in mice with established proteinuria. Thus, our data highlight that targeting cellular iron metabolism with hepcidin represents a promising and a new therapeutic strategy in LN.
Endogenous IL-33 contributes to Treg expansion induced by IL-2/CD25, a novel IL-2 biologic developed for low-dose IL-2 therapy
Authors: Lui Jen Bon P , Malek Thomas R 
Published: January 2023

Abstract
Low-dose IL-2 therapy is being advanced in patients with autoimmunity by its ability to increase Tregs. A drawback of IL-2, however, is its short-half life and potential off-target immuno-stimulatory effects. To circumvent these limitations, we developed a longer-lasting mouse IL-2/CD25 fusion protein with greater selectivity toward Tregs. IL-2/CD25 is more effective than an equivalent amount of IL-2 in controlling diabetes of NOD mice. Furthermore, compared to other tissues, IL-2/CD25 more readily expanded pancreatic Tregs, suggesting a co-factor that acts with IL-2/CD25. We considered IL-33 as a possible candidate as it also promotes Treg expansion. In this regard, low-dose IL-2/CD25 selectively induced the IL-33R (ST2) on NOD pancreatic Tregs. Correspondingly, ST2 was not substantially induced in Tregs with impaired IL-2R signaling. More importantly, IL-33 supported the expansion of wild-type Tregs, but not Tregs with impaired IL-2R signaling. Thus, Treg expansion by IL-33 depends on IL-2. Reciprocally, Treg expansion in the pancreas by IL-2/CD25 required IL-33 because this expansion was suboptimal in IL-33-deficient B6 mice but was maximal when IL-33 was sequentially administered with IL-2/CD25. However, IL-2/CD25 and IL-33 did not act synergistically for Treg expansion in the pancreas of NOD mice, as IL-33 or IL-2/CD25 were similarly effective individually. This result may reflect that other inflammatory mediators or co-factors associated with the NOD pancreas also influence the Treg response to IL-2/CD25 and IL-33. Overall, these data indicate that tissue-derived IL-33 may contribute to the overall outcome of low-dose IL-2 therapy by supporting Treg expansion during therapy of autoimmune diseases.