bluuebunny/crossref_metadata_2025_split huggingface dataset:
Protein Phosphatase 2A (PP2A) is essential for CD8+ T cell mediated anti-tumor immunity
Authors: Rohila Deepak , Lu Linrong 
Published: January 2023

Abstract
PP2A controls a wide variety of cellular pathways responsible for proliferation, growth, differentiation, and apoptosis. However, its role in immune cells, particularly in CD8+ T cells, still remains a mystery. We therefore sought to characterize the influence of PP2A on CD8+ T cell function in anti-tumor response. T cell specific genetic ablation of PP2A in mouse resulted in aberrant anti-tumor activity evidenced by higher tumor volume in both skin melanoma B16F10 and lymphoma E.G7 tumor graft model. Similarly, inhibition of PP2A using small molecule inhibitor LB100 showed similar effect. In addition, lower expression of effector cytokines including IFN-g, TNF-a and Granzyme B was also observed in PP2A-deleted CD8+ T Cells isolated from tumor infiltrate. Importantly, the defect in anti-tumor response is due to deficiency in CD8+ T cell function, which was proven in adoptive transfer experiment, in which CD8+ T cells from both WT and PP2A KO mice into tumor bearing RAG1 KO mice. PP2A KO CD8+ T cells-transferred mice show markedly impaired anti-tumor response. Further mechanism study suggested a possible role of PP2A in regulating CD28 signaling. PP2A KO lead to lower AKT phosphorylation upon CD28 stimulation, along with lower proliferative capability of CD8+ T cells upon stimulation with anti-CD28 and suboptimal anti-CD3 concentration. Thus this study highlights the potentially important role of PP2A in CD8+ T cells and its therapeutic consideration for designing combinatorial immunotherapy.
Identification of a core miRNA signature unique to exhausted cytotoxic CD8+ T cells
Authors: Kiernan Caoimhe H. , Hope Jennifer L , Leon Leticia G. , Zhao Manzhi , van Meurs Marjan , Brouwers-Haspels Inge , van Strien Paulette , Bindels Erik , Hoogenboezem Remco M. , Li Yunlei , Stubbs Andrew P. , Zehn Dietmar , Mueller Yvonne M , Erkeland Stefan J. , Katsikis Peter D 
Published: January 2023

Abstract
Cytotoxic CD8+ T lymphocytes (CTLs) play a major role in protection against chronic viral infections and tumors. Chronic antigen stimulation in both conditions leads to CTL exhaustion and failure of CTLs to efficiently eradicate tumors and virally infected cells. miRNAs are small, non-coding RNA molecules that participate in post-transcriptional gene regulation. This study aimed to determine the core miRNA signature of CTL exhaustion in mouse models of both tumors and chronic infection. Exhausted anti-tumor CTLs were isolated from tumors of mice that received 10^6 mouse mesothelioma cells (AE17sOVA) followed by an adoptive transfer of 10^4 CD45.1+ OT-I cells. Effector anti-viral CTLs were isolated from mice adoptively transferred with 10^4 CD45.1+ OT-I cells or P14 cells and infected with WSN-OVA or LCMV Armstrong respectively. For exhausted anti-viral CTLs, mice were adoptively transferred with 10^4 CD45.1+ P14 cells and infected with LCMV-Cl13. Single cell suspensions were prepared from the different tissues, live CTLs were FACS sorted, RNA isolated and small RNA-seq was performed. Principle component analysis (PCA) of miRNA expression revealed that exhausted anti-tumor CTLs and anti-LCMV Cl13 CTLs cluster apart from effector anti-viral CTLs (WSN-OVA and LCMV Armstrong). Differential expression analysis identified 73 miRNAs commonly shared between the exhausted day anti-tumor CTL and anti-viral CTL. Using RNA sequencing of two different models of exhaustion, distinct RNA profiles common in exhausted CTLs were found. This core miRNA signature can now be used to manipulate the expression of specific miRNA to potentially prevent or revert CTL exhaustion and improve the CTL response against tumors and/or chronic infections.
Differential recruitment and activation of mucosal and systemic ILC for improved vaccine design
Authors: Khanna Mayank , Sheehan Jared , Nichols Olga , Ranasinghe Charani , Ramsay Alistair 
Published: January 2023

Abstract
Innate lymphoid cells (ILC) have emerging roles in immune modulation, tissue homeostasis, and inflammation. ILC may function as innate counterparts of T cells, influencing early host responses against infection or vaccination, potentially modulating development of adaptive immune responses. Little is known, however, of their recruitment and activation following immunization. Here, we investigated the activity of ILC populations after immunization with poxvirus-based vaccine vectors. Our data suggest that fowlpox virus (FPV) and modified vaccinia Ankara strain (MVA) vectors can differentially polarize ILC responses at mucosae. Intranasal (IN) delivery of FPV vectors increased numbers of pulmonary ILC1 and ILC3 subsets expressing IL-17a and IL-22 through 72 hours post-immunization. In contrast, MVA vectors given IN enhanced pulmonary ILC2 numbers and their expression of IL-13. Our previous work indicated that mucosal immunization with these vectors induced high avidity T cell responses associated with transient inhibition of IL-4/13 expression by local ILC2. Since IL-25 has been shown to down regulate IL-12 and promote ILC2/IL-13 expression, we delivered FPV encoding an IL-25 binding protein (IL-25BP) in order to sequester host IL-25 at the site of immunization. While mucosal delivery of FPV-IL-25BP appeared to induce ILC2/IL-13 expression in the lungs, IM delivery reduced ILC2/IL-13 expression, increased IL-17a expression by local ILC1 and ILC3, and enhanced Th1-type adaptive immune responses against co-expressed vaccine antigens. Together, our findings suggest that manipulation of ILC responses by vaccine vectors, co-expressed immune modulators, and delivery route may be exploited for improved vaccine design.
Development of a reverse genetics system to generate vaccine strain of Hepatitis A virus
Authors: Seo Dongwon , Kang Hyeon Jeong , Choi Jung Ah , Seo Sang Hwan , Song Man Ki 
Published: January 2023

Abstract

Objective
Reverse genetic system have been used to generate mutant viruses to investigate the genetic features as well as to produce vaccine strain of various viruses. However, the virus producing system has not been applied to produce vaccine strain of Hepatitis A virus (HAV). Here, we established reverse genetic system to generate vaccine strain of HAV and assessed the generated HAV in terms of immunogenicity.

Methods
To produce HAV, MRC-5 cells were transfected with HAV RNA generated via in vitro transcription. To confirm the virus generation, we performed RT-PCR, Western blot, Rapid amplification of cDNA ends (RACE) and transmission electron microscopy (TEM) analysis using the supernatant of the HAV RNA transfected cells. To assess the immunogenicity of the generated HAV, mice were immunized with the generated and inactivated HAV and performed enzyme-linked immunosorbent assay (ELISA) to assess anti-HAV IgG using sera from the immunized mice.

Results
From the supernatant of HAV RNA transfected cells, HAV-specific RNA and protein were detected. TEM analysis showed that there are two types of HAV particles as shown in a previous report: naked and enveloped HAV, which is one of HAV features. Although seven nucleotide sequences are different in the coding sequence for structural protein compared to those of wildtype vaccine strain of HAV, genomic RNA analysis using RACE showed that the generated HAV have HAV-specific RNA as genome. In the immunogenicity study using mouse, the generated HAV induced HAV-specific IgG and the titer was comparable to wild-type vaccine strain of HAV.

Conclusions
We established the reverse genetic system to produce vaccine strain of HAV and confirmed that the generated HAV can be used as vaccine strain.
Chlamydia recombinant MOMP encapsulated in PLGA 85/15 nanoparticles admixed with Escherichia coli dmLT mucosal adjuvant enhanced protection against a genital challenge in subcutaneously immunized BALB/c mice
Authors: Sahu Rajnish , Joshi Sameer , Norton Elizabeth B. , Singh Shree R. , Dennis Vida A. 
Published: January 2023

Abstract
There is an urgent need to develop a vaccine against Chlamydia trachomatis, the most reported bacterial-sexually transmitted infection globally. We developed a Chlamydia nanovaccine by employing PLGA [poly (D, L-lactic-co-glycolic acid) 85/15 nanoparticles encapsulated with recombinant MOMP (major outer membrane protein) of Chlamydia](named PLGA-rMOMP) that partially protected mice against a genital challenge. Here we hypothesized that admixing PLGA-rMOMP with the Escherichia coli dmLT (double-mutant heat-labile enterotoxin (LT)-R192G/L211A) mucosal adjuvant will improve the protective efficacy. Six groups of BALB/c mice were subcutaneously immunized three-times at two-week intervals with various immunogens and then challenged intravaginally with 105 inclusions forming units (IFUs) of C. muridarum. Vaginal swabs were collected every three days up to three weeks from all mice and analyzed by indirect immunofluorescence to compare the IFUs between groups of mice. Our results showed maximal protection provided in mice immunized with PLGA-rMOMP admixed with dmLT evidently by their lower IFUs as compared with the other mice groups. Moreover, the PLGA-rMOMP adjuvanted mice produced elevated MOMP-specific serum IgG2b (Th1) as compared to IgG1 (Th2) antibodies, which is indicative of an effective Th1 immune response. Interestingly splenic T-cells from these mice also produced enhanced IL-17 and IL-2 MOMP-specific cytokines after the genital challenge in comparison to the other groups of mice. Our results indicate that the E. coli dmLT mucosal adjuvant improves the immunogenicity and the protective efficacy of the PLGA-rMOMP nanovaccine against a genital challenge in mice.
The effective immunization of plasmid DNA vaccine targeted for smallpox virus
Authors: Son Wonrak , Kim Nayoung , Lee Minhoon , Sim Euni , Song Donghyun , Yu Chiho , Song YoungJo , Hur GyeungHaeng , Jeong SeongTae , Hong Sungyoul , Shin YoungKee , Shin Sungho , Choi JunYoung 
Published: January 2023

Abstract
Smallpox, caused by variola virus, could be a bio-terrorism agent although eradicated from the WHO program in the 1980s. DNA vaccine is a third generation vaccine system which contains DNA sequence with specific antigen codes that activates the immune system. DNA vaccines show great potential in inducing immune response with strong triggered-humoral, and cellular immunity. DNA vaccines have emerged as an attractive approach for infectious disease because of rapid manufacture, fast adaptation to newly emerging pathogens and high stability at ambient temperatures. However, since the potency of naked DNA vaccines is limited to immune efficacy, adjuvant incorporation could be a good option to increase the immune response. Several different compound-composed adjuvants are developed to help DNA vaccine efficacy. In other aspects, for inducing a more effective immune response, global vaccine manufacturer focused on targeting dendritic cell (DC) through the DC-specific surface protein, DEC-205. In this study, with plasmid construction modification, we developed a DNA vaccine vector that have effective antigen expression and confirmed immunogenicity against smallpox. For enhancing in vivo immune responses, we screened various adjuvant strategies and observed effective DNA vaccination response with various adjuvant studies and time-pointed reaction. Also, we tested the enhancement of immune response using DNA vaccine containing the smallpox antigen fused with DEC-205 and revealed a th1–th2 cellular immune response. Our results propose how to regulate modified-DNA immunization efficacy within DC targeting stimulation and adjuvant strategy against DNA vaccine development.
Understanding how immune maturity shapes responses to cancer therapy
Authors: Serrano Daniel , Tzelepis Fanny , Chen Andrew , Diallo Jean-Simon 
Published: January 2023

Abstract
Clinical translation of cancer therapies from mice to human remains a constant challenge. Cancer immunotherapy aims develop a sustained long-term adaptive immune response to tumors. This raises the question of whether the typical pathogen-free animal models reflect the human immunological context and cancer pathology. Recent research suggests that exposure of pathogen-free mice to pet shop mice leads to their immunological maturation. We hypothesize that such a model may be more reflective of a human adult context. Specific-pathogen-free (SPF) Balb/c mice were co-housed with pet shop mice for 40 days to generate the “dirty mouse” model. Dirty mice were challenged with Listeria m. infection to test immune maturity. In parallel, CT26LacZ, CT26WT, EMT6 and K7M2 tumor cell lines were used to measure tumor growth in dirty mice. In particular, CT26LacZ tumor model was treated with oncolytic virus to assess the impact of immune maturity over efficacy of therapy. Complementary comparative experiments were performed in SPF mice. Dirty mouse immune response to Listeria challenge was more robust than in clean mice. Immune phenotyping revealed a different distribution of immune populations. A matured immune system impacted the growth of tumors to varying extents, with some models experiencing complete tumor rejection. Finally, oncolytic virus in dirty mice implanted with CT26-LacZ showed an improved efficacy on survival and tumor burden. Immune response to infections and tumor models between SPF mice and dirty mice reveals the potential impact of a mature immune system on immune-therapeutic responses. Therefore, establishment of model that better recreates the state of immune maturity may result in higher chances of translational impact.
Development of a prophylactic vaccine for lung squamous cell carcinoma
Authors: Riolobos Laura , Gad Ekram , Treuting Piper M. , Timms Andrew , Disis Mary Lenora 
Published: January 2023

Abstract
High-grade bronchial dysplasia is a marker for high risk of lung squamous cell carcinoma (SCC). Cancer vaccines targeting dysplasia could prevent the progression to SCC and decrease lung cancer incidence in population at risk. In order to develop a vaccine to prevent lung SCC we need to identify antigens and epitopes within them able to elicit a potent Type I anti-tumor immune response. One caveat to develop a prophylactic vaccine targeting dysplasia is that driver mutations (neo-antigens) are not known. Many mutations appear late in lung cancer and are not shared between patients. However, upregulated non-mutated proteins are needed from the early stages of cancer to support the new proliferation requirements of the cells, are common between patients and are good candidates for a preventive vaccine. Type I CD4+ T cells (Th1) secreting interferon-gamma (IFN-g) are necessary for enhanced function of antigen presenting cells, epitope spreading and activation of cytotoxic CD8+ T cells. However, non-mutated tumor self-antigens frequently induce Type II CD4+ T cells (Th2), which secrete cytokines that inhibit the function of Th1 and CD8+ T cells. To develop a prophylactic lung cancer vaccine we have: (1) searched in public datasets genes that are upregulated in bronchial dysplasia and maintain upregulation in SCC; (2) used web based algorithms for class II epitope prediction followed by functional ELISPOT screening to identified epitopes eliciting IFN-g (Th1) and/or IL-10 (Th2) responses; (3) selected only the Th1 epitopes (excluding the Th2 epitopes) to evaluated efficacy of the vaccines in a chemically induced rodent model of SCC. Preliminary data shows that we can reduce dysplasia frequency in vaccinated mice treated with the carcinogen.
Loss of the Class II Accessory Molecule HLA-DO leads to presentation of lower abundance of DM-resistant epitopes
Authors: Welsh Robin A , Song Nelson , Foss Catherine , Sadegh-Nasseri Scheherazade 
Published: January 2023

Abstract
HLA-DO (DO in human; H2-O in mice) is a highly conserved non-classical MHC class II accessory molecule. Unlike the main Class II peptide editor HLA-DM, expression of DO is restricted to the thymic medulla, B cells and certain dendritic cell subsets. Although DO forms a stable complex with HLA-DM, its biological function remains poorly understood. Previously, our lab provided biochemical evidence in support of a model where DO enhances the binding of DM-resistant peptides, and reduces the binding of DM-sensitive peptides to the HLA-DR1 (DR1) molecules in vitro. Changes in the quantities of either DM-resistant of DM-sensitive peptides could therefore lead to alterations in both the thymic deletion, and peripheral activation of CD4 T cells. Here, using in vivo antigen presentation in an autoimmune as well as infectious model, we found that DO-KO mice present a lower abundance of MOG(35-55)/I-Ab and H5N1-HA(259-274)/DR1 complexes by B cells. Using 2D2 Tg T cells specific for MOG(35-55)/I-Ab as a readout system, we observed a less effective T cell activation upon co-culture of brain recovered APCs from diseased DO-KO mice. When we vaccinated DR1+DO-KO and DR1+DO-WT mice with flu vaccine, we found that HA(259-274)/DR1 specific CD4 T cells had a less avid interaction with B cells presenting HA(259-274)/DR1 from DR1+DO-KO as compared to B cells from vaccinated DR1+DO-WT mice as measured by a novel avidity measurement tool, z-MoviTM.
Identification and characterization of adipose tissue-derived human antibodies with “anti-self” specificity
Authors: Frasca Daniela , Diaz Alain , Romero Maria , Garcia Denisse , Jayram Diya , Thaller Seth , Blomberg Bonnie 
Published: January 2023

Abstract
We have previously shown that the human obese adipose tissue (AT) contributes to increased secretion of adipocyte-specific IgG antibodies in individuals with obesity. We have also shown that this occurs without any exogenous stimulation, because the ongoing process of cell death in the obese AT leads to the release of “self” antigens able to induce chronic stimulation of B cells. Here, we confirm and extend our initial observation on a different cohort of individuals, and we show that also the plasma of obese individuals is enriched in IgG antibodies with specificities for adipocyte-derived antigens. Adipocyte-specific IgG secreted in the obese AT are significantly correlated with those present in plasma. Using immunoprecipitation and mass spectrometry, we have identified these antigenic specificities. The antigens are almost exclusively intracellular or cell-associated, usually not “self” antigens, but they are released by cells dying in the AT. These antigens will be used in protein arrays to screen plasma from individuals with obesity and also with autoimmune diseases. We have indeed preliminary evidence that the plasma of Rheumatoid Arthritis patients is enriched in adipocyte-specific IgG antibodies, suggesting that obesity may contribute to pathogenicity in autoimmune patients. Finally, we show for the first time that not only macrophages but also adipocytes in the obese AT are efficient antigen-presenting cells and stimulate the secretion of IgG autoimmune antibodies. They do so because they express the antigen-presenting molecules CD1d and, to a lesser extent, MHC class II, as well as the co-stimulatory molecule CD86. These results may lead to the development of novel therapeutic strategies to controlautoimmunity.
Insulin-specific regulatory T cell receptor alpha clonotypes restricted to HLA-DQ6 (DQB1*06:02) are shared between blood and islets
Authors: Mitchell Angela M , Alkanani Aimon K , McDaniel Kristen A , Landry Laurie G , Nakayama Maki , Michels Aaron W 
Published: January 2023

Abstract
The major genetic determinant in susceptibility or protection from many autoimmune diseases resides in the human leukocyte antigen (HLA) region. Specific class II alleles (e.g., HLA-DQ8) increase the risk for developing type 1 diabetes (T1D), a prototypical organ specific autoimmune disease, whereas HLA-DQ6 leads to dominant protection. We hypothesize that DQ6 is protective via its ability to present insulin peptides to regulatory CD4+ T cells (Tregs), resulting in anti-inflammatory responses.
We expanded insulin-specific Tregs from peripheral blood mononuclear cells (PBMCs) of DQ6+ non-diabetic individuals using an insulin peptide. Insulin-expanded Tregs were flow sorted, and paired single-cell TCR/RNA-seq (10X Genomics) was performed.
Of 2,096 insulin-expanded Tregs from three DQ6+ individuals, 883 αβTCRs (42%) were present ≥2 times, and we confirmed the antigen specificity of dominant clonotypes. The single-cell RNA-seq data showed 4 clusters of Treg cells based on the expression levels of FOXP3 and IL-2RA. Additionally, we flow-sorted CD4+ T cells from the pancreatic islets of a non-diabetic DQ6+ organ donor and performed single-cell TCR sequencing. Of the 67 CD4+ cells, TCR clonotypes from three cells possessed identical TCRα chains (including CDR3 sequences) with Tregs sequenced from the PBMCs of the separate DQ6+ non-diabetic individuals.
These data indicate the potential presence of insulin-specific Tregs within the pancreatic islets of non-diabetic organ donors. Our findings provide a mechanistic basis for understanding HLA-linked protection from autoimmune diabetes development and have implications for manipulating self-antigen-specific T cell responses to modify the disease course in T1D.
High-salt modulates cellular metabolism of human regulatory T cells
Authors: Kleinewietfeld Markus , Côrte-Real Beatriz F , Hamad Ibrahim 
Published: January 2023

Abstract
High-salt intake has been linked with shifts in the immune cell balance, mainly by promoting proliferation and activity of pro-inflammatory cells, such as T helper 17 (Th17) and M1 macrophages, and by impairing the functions of anti-inflammatory cells such as regulatory T cells (Tregs) and M2 macrophages. However, the precise molecular mechanisms that contribute to this phenomenon are still unknown. The role of metabolic regulation in shaping immune responses has gained increasing attention in recent years and can be greatly influenced by environmental factors such as diet. High-salt intake was previously shown to promote metabolic changes in M2 macrophages by decreasing their mitochondrial oxidative phosphorylation (OXPHOS) and glycolysis necessary for their activation. So we hypothesize that this phenomenon could likely be extrapolated to other cell types such as Tregs. We analyzed changes on the metabolic pathways of human Tregs after high-salt exposure. Our results show significant salt-induced changes in the cellular metabolism of Tregs and since these changes are known to alter suppressive function both in vitro and in vivo, we further hypothesize that the observed metabolic alterations might be linked to the loss of suppressive function seen in human Tregs upon high-salt challenge. Thus, the interference with these pathways may have the potential for targeting Tregs in salt-sensitive diseases.