bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inducible epithelial resistance protects against acute viral infection and subsequent CD8+Tcell dependent lethal Immunopathology
Authors: Wali Shradha , Flores Jose R , Goldblatt David , Tuvim Michael , Dickey Burton F , Evans Scott E 
Published: January 2023

Abstract
Viral pneumonia leads to significant morbidity and mortality worldwide, demanding better understanding of the host immune response to infections for development of novel strategies to prevent respiratory infections. Our group has shown that mice treated with combination of Toll-like receptor (TLR) 2/6 and 9 agonists (Pam2-ODN) are protected against wide range of respiratory pathogens, including viruses. Mice challenged intra-pharyngeally with Sendai virus showed peak viral burden on day 5 and peak mortality on day 11, overlapping with increased lung CD8+Tcells. We hypothesized that mortality is associated with immunopathology caused by CD8+T cells. Pam2-ODN aerosolization one day prior to Sendai challenge reduced lung viral burden and reduced lung CD8+T cells on day 11 and enhanced mouse survival. Depletion of CD8+T cells before infection increased baseline virus susceptibility and enhanced viral replication in vivo, congruent with the known antiviral function of CD8+T cells. Pam2-ODN pre-treatment protected mice against death following viral challenge, even in the absence of CD8+T cells, reflecting antiviral responses induced directly from epithelial cells. Notably, depletion of CD8+T cells eight days after viral challenge also significantly enhanced survival of sham pre-treated mice, indicating rescue from CD8+T cell-mediated lethal immunopathology. Our findings definitively prove that CD8+T cells, although anti-viral in nature, promote lethal immunopathology that can be prevented by Pam2-ODN pre-treatment. Importantly, Pam2-ODN pre-treatment benefitted survival regardless of when CD8+T cells were depleted and may provide an opportunity to protect vulnerable populations against respiratory infections.
Human IFNe and IFNa modulate immune and anti-HIV functions through the differential involvement of IFNa receptors
Authors: Pizutelli Vanessa , Tasker Carley , Sue Lishan , Lu Wuyuan , Chang Theresa 
Published: January 2023

Abstract
Interferon e (IFNe) is a critical innate immune mediator that protects the host against sexually transmitted infections. Although the current paradigm is that IFNɛ is a type I IFN and signals through IFNa receptors (IFNARs), our and other published results show that IFNɛ has unique gene regulatory and immune functions that are distinct from IFNa. IFNɛ is constitutively expressed in the mucosa, and highly abundant in the female reproductive tract. It exhibits superior mucosal immune activity compared to IFNa/b. We have previously shown that that human IFNe induces an anti-HIV state through a mechanism independent of known type I IFN-induced HIV host restriction factors in primary macrophages. IFNɛ induces phagocytosis and reactive oxygen species that contribute to anti-HIV activity. In this study, we determined the involvement of IFNARs in immune modulation and HIV inhibition by human IFNɛ in comparison to human IFNa.
Human IFNɛ elicited a more robust immune response than human IFNa2 in PBMCs and primary macrophages. The immune profiles in response to human IFNɛ were cell-type dependent. Antibodies (Abs) against IFNAR1 or IFNAR2 blocked induction of interferon-stimulated genes (ISGs) by human IFNa2 and murine IFNe. However, anti-IFNAR2 but not anti-IFNAR1 Ab blocked human IFNe-mediated ISG induction. Interestingly, anti-IFNAR1 or anti-IFNAR2 Ab blocked cytokine induction by human IFNa2 but not by human IFNe. Finally, we found that both IFNAR1 and IFNAR2 were involved in anti-HIV activity of human IFNa2 but only IFNAR2 was involved in HIV inhibition by human IFNe. In summary, our results indicate that human IFNe, not a conventional type I IFN, modulates immune and anti-HIV functions through distinct mechanisms from IFNa2.
Novel Vaccine Adjuvants: Exploring the Potential of Rexinoids to Influence Effector T Cell Homing to Mucosal Sites
Authors: Manhas Kavita , Blattman Joseph N , Marshall Pamela A , Wagner Carl E , Jurutka Peter W 
Published: January 2023

Abstract
Vaccines are the most successful defense against infectious diseases. Effective vaccines must target induced immune responses to areas of pathogen entry, a primary site being mucosal surfaces. Certain vitamins can directly impact immune cell migration, indicating potential to function as an adjuvant. We previously showed that all-trans retinoic acid (ATRA), the biologically active form of vitamin A, modifies gut-homing receptor expression on effector T cells upon binding to the retinoic acid receptor (RAR), which joins with the retinoid X receptor (RXR) to function. Giving ATRA during vaccination also resulted in enhanced mucosal protection following viral challenge in murine models. While promising, certain factors limit adding ATRA to a vaccine, including its instability and poor solubility. Rexinoids, synthetic ligands for the RXR, could mimic or better ATRA immune effects by enhancing RAR-mediated transcription. Their improved stability and solubility also allows a greater potential for clinical use. Here we explored the ability of different rexinoids to program effector T cell expression of gut-homing receptors CCR9, α4β7, and CD103. Naïve T cells from transgenic mice were activated with peptide and set concentration of each rexinoid. Flow cytometry showed increased expression of gut-homing receptors in the presence of select rexinoids, indicating their ability to act as an ATRA mimic. A cooperative relationship was also seen between other rexinoids and ATRA as co-culturing with suboptimal doses of each showed enhanced receptor expression, suggesting these rexinoids could potentially act with ATRA normally found in serum. Overall, our findings support the prospective use of rexinoids as adjuvants during vaccination.
In vitro model for assessing the effect of T helper cell cytokines on the barrier function of alveolar type II epithelial cells
Authors: Coello Daniel , Knott Jeffery D. , Porter Edith 
Published: January 2023

Abstract
There are still diseases of global burden for which satisfactory vaccines are unavailable, e.g. tuberculosis. Vaccines against intracellular microbes has relied on the activation of the adaptive T cell response. Our laboratory is interested in investigating whether the innate epithelial host defense could be integrated in a novel approach. In response to airborne pathogens, airway epithelia release antimicrobial peptides like human beta defensin-2 and -3 (HBD2/3), antimicrobial lipids, and chemokines, like IL-8, to attract other immune cells including T helper cells that are integral to the adaptive immune response. We hypothesize that T cell derived cytokines augment the antimicrobial function of airway epithelia and we have developed an in vitro model to generate pilot data. A549 cells, alveolar type II epithelial cells, were grown in air-liquid interface and stimulated with TNF-α & IFN-γ and IL-17 & IL-22, at 5 ng/mL each, for 24 h. IL-8 and HBD2/3 gene expression was quantified by RT-PCR and secretion of antimicrobial peptides was assessed by AU-PAGE and Western blot probing for HBD2/3. Lipid production was assessed with Bodipy and the physical barrier integrity with Phalloidin. IL-8 gene expression was 9-fold elevated when cells were stimulated with TNF-α & IFN-γ. Secretions from stimulated cells appeared to contain more cationic peptides that were not reactive with HBD2/3 polyclonal antibodies. Cellular Bodipy fluorescence was pronounced after IL-17 & IL22 stimulation. Phalloidin staining was consistent with tight junction formation in all samples. This data provides evidence that T helper cells may directly augment epithelial barrier function supporting a novel vaccine design that integrates the epithelial cell response.
EXOSOMES FROM PRIMARY BRONCHIAL EPITHELIAL CELLS INDUCE TOLERANCE IN HUMAN MONOCYTE-DERIVED DENDRITIC CELLS
Authors: Agrawal Anshu , Monterio Clarice , Agrawal Sudhanshu 
Published: January 2023

Abstract
Dendritic cells (DCs) act as sentinels by monitoring the tissue environment for microorganisms and other substances. DCs are also critical modulators of tolerance in the periphery as well as in the mucosa including the respiratory mucosa. DCs exist in close proximity to airway epithelial cells (AECs) and both cells influence the functions of each other. Using a 2D model to study the interaction of human primary bronchial epithelial cells (representative of AECs) with DCs we have recently demonstrated that AECs enhance the immune surveillance capacity of myeloid DCs (mDCs) at homeostasis. Culture of mDCs with AECs resulted in the upregulation of pathogen recognition receptors (PRRs) and downstream signaling molecules which was reflected in the enhanced inflammatory cytokine responses to LPS. In contrast to mDCs, culture of monocyte-derived DCs (MoDCs) with AECs led to a significant inhibition of LPS induced pro-inflammatory responses. Thus it seems AECs induce tolerance in MoDCs but enhance the immunogenicity of mDCs suggesting a division of labor between various DC subsets in the respiratory mucosa. Recent studies have indicated a role of exosomes, the endosomal-derived nano-vesicles in influencing the function of a variety of cells. Therefore we investigated whether exosomes play a role in inducing tolerance in MoDCs. MoDCs were cultured with various concentrations of exosomes before activation with LPS. Results indicate that exosomes from AECs conditioned media increased the secretion of IL-10 from MoDCs compared to control media exosome. The expression of CD80/86 was also decreased. In summary these data indicate that exosomes from AECs may be inducing tolerance in MoDCs in the respiratory mucosa.
Genetic and pharmacological modulation of RORα regulates TH17-driven inflammatory disorders
Authors: Solt Laura A , Wang Ran , Amir Mohammed , Chaudhari Sweena , Campbell Sean , Bassette Molly B , Eliason Amber , Sundrud Mark S , Kamenecka Theodore 
Published: January 2023

Abstract
While RORγt has been well characterized as the lineage defining transcription factor for TH17 cell development, TH17 cells are not absent in Rorc-deficient mice, suggesting other factors may be required. RORα, a close family member of RORγt, is also expressed during TH17 cell development but is considered functionally redundant, thus little is known about its function in TH17 cells. Using mouse models of autoimmunity and chronic inflammation, we show that expression of RORα is required for TH17 pathogenicity. T-cell specific deletion of RORα significantly abrogated the development of experimental autoimmune encephalomyelitis (EAE) and colitis, due to decreased development of TH17 cells and expression of homing receptors required for cells to gain access to sites of inflammation. These results were accompanied by increased Foxp3+T regulatory cells. Using a RORα-selective small molecule that we developed, we found that modulation of RORα activity largely phenocopied our genetic data, inhibiting the development of EAE, colitis, and relapse in a relapsing remitting model of multiple sclerosis. Importantly, treatment with a RORα modulator did not affect thymic cellularity. Finally, modulation of RORα activity inhibited the development of human TH17 cells and pro-inflammatory cytokine expression from subsets of CD4+CCR6+memory T cells. Our results establish that RORα has non-redundant functions to RORγt driving TH17 cell development and identifies RORα as a potential therapeutic target for the treatment of TH17-mediated autoimmunity.
Anti-CD4/CD8 antibody therapy induces Foxo1 transcriptional activity promoting T cell egress from inflamed tissue during type 1 diabetes
Authors: Clark Matthew P , Kroger Charles , Martin Aaron , Tisch Roland 
Published: January 2023

Abstract
Type 1 diabetes (T1D) is considered to be a T cell driven autoimmune disease, for which clinical treatment of T1D has been unsuccessful creating a need for immunotherapeutics to match world-wide rising incidence of disease. Pancreatic inflammation in non-obese diabetic (NOD) mice is primarily T cell mediated resulting in immune destruction of the insulin producing β cells in the pancreatic islets of Langerhans, which induces a hyperglycemic state that requires daily insulin injections to maintain homeostasis. The use of nondepleting antibodies provides a strategy to modulate the function of T cells while avoiding their systemic depletion. Our lab has previously reported that treatment of recent onset diabetic NOD mice with a short-course of both monoclonal anti-CD4 and anti-CD8 (anti-CD4/CD8) antibodies rapidly and indefinitely reversed established T1D. Remission of T1D by anti-CD4/CD8 treatment is accompanied by the egress of T cells from the pancreatic islets and pancreatic draining lymph node. Microarray analysis of anti-CD4/CD8 treated T cells revealed increased activity of the Forkhead box protein O1 (Foxo1) transcriptional axis that controls expression of the blood homing chemokine receptor sphingosine-1-phosphate receptor 1 (S1Pr1). In contrast, T cell activation related genes that promote tissue residency of T cells were down regulated. T cell receptor (TCR) signaling negatively regulates Foxo1 transcriptional activity by phosphorylation via the AKT signaling pathway. Our studies have revealed that dampened TCR signaling shortly after anti-CD4/CD8 treatment results in enhanced Foxo1 transcriptional activity promoting S1Pr1-mediated T cell egress from inflamed tissues providing long-term protection from T1D.
ST8Sia6 generated α2,8-disialic acids mitigate hyperglycemia in multiple low dose streptozocin-induced diabetes
Authors: Belmonte Paul , Shapiro Michael , McCue Shaylene , Rajcula Matthew , Shapiro Virginia Smith 
Published: January 2023

Abstract
Type 1 diabetes is an autoimmune disease that destroys insulin-producing beta cells in the pancreatic islets. Although exogenous insulin replacement therapy is lifesaving, precise blood glucose management remains a significant challenge. Experimental islet transplantation restores normoglycemia, but the lack of available islets and graft rejection present major barriers to widespread adoption as standard therapy. The islet resident macrophage (IRM) is in close contact with beta cells, efficiently presents islet antigen to autoreactive T cells, and is critical to disease occurrence. We have demonstrated that murine IRMs express high levels of Siglec-E, an immune-regulatory receptor that preferentially binds α2,8-disialic acids in cis or in trans. When islets were exposed to streptozocin (STZ)-induced stress in culture, beta cells upregulated ligands for Siglec-E. To determine the biological significance of this, we generated a mouse model where ST8Sia6, the sialyltransferase that produces α2,8-disialic acids and thus Siglec-E ligands, is constitutively overexpressed in beta cells. This increased trans Siglec-E binding affinity to the beta cell surface, and mitigated hyperglycemia in the multiple low-dose STZ model of diabetes. Significant reduction in blood sugar was evident at 14 days post-initial injection of STZ, coinciding with T cell infiltration of islets. T cells from ST8Sia6+ islets were less activated when compared to littermates. This work reveals a novel and important role for Siglec-E in autoimmune diabetes and provides insight into how rational targeting of this immune receptor may instruct islet-residing antigen presenting cells to provide local tolerance signals in the setting of islet transplantation.
Kidney Allograft Recipient Absence of Myeloperoxidase Decreases Donor-Specific Antibody Titers and Attenuates Antibody-Mediated Allograft Rejection
Authors: Fairchild Robert L , Miyairi Satoshi , Dvorina Nina , Valujskikh Anna , Baldwin William M 
Published: January 2023

Abstract
The incidence of antibody-mediated rejection (AMR) in clinical kidney transplants is increasingly observed. We have reported that strong donor-specific antibody (DSA) responses are induced in B6.CCR5−/−mice transplanted with complete MHC mismatched A/J kidney allografts and that NK cells play a critical role in acute injury and graft failure. Since this acute AMR is accompanied by intense macrophage infiltration into the allograft, we tested the role of recipient-derived myeloperoxidase (MPO) production on kidney allograft survival. B6.CCR5−/−and B6.CCR5−/−MPO−/−mice were transplanted with complete MHC mismatched A/J kidney grafts. Allografts were rejected between days 18 and 25 post-transplant in B6.CCR5−/−recipients but not until days 46–54 in B6.CCR5−/−MPO−/− recipients. DSA titers in B6.CCR5−/−MPO−/−recipients were 4–5 fold lower than those induced in the B6.CCR5−/− recipients. There was also a 60% decrease in the number of donor-reactive T cells producing IFN-g in the spleens of the B6.CCR5−/−MPO−/− vs. B6.CCR5−/−recipients on day 7 post-transplant. Despite the extended survival, qPCR analyses indicated slight increases in mRNA encoding TNFa, IL-6 and FasL in kidney allografts on day 14 post-transplant in B6.CCR5−/−MPO−/− vs. B6.CCR5−/−recipients as well as the pro-fibrogenic factors P-selectin and connective tissue growth factor on day 50 post-transplant. These results suggest MPO regulates the magnitude of the DSA and T cell response in kidney transplant recipients and that the decreased DSA titers attenuate acute AMR but induce the indolent development of interstitial fibrosis and glomerular injury that will eventually lead to graft dysfunction and failure at later times.
Human Immune Cells Directly Recognize Xenogeneic MHCs
Authors: Nouri-Shirazi Mahyar A , Katz Haley , Victor Lisa 
Published: January 2023

Abstract
The advancement in tissue typing and the use of immunosuppressive drugs has overcome major complications in organ transplantation, therefore, making this therapy a viable option for irreversible organ diseases. However, organ shortage continues to be the forefront of problems facing clinical transplantation. To compensate for the disparity between organ supply and demand, xenografts have become a promising alternative. Nevertheless, the success of xenotransplants needs further investigation into the contribution of cell-mediated rejection. Here, we explored the direct and indirect contribution of human immune cells in xenorecognition using human and murine in vitro coculture systems. Our data shows that human T cells directly recognized the xenogeneic MHC molecules since blocking of MHCs suppressed their proliferative response and cytokines production, IL-2 and IFN-γ. Furthermore, while B and NK cells alone did not generate significant response, the combination of B and T cells promoted indirect xenorecognition by T cells as evidenced by an increase in their proliferative response. In contrast, the murine immune cells showed limited response to human MHCs. Our study suggests that human T cells have a plasticity to recognize xenogeneic MHCs and contribute to xenograft rejection.
High Stearic Acid diet aggravates aGVHD by promoting Th17 cells through Akkermansia muciniphila
Authors: Ma Shoubao , Yang Bingyu , Gong Huanle , Wu Xiaojin , Wu Depei 
Published: January 2023

Abstract
T-cell metabolism is a promising target for controlling aGVHD. By using GC-MS technique, we previously identified that serum stearic acid/palmitic acid ratio on day 7 was an excellent biomarker to predict aGVHD after allo-HSCT, suggesting an important role of saturated fatty acid intake in the pathogenesis of aGVHD. However, the mechanism remains unknown. By using mouse aGVHD model, here we found that high stearic acid diet (HSAD), but not palmitic acid diet, significantly aggravated aGVHD, and its promoting role was mediated by gut microbiota. 16S rRNA sequencing identified Akkermansia muciniphila (Akk) was obviously increased in HSAD recipient. Add back Akk in WT mice ameliorated aGVHD-related mortality. Furthermore, depleting CD4 T cells significantly rescued HSAD induced aGVHD mortality, whereas depleting CD8 T cells could not. In addition, HSAD recipients of RORgt−/− allografts had improved survival compared with recipients of WT allografts, indicating that Th17 cells were involved in HSAD induced aGVHD. Meanwhile, we found that Th17 cells were significantly increased in aGVHD target tissues in HSAD recipients. Functional studies showed that stearic acid could promote Akk growth in vitro, and extracts from Akk significantly promoted Th17 cells differentiation in vitro, suggesting that Akk increased the proportion of Th17 cells, thereby exacerbating aGVHD.
Inhibition of DNA-PK(cs) Prevents Graft Rejection in a Murine Allograft Skin Graft Study
Authors: Burdine Marie , Harrison David , Waldrip Zachary , Burdine Lyle 
Published: January 2023

Abstract
While the number of patients needing solid organ transplants has steadily increased over the years, the development of safer, more effective transplant drugs has stagnated. Current therapy has been largely successful at thwarting acute cellular rejection; however, antibody-mediated rejection (AMR) remains difficult to treat in the clinic making it the primary cause of graft loss. In this study, we provide proof of principle experiments that DNA-dependent protein kinase catalytic subunit DNA-PK(cs) is a worthwhile target for transplant immunosuppression. In our previous studies, we confirmed that DNA-PK(cs) controls NFAT-induced IL-2 expression, in part, through a mechanism that promotes calcineurin activity by preventing expression of the endogenous calcineurin inhibitor Cabin1. Here, we show that treatment with DNA-PK(cs) inhibitors prevents rejection in a murine skin allograft model by decreasing the production of cytokines including IL2 and IL6 and the infiltration of lymphocytes into the graft. Most importantly, our studies showed that DNA-PK(cs) inhibitors prevented the development of donor-specific antibodies, a driving force behind AMR. These results highlight novel functions for DNA-PK(cs) in the immune system and the potential use of DNA-PK(cs) inhibitors to prevent organ rejection there-bye warranting further investigation.
IL-1β And IL-6 Facilitate T Cell Reconstitution In Heart Allograft Recipients Treated With Anti-Thymocyte Globulin
Authors: Hasgur Suheyla , Fan Ran , Zwick Danny , Valujskikh Anna 
Published: January 2023

Abstract
Antibody-mediated lymphocyte depletion is used in solid organ transplantation. T cell reconstitution after lymphoablation may lead to acute rejection and poor graft outcome. We investigated the mechanisms of T cell reconstitution in heart allograft recipients treated with murine anti-thymocyte globulin (mATG).
B cell depletion with anti-mouse CD20 mAb significantly diminished T cell reconstitution in B6 (H-2b) recipients of BALB/c (H-2d) heart allografts treated with mATG. However, cognate T/B cell interactions were dispensable for T cell recovery suggesting the potential role of B cell derived cytokines. The NanoString gene expression analysis of isolated B cells showed that mATG upregulated IL-1β and IL-6 expression in CD4 T cell-dependent manner. Intracellular flow cytometry confirmed that B cells are the major source of IL-1β and IL-6 following mATG treatment. The proportion of IL-1β secreting B cells was increased in mATG treated compared to untreated recipients, whereas IL-6 producing B cells were not significantly affected by lymphoablation. Neutralization of either IL-1β or IL-6 with mAb significantly decreased T cell recovery and extended heart allograft survival in mATG treated recipients. To further test the role of B cell derived IL-1β, we injected either WT or Caspase-1−/− B cells into B cell deficient μMT mice followed by BALB/C heart transplantation and mATG treatment. Caspase-1 deficiency in injected B cells resulted in significantly lower CD8 T cell recovery.
Our results show the novel functions of IL-1β and IL-6 in regulating homeostatic T cell recovery following mATG treatment. IL-1β and IL-6 neutralization may be a promising strategy for enhancing the efficacy of mATG lymphoablation in transplant recipients.