bluuebunny/crossref_metadata_2025_split huggingface dataset:
Periopathogens Modulate Inflammation by Repressing miR-142-3p and Augmenting miR-155/NFκB axis
Authors: Valverde Araceli Maria , Schaller Samantha , Brandini Daniela Atili , Brambila Maria F , Martinez Gloria , Chapa Gabriela , Li Wei , Wu Christine , Rahat Rani , Siddiqui Hasan , Nares Salvador , Naqvi Afsar 
Published: January 2023

Abstract
MicroRNA (miR) play a critical role in the pathogenesis of immune-mediated diseases. We hypothesize that periopathogens (P. gingivalis [Pg] and A. actinomycemtemcomitans [Aa]) impair host immune response by perturbing miR expression. Periopathogen burden, cytokine and miR expression was quantified by qPCR in healthy and inflamed human gingival biopsies. Human primary CD14+ monocytes were differentiated to M1 or M2 macrophages (MΦ) and challenged with live Pg or Aa to monitor expression profiles of pro- and anti-inflammatory miRs by qPCR. Supernatant levels of cytokines were assessed by multiplex bead assay while, protein levels were quantified by western blot. Murine gingival miRNA and mRNA profiles were validated during disease progression in a liguture-induced periodontitis model and bone loss measured by microCT. Higher periopathogen burden and inflammatory cytokine expression was observed in inflamed gingiva with a higher expression of pro-inflammatory miR-155 and reduced levels of anti-inflammatory miR-142-3p. This corroborates with NFκB activation and downregulation of PU.1 suggesting a distinct role of a miR and transcription factor (TF) axis. miR-155-mediated downregulation of PU.1, a TF of miR-142, suppressed transcription of miR-142, thereby alleviating silencing of IL-6 mediated by miR-142-3p. Our murine ligature model showed progressive bone loss in the presence of Pg and higher expression of cytokine profiles. Local delivery of miR-142-3p showed reduction in IL-6. Periopathogen-mediated activation of miR-155/NFκB axis augments inflammatory signaling by decreasing levels of PU.1 and expression of its target miR-142. This mechanism may be critical in the pathogenesis of periodontal disease.
Supported by R01 DE027980, R21 DE026259, R03 DE027147
Treg donation of sIL-6R to promote intestinal integrity and repair
Authors: Klatt Emily Eileen , Sagstetter Mary , Bamidele Adebowale , Gonzalez Michelle M , Ramos Guilherme Piovezani , Gibbons Hunter R , Wixom Alexander , Hamdan Feda H , Neto Manuel Braga , Faubion William A 
Published: January 2023

Abstract
IBD affects roughly 1.6 million US adults, with its prevalence increasing since the late 1990s. Characterized by unchecked intestinal inflammation, multiple proinflammatory cytokines including IL6 have been implicated as critical factors in this pathology. It has been established that EZH2 is a gene involved in the production of specific histone methyltransferase enzymes, as well as in the differentiation of Treg cells. The association between the Treg master transcription factor, Foxp3 and EZH2 results in the suppressive function of Tregs, with our previous work revealing impairment of this interaction under the influence of IL6 signaling in an inflamed intestinal setting. We investigated this further in a murine preventative T cell transfer model of colitis that involved injecting donor Treg cells that lacked IL6Ra, into immunocompromised mice. Histological scoring of the mice receiving the IL6Rko Tregs revealed a resolved inflammatory milieu and flow cytometric analysis of cells isolated from the spleens and MLNs showed increased Foxp3+ expression when compared to mice that received WT Tregs. Foxp3IL6Rako mice were subjected to a DSS model of colitis to further characterize the absence of the IL6R on Treg function in the intestinal milieu. Exacerbated signs of colitis were observed through histology and colon shortening of the conditional knockout mice when compared to their WT counterparts. We postulate that Tregs could be acting as a donor of sILR to promote intestinal integrity and healing. Further experiments will be conducted to define a role for the IL6R in the development of Foxp3+ cells, along with outlining the instrumental process of sIL6R donation in the intestinal milieu.
Supported by Mayo Clinic Graduate School for Biomedical Sciences
Gut microbiota modulates the development of murine Kawasaki disease vasculitis
Authors: Jena Prasant K , Wakita Daiko , Gomez Angela C , Carvalho Thacyana T , Narayanan Meena , Lee Youngho , Cani Patrice D , de Vos Willem M , Devkota Suzanne , Underhill David M , Chen Shuang , Shimada Kenichi , Crother Timothy R , Arditi Moshe , Rivas Magali Noval 
Published: January 2023

Abstract
Alterations of gut microbiota composition and function influence the development of cardiovascular disease, but the role of these aberrations remains poorly understood in Kawasaki Disease (KD), an acute pediatric vasculitis that targets coronary arteries. Using a murine model of KD vasculitis, we found that germ-free and antibiotic-treated mice displayed reduced inflammation and cardiovascular lesions. Development of KD vasculitis in mice was associated with alterations in the composition of the intestinal microbiota, specifically decreased abundance of Akkermansia muciniphila and Faecalibacterium prausnitzii. Supplementation with live or pasteurized A. muciniphila or F. prausnitzii attenuated the severity of KD cardiovascular inflammation. Oral administration of the short-chain fatty acids acetate or butyrate, which are produced by A. muciniphila and F. prausnitzii respectively, or treatment with Amuc_1100, a purified protein isolated from the outer membrane of A. muciniphila, ameliorated the severity of KD cardiovascular lesions. Reduced development of KD vasculitis in mice receiving either pasteurized A. muciniphila or Amuc_1100 was associated with improvements of gut barrier function. These results reveal an underappreciated gut microbiota-cardiovascular inflammation axis during murine KD vasculitis. Our findings may stimulate the development of novel diagnostic tools and therapeutic strategies that modulate the intestinal microbiota composition and function for KD patients.
Research is supported by the NIH grants R01AI072726 to M.A. and R01HL139766 to M.N.R.
A human ectopic-lymphoid-follicle-on-a-chip for testing vaccines and adjuvants
Authors: Zhai Yunhao , Goyal Girija , Prabhala Pranav , Patil Aditya , Kim Min Sun , Junaid Abidemi , Ku Min Wen , Mahajan Gautam , Bausk Bruce , Gilboa Tal , Lazarovits Roey , Sharma Sanjay , Cohen Limor , Ferrante Thomas , Walt David R , Ingber Donald E 
Published: January 2023

Abstract
New vaccine candidates evaluated in animals can lead to unpredicted toxicities or poor efficacy in human clinical trials due to species-specific differences in immune responses. To address this problem, we developed the human ectopic lymphoid follicle (LF) chip containing primary human immune cells, including autologous blood B and T lymphocytes and antigen presenting cells, cultured in a three-dimensional extracellular matrix (ECM) gel within an organ-on-a-chip microfluidic device. Superfusion via a parallel channel separated by a microporous membrane is required for LF formation and prevents lymphocyte autoactivation. These germinal center-like LFs contain B cells expressing activation-induced cytidine deaminase and exhibit plasma cell differentiation upon activation. Increased basal lymphocyte proliferation is observed in the human LF chip as compared to conventional 2D cultures lacking ECM or perfusion. The human LF chips demonstrated improved antibody responses to split virion influenza vaccination compared to conventional planar 2D cultures, which were enhanced by the addition of a squalene-in-water emulsion (SWE) adjuvant, and this was accompanied by increases in LF size and number. To extend this work, we modelled intramuscular vaccination by co-culturing monocytes with muscle cells in the presence of different vaccines and adjuvants. We found that presence of muscle cells can promote cytokine production, antigen uptake and monocyte activation. Seeding these monocytes into the LF chips with autologous T and B cells could promote cytokine production and LF formation, which extends the LF chips usage for testing intramuscular vaccination.
Supported by grants from the Defense Advanced Research Projects Agency under Cooperative Agreement Number W911NF-12-2-0036, the National Institutes of Health grant UG3HL141797, Bill and Melinda Gates Foundation and the Wyss Institute for Biologically Inspired Engineering.
Anti-CD3 Bi-Fab crosslinking of T cells enables their fratricide by activation induced cell death (AICD) and cytotoxicity
Authors: Pages Diana Gil , Nelson Alfreda , Wang Liangyu , White Tommi , Schrum Adam , Cannon John 
Published: January 2023

Abstract
We showed in the past anti-CD3 Fabs can form stable bivalent non-covalent dimers we called Bi-Fabs, which appear stimulatory to T cells according to the up-regulation of the early activation marker CD69. Now we show that incubation of naïve T cells with anti-CD3 Bi-Fabs in the presence of proper co-stimulation results in their robust stimulation in comparison to mAb and F(ab’)2 species with same specificity. Nonetheless, anti-CD3 Bi-Fab treatment leads to abortive T cell proliferation with impaired accumulation of divided T cells. After detailed studies to dissect the mechanisms of T cell death enabled by Bi-Fab molecules in comparison with anti-CD3 mAb and F(ab′)2 recognizing the same epitope, we observed that anti-CD3 Bi-Fabs promote T cell fratricidal killing via activation induced cell death (AICD) and CD8 cytolytic activity (CTL) more efficiently than the mAb and F(ab′)2 counterparts. While low resolution structural studies indicate that Bi-Fab and F(ab’)2 adopt similar shapes, molecular dynamic simulation shows that F(ab’)2 is a highly flexible molecule, whereas Bi-Fabs are more rigid and of linear conformation, sampling a smaller range of Fab-Fab angles than F(ab’)2. We hypothesize anti-CD3 Bi-Fab is superior as T cell killer stimulatory modality over mAb and F(ab’)2 of equal specificity due to Bi-Fab’s greater rigidity when crosslinking in trans TCR/CD3 molecules located in neighboring T cells. These findings may be applicable into the design of Ig-based immunotherapeutic drugs targeting the TCR/CD3 complex.
Supported by MU start-up funds
CD16 and NKG2D co-clustering facilitates quality of primary NK cell responses
Authors: Sykulev Yuri , Tsygankova Oxana , Anikeeva Nadia , Maskalenko Nicholas , Campbell Kerry 
Published: January 2023

Abstract
Cooperation between CD16 and NKG2D receptors has been demonstrated both in animal and NK cell line models leading to an enhanced cytotoxicity and tumor elimination. However, the mechanisms of the receptor cooperation are not known, and whether the receptors form co-clusters has not been determined either. We exposed human primary polyclonal NK cells with CD16 158V/V-genotype to planar lipid bilayers (PLB) presenting Herceptin and anti-NKG2D Ab in the presence of ICAM-1. NK cells interacting with Herceptin-containing PLBs + ICAM-1 formed mostly unstable synapse, i.e., kinapses, while stimulation with anti-NKG2D alone or Herceptin+anti-NKG2D bilayers + ICAM-1 led to formation of mature synapses showing complete segregation of the receptors from ICAM-1, increased synapse stability, and efficient NK cell cytotoxicity. The cells exposed to PLBs with both anti-NKG2D and Herceptin ligands were mostly confined within the initial contact area while the cells responding to Herceptin alone were significantly more motile. Regardless of stimulatory ligands displayed on PLB, the frequency of adhered cells was very similar, but the degranulation was observed within 40% of kinapses and 80% of mature synapses, suggesting that degranulation efficiency is linked to synapse stability. Importantly, more efficient degranulation was associated with rapid and sustained Ca2+ signaling as opposed to oscillating Ca2+ signaling observed for NK cells showing less efficient degranulation. Our data demonstrated that Herceptin alone functions as a weak agonist, while co-activation of CD16 and NKG2D results in rapid and sustained Ca2+ signaling that mediates stable synaptic interface and efficient cytolytic activity by human NK cells.
Supported by Grant from NIH U01AI148117
Gene therapy increases MHC class-I and T cell Infiltration to promote anti-tumor immune response in 4T1 breast cancer model
Authors: Shi Guilan , Heller Richard 
Published: January 2023

Abstract

Background
Immune checkpoint inhibition (ICI) has led to impressive gains in survival for many patients. However, many cancer patients treated with ICI experience intrinsic or acquired resistance which directly affects efficacy. Decreased levels of MHC class-I molecules on cancer cells and tumor infiltrating lymphocytes (TILs) have been associated with poor clinical outcomes.

Methods
Treatment of breast cancer cell lines with a plasmid encoding IL-12 (pIL-12) and vector pUMVC3 were analyzed using flow cytometry, western blotting and then validated functionally. The 4T1 breast cancer animal model was used to study T cell infiltration into the tumor as well as response following local delivery of pIL-12 with electrotransfer.

Results
Enhanced MHC class-I expression in 4T1 tumor cells, and elevated CD8+ T-cell infiltration into 4T1 tumor tissue were observed after pIL-12 gene electrotransfer. Furthermore, increased capacity to lyse 4T1 cells was observed with cultured cytotoxic T lymphocytes (CTLs) from long-term surviving mice compared to control group. A direct translation was observed in tumor-bearing mice, including attenuating tumor growth and improved tumor-free survival in mice. An immune memory response which protected against rechallenge was induced also.

Conclusion
Our findings suggest that the capacity of an induced endogenous immune response with pIL12 via the recruitment of CD8+ T cells and increased MHC class-I expression in tumor cells results in attenuating the resistance of anti-PD1 and leads to eradication of established, aggressive tumors and generates systemic tumor-specific protective immunity. Combinations of anti-PD-1 with pIL-12 gene therapy may be an effective future therapeutic strategy.
Supported by NCI R01 CA186730
BET inhibitors synergize with anti-PD1 by rescuing TCF1+ progenitor exhausted T cells in Acute Myeloid Leukemia
Authors: Romine Kyle Alexander , Cho Hyun-jun , Kosaka Yoko , Flynn Patrick , Byrd Kaelan H , Coy Jesse L , Newman Matthew T , Scott Jaime , Loo Christopher , Lind Evan F 
Published: January 2023

Abstract
Acute Myeloid Leukemia (AML) is the most common adult leukemia and has a 5-year survival of under 30%. AML is caused by uncontrolled proliferation of myeloid cells resulting from a combination of mutations that affect proliferation, differentiation and epigenetic state. For this reason, drugs targeting epigenetic modifications are being studied in AML. AML cells avoid immune recognition though inhibiting the function of multiple cell types, especially T cells and therefore immune checkpoint blockade presents a promising therapy; however, clinical trials to date have shown very modest efficacy. T cell exhaustion has been shown to be a regulated process involving transcriptional and epigenetic changes. BET proteins, which are chromatin readers, have been implicated in maintaining this exhaustion state. In these studies, we investigated the effects of the BET inhibitor (BETi) JQ1 on T cell exhaustion and checkpoint responsiveness in a murine model of AML and AML patient samples. The AML mouse model bears FLT3-ITD and deletion of TET2 restricted to myeloid lineages and is resistant to anti-PD1 therapy. This mouse model of AML expanded terminally exhausted T cells and impaired proliferative capacity after TCR stimulation. Ex vivo treatment with BETi and anti-PD1 reverses CD8+ T cell exhaustion via rescue of proliferative dysfunction and expansion of more functional precursor exhausted T cells in patient samples and AML splenocytes. Finally, we show that BETi rescues anti-PD1 resistance in vivo and reduces tumor burden in multiple organ sites and enriches CD8+ T cells in the bone marrow. In total, we demonstrate that combining BETi and anti-PD1 therapy in the treatment of AML is a rational strategy to overcome anti-PD1 resistance.
Dissecting molecular mechanisms underlying T cell co-potentiation when targeting the TCR/CD3 complex with anti-CD3 Fab fragments
Authors: Huynh Hien T , Puthanapura Chandrashekara Mahadevappa , Nelson Alfreda D , Becher Laura R E , Abergel Megan , Hu Shuang , Alarcon Balbino , Schrum Adam , Pages Diana Gil 
Published: January 2023

Abstract
A cognate antigen engagement of TCR induces a conformational change in the CD3 complex (CD3Δc) that is required for productive T cell activation. CD3Δc fails when TCRs interact with poorly immunogenic antigens that do not active T cells. Anti-CD3 monovalent Fab fragments bound to the CD3 complex mimic CD3Δc, producing T cell “co-potentiation” when associated with TCRs bound to weak antigens that enhances T cell activation, neither stimulating non-engaged T cells, nor interrupting T cell responses to strong antigens. Previously, we showed T cell co-potentiation can be exploited as a novel immunotherapeutic principle by reducing melanoma burden in B6 mice treated with anti-CD3 Fabs. TCR signaling initiation requires CD3Δc for the exposure of a conserved sequence in the cytosolic domain of CD3ɛ rich in prolines (PRS) that enables the recruitment of adaptor protein Nck. The mutation of cysteines in the CXXC motif at the extracellular domain of CD3ɛ inhibits the conformational change across the plasma membrane that results on PRS accessibility. Here, using two strains of B6 mice that express mutations in either the CXXC or the PRS motifs in CD3ɛ, we study the potential contribution of each motif in T cell co-potentiation. Our results from these studies favor the involvement of the CXXC and PRS motifs in the molecular mechanism supporting the co-potentiation of T cell activation by anti-CD3 Fab fragments.
Supported by MU Start-up funds and NIH grants: NCI, U01 CA244314; NIAID, R01 AI097187
Tumor Adjacent IL2 Cytokine Factories Eradicates Ovarian Cancer in Mice Through Cytotoxic T-cell Activation with Safe and Predictable Dosing in Non-human Primates
Authors: Carmona Guillaume , Nash Amanda , Jarvis Maria , Newman Ryan , Schladenhauffen Jake , Aghlara-Fotovat Samira , Hernandez Andrea , Nouraein Shirin , Mukherjee Sudip , Hecht Andrew , Cui Yufei , Lee Jared , Rios Peter , Zhang David , Xu Chunyu , Sheth Rahul Sheth , Peng Weiyi , Oberholzer Jose , Igoshin Oleg , Jazaeri Amir , Veiseh Omid 
Published: January 2023

Abstract
Recombinant IL-2 has been approved for the treatment of melanoma and renal cancer. However, IL-2 therapy is limited by its short half-life in circulation and the associated toxicities that emerge as a result of high systemic exposure. To overcome these limitations, we developed a localized cytokine LOCOcyteTM platform composed of allogeneic and engineered cells which produce potent immune effector molecules for local delivery with temporal regulation of activity. Administration of AVB-001 in mice and non-human primates demonstrated that IL2 local concentration (IP space) was 100x higher than the systemic concentration. Additional studies in mice demonstrated dose-dependent levels of IL2 in the intraperitoneal cavity in mice. Treatment of peritoneal tumors in mice using a single administration of AVB-001 provided sustained eradication of peritoneal tumors in an ovarian cancer mouse model. Our data in mice confirmed local increases in activation (CD25+CD8+) (8.6 fold) and proliferation (Ki67+CD8+)(3.4 fold) of cytotoxic T cells within the IP space in comparison to sham mice. Finally, a single administration of AVB-001 led to therapeutic levels of IL2 in the IP space in NHP and produced local and systemic T cell biomarker profiles that predict efficacy, AVB-001 was well tolerated by all animals with no abnormal clinical observations during the study. Our findings demonstrate that the LOCOcyteTM platform has a dose-dependent PK/PD effect, is safe and efficacious in animal models and enables itself to deliver a diverse set of cytokines alone or in combination which is presently explored. Combined, our pre-clinical results provide a rationale for testing AVB-001 in ovarian cancer patients.
Novel low-avidity glypican-3 specific CARTs resist exhaustion and mediate durable antitumor effects against hepatocellular carcinoma
Authors: He Yukai , Caraballo-Galva Leidy D , Jiang Xiaotao , Hussein Mohamed S , Zhang Huajun , Mao Rui , Brody Pierce , Peng Yibing , He Aiwu Ruth , Kehinde-Ige Mercy , Sadek Ramses , Qiu Xiangguo , Shi Huidong 
Published: January 2023

Abstract

Background & Aims
Chimeric antigen receptor engineered T cells (CARTs) for hepatocellular carcinoma (HCC) and other solid tumors are not as effective as they are for blood cancers. CARTs may lose function inside tumors due to persistent antigen engagement. The aims of this study are to develop low-affinity monoclonal antibodies (mAb) and low-avidity CARTs for HCC and to test the hypothesis that low-avidity CARTs can resist exhaustion and maintain functions in solid tumors, generating durable antitumor effects.

Methods and results
New human glypican-3 (hGPC3) mAbs were developed from immunized mice. We obtained 3 hGPC3-specific mAbs that stained HCC tumors, but not the adjacent normal liver tissues. One of them, 8F8, bound an epitope close to that of GC33, the frequently used high-affinity mAb, but with ~17 fold lower affinity. We then compared the 8F8 CARTs to GC33 CARTs for their in vitro function and in vivo antitumor effects. In vitro, low-avidity 8F8 CARTs killed both hGPC3high and hGPC3low HCC tumor cells to the same extent as high-avidity GC33 CARTs. 8F8 CARTs expanded and persisted to a greater extent than GC33 CARTs, resulting in durable responses against HCC xenografts. Importantly, compared to GC33 CARTs, there were 5 folds more of 8F8-BBz CARTs in the tumor mass for a longer period of time. Remarkably, the tumor infiltrating 8F8 CARTs were less exhausted and apoptotic, and more functional than GC33 CARTs.

Conclusion
The novel low-avidity 8F8-BBz CART resists exhaustion and apoptosis inside tumor lesions, demonstrating a greater therapeutic potential than high-avidity CARTs.
Development of swine immune reagents for analysis of immune correlates for vaccines, infection, and in biomedical research
Authors: Gourapura Renukaradhya J , Loving Crystal , Kenney Scott , LaBresh Joanna , Patil Veerupaxagouda , Byrne Kristen A , Manirarora Jean , Walker Kristen , Jovan Aquino , Chick Shannon , Dai Chai , Hailstock Taylor , Lunney Joan K 
Published: January 2023

Abstract
The goal of the USDA-NIFA Swine Immune Toolkit Initiative has been to generate priority immune reagents based on inputs from veterinary immunology researchers worldwide, and pipeline them for marketing. So far in our efforts we express soluble proteins in yeast system and then produce panels of monoclonal antibodies (mAbs) using collaborations with commercial partners for protein expression and mAb production. So far we generated several new panels of mAbs reactive to porcine IL-6, IL-13, IFN-γ, IL-17A, IL-28B, CXCL10, and BAFF and screened their reactivity in multiple immune assays. Reactivity tests of labeled anti-IL-6 and anti-IL-13 mAbs for intracellular staining of porcine immune cells using flow cytometry assay are in progress. Our results have confirmed the reactivity of porcine IL-17A, IFN-γ and CXCL10 mAbs. A sensitive sandwich ELISA is now available for IL-17A, IL-13 and CXCL10; other targets are being screened for best mAb pairs for ELISA. Planning has been initiated for the generation of IL-5 and IL-21 mAbs; and use of SLA-I & -II tetramers to identify swine CD4 and CD8 T cells specific for influenza virus peptides. For each target, our goal is to provide the veterinary community with new commercial reagents and standardized assay techniques for their research efforts. Tools and reagents generated by this project will undoubtedly advance our understanding of swine immune responses to disease, vaccine and biomedical research efforts.
Supported by USDA-NIFA AFRI grant # 2019-67015-29815