bluuebunny/crossref_metadata_2025_split huggingface dataset:
Collaboration of therapeutic, systemic IL12p40 with locally released IL12p35 focuses immunity to the tumor microenvironment
Authors: Hill Elizabeth M , Gerber Allison N , Singh Nevil J 
Published: January 2023

Abstract
The cytokine IL-12 was an early candidate for cancer immunotherapy due to its role in stimulating the release of IFN-γ from T and NK cells to promote tumor clearance. Administering IL-12 therapeutically was challenging because systemic treatments led to excessive cytokine storm and inflammation. Developing more targeted ways to focus IL-12 activity to the tumor microenvironment is a significant priority. IL-12 is a heterodimer formed by the subunits, IL-12p35 (p35) and IL-12p40 (p40) which are canonically thought to be made together in the same cell. Although p40 can be released as a monomer, this is inert. We have shown that these p40 monomers however can combine extracellularly with p35 from stromal cells to form functional IL-12 and help control infection. Based on this newly described pathway, we hypothesized that treatment with recombinant p40 monomers would allow it to combine with tumor-released p35 and focus IL-12 formation to the local microenvironment. This would facilitate targeted tumor clearance while limiting systemic immunopathology. To test this hypothesis, we evaluated treatment with p40 monomers in mice challenged with a subcutaneous tumor (MB49). We found that monomeric 40 itself was inert – neither capable of inducing IFN-γ or inhibiting the activity of heterodimeric IL-12. Nevertheless, recombinant p40 monomers helped control tumor growth in p40-deficient mice and increased the amount of CD4 T cells infiltrating the tumor. This suggests that p40 can combine with p35 released in the tumor microenvironment to form functional IL-12. Our data provide evidence for an alternate pathway of IL-12 formation in vivo and suggest that this can be exploited therapeutically to avoid systemic inflammation.
Loss of ADAM17 from macrophages induces ST2+ T regulatory cells that limit obesity-induced metabolic inflammation in mice through changes in both membrane and soluble TNF
Authors: Martin Rebecca , Lownik Joseph Cornelius , Farrar Jared , Way Grayson , Zellner Matthew , Ni Bin , Celi Francesco , Conrad Daniel H 
Published: January 2023

Abstract
Obesity is known to contribute to changes in immune cells, cytokines and metabolism. Secreted TNF (sTNF) levels are directly correlated in obesity with weight gain and morbidity, as well as metabolic dysregulation. ADAM17 cleaves transmembrane TNF (mTNF) into its soluble form. This is well defined, but the mechanism behind the regulation of TNF, the source of TNF, as well as the downstream action of TNF is less clear. Our data showed that mice with ADAM17 deleted from the myeloid lineage (ADAM17LysM) had a significant metabolic advantage as illustrated by an increase in glucose tolerance and decrease in body weight. Immunophenotyping of the fatpad of these mice using high dimensional flow cytometry after high fat diet (HFD) feeding showed an increase in activated ST2+ T regulatory cells (Tregs). Based on these findings, we hypothesized that modulation of TNF on macrophages through loss of ADAM17, helps maintain a healthy metabolic state through recruitment and activation of ST2+ Treg cells that limit metabolic dysregulation. Next, we showed that the accumulation and activation of these Tregs occurred through two separate mechanistic actions of TNF. First, the loss of sTNF production from macrophages decreased soluble ST2 (sST2) production by adipocytes. This loss of sST2 augments ST2+ Treg recruitment and expansion in the fatpad due to increased bioavailable IL-33. These experiments were controlled using a myeloid-specific knockout of TNF. Second, utilizing TNFR blocking antibodies, we demonstrated that ST2+ Treg activation was dependent on macrophage mTNF acting through TNFR2.
Supported by grants from NIH (R56 AI139658 and R01 AI018697)
Characterization of a CLCF1 conditional knock-out mice model
Authors: Laplante Veronique , Rousseau Marine , Fajardo Ernesto , Pasquin Sarah , Lesage Sylvie , Gauchat Jean-François 
Published: January 2023

Abstract
The Cardiotrophin-like Cytokine Factor 1 (CLCF1) is a cytokine of the IL6 family with important pro-neurotrophic and immuno-modulating functions. However, the mechanisms behind CLCF1 activities and CLCF1 properties in pathological models remain poorly understood. This is partly due to a lack of tools for the study of CLCF1 functions in vivo. Indeed, the complete knock-out of CLCF1 in mice is lethal at P1: underdeveloped motor neurons of the face and jaw prevent the pups from suckling. To overcome this obstacle, we obtained a CLCF1 conditional knock-out mice model generated using the CRISPR-Cas9 technology. We then bred these mice with Vav-Cre mice to induce CLCF1 deletion in hematopoietic cells. We are now conducting immuno-phenotyping experiments of the main immune populations in those animals. Considering the activities of CLCF1 that were previously shown in vitro or in vivo using overexpression models, we hypothesize that the knock-out of CLCF1 in immune cells will lead to decreased numbers of B and myeloid cells. This new CLCF1 conditional knock-out mice model will be a potent tool to confirm and further study the activities, mechanisms, and pathological roles of CLCF1 in vivo.
Supported by grants from CIHR (Canadian Institutes of Health Research)
Notch signaling is required for generation of conventional type 1 dendritic cells from human induced pluripotent stem cells
Authors: Ito Fumito , Long Mark D , Kajihara Ryutaro , Matsueda Satoko , Oba Takaaki , Kanehira Kazunori , Liu Song , Makino Kenichi 
Published: January 2023

Abstract
Compelling evidence shows a critical role of conventional type 1 dendritic cells (cDC1) in T-cell mediated antitumor immunity; however, obtaining large numbers of cDC1 is difficult. Theoretically, this limitation can be overcome by using induced pluripotent stem cells (iPSC) that could provide an unlimited source of autologous cDC1. However, generation of cDC1 from human iPSC remains elusive. Here, we hypothesized that Notch signaling would facilitate generation of cDC1 from human iPSC. We found that human iPSC gave rise to CD141+ XCR1+ CLEC9A+ HLA-DR+ cells on OP9 feeder cells expressing the Notch ligand delta-like 1 (OP9-DL1) while the majority of iPSC-derived cells differentiated on OP9 cells were monocytic cells. Single-cell RNA sequencing analyses confirmed the presence of cDC1 as well as identified the heterogeneity of iPSC-derived CD1c+ cells; cDC2A, cDC2B, CD5+ cDC2, and DC3. Inhibition of Notch signaling during co-culture of iPSC-derived CD34+ hematopoietic progenitor cells with OP9-DL1 cells abrogated generation of cDC1, and made expression of CD5, CLEC4A, CLEC10A and CD163 in CD1c+ DC similar to those differentiated on OP9 cells. Notch-activated human iPSC-derived XCR1+ CLEC9A+ HLA-DR+ CD11c+ cells exhibited similar gene expression profile with peripheral blood cDC1, and phagocytotic, T-cell proliferative, and cytokine producing functions. Our study demonstrates that Notch signaling is required for the differentiation of cDC1-like cells, and facilitates generation of various cDC subsets from human iPSC. These findings provide insights into the future development of personalized treatment with unlimited numbers of autologous cDC1 from human iPSC.
This work was supported by National Cancer Institute (NCI) grant P30CA016056 involving the use of Roswell Park’s Flow and Image Cytometry, Bioinformatics, and Genomic Shared Resources. This work was supported by the Melanoma Research Alliance and the Sarcoma Foundation of America (F. Ito), Uehara Memorial Foundation (T. Oba), and National Cancer Institute (NCI) grant, U24CA232979 (M. Long and S. Liu), K08CA197966 and R01CA255240-01A1 (F. Ito).
Stress Induced Eosinophilpoiesis Contributes to Mortality after Pulmonary Resection
Authors: Mei Zhongcheng , Guo Yizhan , Li Dongge , Khalil May , Banerjee Anirban , Kreisel Daniel , Gelman Andrew , Krupnick Alexander 
Published: January 2023

Abstract
Factors affecting mortality after lung surgery remain poorly explored. We performed pulmonary lobar resections(removing □ 1/5 of the total pulmonary volume) from C57BL/6 mice and evaluated quantitative changes in various leucocytes in the remaining lung, blood and spleen. A nearly four-fold increase in eosinophils was evident by day-7 post resection with a gradual return to baseline by day 30. Evaluation of the bone marrow demonstrated no change in common myeloid, granulocyte-monocyte or eosinophil progenitors. However the number of early stage and mature eosinophils doubled after lobar resection suggesting that pulmonary surgery augmented maturation of eosinophils. Neutralization of IL-5 decreased overall eosinophil numbers but the relative differences between resting and post-resection mice remained. Further evaluation of bone marrow demonstrated an increase in IL-25, IL-6, IL-9, KC and MCP-1 after pulmonary resection. We next performed right pneumonectomies in saline or diphtheria toxin-treated iPHIL mice, where diphtheria toxin receptor expression is limited to eosinophils, and noted a 70% mortality in eosinophil sufficient vs. 20% mortality in eosinophil depleted mice (p=.005). Severe hypoxia was noted in eosinophils sufficient vs. deficient mice (pO2 of 54.3+/−5.7 vs. 83.6+/−11.63; p=.04) four hours prior to resection. Taken together out data demonstrate that pulmonary surgery leads to bone marrow-specific maturation of eosinophils and systemic eosinophilia which directly contributes to perioperative mortality. Thus defining factors which may disrupt this process offers the opportunity to improve outcomes after lung surgery.
Supported by PO1 AI116501, RO1 AI145108-01
Maternal immune activation (MIA) impairs neonatal lung ILC2 establishment, function, and airway hyperresponsiveness
Authors: Lopez Diego A , Griffin Aleah , Beaudin Anna E 
Published: January 2023

Abstract
We have recently shown that prenatal inflammation induced by maternal immune activation (MIA) imparts lasting changes to innate immunity by driving the inappropriate expansion and persistence of lymphoid-biased progenitors during fetal life. To test how perturbation during this critical window drives immune dysfunction, we examined underlying changes to lung type-2 innate lymphoid cells (ILC2s) and susceptibility to airway hyperresponsiveness following MIA. MIA induction via a single-low dose injection of poly (I:C) at mid-gestation altered the establishment of a fetal liver (FL) ILC-committed progenitor, increasing cell number and frequency both one- and three-days post MIA. MIA also enhanced lung ILC2 proliferation postnatally, resulting in a robustly expanded ILC2 compartment at postnatal day (P)14 that persisted into adulthood. Concomitant with a greater inflammatory profile, MIA-treated ILC2s were hyperactivated, producing more IL5 and IL13 in-vitro. Lung cytokine levels remained relatively static across early lung development in MIA or saline conditions, suggesting changes to ILC2 proliferation and hyperactivation are cell-intrinsic. Furthermore, hyperactivated ILC2s led to remodeling of the lung immune landscape as observed by expansion of B-, T-, NK-, NKT-cells, alveolar macrophages and eosinophils, but ILC2s were the first immune population to expand in cell number. Altogether, our data suggest MIA imparts lasting changes to lung immunity by perturbing the establishment of committed ILC progenitors in the FL, altering the production of neonatal lung ILC2s poised to hyperactively respond to secondary immune activation, potentially contributing to greater allergic airway hyperresponsiveness.
Supported by grants from NIH/NICHD (T32HD007491
Differential Response of Dendritic Cells and Macrophages to Signals from Airway Epithelial Cells in Humans: Changes with Age
Authors: Agrawal Anshu , Agrawal Sudhanshu , Monteiro Clarice 
Published: January 2023

Abstract
Advancing age significantly enhances the risk of respiratory diseases such as COPD, fibrosis etc. Immune response to respiratory viral infections is also impaired often leading to hospitalization. The underlying mechanisms are not well understood. Airway epithelial cells (AECs) and dendritic cells (DCs) and alveolar macrophages are the primary cells involved in respiratory immunity and tolerance. Since DCs and macrophages exist in close proximity to AECs, the function of both these cells is influenced by AECs. 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) from the circulation at homeostasis. Culture of mDCs with AECs resulted in the upregulation of pathogen recognition receptors (PRRs) and downstream signaling molecules. In contrast to circulatory mDCs, culture of macrophages with AECs led to a significant inhibition of LPS induced pro-inflammatory responses. More importantly, AECs induced the secretion of TGF-β and IL-10 from macrophages even in the absence of LPS stimulation. Thus, it seems AECs induce tolerance in macrophages but enhance the immunogenicity of mDCs suggesting a division of labor between DCs and macrophages in the respiratory mucosa. Investigation into the possible mechanisms for this differential response indicates that metabolites and growth factors from AECs may be responsible for inducing tolerance in macrophages and priming DCs. We also find that the crosstalk between AECs and macrophages and DCs is altered with age which may contribute to the increased susceptibility of elderly to respiratory diseases and infections.
This study was supported by grant from the NIH AG045216 (to AA), and from the National Center for Research Resources and the National Center for Advancing Translational Sciences # UL1 TR000153.
Tobacco smoke activated fibrogenic MARCKS/AXL complex promotes macrophage reprogramming and pulmonary fibrosis
Authors: Yang David Cheng , Zhang Jun , Li Ji-Min , Chu Chih-Wei , Hsu Ssu-Wei , Chen Ching-Hsien 
Published: January 2023

Abstract
Macrophages and tobacco smoke (TS) exposure have been demonstrated to play significant roles in modulating pulmonary fibrosis (PF). However, the mechanisms of how TS exposure modulates pro-fibrotic macrophage polarization and drives lung fibrosis is unclear. In our study, we investigated how TS modulates macrophage polarization and the functional consequences of this polarization. Multicolor flow cytometric data indicated that markers of M2 macrophage polarization were elevated in both human and mouse macrophage cells and tissues upon TS exposure. In addition, multiple primary lung fibroblast cells demonstrated elevated pro-fibrotic markers and aggressive phenotypes upon interacting with TS-exposed macrophage cells in a co-culture system. Elucidation of the signaling pathways activated by TS exposure through a receptor tyrosine kinase array screen revealed AXL receptor as a novel smoke-responsive molecule in macrophage cells. We noted elevated secretion of AXL ligand, Gas6, and AXL activity in TS exposed cells and tissues. Prior work had demonstrated an interaction between MARCKS, a smoke-responsive protein, and AXL in promoting a pro-fibrotic phenotype in lung fibroblasts. Similarly, we observed AXL activity positively correlated with MARCKS phosphorylation in macrophage cells. Pharmacologic and genetic targeting of the MARCKS/AXL signaling complex reduced M2 markers and profibrotic cytokine production in macrophages and reduced fibrotic changes in the co-culture model and in an animal model of smoke-mediated lung fibrosis. In all, our work suggests that the MARCKS/AXL fibrogenic complex is a potential target in attenuating macrophage activity in TS-mediated fibrosis.
Supported by grants from NIH/NHLBI (R01HL146802), DOD DCMRP/PRMRP (PR202411), and UCOP TRDRP (28IR-0061 and T31DT1849).
The role of CD45 in the regulation of B cell subset functions in Atherosclerosis
Authors: Ma Shelby , Moriarty Alina K , Keeter William Coles , Waseem Tayab , Zyskin Alexander , Gauronskas Philip , Mussbacher Marion , Galkina Elena 
Published: January 2023

Abstract
Atherosclerosis is an inflammatory disease of the large and medium size arteries characterized by deposition of oxidized lipids within the vessel. While specific self-antigens are not well characterized, data show that the immune response is involved in atherogenesis. The role of B cells in atherosclerosis is subset specific. FO B cells are proatherogenic, while B1 and MZ B cells serve as protective. B cell receptor signaling is important in B cell activation, differentiation, and functions, while important costimulatory molecules, like CD45, assist in initiating signal propagation. To date, it is unknown whether B cell subsets respond differently to BCR signaling or if CD45-dependent regulation of BCR signaling is subset specific in homeostatic or atherosclerotic conditions. Taking advantage of transgenic mice that express low levels of CD45(CD45L/L), we examined activation of CD45L/L B cells through Ca2+ flux. We show that BCR-induced B cell activation differs between subsets and CD45-dependent modulation of BCR signaling is subset specific. MZ and B1 have high levels of Ca2+ flux, with MZ B cell activation the most affected by CD45 expression. To test the effects of CD45L/L B in atherosclerosis, we performed adoptive transfer of CD45L/L or WT B cells into B cell-deficient atherosclerotic prone mice. After 23–38 wks of western diet feeding, CD45L/L B cell recipients had increased lesion formation despite an increase of protective MZ B cells. This increase in atherogenesis could be due to MZ CD45L/L B cells having reduced activation, disrupting their protective ability. This suggests that the subset-specific modulation of BCR signaling through CD45 plays a role in the regulation of B cell functions in atherosclerosis.
This work was supported by American Heart Association AIREA grant 8AIREA33960546.
Multispectral imaging to detect immune phenotypes associated with the tumor microenvironment in a multi-tissue study: A full automated 7-color mIF immuno-oncology workflow
Authors: Mehra Navi , Patel Bhavika , Allen Stephanie , Boldt Brittney , Ramirez Noah , Mammadova Najiba , Haggerty Agnes 
Published: January 2023

Abstract
Immunotherapy and precision medicine are rapidly developing approaches to cancer therapy. Biomarkers that detect the tumor and tumor microenvironment allow for the development of strategies that accelerate the advancement of treatments to enhance a patient’s immune system. Akoya’s MOTiF™ PD-1/PD-L1 Panel is a validated, multiplex immunoassay enabling detection of the 6 most clinically relevant immuno-oncology and spatial biomarkers: PD-1, PD-L1, FoxP3, CD8, CD68, and PanCK. The MOTiF™ PD-1/PD-L1 Panel was used to analyze the tumor microenvironment and specifically assess immune phenotypes of different types of cancers: non-small cell lung cancer (NSCLC), colon adenocarcinoma, head and neck squamous cell carcinoma (HNSCC), and breast cancer. We demonstrate the utility of Akoya’s MOTiF™ PD-1/PD-L1 panel kit in studying the cellular diversity of various cancers while retaining spatial context. Stained slides were analyzed using the InForm® and PhenoptrReports image analysis platforms to identify and better understand spatial relationships between a variety of simple and complex cell phenotypes. The MOTiF™ PD-1/PD-L1 panel kit provides reproducibility across different tissue types. These data provide insight into the innate and adaptive immune environment for targeted design of new immunotherapies and have implications for improving the treatment paradigm across many tumor types.
Synthetically glycosylated antigens as an inverse vaccine platform to prevent and treat food allergies
Authors: Cao Shijie , Maulloo Chitavi D , Sabados Matthew , Raczy Michal M , Wilson D Scott , Nagler Cathryn R , Hubbell Jeffrey A 
Published: January 2023

Abstract
The only FDA-approved oral immunotherapy for a food allergy provides protection against accidental exposure to peanuts. However, this therapy often causes discomfort or side effects and requires long-term commitment. Better preventive and therapeutic solutions are urgently needed. We have developed a technology for inverse vaccination using glyco-polymerized antigens to induce antigen-specific non-responsiveness. We have demonstrated that following subcutaneous (SC) administration, glyco-polymer conjugates traffic to draining lymph nodes (LNs) and are preferentially internalized by antigen presenting cells, educating the immune system to respond to these antigens in an innocuous way. Here, we tested SC administration of glyco-polymerized β-lactoglobulin (BLG) in a murine model of cow’s milk allergy. Two doses of glyco-polymerized BLG given one week apart (prior to sensitization) prevented an allergic response to BLG upon intragastric challenge as measured by reductions in BLG-specific IgG, IgG1, and IgE production as well as BLG-specific type 2 T helper cell (Th2) responses in vitro. We further explored the therapeutic potential of glyco-polymerized BLG. Unlike unmodified BLG, two SC injections of glyco-polymerized BLG after sensitization did not cause anaphylactic reactions in allergic mice and inhibited cellular Th2 cytokines upon BLG restimulation. Since pre-existing humoral immunity inhibited the therapeutic effect of our glyco-polymerized BLG, we introduced a co-therapy (anti-CD20) to blunt the humoral response and observed ameliorated anaphylactic responses following glyco-polymerized BLG therapy. This platform may provide a potential T cell-modulating strategy to prevent and treat food allergies.
This work was supported in part by seed funding from the Chicago Immunoengineering Innovation Center at the University of Chicago as well as the Food Allergy Fund.
Defining a functionally distinct, lineage dependent B1 cell population in Nippostrongylus brasiliensis infection
Authors: Zellner Matthew , Lownik Joseph Cornelius , Urban Joseph F , Martin Rebecca 
Published: January 2023

Abstract
Nippostrongylus brasiliensis (Nb) is the strongest natural inducer of IgE. Despite this fact, asthma rates have an inverse correlation in countries endemic with parasitic infection. Our lab as well as others have sought to explain this phenomenon with little success. The mechanism behind this relationship is complicated by the fact that two different lineages of B cells, that are difficult to distinguish, are capable of making IgE. These B cell populations, B1 cells and B2 cells, have been classically separated using cell surface markers, but our data shows that these markers do not truly define these populations. Our lab has previously shown that IgE, using classical separation of these B cells, have functional differences in a Nb response. Current work shows that these populations have increased plasticity in a location dependent manner. High dimensional flow cytometry analysis of B cell populations, before and after Nb infection, illustrate this phenomenon. Using a lineage specific mouse model, reconstituted with fetal derived B cells that secrete FLAG-tagged IgE, we demonstrate that fetal derived B1 cells are not responsible for IgE production in Nb infection. However, hematopoietic cells expressing B1 markers are a significant source for this IgE. These data support a functionally distinct B cell population that is hematopoietic derived and has been previously classified as B1 cells.
Supported by grants from the NIH-NIAID (R56 AI139658-01AI)