bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bcl-2 Does Not Rescue T Lymphopoiesis in Interleukin-7 Receptor Transgenic Mice (95.15)
Authors: Van De Wiele C. Justin , Tan Chibing , Teague T. Kent 
Published: January 2023

Abstract
We have previously demonstrated that murine thymocytes down-regulate IL-7 sensitivity between the β-selection and positive selection checkpoints. To assess the consequences of enforced IL-7Rα expression during this developmental phase, we evaluated thymocyte IL-7 responsiveness in transgenic mice expressing the IL-7Rα gene under control of the lck proximal promoter. This model demonstrated that induction of ectopic high-level IL-7Rα expression during thymopoiesis increases the percentage of thymocytes responsive to IL-7 but results in an overall decrease in total thymic cellularity in transgenic mice. Transgenic thymocytes initiated IL-7Rα expression beginning in the DN2 compartment which was maintained through development to naïve peripheral subsets. Interference with thymocyte development became apparent shortly after initiation of transgene expression as there was an ~65% reduction in DN3 thymocytes in transgenic mice. This finding was reminiscent of phenotypes observed in IL-7 signaling-deficient mice insofar as there was a significant loss of pre-T cells, which correlated with a reduction in Bcl-2 expression. Indeed, lower Bcl-2 levels were observed in transgenic DN2 and DN3 thymocytes when assessed directly ex vivo. Additionally, and unlike the IL-7 signaling-deficient models, transgenic restoration of Bcl-2 expression did not correct the thymic hypocellularity induced by IL-7Rα overexpression. These data demonstrate that ectopic overexpression of the IL-7Rα past the pro-T stage interferes with DN3 thymocyte development and inappropriately maintains IL-7 responsiveness in β-selected thymocytes.
IL-23 acts in a novel fashion by antagonizing IL-12 induced IFN-γ secretion (95.16)
Authors: Sieve Amy N. , Meeks Karen D. , Balch Joshua B. , Berg Rance E. 
Published: January 2023

Abstract
Most studies have concluded that interleukin-23 (IL-23) plays a unique role in promoting IL-17 secreting T cells. While some reports have also characterized IL-23 as having redundant pro-inflammatory effects with IL-12, we have instead found that IL-23 can regulate the innate immune response in a novel manner, by antagonizing IL-12 induced secretion of interferon-γ (IFN-γ). Our recent data documents the rapid, innate production of IFN-γ from memory CD8 T cells, as well as NK cells, stimulated with IL-12 and IL-18. When splenocytes are cultured with IL-23, IFN-γ secretion in response to IL-12 is dramatically reduced, as measured by ELISA and intracellular cytokine staining. The impact of IL-23 is most prominent in CD8 T cells, but can also be seen in NK, NK-T, and CD4 T cells. We also show that IL-23 can regulate the induction of IFN-γ by endogenously produced IL-12 from Listeria monocytogenes infected macrophages. Furthermore, IL-23 appears to act directly on purified CD8 T cells to negatively regulate IFN-γ induced by IL-12. We are currently investigating the mechanism by which IL-23 inhibits IFN-γ secretion. In addition, experiments to determine if IL-23 can inhibit other effector functions are ongoing. Our data suggest that IL-23 may be a key player in determining the responsiveness of lymphocytes to IL-12 and their subsequent secretion of IFN-γ.
This research is supported by NIH AI064592.
Characterization of IL-2-stimulatory factor produced from dendritic cells stimulated with a sesquiterpene compound TG651 (95.19)
Authors: RAN LEE YOUNG , KI-HYANG KIM , YOUNG-HEE LEE , HYUNJIN KIM , SUN-A IM , CHONG-KIL LEE 
Published: January 2023

Abstract
Dendritic cells (DCs) have been known to produce cytokines such as IL-2, IL-6, IL-10, IL-12, IFN-α and IFN-γ. None of those cytokines has been shown to activate T cells to produce IL-2 by itself. Here, we show that DCs stimulated with TG651, a sesquiterpene compound, produce a cytokine that induces IL-2 production from T cells. The cytokine, termed as IL-2SF, activated both CD4 and CD8 T cells to produce IL-2. IL-2SF was produced from DCs as a membrane bound form as well as a soluble form. Both form of IL-2SF was produced from DCs within 5 min after TG651 stimulation, and did not require new protein synthesis, although presence of approximately 10% fetal bovine serum was required for optimal production. Soluble form of IL-2SF appeared to be stable at neutral pH, but destroyed readily at alkaline pH when treated at 56°C for 30 min. The molecular weight of IL-2SF was estimated to be 60 KD - 70 KD by gel filtration chromatography. Although the molecular entity of IL-2SF remains to be elucidated, the present study demonstrates that DCs produce a cytokine that induces IL-2 production from T cells.
Differential effects of berberine, baicalein, and triptolide on cytokine production from IL-1β-activated mast cells (95.28)
Authors: Hsieh Chia-Jung , Hall Kenton , Krishnaswamy Guha , Chi David S. 
Published: January 2023

Abstract
Human mast cells are multifunctional cells capable of a wide variety of inflammatory responses. Berberine (BER), baicalein (BAI), and triptolide (TRI) are compounds isolated from traditional Chinese medicinal herbs and have been shown to have anti-inflammatory effects. We examined their effects on the production of inflammatory cytokines from IL-1β-activated human mast cells (HMC-1). Two ml of HMC-1 at 1 x 106 cells / mL were cultured with BER, BAI, and TRI (at predetermined non-toxic concentrations) in the presence or absence of IL-1β (10 ng / mL) for 24 hrs. The supernatants were harvested and assayed for IL-6, IL-8, and MCP-1 by ELISA. The compounds alone did not induce cytokine production from HMC-1. However, BAI (15 and 30 μM) significantly inhibited the production of IL-6, IL-8 and MCP-1 (all p &lt; 0.005) from IL-1β-activated HMC-1 in a dose dependent fashion. TRI (10 and 20 nM) significantly inhibited IL-8 and MCP-1 (p &lt; 0.001 and &lt; 0.05, respectively), but not IL-6 production from IL-1β-activated HMC-1. BER (25 and 50 μM) inhibited the production of MCP-1 (p = 0.02 and = 0.005, respectively), had no effect on IL-6 production, and enhanced the production of IL-8 at 25 μM (p = 0.01) from IL-1β-activated HMC-1. The results show that BER, BAI, and TRI had differential effects on the production of inflammatory cytokines. Since BAI has the most potent inhibitory effect, it may lead to new anti-inflammatory therapy.
(Supported by the Chair of Excellence in Medicine, the Ruth R. Harris Endowment, and RDC of ETSU)
Thymic IL-2R expression is sufficient for production of CD4+CD25+ Foxp3+ T regulatory cells (95.5)
Authors: Bayer Allison L , Yu Aixin , Malek Thomas R 
Published: January 2023

Abstract
CD4+CD25+ FoxP3+ Treg cells critically depend on IL-2, but the contribution of IL-2 to development of Treg cells in the thymus versus controlling their homeostasis in the periphery is unresolved. We addressed this issue using models where IL-2R activity was blocked in the thymus and periphery or only in the periphery. We found that IL-2/IL-2R interaction is active and essential in the thymus by promoting Treg cell expansion and upregulation of Foxp3 and CD25 to normal levels. This thymic signal gave rise to peripheral Treg cells with impaired IL-2-dependent STAT5 activation, yet they persisted in the periphery and suppressed autoimmunity without continual thymic output. Thus, thymic IL-2R function represents one important aspect for Treg cell development. Peripheral Treg cells with impaired IL-2R signaling exhibited slower growth and death rates as assessed by BrdU uptake in vivo and their suppressive activity was slightly lower in vitro. Importantly, by following donor Treg cells in mixed thymic and bone marrow chimeric mice, we found that WT Treg cells dominated peripheral immune tissue compared to Treg cells with impaired peripheral IL-2 signaling. Thus, IL-2 is a dominant mechanism controlling the number of thymic and peripheral Treg cells, although in a non-competitive setting, peripheral Treg cells with minimal IL-2R signaling persist and control autoimmunity.
(Supported by the NIH)
TH17 Augmentation in OTII TCR plus T Cell-Selective Type 1 Sphingosine 1-Phosphate Receptor Double Transgenic Mice (95.9)
Authors: Huang Mei-Chuan , Watson Susan R , Liao Jia-Jun , Goetzl Edward J 
Published: January 2023

Abstract
Sphingosine 1-phosphate (S1P) in blood and lymph controls lymphoid traffic and tissue migration of T cells through signals from the type 1 S1P receptor (S1P1), but less is known of effects of the S1P-S1P1 axis on non-migration functions of T cells. CD4 T cells from a double transgenic (DTG) mouse express OTII TCRs specific for ovalbumin peptide 323–339 (OVA) and a high level of TG S1P1, resistant to suppression by T cell activation. Activated DTG CD4 T cells respond as expected to S1P by chemotactic migration and reduction of OVA-evoked secretion of IFN-gamma. In addition, DTG CD4 T cells stimulated by OVA secrete a mean of 2.5-fold more IL-17 than those from OTII single TG mice with concomitantly higher levels of mRNA encoding IL-17 by real-time PCR and intracellular IL-17 detected by ELISpot assays. OVA challenge of subcutaneous air-pockets elicited influx of more OTII-TCR-positive T cells producing a higher level of IL-17 in DTG mice than OTII control mice. Augmentation of the number and activity of Th17 cells by the S1P-S1P1 axis may enhance host defense against microbes and in other settings increase host susceptibility to autoimmune diseases.
(supported by NIH HL31809)
Complex roles of the CXCR3 chemokines in immune-mediated neurological disease (96.1)
Authors: Campbell Iain L. , Mueller Marcus , Carter Sally , Getts Daniel R. , King Nicholas J.C. 
Published: January 2023

Abstract
The chemokine receptor CXCR3 binds CXCL9, CXCL10 and CXCL11 and promotes the trafficking of activated T-cells. Although these chemokines are highly elevated in the CNS in MS and the animal model, experimental autoimmune encephalomyelitis (EAE), their role is unknown. Here we show that while IFN-γ is the principal mediator of CXCR3 ligand gene expression in the CNS in EAE, the cellular localization of the individual chemokine genes is divergent suggesting functional specialisation of the CXCR3 ligand chemokines. Transgenic mice (GF-) were generated with astrocyte-production of CXCL9, CXCL10 or CXCL11. These mice do not develop neurological disease. In GF-CXCL10 and -CXCL11 mice, modest non-reactive leukocytic infiltrates were seen in the CNS. Thus, these chemokines when produced chronically are a poor stimulus for the CNS recruitment and activation of T-cells. The role of CXCR3 signaling in MOG-induced EAE was examined in CXCR3 KO mice and in GF-CXCL9, -CXCL10 or -CXCL11 mice. CXCR3 KO, GF-CXCL9 and -CXCL11 mice had more severe EAE with widely disseminated demyelinating lesions throughout the CNS. In CXCR3 KO mice with EAE the number of suppressor Treg cells in the brain was reduced. Thus, in EAE, CXCR3 chemokine ligands:have non-redundant functions,are not involved in the initial recruitment of effector T-cells to the CNS, andretain T-cells to the perivascular space and foster Treg cell interactions thereby limiting lesion spread and tissue destruction.
Support: NIH NS044905.
A hemoglobin based oxygen carrier (HBOC-201) has similar effects to conventional resuscitation fluids on cytokine responses in an acute trauma-hemorrhage model (96.12)
Authors: Hutchings Anne , Rodriguez Celina , George Richard , Moon-Massat Paula , Rue Loring , Kerby Jeffrey 
Published: January 2023

Abstract
Although traumatic hemorrhagic shock may be acutely fatal, alterations in the immune response of survivors in the post injury setting may lead to infections, sepsis, multiple organ failure and death. Various fluids, including HBOC-201, have been evaluated as resuscitative fluids for hemorrhagic shock. This study investigated the effects of HBOC-201 on cytokine responses following trauma-hemorrhage. Anesthetized mice (n=8/group) received a 2 cm abdominal incision, and then were hemorrhaged over 30 min through a femoral artery catheter to a mean arterial pressure (MAP) of 25±5 mm Hg. After an additional 1h of sustained hypotension, animals were resuscitated with lactated Ringer’s (LR), shed blood, or HBOC-201 and allowed to recover for 24h. Peripheral blood and spleens were collected for analysis. Survival rates were equal. HBOC-201 significantly increased post-resuscitation MAP compared to LR, but had little effect on serum IL6, IL10, IL12, IFNγ, TNFα or MIP1α levels compared to other groups at 24h post-resuscitation. There were no significant differences between groups in cytokine production by splenocytes stimulated for 24h in vitro with LPS. These results suggest that low volume resuscitation with HBOC-201 is superior to LR for maintenance of blood pressure and has no obvious effects on peripheral blood or splenocyte cell function with regard to cytokine production.
Supported by Biopure Corporation, Cambridge, MA.
IL-17 receptor-deficient mice have increased resistance to otherwise lethal influenza infection (96.14)
Authors: Crowe Christopher , Kolls Jay 
Published: January 2023

Abstract
IL-17 is a CD4+ T cell-derived cytokine that has been previously been shown to play a role in the immune response to several different lung infections. Specifically, it has been shown to induce neutrophil recruitment and inflammation in response to a variety of pathogens, including Pseudomonas aeruginosa, Klebsiella pneumoniae, and Mycoplasma pneumoniae. Additionally, it has been shown that IL-17 is upregulated in respiratory syncitial virus (RSV) infection, but its role in other viral pulmonary infections remains poorly described. In order to investigate the role of IL-17 in influenza infections, BALB/c mice lacking the IL-17 receptor were challenged intranasally with a lethal dose of influenza A/Puerto Rico/8/34 (H1N1). The mice were monitored for weight loss and signs of clinical illness on a daily basis. When compared to BALB/c controls, the IL-17 receptor knockout mice exhibited delayed weight loss and had higher survival rates. Additionally, the control mice exhibited higher levels of protein and lactate dehydrogenase in the BAL fluid, markers of acute lung injury, during the course of infection. These data suggest that the effects of IL-17R signaling contributes to lung injury and the pathology of a lethal influenza infection in a murine model.
Neuronal Nitric Oxide Synthase alters inflammatory response after endotoxin challenge (96.16)
Authors: Khaibullina Alfia , Besch Virginia , Middleton LaShon , Quezado Zenaide 
Published: January 2023

Abstract
We have recently shown that mice deficient in the neuronal isoform of NOS (nNOS−/−, KO) have higher mortality compared to wild type (nNOS +/+, WT) in a polymicrobial sepsis model. We sought to determine what effect the genetic deficiency of nNOS had on LPS-induced expression of cytokines. KO and WT mice were injected with LPS (i.p.) and sacrificed 6 or 24 h later. Peritoneal lavage cells (PLCs) were counted and profiled for cell type. RNA from spleens and PLCs was isolated and the relative expression of cytokines was determined by real-time PCR. After LPS challenge, KO mice had higher mortality (100% vs. 40% at 40 mg/kg), higher number of PLCs at 6 and 24h, higher proportion of macrophages at 6h and higher neutrophils counts at 24h compared to WT. In splenocytes, relative expression of proinflammatory cytokines at 6 hours was lower in KO (TNFα=0.18, IL1β=0.05) compared to WT mice (TNFα= 12.1, IL1β =2.9, p ≤ 0.002). Conversely in PLCs, KO mice had significantly higher IL-1β mRNA levels compared to WT (251 vs. 0.64, p ≤ 0.008). These results suggest that KO mice have altered neutrophil trafficking to peritoneum, do not mount a global inflammatory immune response, and exhibit aberrant local immune response after LPS challenge. These differences may explain increased mortality in knockout mice after LPS challenge and polymicrobial sepsis and indicate that nNOS has an important role in immune response during sepsis.
Manipulaion of TNFalpha delays loss of CD28 expression in human CD8 T cells (96.19)
Authors: Parish Stanley T , Fauce Stanley R , Effros Rita B 
Published: January 2023

Abstract
The CD28 costimulatory receptor is essential for T cell activation, glucose metabolism, and optimal telomerase activity. Increased proportions of CD8+CD28 T cells, seen in HIV infection and aging, correlate with decreased vaccine responsiveness and early mortality. CD28 expression is also lost in chronically stimulated cultures of CD8 T cells as they approach replicative senescence, a state of irreversible cell cycle arrest, associated with shortened telomeres and increased levels of TNFα. Based on the link between TNFα and the CD28 gene promoter, we sought to investigate the effects of manipulating TNFα on replicative senescence. Human T cells were repeatedly stimulated with an allogeneic cell line in the presence of either a neutralizing antibody (anti-TNFα) or a TNFα receptor 1 inhibitor (rhuTNFR:Fc). CD28 expression and gene transcription were monitored by flow cytometry and RT-PCR, respectively. Chronic exposure of antigen-stimulated CD8 T cells to rhuTNFR:Fc or anti-TNFα delayed the loss of CD28 expression. Even after completing 17 population doublings, the treated cultures contained &gt;20% more CD28+ cells; CD28 gene transcription was similarly elevated. Finally, telomerase activity was increased by as much as 6-fold in the treated cultures. Manipulation of TNFα, therefore, may be a useful clinical approach to delay accumulation of senescent CD8 T cell in vivo.
Supported by NIH AI060362 and AG023720
CCL25-CCR9 axis role in ovarian cancer progression (96.2)
Authors: Johnson Erica , Johnson-Holiday Crystal , Singh Shailesh , Singh Udai , Partridge Edward , Lillard James 
Published: January 2023

Abstract
Ovarian cancer (OvCa) is the most lethal among gynecologic malignancies. Recently, chemokines have been shown to function in non-random tumor-cell metastasis to distant organs in a fashion that is similar to chemokine-directed lymphocyte migration. Also, the interactions between CCR9 and its ligand, CCL25, have been implicated in leukocyte trafficking to the small intestines, a frequent metastatic site for OvCa. Previously, our laboratory has shown that ovarian cancer cells strongly express CCR9. We have also shown that CCL25 induces the chemotaxis of OvCa cell lines and increases the invasive potential of OvCa cells. Recent studies in T cells, have shown that CCR9-CCL25 interaction can induce anti-apoptotic pathways, namely through Akt/protein kinase B activation, which is PI3K- and G?i protein-dependent. Activated Akt phosphorylates multiple down stream targets that are involved in cell survival signals, which include glycogen synthase kinase-3? and forkhead transcriptional factor. Cisplatin is a commonly used anticancer drug for the treatment of OvCa. It has been shown that the anti-apoptotic signals of the PI3K-Akt survival pathways are involved in tumor cell cisplatin resistance. Here we characterize the molecular mechanisms of CCL25-CCR9 cell-signaling cascades involved in ovarian cancer survival and show that CCL25-CCR9 interactions mediate cisplatin survival through the activation of anti-apoptosis pathway. These studies will provide novel information regarding the cellular and molecular mechanisms, following CCL25-CCR9 interaction, that modulate ovarian cell metastasis and cisplatin resistance.
Abnormal innate immune response and Th1/Th17 induction in murine and human inflammatory bowel diseases (96.22)
Authors: Kobayashi Taku , Kamada Nobuhiko , Okamoto Susumu , Hisamatsu Tadakazu , Inoue Nagamu , Hibi Toshifumi 
Published: January 2023

Abstract
Recently, a lot of papers concerning IL-23/17 axis in some inflammatory conditions have been reported. IL-12p70/IL-23 are considered to regulate Th1/Th17 reciprocally in vitro, however, in vivo, both IFN-γ and IL-17 were reported to be upregulated in murine and human inflammatory bowel diseases (IBD). We previously reported the excess production of IL-12 and IL-23 by innate immune system in IL-10 deficient mice (Kamada N et al. J Immunol 2005; 175(10)) and in the present study, we have tried to unravel the role of innate immune system in induction of Th1/Th17 in murine colitis and human IBD. Colonic lamina propria CD4+ T cells of colitis mice were examined by intracellular cytokine staining and ELISA, and showed the upregulated production of both IFN- γ and IL-17. Similarly, these cytokines from CD4+ cells isolated from inflamed mucosa of human IBD are also increased in relation to upregulated IL-12/23. As expected, both macrophages and dendritic cells from IL-10 deficient mice produced larger amount of IL-12 and IL-23 than WT mice. Furthermore, macrophages and dendritic cells from IL-10 deficient mice could effectively stimulate both IFN- γ and IL-17 production in vitro. These results indicate that abnormal innate immune response stimulates both Th1 and Th17 simultaneously in murine and human inflammatory bowel disease.
IL1 is Required for CD8 T Cell Stimulation of Endothelial Cells to Produce KC in Contact Hypersensitivity (96.27)
Authors: Kish Danielle D. , Fairchild Robert L 
Published: January 2023

Abstract
Contact hypersensitivity (CHS) is a CD8 T cell-mediated response to cutaneous sensitization and challenge with hapten. Our recent studies indicate that hapten-primed CD8 T cells induce CXCL1/KC production in response to hapten challenge, which directs subsequent neutrophil infiltration into the challenge site and these infiltrating neutrophils then recruit effector CD8 T cells into site to mediate the swelling response. Keratinocyte-hapten contact induces IL1 production, which in turn also stimulates CXCL1/KC production. This study investigated the role of IL1 in the elicitation of CHS. IL1R−/− mice had decreased priming of effector CD8 T cells and CHS swelling responses to DNFB sensitization and challenge than wild-type mice. Production of CXCL1/KC following challenge of sensitized IL1R−/− mice was near background levels. Flow cytometry analyses of cell suspensions prepared from the challenged skin revealed decreased neutrophil infiltration in IL1R−/− vs. wild-type mice. Transfer of wild-type hapten-primed CD8 T cells to naïve wild-type recipients induced CXCL1/KC production in response to hapten challenge whereas transfer to naive IL1R−/− mice did not. Transfer of CD8 T cells from sensitized IL1R−/− mice did not induce CXCL1/KC production in wild-type mice. Injection of LPS i.v. stimulated endothelial cells in the skin of the IL1R−/− mice to produce KC indicating the ability of the IL1R−/− endothelial cells to produce the chemokine. We hypothesize that IL1 signaling is required for optimal CD8 T cell priming to hapten as well as for hapten-primed CD8 T cell interaction with endothelial cells in the challenge site to induce CXCL1/KC production and the CHS response.