bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bilirubin Nanoparticle as an anti-inflammatory therapy for graft versus host disease
Authors: PAreek Sumedha , Zhang Mao , Trujillo-Ocampo Abel , Kim Jinyong , Jon Sangyong , Im Jin Seon 
Published: January 2023

Abstract
Graft versus host disease (GvHD) caused by alloreactive donor lymphocytes is a fatal complication of hematopoietic stem cell transplant (HSCT). Conditioning regimen consisting of chemotherapy and/or radiation given prior to HSCT can cause the initial tissue damage, which triggers the cross-presentation of alloantigens to the donor immune cells. Previous studies have shown that water-soluble PEGylated bilirubin nanoparticles (BRNP) selectively accumulate at the site of inflammation and prevent further tissue damage through scavenging reactive oxygen species. Therefore we, hypothesized that BRNP treatment after conditioning regimen can reduce GvHD by abating the initial tissue damage in HSCT, and investigated therapeutic efficacy of BRNP using murine GVHD model. Sublethally irradiated recipient mice (Balb/c) were infused with 4×106 bone marrow and 1×106 splenic T cells from donor mice (C57/B6) on day 1, and with or without BRNP (10mg/kg) on days 0, 2, and 4. GvHD symptoms were monitored for 60 days, and scored for changes in fur, skin, posture, activity, and weight. Untreated recipient mice (n=9) developed significantly worse GVHD (mean GVHD score=4.667) compared to BRNP treated recipient mice (n=9, mean GVHD score=1.556) (p=0.0028). This translated into the significantly better survival of BRNP treated mice with day 60 survival of 65% as compared to the untreated recipient mice with day 60 survival of 11% (p=0.03). In summary, we show that BRNP treatment can ameliorate clinical GvHD symptoms and improve survival in murine GVHD model. In the future, we plan to investigate the role of reactive oxygen species in GVHD and mechanism of action on anti-GVHD effects by BRNP.
Donor-specific antibody mediates chronic, but not acute, kidney allograft rejection in the absence of natural killer cells
Authors: Kish Danielle , Yagisawa Takafumi , Dvorina Nina , Valujskikh Anna , Baldwin William M. , Fairchild Robert L. 
Published: January 2023

Abstract
Dysregulated donor-specific antibody (DSA) responses are induced in B6.CCR5−/− mice transplanted with complete MHC mismatched kidney allografts and NK cells play a critical role in acute graft injury and failure. The current study investigated the consequence of high DSA titers on graft outcomes in the presence vs. absence of NK cell activation within the kidney graft. B6.CCR5−/− recipients of complete MHC mismatched A/J allografts or semi allogeneic (A/J x B6)F1 kidney grafts responded with equivalent serum DSA titers that reached peak by day 14 post-transplant. B6.CCR5−/− recipients rejected A/J allografts between days 15–30, whereas B6 isografts and (A/J x B6)F1 semi allogeneic grafts survived past day 65. On day 7 post-transplant NK cell infiltration into A/J allografts was composed of distinct Ly49b+ cell populations expressing low and high levels of NK1.1 and both NK1.1high and NK1.1low cells increased in A/J allografts with time to rejection. In contrast, NK cell infiltration into (A/J x B6)F1 grafts on day 7 post-transplant was composed entirely of NK1.1high cells that decreased to background/isograft numbers thereafter. (A/J x B6)F1 grafts at day 65 post-transplant had severe chronic injury with prominent interstitial fibrosis, glomerulopathy and arteriopathy that was accompanied by gene expression of profibrogenic factors. These results indicate that NK cells synergize with DSA to cause acute kidney allograft failure whereas high titers of DSA in the absence of NK cell activation cannot cause acute antibody-mediated rejection but induce the indolent development of interstitial fibrosis and glomerular injury that leads to graft dysfunction and failure at later times after transplantation.
Novel Consistently Ex-vivo Expanded CD8+CD25+FoxP3+Treg Cells Prolongs the Allogeneic Islet Survival
Authors: Kim Jung-Sik , Han Kyeo-Rae , Park Chung-Gyu 
Published: January 2023

Abstract
In addition to CD4+ regulatory T cells(Tregs), adoptive transfer of CD8+CD25+FoxP3+T cells is emerging as an alternative adjunctive therapy to diminish current reliance on lifelong, nonspecific immunosuppression after transplantation. Here we evaluated the possible therapeutic application of consistently ex-vivo expanded mouse CD8+CD25+FoxP3+T cells to prevent the rejection of allogeneic islets.
Around 80% of mouse CD8+CD25+T and CD4+CD25+T cells expressed FoxP3+T cells in spleen. Higher level of FoxP3 was expressed in CD8+Tregs compared to CD4+Tregs. Ex-vivo enriched CD8+CD25+Treg cells were successfully and consistently expanded to 600–1,000 times from the initial culture for 12 days with optimized culture conditions. High concentration of IL-2 (&gt;500units) and TGF-beta together with aCD3-and aCD28 stimulation were essential to the massive ex-vivo expansion. Expanded Tregs showed 99% of FoxP3 expression and expressed high level of CD27, CD47 and Lag-3 in their surface. Very high amount of Granzyme B and minor amount of IFN-r and IL-17 were xpressed. Expanded CD8+Tregs successfully inhibits the proliferation and activation of CD4+T cells in vitro. In the allogenic islet transplantation model (Balb.Cà B6), Single injection of ex-vivo expanded CD8+Tregs prolonged islet allograft survival more than 100 days after the reconstitution with CD90.2+T cells in immunodeficient B6 rag2(−/−) mice by inhibiting antigen-specific IFN-r producing cell generation. In this study, we proved that ex-vivo expanded CD8+Treg retain regulatory activity and can protect islet allograft rejection by single administration. These implies that ex-vivo expanded CD8+Tregs has clinical relevance of an adjunctive cellular therapy.
In vitro characteristics and in vivo outcome of α1,3-galactosyltransferase gene-knockout miniature pig cornea in full-thickness corneal xenotransplantation using nonhuman primate
Authors: Yoon Chang Ho , Choi Se Hyun , Lee Hyun Ju , Kim Hong Pyo , Kang Hee Jung , Kim Jong Min , Choi Hyuk Jin , Park Chung-Gyu , Choi Kimyung , Kim Hyunil , Ahn Curie , Kim Mee Kum 
Published: January 2023

Abstract
We investigated the biofunctional profiles of α1, 3-galactosyltransferase gene knockout (GTKO) miniature pig cornea and its outcome in pig-to-rhesus full thickness corneal xenotransplantation (FTCX). Ten eyes of GTKO miniature pigs (n = 5) were evaluated for biofunctional characteristics. Five Chinese rhesus macaques underwent FTCX (mean donor age: 11.3 mo). Systemic tacrolimus, basiliximab, and IVIG were administered as programmed schedules. T cell subsets, antibodies (Ab) in blood, and C3a in aqueous humor (AH) were evaluated. The results were compared with those of the wild type miniature pig corneal grafted group (n = 4, all grafts survived &gt; 24 weeks, using anti-CD154 Ab) as controls. Mean central corneal thickness was 763 μm. Na+/K+ ATPase was well expressed 7 days after preservation. Mean endothelial cell density was 4729/mm2, and was maintained &gt;2800/mm2 through 7 days. Mean doubling time was 30.4 hour which was comparable to that in SNU miniature pigs (30.7 hour, p = .413). Median graft survival was 72 days. Regulatory T cells at rejection decreased from baseline (p = .043) and were lower than those of controls at postoperative (postop) 24 weeks (p = .014), however the other T cell subsets did not change. At rejection, anti-non-αGal Ab increased from baseline (p = .043), however Anti-αGal Ab did not. C3a in AH at postop 4 weeks and at rejection increased from baseline (all p = .043). At postop 4 weeks, AH and plasma C3a were higher than those of controls (p = .016 and .036, respectively). The biofunctional characteristics of GTKO miniature pig cornea are feasible for FTCX. Rejection of GTKO pig-to-rhesus FTCX is related with non-αGal Ab, complement and decrease of regulatory T cells. Addition of B cell suppressing regimen should be considered.
Tc17 cells mediate gut GVHD via IL-22 and dysbiosis
Authors: Song Qingxiao , Wu Xiwei , Zeng Deye , Jenq Robert R. , Van den Brink Marcel R. M. , Riggs Arthur D. , chen Yuanzhong , Zeng Defu 
Published: January 2023

Abstract
Gut GVHD is critical for determining the outcome of allogenic hematopoietic cell transplantation (HCT). Damage of Paneth cells and loss of RegIIIg can lead to dysregulation of intestinal microbiota and gut GVHD pathogenesis. RegIIIg can be produced by Paneth cells and intestinal epithelial cells, and its secretion is augmented by IL-22. However, the circumstances leading to RegIIIg dysregulation and dysbiosis remain largely unknown. Here we show that, in a MHC-mismatched model of C57BL/6 donor to BALB/c recipient, one injection of depleting anti-CD4 effectively prevents acute gut GVHD in recipients given wild-type (WT) transplants but has no effect in recipients given IFN-g−/− transplants. Interestingly, PD-L1−/−recipients given IFN-g−/− transplants showed spontaneous recovery of the disease. Recipients given IFN-g−/− transplants showed expansion of Tc17, IL-22 but not IL-17 from Tc17 was required for the disease induction. Furthermore, host tissue PD-L1 augmented expansion of Tc17 as well as CX3CR1+ DC, CX3CR1+ DC mediate bacteria clearance and prevent bacteria translocation, at the same time, CX3CR1+ DC produce IL-18 BP, which limit Tc17 expansion through IL-18 signaling. In addition, recipients given IFN-g−/− transplants showed no damage of Paneth cells but markedly increased RegIIIg. Gut GVHD in recipients had significant reduction of Barnesiella in feces and significant increase of E. Coli and Lactobacillus. Taken together, these results indicate that 1) in the absence of donor IFN-g, host-tissue PD-L1 mediates expansion of Tc17 through IL-18 signaling and dysregulation of IL-22 functional effect; 2) Dysregualtion of IL-22 from Tc17 cell can lead to dysbiosis and gut GVHD in the absence of Paneth cell damage.
Development of a murine intestinal explant model to examine immune mechanisms of graft versus host disease
Authors: Gao Chunxu , Hitchcock Shannon , Deutsch Heather , Rao Tadimeti , Amuzie Chidozie , Malaviya Ravi 
Published: January 2023

Abstract
Graft-versus host disease (GvHD) is a major course of mortality and morbidity in transplant recipients. Cytokine signaling plays a key role in the development of GvHD. To study the immune mechanisms relevant to intestinal pathology associated with GvHD, we developed a murine ex vivo model on intestinal explants derived from Xeno-GvHD to characterize mouse and human cytokine secretion and signaling. Intestinal tissues from Xeno-GvHD induced by transfer of human PBMC to irradiated NSG mice and control naïve mice were dissected and cultured. Histological analysis of the explants and measurement of lactate dehydrogenase (LDH) in the ex vivo culture system indicate that intestinal explants remain viable in culture for up to 72 hours, with an increased cytotoxicity developing over this time. Release of mouse proinflammatory cytokines including RANTES, IL-6, KC, and GM-CSF were significantly higher in GvHD explants compared to naïve controls. Among the human cytokines, RANTES was a key cytokine detected only from Xeno-GvHD explants. The patterns of cytokine/chemokine release were shown to be consistent in the ex vivo culture over time and on explants from different sites of intestine including ileum and colon. Furthermore, histochemical analysis of GvHD explants revealed much stronger immunoreactivity for RANTES compared to naïve explants. Overall, our results indicate inflammatory milieu in the intestines of GvHD mice leading to sustained production of proinflammatory cytokines in the ex vivo culture. These features indicate that the explant system is a relevant model for studying the mechanism(s) of inflammatory responses in intestinal GvHD.
Activation of host p53 enhances anti-tumor immunity by driving differentiation of immunogenic myeloid DCs in tumors, while simultaneously protecting against collateral autoimmunity against shared self-tumor antigens.
Authors: Sharma Madhav D. , Munn David H. 
Published: January 2023

Abstract
During immunotherapy, we show that large numbers of immunogenic CD103+ DCs can arise in murine tumors via direct differentiation myeloid-lineage (Ly6c+ monocytic) precursors. These “dual-phenotype” Ly6c+CD103+ DCs differentiate from a precursor population of monocytic myeloid-derived suppressor cells (MDSCs) in tumors. Differentiation was controlled by inflammation-induced activation of the transcription factor p53 in MDSCs. Targeted deletion of myeloid p53 selectively ablated the Ly6c+CD103+ DC population, which resulted in abrogation of all response to multiple forms of tumor immunotherapy. Pharmacologically elevating the level of p53 expression in host cells, by administering the p53-agonist (MDM2-inhibitor) drug nutlin-3a, caused a sustained increase in Ly6c+CD103+ DCs in tumors during immunotherapy, and markedly enhanced anti-tumor immunity. Conversely, however, when a nominal tumor antigen (OVA) was also shared by a critical host tissue (Rip-mOVA mice), elevation of host p53 using nutlin-3a was potent immunosuppressive and protective against lethal autoimmunity outside of the tumor, while still driving enhanced immunity – against the same antigen – within the tumor itself. Thus, p53-agonist drugs may represent a novel class immunotherapy agents, with high discrimination between tumor-associated antigens and collateral autoimmunity.
HMGN1 and R848 synergistically activate DCs by multiple signaling pathways
Authors: Alam Md Masud , Yang De , Trivett Anna , Oppenheim Joost J. 
Published: January 2023

Abstract
HMGN1 (High-mobility group nucleosome-binding protein 1, N1) is a non-histone DNA binding protein that acts extracellularly as Th1-polarizing alarmin. N1 promotes dendritic cell (DC) recruitment and maturation primarily via TLR4. Initially various other TLR ligands were tested in combination with N1 to identify the most stimulatory combination. This showed that R848, a synthetic TLR7/8 agonist in combination with N1 synergistically stimulated DC maximally. However, it remained unclear how N1 and R848 synergistically activated human DCs. Here, we show that co-stimulation with N1 and R848 in human monocyte-derived DCs (MoDCs) resulted in marked upregulation of DC surface expression of CD80, CD83, CD86, and HLA-DR, as well as synergistic production of pro-inflammatory cytokines including IL-12p70, IL-1β and TNFα. This combination synergistically activated NF-κB and multiple MAPKs including SAPK/JNK, c-Jun, CREB and p38 involved in pro-inflammatory signaling pathways. Moreover, N1 and R848 also synergistically increased nuclear translocation of interferon (IFN) regulatory transcription factors (e.g. IRF3 and IRF7) and promoted the expression of type I IFN genes including IFNα2, IFNα4 and IFNβ1. Similar signaling pathways were also be induced in mouse bone marrow-derived DCs (BMDC). Overall, our findings show that 1) N1 synergizes with R848 in activating human and mouse DCs and 2) the synergistic effect based on various intracellular signaling events culminated in the activation of multiple transcriptional factors. These findings have important implications for future clinical trials since N1 and R848 synergistically promoted optimal Th1-lineage immune response mediating tumor rejection in mice.
In situ vaccination improves efficacy of PD-1 blockade in unresponsive lymphoma tumors through induction of a highly efficient cross-presenting dendritic cell subset expressing TLR3
Authors: Hammerich Linda , Dhainaut Maxime , Keler Tibor , Davis Thomas , Salazar Andres M. , Brown Brian D. , Brody Joshua D. 
Published: January 2023

Abstract
B cell lymphomas are generally incurable, with low response rates to anti-PD1 therapy. Here, we tested the combination of in situ vaccination with PD-1 blockade in a pre-clinical mouse model.
A20 lymphoma-bearing mice were treated with a PD1 blocking antibody with or without an in situ vaccine combining intratumoral (it) injections of FMS-like tyrosine kinase-3 ligand (Flt3L), poly-ICLC (pIC) and local irradiation (XRT) of the tumor.
Untreated lymphoma tumors contained few dendritic cells (DC) and anti-PD1 treatment did not induce tumor regression. Flt3L treatment resulted in a dramatic increase of IRF8+TLR3+ DC in the tumor. XRT of A20 cells induced activation of Flt3L-treated splenic DC in vitro and local XRT of the tumor in vivo induced expression of CD103 on TLR3+ DC. Local XRT also enhanced uptake of dying tumor cells by it DC. Importantly, tumor antigens were taken up mainly by CD103+ DC and not CD103-DC. Accordingly, CD103+ DC isolated from the tumor induced proliferation of tumor-specific CD8+ T cells more efficiently than CD103-subsets. Combination of Flt3L with XRT and pIC induced tumor-reactive T cells, and delayed tumor growth and improved survival in 40% of mice. In situ vaccination also increased expression of PD-L1 on tumor cells and it DC. Consistently, combination of in situ vaccination with anti-PD1 led to complete tumor regression and increased long-term survival in the majority of mice. PD1 blockade also increased the number of tumor-reactive T cells and depletion of CD8+ T cells abrogated the anti-tumor effect.
In conclusion, in situ vaccination combining Flt3L, local XRT and pIC improves efficacy of anti-PD1 in checkpoint-unresponsive lymphoma tumors through induction of a highly efficient cross-presenting DC subset.
Cell death-inducing reagents activate cancer immune surveillance via inducing Interleukin-17D in macrophages
Authors: Seelige Ruth , Bui Jack D. 
Published: January 2023

Abstract
We have previously established a role for the cytokine IL-17D during cancer immune surveillance in that its cancer-intrinsic expression recruits NK cells into the tumor microenvironment, thereby leading to cancer regression. We additionally showed that the transcription factor Nrf2 regulates IL-17D and can itself be induced by the chemical tBHQ. Here, we sought to investigate what other endogenous or exogenous signals induce IL-17D not only in cancer cells but also in macrophages that are abundant in the tumor microenvironment. By testing different agonists in vitro, we found that all reagents that led to increased cell death (such as chemotherapy) also induced IL-17D. This effect was moderate in cancer cells but more profound in macrophages and depended on Nrf2. Interestingly, the expression of IL-17D correlated with type I Interferon expression, suggesting that these molecules function in the same pathway. Additionally, the only reagent that induced IL-17D without leading to cell death was poly (dA:dT), a nucleic acid known to induce intracellular DNA sensing pathways such as the Absent in melanoma (AIM)2 or the cGAS/Stimulator of IFNs (STING) pathway. This suggests that ectopic nucleic acids (likely resulting from cell death) are inducers of IL-17D. Remarkably, co-injecting bone marrow-derived macrophages (BMDMs) that had induced IL-17D together with cancer cells increased the number of NK cells in tumors and decreased cancer growth. This effect was dependent on IL-17D because it was abolished if IL-17D deficient BMDMs were injected. We hypothesize that the IL-17D pathway evolved to sense ectopic DNA, likely present due to cell death, and that boosting this pathway in cancer cells or macrophages could serve as cancer immune therapy.
Cell-cell fusion as a mechanism of DNA exchange in cancer
Authors: Searles Stephen , Santosa Endi , Bui Jack D. 
Published: January 2023

Abstract
Cell-cell fusion is a common biological phenomenon involved in both healthy and disease conditions. Fusion occurs when two or more cells combine their plasma membranes and become a single hybrid cell that retains certain genetic information from each parent cell. Here, using a Cre-loxP-based method initially developed to investigate extracellular vesicle targeting, we found that cancer cells spontaneously and rapidly deliver DNA to non-cancer cells in vitro via a cell-cell fusion event. We found that B16 melanoma cells and progressor and regressor fibrosarcomas were capable of spontaneously fusing with immune cells (macrophages) and non-immune cells (fibroblasts) in vitro, with varying degrees of efficiency. Further analysis of hybrid clones resulting from fusion between B16 melanoma cells and mouse embryonic fibroblasts showed that these cells were aneuploid and possessed enhanced clonal diversity and chemoresistance compared to non-hybrid melanoma cells. We also observed cell-cell fusion to occur in vivo between melanoma cells and non-cancer cells of both hematopoietic and non-hematopoietic origin. These findings suggest that cell-cell fusion occurs in murine cancer models and show that this mechanism has the potential to cause massive genomic alterations that are observed in cancer. Furthermore, these findings somewhat contradict recent publications suggesting that the Cre-loxP method measures only extracellular vesicle-mediated intercellular communication.
MyD88-stimulated T cells acquire resistance to MDSC-mediated suppression
Authors: Ciavattone Nicholas G. , Parker Dominique , Joseph Ann Mary , Davila Eduardo 
Published: January 2023

Abstract
The immunosuppressive tumor microenvironment presents a significant challenge to developing effective T cell-based cancer immunotherapies. The heterogeneous population of myeloid-derived suppressor cells (MDSCs) is a major contributor to the suppressive tumor microenvironment in many cancers. Generating tumor-reactive T cells with the capacity to resist MDSC-mediated suppression would help facilitate the production of potent anti-tumor T cell therapies. The activation of the Toll-like receptor-Myeloid differentiation primary response 88 (TLR-MyD88) signaling pathway in CD8+ T cells enhances cell proliferation, cytotoxic function, and survival. Our studies show that TLR-stimulated T cells are resistant to MDSC-mediated suppression from multiple cancer cell lines. MyD88-activated CD8+ T cells co-cultured with tumor-derived MDSCs displayed enhanced proliferation and cytokine production over control T cells. This is recapitulated using synthetic co-receptor CD8a-MyD88 engineered T cells which exhibit greater potential for proliferation, even in the presence of MDSCs. Our future objectives are to understand the mechanisms by which MyD88 stimulated T cells gain resistance properties and observe this phenomenon in vivo. We look to utilize these observations to augment antitumor immune responses.