bluuebunny/crossref_metadata_2025_split huggingface dataset:
The p38 MAPK inhibitor SB203580 abrogates TNF-mediated expansion of regulatory T cells
Authors: Chen Xin , He Tianzhen 
Published: January 2023

Abstract
CD4+FoxP3+ regulatory T cells (Tregs) represent a major cellular mechanism of immune evasion of tumor. There is compelling evidence that TNF is able to activate and expand Tregs through TNFR2. However, it remains unclear which TNFR2 signaling pathway(s) is required for the activation of Tregs. In this study, we determined the identity of the TNFR2 signaling components by using specific small molecule inhibitors. To this end, MACS purified CD4+ T cells from WT C57BL/6 mice were labeled with CFSE and stimulated with TNF in the presence of IL-2. The proliferation of Tregs was analyzed by FACS, gating on FoxP3+cells. Treatment with the p38 MAPK inhibitor SB203580, rather than NF-κB inhibitors, abrogated TNF-mediated expansion of Tregs. In addition, administration of SB203580 blocked LPS-induced expansion of Tregs and up-regulation of TNFR2 expression on Tregs in vivo. Importantly, treatment with SB203580 potently inhibited lung metastasis of mouse 4T1 breast cancer, which presumably resulted from reduction of Treg activity in tumor bearing mice. Therefore, our data suggest that activation of p38 MAPK signaling pathway is required for TNF-mediated proliferative expansion of Tregs. Targeting p38 MAPK pathway may represent a novel strategy to enhance the efficacy of cancer immunotherapy through suppression of Treg activity.
[Supported by the University of Macau Research Grants (MYRG2016-00023-ICMS-QRCM) and a grant from the Science and Technology Development Fund (FDCT); Macao S.A.R. (014/2015/A1)]
Engraftment of Human Immune System and Human Tumor Xenograft in Murine Models: Applications in Oncology Drug Discovery
Authors: Tanaka Azusa , Connerney Mona 
Published: January 2023

Abstract
The development of animal models capable of mimicking human immune responses and function as a host of human tumor xenograft are crucial to study the safety and efficacy of immune checkpoint inhibitors, functionality of CAR-T therapies, and immuno-oncology treatment including gene and cell therapies. Mice reconstituted with human hematopoietic stem cells (HSC) are able to stably differentiate and repopulate human immune cells sufficient to model many aspects of tumor immunity. Taconic’s CIEA NOG mouse ® (NOG) portfolio is one of such powerful tools: capable of accepting human HSC which develops into multiple immune cells including T cells, as well as ideal hosts for human tumor xenografts due to its hyper immune deficiency caused by IL-2Rg mutation and NOD/Scid background.
In partnership with the Central Institute for Experimental Animals (CIEA), Taconic has developed a variety of next generation immune-deficient NOG mice that are suitable and highly effective at modeling the diverse mechanistic functions of human immunity as well as being the host for human tumor xenograft including patient derived xenograft (PDX). This presentation will focus on the current state of human immune system engraftment models (huNOG and huNOG-EXL) and utilization of wide range of NOG mice portfolio as the next generation models that diversify the mechanistic functionality of immune engraftment and recent advances in modeling human immunity in immuno-oncology applications.
MyD88-dependent activation of protein kinase D1 is essential for Group B streptococci-induced proinflammatory responses
Authors: Yi Ae-Kyung , Yoon Tae Won , Upadhyay Kirtikumar , Park Jeoung-Eun , Kim Young-In , Metcalfe Victoria , Halder Priyanka , Talati Ajay J. 
Published: January 2023

Abstract
Group B streptococcus (GBS) causes life-threatening diseases, like pneumonia, septicemia and meningitis, in neonates. It has been reported that the pathogen-associated molecular patterns in GBS are recognized by pattern-recognition receptors, including Toll-like receptors (TLR), in the host innate immune cells. This recognition results in the cascades of inflammatory reactions. However, the molecular basis of GBS-induced proinflammatory responses are not yet completely elucidated. Protein kinase D1 (PKD1) has been identified as one of key signaling intermediaries that plays an indispensable role in proinflammatory responses mediated by TLRs. However, it is currently unknown whether PKD1 is activated, and contributes to proinflammatory responses induced by GBS. In the present study, the role of PKD1 in the proinflammatory responses to GBS is investigated. We found that both live and antibiotic-killed GBS activate PKD1 in human and murine cells. This PKD1 activation is dependent on the TLR signaling adaptor MyD88 and its downstream kinase IRAK1, but not TRAF6. Inhibition of PKD using a pharmacological inhibitor revealed that PKD1 is indispensable for GBS-mediated expression of various cytokines/chemokines in vitro and in vivo. Furthermore, systemic administration of PKD inhibitor protects D-galactosamine-sensitized mice from shock-mediated death caused by antibiotic-killed GBS. Our findings imply that PKD1 is one of the critical factors that play a regulatory role in GBS-induced proinflammatory reactions, and inhibition of PKD1 activation together with antibiotic treatment in GBS-infected neonates might be an effective way to control GBS diseases.
Molecular Basis Of Tick Saliva Proteins Effects On Macrophage Function
Authors: Bakshi Mariam , Mwangi Waithaka , Mulenga Albert 
Published: January 2023

Abstract
Ticks are medically important ectoparasites and transmit many tick borne diseases. Tick feeding and transmission of TBD agents start with the tick penetrating host skin and is aided by immune-modulatory tick salivary components. Macrophages are critical early responders in host defense after neutrophils and have profound effect on host defense mechanisms against ticks. Combatting the inflammatory mechanisms directed by macrophages is one task that ticks face during feeding. To address the possibility that ticks affect macrophage function, we previously screened effects of 15 recombinant TSPs. This analysis found pro-inflammatory and anti-inflammation TSPs. Of significant interest anti-inflammation rTSPs reversed pro-inflammation rTSP activation of macrophages. To investigate the molecular basis of this response, we carried out a systematic examination of the transcriptional gene expression in these macrophages in response to the rTSPs. We found a remarkable program of gene expression induced/suppressed specifically by pro- and anti-inflammatory rTSPs. An intricately choreographed expression pattern devoted to immunity, locomotion, response due to stimulus and adhesion was a prominent feature of this response. Our finding indicate that meshwork of host molecules respond to pro- and anti-inflammation rTSPs. Our analysis reveal a complex and intimate relationship between host and ticks.
Functionality of tissue-resident CD8+ T-cells in the human gastrointestinal tract during chronic HIV-1 infection.
Authors: Kiniry Brenna , Ganesh Anupama , Hunt Peter W , Somsouk Ma , Deeks Steven G , Shacklett Barbara L 
Published: January 2023

Abstract
The gastrointestinal tract (GIT) is an important site of HIV transmission and pathogenesis. Preventing or curing HIV infection will likely entail maintenance of protective anti-HIV immune responses in the GIT. Tissue resident memory T-cells (TRM) are long-lived, non-recirculating memory T-cells localized in tissues including the GIT. Positioned near sites of infection, TRM have protective efficacy against several pathogens. The precise role CD8+ TRM in HIV infection is not known. Blood and GIT CD8+ TRM cells from HIV+ and HIV- adults were identified by flow cytrometry using CD45RO, CD69, CD103, S1PR1, T-bet, and Eomesodermin (Eomes). Functionality was assessed in a 6-hour ex vivo stimulation assay. Cells were stimulated with DMSO, Gag, or Staphylococcal Enterotoxin B, and stained for CD107a, IFNγ, Mip-1β, TNFα, and IL-2. The frequency of CD103+CD69+S1PR1− CD8+ T-cells was significantly greater (P=0.005) in gut compared to blood (medians 54.8% and 0.10% respectively), indicative of tissue residency. CD45RO+ cells in this subset were classified as TRM. Consistent with murine studies, CD8+TRM were mainly T-betLowEomesNeg; however a population of EomesHigh CD8+ TRM emerged in HIV+ individuals not on antiretroviral therapy. Although polyfunctional Gag-specific CD8+ TRM responses (CD107a+, IFNγ+, Mip-1β+) appeared strongest in participants who control HIV without therapy (“controllers”), they constituted a smaller proportion of the total rectal Gag-specific CD8+ T-cell response compared to viremic participants not on ART (approximately 15% and 29% respectively). In summary, a large subset of CD8+ T-cells in rectal mucosa are TRM and likely play an important role in HIV-1 containment.
Itk is required for Th9 differentiation via TCR-mediated induction of IL-2 and IRF4
Authors: Gomez-Rodriguez Julio Washington , Meylan Francoise , Handon Robin , Hayes Erika T , Anderson Stacie , Kirby Martha R , Siegel Richard , Schwartzberg Pamela L. 
Published: January 2023

Abstract
A major function of the immune system is to mount specific responses to pathogens, while minimizing self-reactivity. To help orchestrate this, CD4+ T cells differentiate into distinct effector cell populations. Among these, T helper 9 (Th9) cells produce IL-9, a cytokine important for the eradication of type-II-inducing pathogens; however IL-9 has been also implicated in allergic asthma and autoimmunity. We show that Itk, a mediator of T-cell receptor signalling required for Th2 immune responses and the development of asthma, is a positive regulator of Th9 differentiation in vivo and in vitro. In a papain-induced model of allergic lung disease, Itk-deficient mice have reduced pulmonary inflammation and decreased IL-9 production by T cells and innate lymphoid type 2 cells (ILC2), despite normal early induction of ILC2s. In vitro, naïve Itk−/− CD4+ T cells essentially fail to produce IL-9 and have reduced levels of IRF4, a critical transcription factor for effector T-cell function. Both IL-9 and IRF4 expression are rescued by either IL-2 or expression of constitutively-active STAT5, but not NFATc1. We further find that STAT5 binds the Irf4 promoter, indicating one mechanism by which IL-2 can rescue weakly-activated T cells. Interestingly, while naïve WT CD4 T cells cultured in presence of the Itk inhibitor do not produce IL-9, addition of IL-2 leads to paradoxically high levels of IL-9, suggesting complex effects of Itk inhibitors that may prevent their use as therapeutics. Thus, inhibiting Itk as a therapeutic strategy for treating or preventing symptoms of asthma and other diseases in which IL-9 participates still remains an important and complex question.
The role of CD40 in calcitonin gene related protein (CGRP) mediated antiviral effect in glial cells
Authors: Grlickova-Duzevik Eliza , Cao Ling 
Published: January 2023

Abstract
LP-BM5 is a murine retrovirus that induces peripheral neuropathy and AIDS like immune deficiency in B6 mice. Upon infection, primary sensory neurons in the spinal cord express elevated levels of calcitonin gene related protein (CGRP). Previously, we showed that CGRP induced reduction of LP-BM5 viral loads in primary mixed glial cells. The purpose of this current study was to investigate possible CGRP downstream targets that may contribute to its antiretroviral response. We hypothesized that CGRP decreases the viral load in glial cells by modifying the CD40 expression; increasing in CD40 signaling may promote production of cytokines and chemokines with antiretroviral properties. Using CD40 knockout mice we showed that CD40 is required for CGRP mediated antiretroviral response in mixed glia. In microglial cell line, CD40 expression was elevated after treatment with CGRP. Similar effect was seen in microglia from spinal cord derived mixed glial cultures. While LP-BM5 increased microglial CD40 expression gradually over time up to 7 days post-infection in primary mixed glia, CGRP caused an early elevation (24 hr post-infection) of microglial CD40 expression in microglia and stayed relatively steady levels 7 days after infection. CGRP also promoted chemokine CXCL1(KC) production in mixed glia. Our data suggest that activation of CD40 mediated signaling may result in increased production of chemokines that may interfere with retroviral replication. Future studies will be directed to specifically delineate CD40 downstream target involved in antiretroviral response in glial cells.
Toll-like receptor 9 is required for the maintenance of CD25+FoxP3+CD4+ Treg cells during Listeria monocytogenes infection
Authors: Dolina Joseph S , Schoenberger Stephen P 
Published: January 2023

Abstract
It has long been appreciated, but not understood, that the CD8+ cytotoxic T lymphocyte (CTL) dependence on CD4+T cell help (Th) is conditional; needed for some immunogens but not others. One explanation for this phenomenon envisions Th requirement as an intrinsic property of the pathogen itself rather than its introduction to the immune system. Here we show that dependence of the optimal CD8+ T cell response to Listeria monocytogenes (Lm) on CD4+ T cells is a function of the immunogen dose used for priming, with low dose Lm (LD; 50 or 103 CFU WT or ΔActA, respectively) inducing a primary antigen-specific CTL response profoundly dependent on CD4+ Th cells while that induced by high dose Lm (HD; 4×103 or 106 CFU WT or ΔActA, respectively) is significantly inhibited by CD4+CD25+FoxP3+regulatory T cells (Treg). The Th-independence of HD immunization is not overcome by additional antigen but instead involves the inflammatory response to more bacteria. Evaluation of various TLR pathways as the relevant sensing mechanism showed that TLR2 is required for CTL responses to LD immunization, and that HD immunization in the absence of TLR9 results in a simultaneous loss of CD25+FoxP3+CD4+ Treg cells and increase in conventional CD4+ Th cells and CTLs. Our data thus reveal that the CTL response to the same pathogen is determined by distinct roles for CD4+ T cells as helpers versus regulators based on immunogen dose and demonstrate a previously undescribed role for TLR9 in the regulation of CD4+ Th and Treg cells.
Human T follicular regulatory cells (Tfr) able to inhibit antibody production are enriched at the lymph node T-B border and rare in germinal centers
Authors: Sayin Ismail , Radtke Andrea J , Vella Laura A , Buggert Marcus , Jin Wenjie , Wherry E. John , Germain Ronald N , Betts Michael R , Herati Ramin S , Canaday David H 
Published: January 2023

Abstract
T follicular helper (Tfh) cells are a subset of CD4+ T cells that are critical for the formation of germinal centers (GCs) and the development of high affinity, class-switched antibodies. Tfh cells are differentiated in a Bcl6-dependent manner and are characterized by expression of the inhibitory receptor PD-1 along with CXCR5, a chemokine receptor that directs these cells to the follicle. More recently, T follicular regulatory cells (Tfr) have been shown to express regulatory T cell (Treg) markers and suppress humoral immunity. To date, much of our understanding of the function and distribution of Tfh and Tfr cells has been obtained from animal studies. Using highly multiplexed, quantitative imaging and functional assays, we examined the spatial distribution and suppressive function of human Tfr cells obtained from mesenteric lymph nodes. Here, we report that the majority of Tfr cells reside at the border between the T cell zone and B cell follicle and are rare within the GC, being far outnumbered by Tfh cells. In contrast to Tfh cells, Tfr cells expressed low to undetectable levels of PD-1 in situ yet potently suppressed antibody production in vitro. These findings reveal that Tfh and Tfr cells are spatially segregated within human lymph nodes and caution against using PD-1 expression levels as a phenotypic marker for human Tfr cells.
This research was supported in part by NIH R01, VA, and Intramural Research Program of the NIH, NIAID.
Outcomes of congenital Zika disease depend on timing of infection and maternal-fetal immunophysiological conditions
Authors: Chen Jinling , Liang Yuejin , Yi Panpan , Azar Sasha , Rossi Shannan , Sun Jiaren 
Published: January 2023

Abstract
Here, we examined outcomes of congenital Zika disease in mice at different times of infection. Female mice lacking interferon alpha receptor (IFNAR−/−) were crossed to their male counterparts (KO×KO), and were subsequently infected with ZIKV at embryonic day (E) 5.5, E10.5 and E15.5, respectively. Dams infected with ZIKV at E5.5 had nearly 100% fetal demise. Dams exposed to ZIKV at E10.5 exhibited fetal demise and IUGR, whereas infection at E15.5 did not result in these clinical outcomes. To gauge the effect of IFN signaling at the maternal-fetal interface on the pregnant outcomes, we established a mouse model in which IFNAR−/−female mice were susceptible to ZIKV infection and their offspring were capable of type I IFN signaling due to a heterozygous breeding scheme (KO×WT). No viral burden was detected in multiple organs; also, no fetal demise was observed in the KO×WT group when they were infected at mid- and late pregnancy. The immune resistance in the heterozygous group was accompanied by high expression of IFN-stimulated genes (ISGs). Type III IFN also induced ISGs similar to type I IFNs through a distinct receptor. To gauge the effect of type III IFN on the host resistance to ZIKV and pregnant outcomes, we i.p. treated the pregnant mice with IFN-λ prior to and after ZIKV infection at E5.5, E10.5, and E15.5. We found that IFN-λ treatment reduced viral burden in maternal and fetal organs, upregulated ISG expression, and relieved fetal growth restriction at mid pregnancy. Thus, our data demonstrated that IFN signaling at the maternal-fetal interface is critical for the outcomes of pregnancy, especially at the mid pregnancy stage, during maternal ZIKV infection. IFN-λ inhibits ZIKV infection and confers protection to infected fetuses.
Topical exposure to the quaternary ammonium compound didecyldimethylammonium chloride augments dermal type 2 innate lymphoid cell populations
Authors: Shane Hillary L , Marshall Nikki B , Lukomska Ewa , Long Carrie M , Anderson Stacey E 
Published: January 2023

Abstract
Didecyldimethylammonium chloride (DDAC) is a dialkyl-quaternary ammonium compound used in commercial and industrial products for its antimicrobial properties. Clinical reports suggest that topical exposure to DDAC can resultin immediate and delayed type hypersensitivity, and recently our laboratory has confirmed that DDAC is a strong non-IgE mediated sensitizer in mice at relevant human exposure concentrations. Here, we further investigate the mechanism of DDAC induced hypersensitivity in mice, focusing on dermal immune responses. Gene expression analysis shows significant reduction in epithelial Cdh1, and increases in Tslp and Ccl17 following DDAC application to the skin, indicating that dermal type 2 innate lymphoid cells (ILC2s) may be involved in DDAC-induced hypersensitivity. Phenotypic analysis of dermal ILC2s shows that these cells are rapidly activated with increased expression of CD25, ICOS, and KLRG1 occurring by 24 hours post DDAC exposure. Increases in ILC2 number were observed following 2 weeks of DDAC exposure and found to persist for up to 2 weeks in the absence of DDAC and Tslp. Additionally, the phenotype of these cells suggests a potential for longevity (reduced KLRG1 and increased CD127) and enhanced activity (increased ICOS), which may influence future sensitization events. ILC2s have been previously implicated to play a role in IgE-independent allergic responses, but to our knowledge have not been investigated in chemical allergy. These results indicate that skin resident ILC2s may play a role in the development of the hypersensitivity response to DDAC and raise concerns about the use of this chemical and other quaternary ammonium compounds that may elicit similar effects.
The formation of memory-like innate lymphoid cells 2 in allergic asthma
Authors: Jing Xiuli , Wang Meixiang , Zhuang Qiannan , Zhao Bin , Liu Jianguo , Yi Shuying , Song Wengang , Tang Hua 
Published: January 2023

Abstract
Numerous studies have been suggested that ILC2s play a critical role in the initiation of allergic asthmatic airway inflammation, but their property and role in chronic allergic lung inflammation has not been well established. Our study found that purified allergen-experienced ILC2s produce more cytokines than naive ILC2s when stimulated by suboptimal amounts of IL-33 and IL-2 in vitro. Furthermore, upon allergen intranasal re-challenge, allergen-experienced ILC2s proliferate more vigorously and produce much greater amounts of type 2 cytokines than naive ILC2s. These results suggested that allergen-experienced ILC2s showed some memory-like properties in allergic asthma. To further characterize these memory-like ILC2s, we analyzed the expression of some activation surface markers, such as KLRG1, CD62L, CD69, ST2, ICOS, NKG2D, CD44, CD25 and CD122 on ILC2s in the lungs of naive and IL-33-treated mice at different time point. The results showed that during the initial expansion phase, IL-33 treatment resulted in a rapid increase in the number of CD25− ILC2s cells, which peaked at d6. And then the CD25− ILC2 population in the lung started to contract, the number of CD25− ILC2s cells was rapidly declined at d14, a time point that the numbers of CD25+ ILC2s reached their peak. Both of CD25− and CD25+ ILC2s rapidly declined at d30, however, the numbers of CD25+ ILC2s was much higher than CD25− ILC2. Thus, these results suggested that allergen-experienced ILC2s could differentiate into two different memory potential ILC2 subsets, namely CD25− short-term surviving effector ILC2s cells an CD25+ long-term surviving memory ILC2s cells. This work was supported by grants (2015CB943203, 31300730, 81272315, 2016GCC09)
Aortic B cells: Identifying antibodies specific for atherosclerosis in mice
Authors: Hutchinson Mark A , Maul Robert W , Gearhart Patricia J 
Published: January 2023

Abstract
Atherosclerosis is characterized by the development of arterial plaques containing lipid deposits and immune cells, such as B cells. Once recruited to the site of inflammation, B cells become activated and form germinal centers within the adventitia of the artery adjacent to the plaque. To determine if these B cells produce antibodies specific for atherosclerosis, abdominal aortas were collected, and germinal center B cells were isolated by flow cytometry. We characterized their heavy and light chain repertoires by next generation sequencing to compare non-diseased (C57BL/6) and diseased (ApoE−/−) tissues. Furthermore, the aortic repertoires were compared to the germinal center repertoires from spleens taken from the same mice to address whether local (aortic) and systemic (spleen) B cell responses were similar. Preliminary results suggest that all the splenic repertoires were similar, independent of diet or disease. However, aortic repertoires from ApoE−/− mice indicated that some VH and Vk genes were overexpressed, suggesting selection for particular antigens. We therefore performed single cell analysis and identified over 100 VH and Vk gene pairs, with different frequencies of expression and somatic hypermutation. Antibody gene pairs will then be cloned and expressed in culture using HEK293A cells. The purified recombinant antibodies will be used to identify antigens specific to atherosclerosis by protein array. These results may be useful in creating potential vaccines for its prevention, as well as treatment via antibody immunotherapy.