bluuebunny/crossref_metadata_2025_split huggingface dataset:
Unveiling the immunological attributes of chicken tumor necrosis factor-α (TNF-α) using new sets of monoclonal antibodies specific for poultry TNF-α
Authors: Lu Mingmin , Lee Youngsub , Lillehoj Hyun S 
Published: February 2023

Abstract
Tumor necrosis factor-α (TNF-α) is a type II transmembrane protein with either membrane-bound or soluble forms and is a prototypical member of TNF superfamily. TNF-α is a pleiotropic cytokine associated with the regulation of systemic inflammation and host defense. Chicken TNF-α (chTNF-α) is a long missed avian ortholog and the immunological properties of chTNF-α remains largely unknown compared to the mammalian counterpart. Here, we report on the functional characterization and immunomodulatory roles of chTNF-α using a panel of new anti-chTNF-α mouse monoclonal antibodies (mAbs) that we developed. A chTNF-α-specific antigen-capture ELISA was developed using compatible mAb partners via the screening and validation of ten different mAbs (3G11, 4C4, 4F3, 6A52, 6E6, 10E8, 12G6, 12H7, 14H2 and 15G7). Employing 3G11 and 12G6 as capture and detection antibodies, respectively, the levels of native chTNF-α in the circulation of Clostridium perfringens-, Eimeria- or dual C. perfringens/Eimeria-infected chickens were determined. Furthermore, intracellular expressions of chTNF-α in primary cells or cell lines derived from chickens were validated in immunocytochemistry and flow cytometry assays using both 3G11 and 12G6 mAbs. Importantly, both 3G11 and 12G6 neutralized chTNF-α-induced nitric oxide production in chicken HD11 cells in vitro. Collectively, our results provide a better understanding of the functional characteristics of chTNF-α and these anti-chTNF-α mAbs will serve as valuable immunological tools for the fundamental and applied studies in avian species.
Active Influenza D coinfection reduces Influenza A disease severity in mice
Authors: Robinson Ema , Shepardson Kelly M. , Skelton Raegan M. , Hernandez Emiliano , Huber Victor C. , Rynda-Apple Agnieszka 
Published: February 2023

Abstract
Influenza D virus (IDV), the only member of the fourth genus of the Orthomyxoviridae family, primarily infects bovines. While no symptoms have been identified in humans, studies find seropositivity rates up to 95% among people with close contact to cattle. This suggests that IDV may cocirculate in human populations with Influenza A viruses (IAV). Clinical data indicate that coinfections with some mild respiratory viruses, such as rhinovirus, can reduce IAV severity. The current study investigates whether and how the immune response to preceding IDV infection affects the outcome of subsequent IAV. Previously we reported that IDV stimulates strong IFN-β production in mice and in human epithelium (A549 cells). We extend these findings by showing that IDV productively infects respiratory tract of wild-type (WT) mice and induces recruitment of innate and adaptive immune cells to the lungs. IDV-IAV coinfected mice exhibit a stronger IFN-β response early in IAV infection compared to mice infected with IAV alone. Consistently, IDV-infected WT mice showed less weight loss and improved survival after IAV. To better understand the role of type I IFN signaling in host response during IDV and IAV coinfection, we used mice deficient in either subunit of type I IFN receptor (Ifnar1−/−, Ifnar2−/−). While IDV-infected WT mice exhibit only slight weight loss occurring mid-infection, Ifnar1−/− mice exhibit dramatic weight loss mid-infection but recover fully within 3 days. In contrast to its effect in WT mice, IDV exacerbated IAV-induced mortality and lengthened the duration of symptoms in Ifnar1−/− and Ifnar2−/− mice. Our results show that active IDV infection elicits a type I IFN-dependent protection against IAV-associated weight loss and mortality.
Endogenous Type I interferon signaling plays a pivotal role in restricted SARS-CoV-2 infection in human ACE2 transgenic mice
Authors: Kempaiah Kempaiah Rayavara , Drelich Aleksandra , Hsu Jason C , Tat Vivian , Tseng Chien-Te K 
Published: February 2023

Abstract
Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has become pandemic, resulting in severe morbidity and mortality around the world. COVID-19 pathogenesis includes lung inflammation, cytokine storm and organ failures, but the mechanism underlying the disease remains to be studied. To determine what role(s) the well-characterized type I interferon (IFN-I)-mediated innate antiviral responses might play against SARS-CoV-2 infection, we treated hACE2 transgenic (Tg) mice, AC70 line, with mouse IFN-I receptor-specific monoclonal antibody prior to challenge with 106TCID50 of SARS-CoV-2 (US-WA-1/2020). Both IFN-blocked and-unblocked animals developed clinical signs of disease with body weight loss from day 3 and 100% mortality between 4 and 5 days post-infection with SARS-CoV-2 at high dose (106 TCID50). Although live virus was detected in the lungs and brain of both groups, live virus was detected only in the kidneys, liver, spleen, heart, and gastrointestinal tract of the IFN-blocked group. Elevated expression of proinflammatory cytokines and interferons including IFN-λ were detected in all organs tested in both groups. Immunostaining for SARS-CoV-2 spike protein along with profound histopathological observations in the lungs and brain of both groups show severe COVID-19 disease pathogenesis. Our results show that AC70 Tg mice are highly permissive to SARS-COV-2 infection and develop severe COVID-19. Furthermore, we show type I IFN-mediated signaling pathways play a critical role in restricting viral spread and are ideal for development of medical countermeasures (MCMs) against COVID-19.
House dust mite proteins reduce the spread of Respiratory Syncytial Virus in the BEAS-2B bronchial epithelial cell line.
Authors: Rife Maria , Freiman Andrew , Hillyer Philippa , Rabin Ronald L 
Published: February 2023

Abstract
House dust mites (HDM) are a predominant causative agent of airway hypersensitivity and asthma. The HDM group 1 allergens include Der p 1, a cysteine protease that contributes to sensitization and symptom exacerbation. Respiratory syncytial virus (RSV) infection of infants and small children can lead to severe pneumonitis. RSV activation of TLRs and RIG-I-like receptors induces proinflammatory cytokines and chemokines, which exacerbate disease, and type I/III interferons (IFNs) which induce expression of antiviral IFN stimulated genes (ISGs). Since RSV in children is coincident with HDM exposure, we used BEAS-2B human bronchial epithelial cells to explore the effects of HDM on RSV infection
BEAS2B cells were exposed to HDM extract or Der p 1 alone before, during, or after RSV infection. Surprisingly, HDM extract or Der p 1 decreased RSV infection in a dose and time-dependent manner. In a viral entry assay, HDM extract reduced the area of foci, rather than their number, suggesting that one or more HDM proteins attenuate cell to cell spread. Preliminary experiments point to a role for the cysteine protease activity of Der p 1. We also measured expression of a panel of representative proinflammatory mediators and ISGs. Compared to RSV alone, Der p 1 increased expression of some proinflammatory mediators, while HDM extract decreased expression of a panel of ISGs. In the absence of RSV, HDM extract increased proinflammatory mediators while Der p 1 did not affect gene expression. These results were unchanged following heat treatment at 65°C for 1 h. Elucidating mechanisms by which HDM proteins enhance protection against RSV may reveal novel antiviral mediators that locally control RSV infection.
Murine Th9 cell-mediated allergic inflammation shows different steroid responsiveness depending on target organs
Authors: Miura Kento , Koyama Teidai , Yamasaki Norimasa , Ogata Sawako , Saeki Mayumi , Hiroi Takachika , Mori Akio , Kaminuma Osamu 
Published: February 2023

Abstract
Th9 cell has been implicated in the development of allergic inflammation, though the sensitivity of Th9 cell-mediated responses to steroid therapy remains unclear. Here, allergen-induced bronchial and nasal inflammatory responses and their steroid responsiveness were comparatively investigated in ovalbumin (OVA)-specific Th9 cell-transferred mice. BALB/c mice were transferred with in vitro-differentiated Th9 cells and challenged by intratracheal or intranasal injection of OVA. The significant nasal hyperresponsiveness accompanied by nasal infiltration of eosinophils as well as allergen-specific T cells was induced in Th9 cell-transferred mice upon intranasal allergen challenge. These responses were strongly suppressed by the treatment with dexamethasone. Substantial accumulation of eosinophils in the lungs and bronchial hyperresponsiveness (BHR) were induced by intratracheal OVA challenge. In contrast with the nasal responses, neither cellular infiltration in the lungs nor BHR in Th9 cell-transferred mice was affected by dexamethasone treatment. In addition to the possible contribution of Th9 cells to the development of allergic inflammation in various tissues, different steroid responsiveness of Th9 cell-mediated responses among target organs was suggested.
Glycogen synthase kinase 3 (GSK3) is a critical regulator of allergen-mediated mast cell activation
Authors: Crozier Robert W.E. , Hamstra Sophie I. , Fajardo Val A. , MacNeil Adam J. 
Published: February 2023

Abstract
Mast cells are granulated immune sentinels responsible for perpetuating allergic inflammatory events. Allergen-induced FcɛRI signaling leads to the activation of MAPK, PLC and PI3K/Akt signaling cascades, which are responsible for mediating a biphasic mast cell response, characterized by degranulation in the early phase and sustained release of pro-inflammatory mediators in the late phase. Glycogen synthase kinase 3 (GSK3) is a constitutively active serine/threonine kinase and a major convergence point for several highly conserved signaling cascades, including the PI3K/Akt pathway. Due to its central role in regulating downstream pro-inflammatory signals, GSK3 has become a therapeutic target in inflammatory pathologies, however, the role that GSK3 plays in allergen-induced FcɛRI signaling has yet to be characterized. Thus, the objective of this study was to determine the functional role of GSK3 in allergen-activated mast cells. Sensitized murine bone marrow-derived mast cells were incubated with the GSK3 inhibitor CHIR99021 and stimulated with allergen. CHIR99021 treatment significantly inhibited degranulation dose-dependently to 63% (10 μM, p&lt;0.001) and 43% (20 μM, p&lt;0.001) of the control. Release of cytokines TNF (p&lt;0.01), IL-6 (p&lt;0.001), IL-13 (p&lt;0.01) and chemokines CCL1 (p&lt;0.001), CCL2 (p&lt;0.05) and CCL3 (p&lt;0.001) were inhibited following treatment at 20 μM. Finally, inhibition of GSK3 was found to significantly reduce downstream phosphorylation of JNK (10 & 20 μM, p&lt;0.05), while ERK and p38 remained unaffected. These results are the first to characterize GSK3 as a central regulator of allergen-induced FcɛRI signaling, making it an intriguing target to attenuate mast cell functionality in pathological contexts.
GC-Tfh cell induction by Langerhans cells in steady-state is antigen-dose dependent and IL-6 independent.
Authors: Bouteau Aurélie , Zurawski Sandra , Zurawski Gerard , Igyártó Botond Z 
Published: February 2023

Abstract
We previously compared the contributions of different skin dendritic cell subsets in the induction of humoral immune responses. We found that antigen delivery without adjuvants to Langerhans cells (LCs) through the c-type lectin receptor Langerin, unlike cDC1, induces germinal center T follicular helper cells (GC-Tfhs), GC B cells, and high antibody titers.
We also showed that targeting LCs with higher antigen dose induced significantly lower B cell responses and antibody titers. To better understand the role of antigen dose in this response, we assessed the T cell responses following targeting of LCs with different amounts of antigen. In accordance with the B cell responses, we found that GC-Tfh cell formation was also antigen dose-dependent. Using two-complementary approaches targeting IL-6-deficient LCs and targeting LCs in mice treated with neutralizing anti-IL-6 antibody, we showed that LCs induced GC-Tfh cells’ formation is an IL-6 independent fashion.
In summary, this study shows that targeting LCs at steady-state induces the formation of GC-Tfh cells through an antigen-dose dependent and IL-6 independent mechanism.
Role of PRMT5 on splicing events in T cell biology
Authors: Sengupta Shouvonik , West Kelsi O , Agosto-Rosa Laura , Lynch Kristen W , Watson Robert , Patrick Kristin , Guerau-De-Arellano Mireia 
Published: February 2023

Abstract
Protein Arginine Methyltransferase (PRMT) 5 catalyzes symmetric dimethylation of arginine, a post-translational modification involved in cancer and embryonic development. The mechanisms by which PRMT5 modulates T helper (Th) cell activity are still not completely understood. Here we find that PRMT5 in T cells promotes spliceosome component methylation and influences splicing in resting and activated cells. RNAseq of resting and activated wildtype (WT) and PRMT5-knockout (KO) CD4 T cells show the breadth of pathways affected by knockout of PRMT5. Importantly, metabolic processing, signaling, cell cycle and protein localization were affected. We also used a novel software, Modeling Alternative Junction Inclusion Quantification (MAJIQ), to extract information on the classes of alternative splicing (AS) events – Exon skipping, Intron retention, Alternative 5′ splice site and Alternative 3′ splice site. In our samples, we noticed a bias towards exon skipping. We took a closer look at the number of alternative junctions (AJ) arising from these events and the location of the local splice variants (LSV). Analysis of WT and KO T cells showed 9071 AJs upon T cell activation and 2136 AJs dependent on PRMT5. This work demonstrates that PRMT5 modulates the gene expression splicing profile of T cells.
Depletion of microRNA-183-96-182 miRNA cluster in lymphocytes suppresses anti-dsDNA autoantibody production and IgG deposition in kidney in C57BL/6-Faslpr/lpr mice
Authors: Wang Zhuang , Heid Bettina , Ren Jingjing , Edwards Michael R , Cecere Thomas E. , Lu Ran , Khan Deena , Kirsch David , Reilly Christopher M. , Dai Rujuan , Ahmed S. Ansar 
Published: February 2023

Abstract
The miR-183-96-182 (miR-183C) is a highly conserved miRNA cluster among species. Our previous work found a significant upregulation of miR-183C in the splenic cells of three different murine models of systemic lupus erythematosus (SLE). Current studies revealed that miR-183C miRNAs are critically involved in immunity and autoimmunity.
In this study, we found that inhibition of miR-182 alone or miR-183C in vitro with antagomirs significantly reduced lupus-related inflammatory cytokine IFN-γ and IL-6 in activated splenocytes from MRL or MRL/lpr mice. To further characterize the pathogenic role of miR-182 and miR-183C in lupus in vivo, we developed B6-lpr mice with conditional depletion of miR-182 or miR-183C in CD2+ lymphocyte. We found that depletion of either miR-182 or miR-183C in the lymphocytes of B6/lpr mice had no obvious effect on T and B cell development as similar percentage of CD4+. CD8+, CD19+, as well as Tregs, follicular helper T (TFH), germinal center B (GCB), and plasma cells were observed in the miR-182−/− and miR-183C−/− and their respective control. Importantly, we observed a significant reduction of serum anti-dsDNA autoantibodies in miR-183C−/− mice when compared to age-matched controls and the B6/lpr mice with miR-182 or miR-183C deficiency have significantly reduced IgG deposition in the kidneys. Meanwhile, there was reduced IFN production in ex vivo activated splenocytes from the knockout mice. Furthermore, we demonstrated that miR-182 and miR-183C regulated the inflammatory response in splenocytes via targeting forkhead box O1 (Foxo1). Together, our data suggest a potential therapeutic effect of targeting miR-183C in lupus.
Interleukin-2-Inducible T-cell Kinase (ITK) signaling regulates non-canonical regulatory T cells
Authors: Mammadli Mahinbanu , Harris Rebecca , May Adriana , Gentile Teresa , Karimi Mobin A 
Published: February 2023

Abstract
Regulatory T cells (Tregs) play an indispensable role in maintaining immunological tolerance to self-antigens and foreign antigens in suppressing excessive immune responses deleterious to the host by exerting a suppressive program towards a wide variety of target immune cells.
Here we showed that ITK signaling pathways control the increased frequency of “Non-Canonical” CD4+CD25−FoxP3+ Tregs (ncTregs) population as well as canonical Tregs CD4+CD25+FoxP3+ expression significantly. Using several animals models we demonstrated that transplanting ncTregs and cTregs from ITK deficient mice ameliorated WT T cells mediated Graft versus Host Disease (GVHD). Furthermore, our data showed that adaptively transferring ncTregs and cTregs from ITK deficient mice significantly reduces proinflammatory cytokine production and T cell proliferation in GVHD induced recipient mice. Furthermore, our data uncover that adaptatively transferring ncTregs and cTregs from ITK deficient mice to GVHD induced recipient mice reduced tissue damage which induced by WT T cells. Finally, we have recently developed a novel peptide inhibitor which enhance Tregs expression in both mice and human. This novel peptide inhibitor also reduced cytokine production in mice and primary GVHD patient samples. Our data demonstrated that targeting specifically SLP76:ITK signaling could be used as a therapeutic strategy not only limit to GVHD but also treat multiple T cells mediated autoimmune disorders.
CD22/Lyn-mediated suppression of trophoblast antigen-specific B cells underlies fetomaternal tolerance
Authors: Rizzuto Gabrielle , Brooks Jeremy , Fisher Susan , Zikherman Julie , Erlebacher Adrian 
Published: February 2023

Abstract
The placenta releases allogeneic trophoblast antigens into maternal circulation during pregnancy. Maternal antigen presenting cells present these antigens to T cells but fail to induce strong immunogenic responses, even in the presence of danger signals. While this observation illustrates the phenomenon of fetomaternal tolerance, its cellular and molecular basis remains largely unknown. By studying the pathways mediating adaptive immune responses to a surrogate trophoblast antigen in mice, we found that maternal B cells, and not regulatory T cells, directly underlie the impairments in CD4 T cell activation. Trophoblast-antigen specific B cells, rather than dendritic cells, are the main presenters of trophoblast antigen to CD4 T cells. The B cells are themselves profoundly suppressed, thus limiting their ability to activate cognate CD4 T cells. Mechanistically, B cell suppression occurs as the result of CD22/Lyn inhibitory signaling, likely induced by sialic acid residues that coat the trophoblast-derived antigen. These results not only identify maternal B cells as key players in fetomaternal tolerance but also provide a basis for further work on B cell responses during healthy and pathologic pregnancies. More broadly, these results suggest that exploiting antigen-intrinsic properties such as sialylation may provide new therapeutic ways to ignite or inhibit antigen-specific responses outside the context of pregnancy.
Therapeutic Vaccine (TheraVac) in combination with cGAMP cures more differentiated melanomas in mice
Authors: Alam Md Masud , Yang De , Tross Debra , Oppenheim Joost J 
Published: February 2023

Abstract
We have identified a combination of immunotherapeutics, termed TheraVac, capable of curing multiple large established mouse tumors, but it failed to cure melanoma in mice. TheraVac consists of an immunostimulating arm containing an agonist (HMGN1) for TLR4 and an agonist (R848) for TLR7/8 that synergize to activate tumor-infiltrating dendritic cells (DCs) and upregulate the polarization of Th1 immune responses. The second arm uses an immune checkpoint blockade inhibitor, αPDL-1, to diminish tumor associated immunosuppression. Here, we investigated supplementation of TheraVac by a STING agonist, cGAMP, because they synergize in activating DC’s and produced more immunostimulating IL-12p70 and TNFα cytokines. We therefore treated five different tumor types including melanin producing B16F1, M3, and M4 mouse melanomas that are homologous to human melanomas with intratumoral delivery of cGAMP and TheraVac. This eradicated 80% of the B16F1, 80% of the M3 and 60% of the M4 tumors, but still failed to cure the non-melanin producing undifferentiated M1 and M2 tumors. Biopsies of M3 tumor treated with TheraVac plus cGAMP showed a significant increase of CD8+ CTL’s infiltrating into the tumor. Importantly, there was a marked increase of both CD4 and CD8 memory T cells and generation of functional tumor specific CTLs in tumor-draining lymph nodes. The resultant tumor-free mice were selectively resistant to subsequent challenge with the same tumors, indicating long-term tumor specific protective immunity. Overall, our findings have important implications for clinical trials with a combination of immunotherapeutics to cure melanin producing human melanomas, without the need for exogenous tumor antigens and no obvious toxic effects in mice.
αPD-1 and extended half-life IL-2 combination therapy clears established GL261 gliomas in an MHC class I independent fashion
Authors: Tritz Zachariah Peter , Ayasoufi Kathy , Malo Courtney Sarah , Himes Benjamin , Khadka Roman , Yokanovich Lila T , Goddery Emma , Fain Cori , Hansen Michael , Jin Fang , Wang Chensu , Moynihan Kelly , Irvine Darrell J , Wittrup K Dane , Parney Ian , Johnson Aaron J 
Published: February 2023

Abstract
Glioblastoma multiforme (GBM) is a deadly CNS malignancy with an average survival of ~12 months post diagnosis despite aggressive treatment. Immunotherapy approaches for GBM have been attempted but the unique immune environment of the brain and the immunosuppressive features of GBM complicate these efforts. Even αPD-1 checkpoint blockade, highly useful in the context of melanoma, has proved ineffective in the treatment of GBM. In order to promote the efficacy of αPD-1 against the GL261 murine model of GBM, we supplemented this therapy with an engineered IL-2 cytokine. IL-2 therapy has been used clinically for metastatic melanoma and renal cancer, but its short half-life in circulation necessitated dangerously high doses to keep the drug concentration sufficiently high. Fusing IL-2 to the mouse serum albumin molecule (MSA-IL-2), extends the cytokine’s half-life. While neither αPD-1 nor MSA-IL-2 are effective monotherapies against the GL261 model tumor, we have found this combination immunotherapy to provide durable tumor clearance and to create immunologic memory capable of resisting GL261 rechallenge. Most strikingly, this combination therapy cleared established GL261 tumors even in mice incapable of MHC class I restricted antigen presentation. Instead, therapeutic efficacy was abrogated by the depletion of CD4 T cells. Independence of MHC class I restricted antigen presentation makes this combination immunotherapy using αPD-1 checkpoint blockade both highly novel and translatable to clinical use.