bluuebunny/crossref_metadata_2025 huggingface dataset:
Antitumor effects of anti-CD137 mAb are influenced by Fc: Fcγ receptor interactions (156.5)
Authors: Sallin Michelle , Lin Wei , Chapoval Andrei , Schulze Dan , Strome Scott
Published: January 2023

In various preclinical models, agonist antibodies directed against CD137 (4-1BB) enhance the antitumor immunity. However, the contribution of the Fc:Fc receptor interactions to the process of anti-CD137 mediated tumor rejection is unknown. In this study, we provide early evidence that treatment of tumor bearing mice lacking all activating Fcγ receptors (Fcγ chain deficient mice) with the anti-CD137 mAb, 4-1BB2A, improves the survival compared to wild type mice. When Fcγ chain deficient mice and wild type are challenged with EL4E7 tumor cells, tumors were allowed to grow to an approximate size of 10mm by day 7 or 12mm by day 10. When the mice are treated with anti-CD137 mAb, the Fcγ chain deficient mice had increased survival compared to wild type mice even bearing ten day established tumors. Similarly, improved survival is seen in FcγRIII deficient mice bearing ten day established tumors after treatment with anti-CD137 mAb. While still preliminary, our observations suggest that strategies to modify Fc:FcR interactions may improve the therapeutic efficacy of anti-CD137 based cancer immunotherapy.
Muramyl dipeptide potentiates the ability of lipoteichoic acid to induce cyclooxygenase-2 expression in murine macrophages (157.11)
Authors: Ahn Kibum , Jeon Jun Ho , Park Ok-Jin , Yun Cheol-Heui , Han Seung-Hyun
Published: January 2023

Gram-positive bacterial cell wall contains lipoteichoic acid (LTA) and thick peptidoglycan (PGN) layers, both of which are the major virulence factors containing proinflammatory potential. In order to examine if LTA together with PGN synergistically induces inflammatory responses, we investigated the ability of staphylococcal LTA to induce cyclooxygenase-2 (COX-2) expression in the presence of muramyl dipeptide (MDP), the minimal structural unit of PGN with inflammatory potential, in murine macrophages. Although MDP failed to induce COX-2 expression, LTA alone was able to induce COX-2 expression and MDP remarkably enhanced LTA-induced COX-2 expression at both protein and mRNA levels. Up-regulation was also observed in the production of prostaglandin E2, which is mediated by COX-2. Notably, COX-2 expression, induced by LTA plus MDP treatment, was down-regulated by inhibitors for MAP kinases (ERK, p38 and JNK/SAPK), PI3 kinases, and reactive oxygen species, but not by inhibitors for PKC or PKA. Transient transfection and luciferase reporter gene assays demonstrated that MDP also enhanced the LTA-induced activation of transcription factors, NF-κB, CRE and AP-1, all of which are involved in the transcriptional activity of COX-2. Conclusively, these results suggest that MDP potentiates LTA-induced inflammatory responses by enhancing the expression of COX-2 in macrophages.
A novel mechanism for inhibition of IRF-3-dependent gene expression by Human Respiratory Syncytial Virus NS1 protein (158.19)
Authors: Bao Xiaoyong , Ren Junping , Liu Tianshuang , Garofalo Roberto , Casola Antonella
Published: January 2023

Human respiratory syncytial virus (RSV), a leading cause of respiratory tract infections in infants, inhibits type I interferon (IFN)-dependent signaling, as well as IFN synthesis. RSV nonstructural protein NS1 plays a significant role in this inhibition, however the mechanism(s) responsible are not fully known. The transcription factor interferon regulatory factor (IRF)-3 is essential for viral-induced IFN-β synthesis. In this study, we found that NS1 protein inhibits IRF-3-dependent gene transcription in constitutively active IRF-3 overexpressing cells, demonstrating that NS1 directly targets IRF-3. NS1 associates with IRF-3 and inhibits IFN-β transcription by blocking IRF-3 binding to the IFN-β promoter, without affecting viral-induced IRF-3 phosphorylation and nuclear translocation, thereby identifying a novel molecular mechanism by which RSV inhibits IFN-β synthesis.
Regulation of human paramyxovirus pathogenesis by pulmonary dendritic cells (158.4)
Authors: Guerrero-Plata Antonieta , Wakamatsu Nobuko , Lancelin Wallissa
Published: January 2023

Human metapneumovirus (hMPV), is an important paramyxovirus that is associated with clinical syndromes ranging from mild illnesses, such as common cold, to more devastating conditions, such as bronchiolitis and pneumonia. hMPV has worldwide distribution and high incidence in children, elderly and immunocopromised patients. Understanding the immune mechanisms that regulate hMPV pathogenesis is important clinically for the development of new treatment strategies or novel vaccines for hMPV infection. Respiratory tract dendritic cells (DC) play a fundamental role in the immune system by bridging both innate and adaptive immunity. Therefore, we examined the role of plasmacytoid DC (pDC) in hMPV pathogenesis in an experimental mouse model of infection. Depletion of pDC resulted in an increased viral replication in the lungs from 4.3±1.3×104 to 1.7±6.1×105. However, no change in IFN-α/β production was observed. In addition, pDC-depleted mice demonstrated an exacerbated mucus production and pulmonary inflammation (>60%). Interestingly, the balance of lung DC was altered by the depletion of pDC as the recruitment of classical DC (cDC) into the lungs of infected mice was significantly incremented from 9.9±1.2×105 to 1.7±1.6×106. Finally, the production of CCL20, a DC-attracting chemokine, was also increased after depletion of pDC (>60%). In conclusion, these novel findings demonstrate that pDC play a protective role during hMPV infection by modulation of local immune responses.
Maintenance of peripheral naive T cells: a mouse-man divide (159.1)
Authors: Vrisekoop Nienke , Den Braber Ineke , Mugwagwa Tendai , Westera Liset , Mogling Ramona , De Boer Anne Bregje , Willems Neeltje , Schrijver Elise , Spierenburg Gerrit , Gaiser Koos , Mul Erik , Otto Sigrid , Ruiter An , Ackermans Mariette , Miedema Frank , Borghans Jose , De Boer Rob , Tesselaar Kiki
Published: January 2023

Insights from T-cell dynamic studies in mice are widely extrapolated to men and vice versa, while no formal proof exists that such extrapolations are justified. At the same time, there is a lot of controversy about the dynamic properties of T cells, including life span and the relative contributions of thymus output and peripheral proliferation to the maintenance of the naive T-cell pool during aging. We have investigated the hypothesis that part of this controversy is due to improper extrapolation of dynamic parameters of T cells from mice to men. By combining in vivo kinetic labeling using deuterated water, thymectomy experiments, analysis of TRECs, the expression of the thymic proximity marker CD31, and mathematical modeling, we have quantified the contributions of de novo T-cell production by the thymus and peripheral naive T-cell division to life-long maintenance of the naive T-cell pool in both mice and men under normal physiological conditions. Our results show that mice and men are incomparable with respect to naive T-cell maintenance. While in mice - throughout their life time - the naive T-cell pool is almost exclusively sustained by thymus output, the maintenance of the adult human naive T-cell pool occurs almost exclusively through peripheral T-cell division. These findings put serious constraints on the use of mouse experiments to understand immunological processes such as T-cell reconstitution in lymphopenic patients or the effects of aging in healthy humans.
B cell autonomous function of Bim in B cell apoptosis and tolerance (159.10)
Authors: Wright Jacqueline , Castro Iris , Zinkel Sandra , Khan Wasif
Published: January 2023

Splenic immature B cells traverse through the early transitional (T1) and late transitional (T2) stages to produce the mature B cell repertoire. BCR specificity-based selection ensures elimination of pathogenic B cells by apoptosis, receptor editing or anergy. These processes are tightly regulated by anti- and pro-apoptotic members of the Bcl-2 family. Conventional gene deletion of the pro-apoptotic member Bim lead to prolonged life-span of autoreactive B cells, defective anergy and production of autoantibodies. Whether these pathophysiological conditions arise as a result of B cell autonomous function of Bim remains unclear. We have found that T1 cells express higher levels of Bim and display increased susceptibility to BCR-induced apoptosis than more mature B cells. These properties are shared by BCR signaling defective Bruton’s tyrosine kinase (Btk) deficient B cells. We investigated the B cell-intrinsic role of Bim in the regulation of B cell maturation and survival in mice carrying floxed Bim and CD19-Cre in Btk-sufficient or deficient background. B lineage-specific deletion of Bim increased the B cell compartment through selective expansion of mature B cells and significantly restored mature B cells in Btk-mutant mice. These studies demonstrate a B cell autonomous role for Bim in BCR-dependent regulation of B cell homeostasis. Analysis of these mice in the context of BCR transgene will reveal the B cell autonomous function of Bim in tolerance and autoimmunity.
Role of antigen persistence and dose for CD4+ T-cell exhaustion and recovery (159.7)
Authors: Obst Reinhard , Han Shaobo , Asoyan Ayuna , Rabenstein Hannah , Nakano Naoko
Published: January 2023

It is currently not understood how some chronic infections exhaust antigen-specific T cells over time and how different pathogen components contribute to the exhausted state. Here, we dissected the behavior of primed CD4+ T cells exposed to persistent antigen using an inducible transgenic mouse system that allowed us to control antigen presentation as the only experimental variable independent of persistent inflammation and disease progression. In this system presentation of an MHC-II presented antigen is restricted to dendritic cells so that confounding B, CD8+ T, or innate cell responses are not triggered. When antigen presentation was extended beyond the expansion phase, primed CD4+ T cells survived, but exhibited reduced memory functionality in terms of their proliferative capacity and cytokine expression potential. The effect was antigen dose and time dependent, not associated with increased PD-1 expression or reduced calcium influx, but with impaired Jun phosphorylation in response to TCR engagement. Upon antigen removal, the cells regained the ability to proliferate, but remained unable to produce high levels of IL-2 and TNF-α. These data show that persistent antigen at a high dose rapidly induces a dysfunctional state in CD4+ T cells that is only partially reversible upon antigen removal. At a low dose, however, antigen has to persist longer before T cell impairment becomes detectable.
Oral L. monocytogenes infection preferentially imprints memory precursor CD8 T cells for gut retention (161.10)
Authors: Sheridan Brian , Pham Quynh-Mai , Lee Young-Tae , Cauley Linda , Lefrançois Leo
Published: January 2023

The intestinal mucosa contains unique sites designed for antigen presentation and promotion of primary and memory CD8 T cell responses. Thus, generation of adaptive immunity within distinct tissues likely results in T cell responses with specific cellular outcomes. Here, we utilized an oral Listeria monocytogenes (LM) infection model in which internalin A is modified to promote invasion of murine intestinal epithelial cells. Oral infection induced a robust CD8 T cell response. In peripheral nonmucosal tissues, the majority of LM-specific CD8 T cells are short-lived effector cells (CD127- KLRG1+). Surprisingly, the majority of effector CD8 T cells in the intestinal mucosa are memory precursor effector cells (MPEC; CD127+ KLRG1-) early following oral infection. This effect appears to be due to preferential infiltration and retention of MPECs in intestinal tissues. CD103 (integrin αE), which is thought to be vital for retention of intestinal epithelial lymphocytes within the epithelium, was almost exclusively expressed by MPECs. The generation of distinct CD8 T cell subsets uniquely tailored to respond to intestinal pathogens early following infection may be a hallmark of oral challenge. Indeed, CD8 T cells that infiltrate the intestinal mucosa following intranasal influenza virus infection are early effector cells (CD127- KLRG1-), but failed to be maintained long-term, demonstrating the importance of intestinal priming for generating robust mucosal memory CD8 T cells.
Global gene expression analysis of Act1’s effects in airway epithelial cells (161.17)
Authors: Velichko Sharlene , Anderson Johnathon , Ryan Stephanie , Wu Reen
Published: January 2023

Act1/CIKS is an intracellular protein that has been shown to play an important role in mediating IL-17A and IL-25 signaling effects. Recently, defects in Act1 function and/or expression has been implicated in inflammatory disease, such as psoriatic arthritis and atopic dermatitis. We have found that the modulation of Act1 expression levels in human airway epithelial cells changes the expression levels of some genes, in the absence of cytokine stimulation. RNAseq is a powerful new technique to quantitatively measure changes at the transcriptome level. Here we describe the use of RNAseq to globally analyze the effects of modulating Act1 expression in human airway epithelial cells.
MyD88-dependent TLR signals are necessary and sufficient to confer dendritic cells with gut-specific imprinting properties. (161.2)
Authors: Wang Sen , Villablanca Eduardo , Gomes Daniel , Nguyen Deanna , Mizoguchi Emiko , Kagan Jonathan , Reinecker Hans-Christian , Hacohen Nir , Nagler Cathryn , Xavier Ramnik , Rossi-Bergmann Bartira , Chen Yi-Bin , Blomhoff Rune , Snapper Scott , Mora Rodrigo
Published: January 2023

Lymphocyte migration is at the heart of normal and pathological immune responses in the intestinal mucosa. We and others have shown that, in addition to activating lymphocytes, gut-associated dendritic cells (GALT-DC) can metabolize dietary vitamin A into all-trans retinoic acid (RA), which is required for inducing gut-tropic lymphocytes and IgA antibody-secreting cells (IgA-ASC). GALT-DC from mice deficient in the intracellular signaling adaptor MyD88, which lack most Toll-like receptor (TLR) signals, were significantly impaired in their capacity to induce gut-homing T cells and IgA-ASC and expressed low levels of retinal dehydrogenases (RALDH), which are critical enzymes for RA biosynthesis. MyD88-/- mice were also impaired in generating gut-tropic T cells upon immunization and exhibited low numbers of intestinal IgA-ASC. Pre-treatment of murine spleen-DC and human monocyte-derived DC with a TLR1/2 agonist was sufficient to induce RALDH and to confer these extra-intestinal DC with the capacity to induce gut-homing lymphocytes and IgA-ASC via a mechanism dependent on MyD88 and c-JNK/MAPK. Thus, MyD88-dependent TLR signals are both necessary and sufficient to educate DC with gut-specific imprinting properties.
Secretory IgA: coordinating the capture and sampling of enteric pathogens (161.20)
Authors: Mantis Nicholas , Forbes Stephen , Amarasinghe Jayaleka , Reid Richelle , Park Chae Gyu , Corthesy Blaise
Published: January 2023

Secretory IgA (SIgA) antibodies directed against the O-antigen (O-Ag) component of lipopolysaccharide are the primary determinants of mucosal immunity to enteric bacterial pathogens, such as Shigella flexneri and Salmonella enteric serovar Typhimurium. In this report, we demonstrate that monoclonal IgA Abs against the O-Ag are effective inhibitors of the bacterial type 3 secretion systems (T3SS) required for bacterial entry into intestinal epithelial cells. While these Abs did not affect bacterial viability, they did transiently de-energize both S. flexneri and S. Typhimurium, possibly accounting for the “paralysis” that has been observed in the case of S. Typhimurium, following Ab treatment. O-Ag specific IgA Abs also elicited long lasting ultrastructural changes in the bacterial outer membranes, suggesting that S. flexneri and S. Typhimurium made undergo specific adaptations in response to SIgA. These findings may have implications for understanding immune surveillance in the gut. We have shown that Peyer’s patch M cells transport SIgA and SIgA-antigen complexes from the intestinal lumen to underlying dendritic cells (DC), and that the uptake of SIgA-immune complexes by DCs may be mediated by the mannose receptor and/or DC-¬SIGN. The fact that SIgA may be capable of transiently rendering S. flexneri and S. Typhimurium avirulent suggests a mechanism by which the normally pathogenic bacteria can be sampled safely by the mucosal immune system.
TLR2-mediated IL-27 and chemokine production by respiratory epithelial cells promotes bleomycin-induced pulmonary fibrosis by inhibiting IL-17 production and recruiting immune cells, respectively (162.7)
Authors: Kim Hye sung , Chung Doo Hyun
Published: January 2023

Idiopathic pulmonary fibrosis (IPF) is a chronic disease that is characterized by destruction of the lung parenchyma. Toll-like receptor 2(TLR2) is a key innate mediator by sensing tissue injury. Recently reported that TLR2 promote bleomycin-induced pulmonary fibrosis(BIPF), but the mechanism remains unclear. To address this, we compared TLR2 knockout(KO) and wild type(WT) mice in the BIPF model. TLR2 KO mice attenuated BIPF in terms of inflammation, and hydroxyproline levels. The levels of IL-17 and-producing CD4+ and γδ T cells in bronchoalveolar lavage fluid of TLR2 KO mice were higher than those of WT mice. Moreover, IL-17 blockade in TLR2 KO mice increased the hydroxyproline and TGF-β levels. Using Bone-marrow chimeric mice revealed that TLR2 expressed on non-hematopoietic cells rather than immune cells reduced IL-17 production but enhanced the levels of hydroxyproline. Moreover, upon stimulation with TLR2 ligands, respiratory epithelial cells from WT mice produced IL-27, CC, and CXC chemokines, depending on IRF complex and MyD88. Blockade of IL-27 in WT mice enhanced IL-17 production, resulting in reducing amounts of hydroxyproline, whereas injection of chemokine cocktail to TLR2 KO mice enhanced recruitment of inflammatory cells into the lungs. Taken together, TLR-2 mediated production of IL-27 and chemokines by respiratory epithelial cells inhibits IL-17 production and recruits inflammatory cells into the lungs, respectively, which results in promoting BIPF.
Notch signaling promotes MUC5AC expression through epidermal growth factor receptor-extracellular regulated kinase pathway (163.1)
Authors: Kim Su Hwan , Kang Jin Hyun , Uhm Tae Ki , Kim Do Jin , Chung Il Yup
Published: January 2023

Airway mucus hypersecretion and goblet cell hyperplasia plays an important role in the pathogenesis of several major airway inflammatory diseases. MUC5AC is a major component of the mucus produced by airway epithelial cells and considered to be a marker of mucus cell hyperplasia. In the study we investigate the involvement of Notch signaling in EGF-mediated expression of MUC5AC in human epithelial NCI-H292 cells. EGF stimulated the generation of Notch intracellular domain (NICD) in a RBP-Jκ-dependent manner. Treatment with γ-secretase inhibitor (GSI) or introduction of small interfering RNA directed against Notch1 reduced EGF-induced expression of MUC5AC mRNA and protein. The inhibitory effect of both GSI and Notch1 silencing on MUC5AC expression was accompanied by reduced generation of NICD and reduced expression of a Notch downstream target, Hes-1. Blockage of the Notch signaling with the two agents resulted in a decrease in ERK phosphorylation induced by EGF stimulation. ERK inhibitor inhibited MUC5AC production. These results indicate that ERK activation is necessary for Notch signaling to regulate the EGFR-mediated MUC5AC expression. Collectively, these results suggest that Notch signaling regulates the EGF-induced expression of MUC5AC through modulation of ERK activity.
Genetic analysis of arginase 1 in Th2-dominated lung inflammation (163.12)
Authors: Barron Luke , Smith Amber , El Kasmi Karim , Qualls Joseph , Huang Xiaozhu , Wynn Thomas , Murray Peter
Published: January 2023

Arginase 1 (Arg1) expression in myeloid lineage cells, and especially alternatively-activated macrophages, increases dramatically in the lungs of mice with acute and chronic Th2-driven inflammation. Arg1 has been proposed to contribute to asthma pathogenesis by regulating at least four pathways: inhibiting the production of nitric oxide by substrate competition, regulating fibrosis and polyamine supply, regulating the bioavailability of arginine in the lung, and inhibiting T cell proliferation. Here we used mice lacking Arg1 in myeloid lineage cells, to investigate the contribution of Arg1 to asthma pathophysiology and lung inflammation. In six distinct model systems encompassing acute and chronic Th2-mediated inflammation, we observed neither a pathogenic nor protective role for myeloid-expressed Arg1 in any measured parameter of lung pathophysiology. Collectively, our results argue that (i) attempts to inhibit arginases in the lung by small molecule arginase inhibitors or RNAi should be examined with caution and (ii) whether macrophages use Arg1 to limit Th2 responses depends on the organ and type of infection or injury.
Cys-leukotrienes mediate lung dysfunction but not histopathologies in an IL-5/Eotaxin-2 double transgenic mouse model of severe asthma (163.13)
Authors: Ochkur Sergei , Protheroe Cheryl , Li Wen , Colbert Dana , Zellner Katie , Lee James , Lee Nancy
Published: January 2023

Products of the 5-lipoxygenase (5-LO) pathway (i.e., cys-leukotrienes and LTB4), are contributors to lung inflammation, airway remodeling, and hyperresponsiveness associated with allergic respiratory inflammation. Eosinophils are capable of leukotriene synthesis/secretion and chemotactically respond to these compounds. However, the in vivo role of eosinophils in leukotriene mediated events during asthmatic airway inflammation is not defined. To study the link between eosinophils and leukotrienes we used the I5/E2 double transgenic mouse model of severe asthma, over-expressing IL-5 systemically and eotaxin-2 in airways, in which lung pathologies are entirely dependent on the presence of eosinophils. We show that airway levels of cysteinyl leukotrienes and LTB4 are significantly elevated in I5/E2 mice. Crosses of I5E2 mice with 5-LO knockout animals abolished AHR in the I5/E2 parental model without significant effects on lung histopathology. Administration of a cys-leukotriene receptor antagonist (Montelukast®) to I5/E2 mice replicates the effects of the total leukotriene loss in I5/E2/5-LO-/- animals. In contrast, I5/E2 mice crossed with BLT-1-/- mice which are deficient of the LTB4 receptor fail to display effects on the pulmonary pathologies occurring in the I5/E2 parental model. These data demonstrate a unique role for cys-leukotrienes in the development of AHR independent of lung remodeling events occurring in this transgenic model of severe asthma.
TSLP augments Th9-mediated allergic inflammation (163.23)
Authors: Yao Weiguo , Zhang Yanlu , Kaplan Mark , Zhou Baohua
Published: January 2023

Thymic stromal lymphopoietin (TSLP) is an epithelial cell derived cytokine important for the initiation and development of Th2-driven allergic inflammation in asthma and atopic dermatitis. The role of TSLP in Th9-mediated inflammation has not been identified. In this study, we investigated the role of TSLP on Th9 differentiation and IL-9 production both in vitro and in vivo. Our data showed that Th9 cells expressed greater amounts of TSLP receptor than Th2 cells, and addition of TSLP to Th9 cultures increased IL-9 production in vitro. Lung-specific transgenic expression of TSLP stimulated IL-9 production in BALF and lung. To further characterize the role of TSLP on Th9-mediated allergic inflammation in vivo, we transferred OVA-differentiated Th2 or Th9 DO11.10 CD4+ T cells to wild type recipient followed by OVA or OVA+TSLP challenge. Our data showed that challenge with OVA+TSLP, but not with OVA alone increased IL-9 production and allergic inflammation in mice injected with Th9 cells, with less effect on Th2 cells. Therefore, Th9 cells can mediate TSLP-mediated exacerbation of allergic airway inflammation. These results suggest that, while both Th2 and Th9 promote allergic inflammation, Th9 cells along are sufficient to respond to TSLP for increased allergic inflammation. Our findings provide a potential role of TSLP on the development of Th9-mediated allergic inflammation.
The role of the keratinocyte in chemical-induced adaptive immune responses (163.4)
Authors: Summerfield Vicki , Maxwell Gavin , White Andrew , Chain Benjamin
Published: January 2023

Allergic contact dermatitis results from the induction of an adaptive immune response to a chemical allergen following repeated skin exposure. The initiation of this response requires the production of an inflammatory microenvironment, created in part by the response of epidermal keratinocytes to chemical exposure. The molecular pathways that regulate inflammatory mediator release and the function of keratinocyte-derived mediators in dendritic cell (DC) migration and maturation have not yet been fully characterised. We have established that HaCaT immortalised human keratinocytes (our experimental model for epidermal keratinocytes) transcribe and release IL-6, IL-8, IL-1β and GM-CSF in response to our model skin allergen (1-chloro-2, 4-dinitrobenzene). A global transcriptomic analysis was performed to investigate the molecular mechanisms behind the inflammatory mediator release, which uncovered significant differential expression of 1728 genes, many of which map to components of stress induced pathways such as NF-κB and Nrf-2. Investigation of upstream signal transduction cascades revealed activation of the MAPK pathways, specifically phosphorylation of p38 and JNK, and some evidence of NF-κB translocation. The mechanism by which chemical allergens trigger this response remains unknown, however experiments are ongoing to characterise the role of reactive oxygen species production in disease induction.
Prostaglandin I2-IP signaling promotes the generation of IL-17-producing γδ T cells that associate with the airway epithelium during allergic lung inflammation (163.5)
Authors: Ferrini Maria , Roberts Kevan , Jaffar Zeina
Published: January 2023

γδ T cells rapidly produce cytokines that eliminate pathogens, regulate inflammation and promote homeostatic functions in response to tissue stress and environmental insult. Here, we demonstrated that an innate IL-17-producing γδ T cell response developed in tandem with allergic lung inflammation and that these cells expressed the αEβ7 integrin and are closely associated with the airway epithelium. Although PGI2-IP receptor signaling downregulated allergic pulmonary inflammation, it promoted the emergence of intraepithelial IL-17-expressing γδ T cells in the airways. In keeping with these findings, a pronounced reduction of IL-17-producing γδ T cells was observed in the thymus, as well as lungs, of IP deficient mice implying a key role for PGI2 in programming IL-17 production by natural γδ T cells. Consistently, iloprost, a stable analog of PGI2, augmented IL-17 production by γδ T cells. Moreover, the Vγ4+ subset of γδ T cells in the airways served to limit the allergic inflammation. Overall, these findings reveal that PGI2 plays a key immunoregulatory role by promoting the programming of innate intraepithelial IL-17-producing γδ T cells.
Curcumin attenuates the scurfy-induced immune disorder (164.14)
Authors: Lee Gihyun , Nam Ji-Young , Shin Minkyu , Hong Moochang , Bae Hyunsu
Published: January 2023

The scurfy mutant mouse is the genetic and phenotypic equivalent of the single-gene human autoimmune disease immunodysregulation, polyendocrinopathy, enteropathy, X-linked (IPEX). Scurfy mice have a deletion in the forkhead domain of Foxp3, fail to develop thymic-derived Foxp3+ regulatory T cells, and develop a fatal lymphoproliferative syndrome with multiorgan inflammation. Curcumin(1,7-Bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione) is a naturally occurring polyphenolic phytochemical isolated from the rhizome of the medicinal plant Curcuma longa. It has profound anti-inflammatory activity and been traditionally used to treat inflammatory disorders. Recent studies have shown that curcumin ameliorates multiple sclerosis, rheumatoid arthritis, psoriasis, and inflammatory bowel disease by regulating inflammatory cytokines such as IL-1, IL-6, IL-12, TNF-α and IFN-γ and associated JAK-STAT, AP-1, and NF-κB signaling pathways in human or animal models. In this study we have examined the remedial value of curcumin on the condition of scurfy mice. In vivo treatment with curcumin significantly prolonged survival time and reduced clinical severity. In addition, it decreased inflammatory cytokines and CD4+ T cells proliferation owing to diease. These findings highlight the fact that curcumin inbibits symptoms arose from Foxp3 hypoplasia and suggest its use in the treatment of IPEX.
p38 MAP kinase controls pathogenic T cell responses in experimental allergic encephalomyelitis (164.8)
Authors: Krementsov Dimitry , Noubade Rajkumar , del Rio Roxana , Nagaleekar Viswas , Saligrama Naresha , Sabio Guadalupe , Davis Roger , Rincon Mercedes , Teuscher Cory
Published: January 2023

Multiple sclerosis (MS) is an autoimmune disease characterized by progressive demyelination of the central nervous system (CNS). Therapies with limited efficacy are currently available for MS. The presence of autoreactive CD4 T helper (Th)1 and Th17 cells in the CNS is thought to be important for MS pathogenesis. Here, we show that pharmacological inhibition of p38 MAP kinase (MAPK) by SB203580 (SB) significantly ameliorates clinical EAE elicited by immunization of C57BL/6J mice with MOG35-45 (chronic EAE) and SJL/J mice with PLP135-151 (remitting-relapsing EAE). While SB treatment of actively immunized animals did not prevent immune infiltration of the CNS, it reduced production of IL-17, but not IFNγ by antigen-specific Th cells. Interestingly, when EAE was induced by passive transfer of encephalitogenic Th1-polarized cells, administration of SB to recipients also resulted in prevention of disease, along with reduced CNS infiltration and antigen-specific IFNγ and IL-17 production. Importantly, we show that genetic manipulation of p38 MAPK activity specifically in T cells was sufficient to alter EAE progression and IL-17 production. Taken together, our results indicate that p38 MAPK activation in Th1 and Th17 cells controls their pathogenic functions in autoimmune inflammatory disease of the CNS, suggesting that pharmacological targeting of components of this pathway specifically in T cells may be useful in the treatment of MS.
Promotion of breast cancer development via Th2 polarization by TSLP (165.2)
Authors: Xu Kangling , Pedroza-Gonzalez Alexander , Wu Te-Chia , Aspord Caroline , Shafer Sasha , Marches Florentina , Gallegos Michael , Burton Elizabeth , Hori Toshiyuki , Tanaka Yuetsu , Zurawski Sandra , Zurawski Gerard , Bover Laura , Liu Yong-Jun , Banchereau Jacques , Palucka A. Karolina
Published: January 2023

Inflammation can promote both oncogenesis and tumor rejection. Chronic inflammation promotes cancer cell survival and metastasis, while acute inflammation triggers cancer cell destruction. Chronic inflammation is often linked with the presence of type 2-polarized macrophages (M2), which are induced by TH2 cytokines, IL-4 and IL-13. Human breast cancer tumor microenvironment displays features of Th2 inflammation that promote tumor development. The underlying molecular mechanisms are unknown. Here we show that breast cancer cells produce cytokine TSLP that induces DCs to express OX40L in vitro. OX40L+ DCs were found in primary breast cancer tumor infiltrates. These DCs drive IL- 13+TNFα+IL-10negCD4+ inflammatory Th2 cell development in vitro. TSLP-neutralizing antibodies block the upregulation of OX40L by DCs induced by breast cancer cells, thereby blocking their capacity to induce inflammatory Th2 cells in vitro. Antibodies neutralizing TSLP, OX40L, or TSLPR inhibit tumor development and IL-13 secretion in the xenograft model. Furthermore,PAR2-signaling is involved in TSLP regulation in cancer cells. Thus, TSLP-driven inflammation might contribute to breast cancer pathogenesis and development via Th2 polarization; and TSLP production by cancer cells is regulated via PAR2-signaling.
Sub-lethal irradiation of human carcinoma cells modulates expression of important modulators of effector CTL anti-tumor activity (165.20)
Authors: Garnett Charlie , Ifeadi Victoria
Published: January 2023

Sub-lethal doses of radiation can alter the phenotype of target tissue by modulating gene products that may make tumor cells more susceptible to T-cell-mediated immune attack. Previously, we demonstrated that colorectal cancer lines responded to radiation by up-regulating surface expression of 5 CTL relevant proteins (Fas, ICAM-1, CEA, MUC and MHC-I). The present study was designed to determine the extent of CTL relevant changes induced by radiation in human colorectal carcinoma cells. Here, SW620, HCT116, and Widr cells were examined for their response to various sub-lethal doses of radiation (0-15 Gy). Experiments quantified changes in the expression of genes that could result in enhanced CTL activity against irradiated tumor cells. One to seven d post-irradiation, changes in expression of other death receptors (DR4, DR5, LTβR and TNF-R1), as well as expression of other molecules involved in T cell mediated lysis of target cells (c-FLIP, bcl-xL, and survivin) were examined. All cell lines altered expression of one or more of these molecules post-irradiation. Furthermore, we found that the expression of effector co-stimulatory molecules, such as 41BBL, ICOSL and OX40L, was also altered. Altered expression of these gene products may enhance the tumor cells’ susceptibility to T-cell-mediated immune attack. Overall, the results of this study suggest that non-lethal doses of radiation can be used to make human tumors more amenable to immune system attack.
Innate immunity, but not adaptive immunity, induced by a therapeutic oral vaccine of colorectal cancer is crucial for tumor elimination in APCMin/+ hCEA mice (165.23)
Authors: Zhong Zhenyu , Zhai Yougang , Qiao Liang
Published: January 2023

We developed a colorectal cancer vaccine, comprised of a human CEA gene-containing plasmid packaged inside the bovine papillomavirus like particles (VLP-CEA vaccine). Oral immunization of APCMin/+ hCEA mice (a model of colorectal cancer) with VLP-CEA vaccine greatly reduced the tumor number compared with those received PBS. Surprisingly, CEA-specific adaptive immunities are dispensable for tumor elimination as similar tumor number reductions are seen in the APCMin/+ or APCMin/+ hCEA mice that are immunized with either VLP-CEA or VLP-GFP (VLPs packaging a GFP-containing plasmid). Consistently, we found that CTL responses are not required for tumor eradication since depleting CD8 T cells prior to vaccinations didn’t impair the vaccine efficacy. We confirmed that the hCEA-specific cellular and antibody responses were generated after vaccination with VLP-CEA but not with VLP-GFP. Importantly, we found that VLP-CEA and VLP-GFP are both able to activate the NLRP3 inflammasome which potentially facilitates colorectal tumor eradication. Thus, our data indicate that the innate immunity, but not adaptive immunity, is required to abolish colorectal tumorigenesis, and the understanding of the immune mechanism by which pseudovirus vaccines eliminate colorectal tumorigenesis will further our knowledge for designing vaccines for other cancers.
The role of Type I interferons in antitumor responses: implications in radiation therapy (165.30)
Authors: Lim Joanne , Gerber Scott , Lord Edith
Published: January 2023

Interferon-alpha (IFN-α) is clinically approved for treatment of melanoma. Besides having direct antitumor effects, type I IFNs (IFN-α/β) can also enhance IFN-γ responses in cells. Previously, we found that local treatment with a single high dose of radiation induced an increase of IFN-γ within the tumor microenvironment, and this increase is important for radiation-induced antitumor responses. Since the interplay between IFN-α/β and IFN-γ has been shown to be important in several immunological processes, we hypothesize that the lack of type I IFN signaling will result in poor radiation-induced IFN-γ responses and as a consequence, a lack of tumor control. To test our hypothesis, we first compared the growth of B16 tumors between wild type (WT) mice and IFN-α/β receptor knockout (KO) mice. Interestingly, tumor growth was faster in KO mice than WT mice, suggesting that type I IFNs play a role in antitumor immunity. In mice that were treated with 15Gy radiation, the tumor burden reduction observed in KO mice was comparatively smaller than that in WT mice. Corresponding to lower reduction in tumor growth after radiation treatment, KO mice have limited levels of radiation-induced intratumoral IFN-γ and number of tumor-infiltrating CD8+ T cells compared to that in radiation-treated WT mice. Taken together, our data suggest that type I IFN signaling within host cells is important for antitumor immunity, including IFN-γ-mediated responses induced by radiation treatment.
Role of the AIM2 inflammasome in nasopharyngeal carcinoma (165.35)
Authors: Chen Lih-Chyang , Wang Li-Jei , Tsang Nang-Ming , Ojcius David M. , Chang Yu-Sun
Published: January 2023

Inflammation is a hallmark of cancer, especially in the infection-associated cancer. IL-1β, a major proinflammatory cytokine in tumor milieu, is regulated by inflammasome in response to pathogen and damage. Nasopharyngeal carcinoma (NPC) is a cancer prominent in Taiwanese population and is closely associated with Epstein-Barr virus (EBV) infection. We have examined the AIM2 inflammasome expression in Epstein-Barr virus (EBV)-associated NPC tissues. We found that the AIM2 inflammasome is overexpressed in tumor cells and significantly correlated with better local-recurrence- and disease-free survival of NPC patients after treatment. In addition, the AIM2 inflammasomes was induced by dsDNA in NPC cells, which could be inhibited by caspase inhibitor. The new findings in this study may open a new direction for future therapy or drug development for cancer by inflammation modulation.
Dendritic cell generation from PBMC obtained from jaundiced patients with pancreatic adenocarcinoma — a potential role for STAT-1? (165.48)
Authors: do Carmo Joao Paulo , Treglia Marisa , Jukemura Jose , Matheus Andre , Rodrigues Joaquim Jose , da Cunha Jose Eduardo , Machado Marcel , Bergami-Santos Patricia , Barbuto Jose Alexandre
Published: January 2023

Pancreatic adenocarcinoma (PAdc) is an aggressive malignancy with poor prognosis, urging for new therapeutic strategies. DC-based vaccination is one of such promising approaches since DC are the most potent APCs. However, in cancer patients, DC generation and function may be deficient. Here, we describe the in vitro generation of DC from jaundiced patients with PAdc and also the effect of jaundiced plasma (JP) in DC generation. PBMC from 16 patients and 16 healthy donors were separated and adherent cells were cultured with GM-CSF, IL-4 and TNF-α. Cultures were supplemented with 10% JP or normal plasma (NP). Non-adherent cells were harvested at day 7, labeled and analyzed by flow cytometry. Patients’ cells, cultured in 10% JP, had significantly lower expression of CD11c, CD80, CD86 and HLA-DR. Healthy donors’ cell culture supplementation with JP decreased the percentage of HLA-DR+, CD11c+ and CD86+ cells, but not to the same extent as that of patients’ cells. When patients’ PBMC were cultured in NP, there was a significant increase in HLA-DR and a tendency to increase CD86 expression. Preliminary data suggest that these alterations may be related to low levels of phosphorylated STAT-1 in patients’ DC after activation. Altogether, these data demonstrate an impairment in PAdc patients’ PBMC ability to differentiate into DC in vitro, a phenomenon to be considered when designing DC-based vaccination strategies for these patients.
Multi-epitopic anti-tumor T cell responses and efficient Trastuzumab-mediated ADCC promote a favorable clinical outcome in HER2+ breast cancer patients after neoadjuvant chemotherapy (165.8)
Authors: Dolcetti Riccardo , Martorelli Debora , Muraro Elena , Miolo Gianmaria , Lombardi Davide , Massarut Samuele , Perin Tiziana , Talamini Renato , Turchet Elisa , Comaro Elisa , Carbone Antonino , Veronesi Andrea , Crivellari Diana
Published: January 2023

The ability of drugs such as Trastuzumab and taxanes to act through immunomediated and/or immunomodulatory mechanisms suggest that anti-tumor immune responses may influence the clinical outcome of breast cancer patients. We have enumerated circulating CD8+ T cells specific for a broad spectrum of tumor-associated antigens (her2, muc-1, mammaglobin-A, trag-3, survivin, bcl-xL) in 13 patients with locally advanced breast cancer (6 HER2+ and 7 HER2-). At diagnosis, increased numbers of CD8+ T cells specific for all 13 HLA-A*0201 epitopes were observed in the whole series, with a markedly higher extent in HER2+ cases. This subset of patients retained an immunological profile similar to that of healthy women, whereas HER2- cases showed decreased circulating CD3+ T lymphocytes, higher numbers of regulatory T cells and alterations in serum cytokine levels (decreased IL-2, IL-6, and IL-8). Notably, patients with complete pathologic response (pCR), unlike those showing partial responses, retained high numbers of epitope-specific CD8+ T cells throughout the neoadjuvant chemotherapy (NC) treatment, particularly versus survivin and mammaglobin-A epitopes. Moreover, a higher efficacy of Trastuzumab-mediated ADCC in vitro correlated with the induction of pCR in HER2+ cases. These findings emphasize the crucial role of host anti-tumor T cell responses in promoting a favourable clinical outcome in the setting of neoadjuvant chemotherapy for breast cancer, particularly in HER2+ cases.
The infusion of suicide gene-modified donor T cells after hematopoietic stem cell transplantation prompts thymic renewal in adult patients by an IL-7 dependent mechanism (169.4)
Authors: Oliveira Giacomo , Vago Luca , Noviello Maddalena , Soldati Corrado , Ghio Domenico , Brigida Immacolata , Aiuti Alessandro , Lupo Stanghellini Maria Teresa , Peccatori Jacopo , Bondanza Attilio , Del Maschio Alessandro , Bordignon Claudio , Ciceri Fabio , Bonini Chiara
Published: January 2023

The infusion of donor lymphocytes genetically modified to express the HSV-Thymidine kinase (Tk) suicide gene after T cell-depleted Hematopoietic Stem Cell Transplanation (HSCT) provides protection against pathogens and disease recurrence, while allowing complete GvHD control. In the TK007 phase I/II clinical trial 28 adults with hematologic malignancies received purified Tk-transduced cells after T cell-depleted haploidentical HSCT, and 22 experienced a rapid immune reconstitution. Ten patients had GvHD that was controlled by ganciclovir (Lancet Oncol. 2009). Since the newly reconstituting T cells were mostly Tkneg, we hypothesized that Tk cells, which are necessary to achieve this effect, might act by prompting T cell development from graft progenitors. Patients treated with Tk cells showed prompt recovery of Tkneg naïve T cells. In particular, the CD4+ naïve subset was highly enriched in CD31+ cells, bona fide recent thymic emigrants. Accordingly, after Tk cell add-backs we documented an increase in single joint T cell Receptor Excision Circles counts and in thymic volume, as evidenced by chest CT scans. Serum levels of IL-7 markedly rose after Tk cell add-backs, suggesting that gene-modified cells may act by inducing IL-7 release, in turn supporting thymopoiesis. Taken together, our data show that the infusion of Tk cells boosts the function of adult thymus, prompting the recovery of a polyclonal, fully competent, T cell repertoire by an IL-7 dependent mechanism.
Combination of IL-2 and anti-TGF-β increases natural killer cell reconstitution after hematopoietic stem cell transplantation (169.7)
Authors: Alvarez Maite , Murphy William
Published: January 2023

Despite the efficacy of hematopoietic stem cell transplantation (HSCT) as a cancer therapy, cancer relapse remains a significant complication. Natural killer (NK) cells are the first lymphoid population to recover after HSCT and can kill transformed or virally-infected cells without prior sensitization. Therefore, accelerating NK cell reconstitution may improve HSCT therapy. We utilized the combination of administering the stimulatory cytokine, IL-2 concurrently with the blockade of the immunosuppressive cytokine, transforming growth factor-beta (TGF-β) as a possible therapy to accelerate NK cell reconstitution after syngeneic HSCT. Resting C57BL/6 mice were treated with daily doses of IL-2 (2x105 IU) and anti-TGF-β (120ug, clone 1D11) every other day for one week. Although combinatorial therapy did not significantly expand NK cell numbers in comparison with IL-2 monotherapy, NK cytotoxicity, as measured by the ability to lyse Yac-1 tumor cells, was significantly greater. However in a HSCT model, combinatorial therapy administered in C57BL/6 mice starting at day 11 post-HSCT did significantly increase both the percentage of activated NK cells and NK tumor lytic ability compared with IL-2 alone. Additionally, mice showed no outward evidence of toxicity during administration of therapy. Together, these results suggest that this combination regimen improves reconstitution of NK cells after HSCT which may lead to better anti-viral and anti-tumor responses.
Evolutional conservation of cytosolic antiviral activity with type I interferon mediated by the retinoic acid-inducible gene I-like receptor pathway in teleosts (170.11)
Authors: Hikima Jun-ichi , Ohtani Maki , Kondo Hidehiro , Hirono Ikuo , Jung Tae-Sung , Aoki Takashi
Published: January 2023

The retinoic acid-inducible gene-I-like receptor (RLR) family, one of the pattern recognition receptors (PRRs), is important for the cytosolic immune response to recognize viral RNAs as pathogen-associated molecular patterns (PAMPs). RLR family members MDA5 (melanoma differentiation-associated gene 5) and LGP2 (laboratory of genetics and physiology 2) enhance gene expression of type-I interferon (IFN-I) and inflammatory cytokines through the IPS-1 (mitochondrial adaptor IFN-β promoter stimulator-1) cascade. In this study, MDA5, LGP2 and IPS-1 were cloned from the kidney of the Japanese flounder, Paralichthys olivaceus, and their phylogenetic analyses showed their conservation to those of higher vertebrates. MDA5 and LGP2 mRNA expression were strongly up-regulated by poly I:C stimulation or VHSV infection, whereas IPS-1 mRNA expression was not. To reveal the antiviral activity of these molecules, expression vectors were constructed and used in an antiviral assay against VHSV and HIRRV using the HINAE (Hirame natural embryonic) cell line. Viral replications were visibly repressed by overexpression of the MDA5, LGP2 and IPS-1 genes. The antiviral activities were attributed to the up-regulation of type I interferon (IFN) and IFN-inducible genes (Mx and ISG15). These results indicate that MDA5, LGP2 and IPS-1 are capable of causing the inhibition of viral replication through the induction of type I IFN and IFN-inducible genes.