bluuebunny/crossref_metadata_2025_split huggingface dataset:
Mutation of Tgfbr1 leads to non-hematopoietic defects that drive early onset eosinophilic inflammation in the esophagus
Authors: Laky Karen , Kinard Jessica L , Guerrerio Anthony , Li Min Jenny , Guerrerio Pamela 
Published: January 2023

Abstract
Eosinophilic esophagitis is a chronic disease characterized by tissue-restricted eosinophilia, epithelial basal cell hyperplasia, and esophageal dysfunction. There are three endotypes of eosinophilic esophagitis which are classified based upon the age of disease onset, degree of fibrostenosis, and gene expression patterns. Genetic studies have identified variants that correlate with an increased likelihood to develop eosinophilic esophagitis. Among the top ten variants, nearly all occur in genes expressed by epithelial cells, and approximately one-third encode proteins involved in the transforming growth factor beta (TGFβ) signaling pathway. The pathophysiologic mechanisms underlying these genetic risk factors are poorly understood. Using mice engineered to have an autosomal dominant loss-of-function mutation in the kinase domain of Tgfbr1 we demonstrate that altered TGFβ signaling in non-hematopoietic cells is necessary and sufficient to cause disease that clinically, immunologically, histologically, and transcriptionally recapitulates human eosinophilic esophagitis endotype 2. Tgfbr1 mutation impairs esophageal epithelial cell homeostasis resulting in tissue-restricted increases in proteins that act directly, indirectly, and synergistically to drive accumulation and activation of eosinophils, mast cells, and type 2 innate lymphoid cells (ILC2). These data suggest new ways to approach the prevention and treatment of eosinophilic esophagitis.
Comparison of the suppressive function of alloantigen-reactive Treg cells and non-stimulated Treg cells in allogeneic skin transplantation
Authors: Kim Yong-Hee , Shevach Ethan M. 
Published: January 2023

Abstract
Adoptive cellular therapy with Treg cells is being evaluated in numerous clinical trials in GvHD and organ transplantation. Cellular biotherapy with Treg cells requires ex vivo expansion. While alloantigen stimulated Treg cells have been used in several studies, very few studies have directly compared the efficacy of alloantigen-reactive with non-stimulated Treg cells in vitro and no studies have compared their efficacy in vivo in organ transplantation. In this study, we obtained enriched population of alloantigen-reactive Treg by stimulating proliferation dye-labeled C57BL/6 CD4+Foxp3+CD25+ Treg cells with allogeneic H-2bm12 dendritic cells in the presence of IL-2. After 5~7 days, the alloantigen-reactive Treg cells and resting Treg cells were separated by FACS. Treg cells that had proliferated in response to alloantigen stimulation were more potent suppressors of the reactivity of C57BL/6 CD4+Foxp3− T cells to H-2bm12 stimulators than non-reactive Treg cells. In contrast, both Treg populations equally suppressed the response of C57BL/6 CD4+Foxp3− T cells to 3rd party BALB/c dendritic cells. Both alloantigen-reactive and non-reactive Treg cells were co-transferred with C57BL/6 CD4+Foxp3− T cells (1:2 ratio) to immunodeficient recipients followed by transplantation of allogeneic H-2bm12 and 3rd party BALB/c skin. Recipients of alloantigen-reactive Treg cells demonstrated delayed rejection (26 days -&gt; 49 days) of H-2bm12 skin, but normal rejection of 3rd party skin. Taken together, this protocol offers a simple, rapid method for enrichment of alloantigen-reactive Treg with enhanced antigen-specific suppressive function in vitro and in vivo.
This research was supported by the Intramural Research Program of NIAID, NIH.
p40 Homodimers Indirectly Induce Endogenous Donor-Reactive Memory CD8 T Cell Proliferation within High-Risk Allografts via IL-15 signaling
Authors: Fairchild Robert L , Valujskikh Anna , Tsuda Hidetoshi 
Published: January 2023

Abstract
Increasing cold ischemic storage (CIS) time from 0.5 to 8 hrs prior to transplant promotes the increased proliferation and effector functions of endogenous donor-reactive memory CD4 and CD8 T cells infiltrating mouse cardiac allografts 24–48 hrs after transplant. This increased mCD8 T cell proliferation within high-risk allografts requires mCD4 T cell help via CD40-CD154 interactions with graft dendritic cells to induce IL-12p40 homodimer (p40 HD) production required for this mCD8 T cell proliferation. This study tested if p40HD directly or indirectly stimulate the mCD8 T cell proliferation within high risk allografts. While endogenous mCD8 T cells within highly ischemic allografts expressed both IL12Rβ1 and β2, most proliferating CD8 T cells did not express the IL12Rβ1 needed for p40 HD mediated signaling. Moreover, endogenous mCD8 T cells isolated from allografts did not proliferate ex vivo when cultured with p40 HD. qPCR analysis indicated increased mRNA expression of IL2Rα (CD25), β(CD122) and IL15 Rain infiltrating mCD8 T cells purified from allografts subjected to prolonged vs. minimal CIS on day 2 post-transplant. ELISA indicated longer CIS time markedly increased IL-12p40 and IL-15 protein in the allografts and these increases were dependent on recipient CD4 T cells. Blocking of p40 or IL-15 signaling with anti-p40 or anti-CD122 mAb inhibited endogenous mCD8 T cell proliferation within high-risk cardiac allografts and extended the survival of the high-risk allografts from day 18 to day 54 in CTLA-4Ig conditioned recipients. These results suggest that the p40 HD stimulate proliferative cytokines such as IL-15, which directly stimulate endogenous donor-reactive mCD8 T cell proliferation within the higher-risk allografts.
Exopolysaccharide from a commensal bacterium improves outcome of graft versus host disease by decreasing donor T cell activation
Authors: Kalinina Olga , Talley Sarah , Paik Wonbeom , Campbell Edward M , Knight Katherine L 
Published: January 2023

Abstract
We previously showed that exopolysaccharide (EPS) produced by a commensal bacterium, Bacillus subtilis, induces alternatively-activated, anti-inflammatory M2 macrophages that protect mice from T cell-dependent inflammatory disease, induced by the enteric pathogen Citrobacter rodentium. Here, we show that EPS induces clinical improvement in another T cell-mediated disease, graft versus host disease (GVHD). GVHD is a common complication of allogenic bone marrow transplantation and is caused by the activation of alloreactive donor T cells. To monitor the inflammatory response of allogeneic bone marrow cells in recipient mice, we used donor cells from transgenic mice that express a luciferase-based biosensor that is cleaved by caspase-1, a protein activated during inflammation. We found that donor inflammatory responses and T cell activation was decreased in EPS-treated mice. Our previous data indicate that EPS does not have a direct effect on T cells. To identify the cell type affected by EPS, we tested the activity of EPS on bone marrow-derived dendritic cells (BMDCs) and found that these cells have upregulated expression of the inhibitory molecule, PD-L1, and the cells inhibited alloreactive T cell activation and proliferation in mixed lymphocyte reactions (MLR). We also found that in vivo injection of EPS resulted in the upregulation of PD-L1 expression on splenic dendritic cells. We conclude that EPS ameliorates clinical manifestations of GVHD by decreasing the activation of donor T cells, and we suggest that EPS affects T cells by inducing inhibitory dendritic cells.
Human monocytes educated with exosomes from TLR4 primed mesenchymal stem cells treat acute radiation syndrome by promoting hematopoietic recovery
Authors: Forsberg Matthew H , Kink John , Hematti Peiman , Capitini Christian M. 
Published: January 2023

Abstract
Total body irradiation is often used as a conditioning regimen for bone marrow transplants but can cause life threatening damage to host tissues especially the bone marrow. Developing a cellular therapy that can protect the bone marrow from acute radiation syndrome and stimulate hematopoiesis is a priority for patients exposed to therapeutic or even accidental radiation injury. In this study, exosomes derived from MSCs stimulated with the TLR4 agonist lipopolysaccharide (LPS) were used to alternatively activate human monocytes, termed LPS EEMos, as a potential novel radioprotective cellular therapy. LPS EEMos expressed higher levels of PD-L1 (p&lt;0.0001), and lower levels of CD16 (p&lt;0.01), CD86 (p&lt;0.01), and CD206 (p&lt;0.0001) by flow cytometry compared to monocytes educated with exosomes from unstimulated MSCs (EEMos). Using qPCR, increased gene expression in LPS EEMos of IL-10 (p&lt;0.05), IDO (p&lt;0.001), FGF2 (p&lt;0.05), IL-15 (p&lt;0.05), and IL-6 (p&lt;0.0001) were detected compared to EEMos. Using a xenogeneic radiation injury model, infusion of human LPS EEMos 4 hours after lethal radiation led to reduced clinical scores and an increased survival at 40 days post-infusion, as compared to infusions of PBS, EEMos, and monocytes alone, all of which led to worse clinical scores and 0% survival with uniform death by 20 days (p&lt;0.05). Complete blood cell counts in LPS EEMo recipients showed leukocyte, erythrocyte and platelet counts equivalent to non-irradiated mice, demonstrating complete restoration of hematopoiesis. Infusion of LPS EEMos may be a useful strategy to protect the bone marrow from acute radiation syndrome by expression of anti-inflammatory molecules and cytokines that promote hematopoiesis/engraftment.
Extracellular mitochondria Released from Hepatic Ischemia Reperfusion Induce Macrophage activation both in vitro and in vivo
Authors: Hu Qianni , Haidl Ian D. , Marshall Jean S , Alwayn Ian P.J. 
Published: January 2023

Abstract

Background
During liver transplantation ischemia and reperfusion injury (IRI), an immune driven inflammatory response provoked by cellular oxygen deprivation, is unavoidable. The inflammatory responses, resulting from acute oxidative stress and consequent hepatocellular death during the early reperfusion phase, cause the release of endogenous danger signals. Recent studies show that the release of free mitochondria (FM) from deceased organ donors is directly linked with functionally important immune activation and early allograft dysfunction (Pollara, et al., 2018). Organs from donors after cardiocirculatory death (DCD) undergo a prolonged warm ischemia process compared with organs from donors after neurological determination of death (NDD), and therefore have more severe IRI. Herein, we investigated the roles of FM in liver IRI, particularly as related to macrophages.

Methods
Liver biopsies were assayed by TEM to assess for FM release. FM were co-cultured with macrophages (cell line, bone marrow derived macrophages, and primary Kupffer cells), and injected i.p. in vivo to explore the recruitment and activation of macrophages. Inflammatory mediators were tested by luminex.

Results
FM levels were significantly higher in IRI. Co-culture of FM with macrophages significantly increased inflammatory mediator release but this process was not Formyl Peptide Receptor 1 dependent. The injection of FM induced significant accumulation of monocyte-derived macrophages and activation of macrophages in the peritoneum.

Conclusion
Our results indicate that IRI can elicit significantly increased FM, and that FM from hepatic IRI can attract and activate macrophages, which may influence the outcome of liver transplantation.
Two ICD Inducers are better than one for induction of anti-tumor immunity in cold carcinomas
Authors: Jessup J. Milburn , Kabbout Mohamed , Henderson Paul T. , Hewitt Stephen , Mattoo Abid R. 
Published: January 2023

Abstract

Background
Cytotoxic agents and viruses cause Immunogenic Cell Death (ICD), which initiates a primary immune response in tumor-bearing hosts by releasing cellular antigens with immune chaperones. We postulate that two ICD inducers are better than one in immune competent mouse models of colorectal and breast cancer.

Methods
Approximately 6 mm diameter CT26 rectal or 4T1 breast carcinomas were established in groups of 5–7 BALB/c mice and injected once intratumorally (IT) with oxaliplatin (OX) mixed with a human adenovirus AdV optimized for calreticulin externalization in 3-D spheroids of human carcinomas. Tumor volumes were measured every other day and rate of growth analyzed by Rate-Based T/C Ratio at 19–21 days. Tumor-Infiltrating Lymphocytes (TIL) and nodal cytotoxicity were assessed 11 days after treatment.

Results
In 3 experiments with OX-AdV IT in PBS buffer the T/C ratios were 0.17–0.29 those of the volume of PBS IT injected mice. The T/C Ratio for OX-AdV IT in D5W (used to stabilize the OX) was 0.10 of the IT control volume with 0.32–0.40 for OX or AdV alone and with all groups different by P&lt;0.05 by 2-way ANOVA. OX-AdV IT increased TIL by 4-fold and decreased CD8+PD-1+EOMES+ from 79% to 19% with a dramatic influx of CD8+PD-1-Eomes- T cells compared to PBS IT controls. Draining node lymphocytes were cytotoxic to CT26 cells from OX-AdV IT mice while lymphocytes from PBS IT mice were not. OX-AdV IT in D5W caused 40% of mice to be complete responders that later rejected a contralateral challenge of CT26.

Conclusions
The buffer for IT treatment with OX is critical. The two ICD inducer formulation with D5W is promising as a checkpoint therapy sensitizer for carcinomas that lack TIL or resist checkpoint inhibitor therapy like colorectal carcinoma.
Optimization of the T2 HLA-A2 shift assay for testing of the biological activity of immunotherapies
Authors: Tram Cynthia , Hrytsenko Olga , Stanford Marianne 
Published: January 2023

Abstract
T2 is a lymphoma-derived cell line that express low amounts of HLA-A2 on the cell surface due to TAP deficiency and can only present exogenous peptides. Binding of exogenous peptides to HLA-A2 stabilizes the HLA-A2- peptide complexes and can be detected using immunofluorescence staining. The T2 cell line is commonly used for identification and analysis of novel HLA-A2-restricted CTL epitopes in the assay known as T2 shift. In this study we evaluated parameters that may influence the T2 shift assay performance and considered new applications of this method for testing of the biological activity of peptide-based immunotherapies. Method optimization was performed using HLA-A2-restricted peptide SurA2.M alone and in the context of a peptide-based immunotherapy drug, DPX-Survivac. DPX-Survivac is a lipid nanoparticle-based formulation that contains five peptides aiming to elicit a CTL response against tumor cells presenting the survivin peptides on HLA class I molecules. Results indicated that performance of the T2 shift assay was highly sensitive to cell passaging intervals, pH variations, cell concentrations and β2-microglobulin concentrations. Discrimination of the T2 cells by either size or granularity did not affect the results for the T2 shift assay. We determined the linear response range for the SurA2.M peptide to be 30-110 μg/mL in the T2 shift assay. The T2 shift assay can detect SurA2.M within the DPX-Survivac matrix. In conclusion, we found that optimal culturing conditions of T2 cells are critical for the T2 shift assay reproducibility and linearity and should be carefully considered if the assay is to be used for quantitative or semi-quantitative evaluation of the peptide activity alone or as a part of an immunotherapeutic drug.
Vasoactive intestinal peptide signaling – a novel immune checkpoint in pancreatic ductal adenocarcinoma
Authors: Ravindranathan Sruthi , Li Yiwen , Wang Shuhua , Zaidi Mohammad , Lesinski Gregory B , El-Rayes Bassel , Waller Edmund K 
Published: January 2023

Abstract
Pancreatic ductal adenocarcinoma (PDAC) is projected to become the second leading cause of cancer related death in the U.S. by 2030. Being completely refractory to checkpoint therapies, management of PDAC remains an unmet clinical need. Intriguingly, the human protein atlas (THPA) shows elevated mRNA expression of vasoactive intestinal peptide (VIP), a neurotransmitter in exocrine pancreatic cancers. Interestingly, VIP also has immunosuppressive properties leading to decreased T cell proliferation and enhanced expansion of myeloid derived suppressor cells (MDSCs) and regulatory T cells (Tregs). Thus, we are investigating the role of VIP in PDAC progression and the effect of inhibiting VIP signaling for improved responsiveness to immunotherapy.
We found that consistent with THPA, cell culture supernatants collected from murine PDAC cell lines had significantly higher levels of VIP as compared to melanoma and breast cancer cells. Similarly, elevated levels of VIP were observed in mice bearing PDAC tumors, and in plasma from human PDAC patients, when compared to naïve mice and healthy volunteer samples, respectively. Treatment of immune competent mice bearing PDAC tumors (MT5 or KPC cell lines) with the combination of a VIP antagonist (VIPhyb) and anti-PD1, produced complete and durable regression of tumors in 20% of the mice and an overall delayed tumor progression in both tumor models. Upon analysis of the tumor tissues sections via immunohistochemistry, significant infiltration of CD8+ T cells was observed in mice treated with the combination of VIPhyb and anti-PD1. Further experiments are underway to understand the mechanism of action of VIP antagonist and to devise methods to improve its efficacy in-vivo.
DeepTM IL-12 Priming enhances tumor-specific T cell efficacy and inflammatory activity in tumors without eliciting systemic toxicities
Authors: Sauer Karsten , Chang De-Kuan , Ahmad Gulzar , Nardozzi Jonathan , Sackton Kate , Lyons Jesse , Andresen Thomas , Jones Douglas 
Published: January 2023

Abstract
Interleukin-12 (IL-12) is a potent pro-inflammatory cytokine that augments anti-tumor immune responses by promoting CD4 T cell Th1 differentiation, increasing CD8 T cell and NK cell cytotoxicity, inducing MHC expression on antigen presenting cells, and reprogramming myeloid-derived suppressor cells. However, the clinical utility of IL-12 has been limited by systemic toxicities. DeepTM IL-12 is a fusion protein between IL-12 and a Fab antibody against CD45, an abundant surface receptor on T cells. This enables tethering of IL-12 onto T cells prior to adoptive cell transfer (ACT). We report here the superior safety and efficacy of ACT with tumor-specific Deep IL- 12 Primed T cells which carry surface-tethered IL-12.
The PMEL/B16-F10 cancer model utilizes adoptively transferred PMEL CD8 T cells that contain a TCR specific for the gp100 antigen expressed in B16-F10 melanoma cells. Deep IL-12 tethering to PMEL T cells significantly improved tumor growth inhibition and survival in tumor-bearing mice compared to PMEL T cell therapy alone or combined with systemically delivered IL-12. Doses up to 40 million Deep IL-12 Primed T cells did not result in overt toxicities. Thus, in contrast to systemically delivered free IL-12, T cell tethered IL-12 is well-tolerated. Deep IL-12 Primed PMEL T cells additionally resulted in elevated and sustained induction of IFNγ in the tumor microenvironment. In contrast, IFNγ induction in serum was transient and returned to baseline levels within four days of ACT. Further toxicological analyses will be discussed.
In conclusion, ACT with Deep IL-12 Primed T cells delivers a favorable therapeutic profile by promoting T cell function selectively in the tumor microenvironment while limiting systemic IL-12 exposure.
Impact of HIV-1 infection and antigen class on T follicular helper (TFH) cell responses to pneumococcal polysaccharide-protein conjugate vaccine (PCV-13)
Authors: Jha Vibha , Nicholson Lindsay K , Gardner Edward M , Rahkola Jeremy T , Pratap Harsh , Janoff Edward N 
Published: January 2023

Abstract
Pneumococcal infections are common and serious complications of HIV-1 disease. Prevention has been compromised by the limited magnitude and quality of antibody responses to T cell-independent (TI-2) pneumococcal capsular polysaccharides (PPS). The PCV-13 vaccine conjugates PPS to a T cell-dependent (TD) protein (diphtheria toxoid; DT). We investigated the differential response to PPS and DT by antibody-secreting B cells (ASC) after immunization with PCV-13 in newly-diagnosed healthy HIV+ and control adults. The numbers of PPS-specific IgG ASC increased significantly and similarly in HIV+ and controls 1 week after vaccination. However, DT-specific IgG ASC increased significantly in controls but not HIV+ subjects. To determine the cellular basis of these disparate responses to DT and PPS, we characterized the frequency and activation of T follicular helper (TFH) cells, the predominant T cell subset providing B cell help. The percentages of and intracellular IL-21 in TFH cells were unchanged with vaccination. However, expression of ICOS, which sustains TFH function and phenotype, increased significantly after vaccination among controls, but not the HIV+ group. Increases in ICOS+ TFH correlated with changes in TD DT-specific IgG ASC in controls but not in HIV+. In contrast, ICOS expression did not correlate with TI-2 PPS-specific ASC in either group. Levels of TFH differentiating factors IL-12 and IL-6 increased post vaccination only in controls. In summary, although TI-2 PPS-specific responses were similar in both groups and independent of TFH activation, TFH ICOS-expression was most closely related to responses to TD DT, which were compromised even in recently diagnosed healthy HIV+ adults.
Skin recirculating B cell subsets in a model of lymph cannulation in sheep
Authors: Hunka Julia , Riley John T. , Debes Gudrun F. 
Published: January 2023

Abstract
The skin is a key barrier organ and cells of the skin immune system are integral to skin immunity and inflammation. While the trafficking of skin T cells is well defined, less is known about B cells that reside in and traffic through skin. Here we studied skin-associated B cells and their homing receptors using lymph vessel cannulation in sheep. Afferent lymph draining the skin was collected and lymph-borne B cells were compared to B cells from skin, blood, skin and mesenteric lymph nodes, mesenteric efferent lymph, and spleen using flow cytometry. The skin-homing molecule α4β1-integrin as well as activated β1-integrin were upregulated in B cells in skin-draining lymph and skin compared with lymph nodes, suggesting that α4β1 integrin is key to B cell recirculation through and/or homing to skin. Of all tissues analyzed, B cells from skin exhibited the highest frequency in skin-homing molecule E-selectin ligand expression. Among afferent lymph B cells, the CD21− B cell subset showed the highest expression of E-selectin ligand. As recent studies showed a functional endocannabinoid system in the skin, we assessed chemotaxis of skin-recirculating B cells toward 2-arachidonoylglycerol (2-AG), a ligand for cannabinoid receptor 2 (CB2). Interestingly, CD21+ B cells from skin draining lymph were highly enriched in responsiveness to 2-AG as well as CCL20, a skin-expressed chemokine. We propose that α4β1 integrin is integral to all B cell homing to skin and that CD21 expression delineates skin-recirculating B cell subsets with distinct trafficking receptor usage, while CD21− B cells use E-selectin ligand, CD21+ B cells employ receptors for 2-AG and CCL20 to recirculate through skin.