bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD317-mediated cell cytoskeleton regulation as a novel means of tumor immune evasion
Authors: Zhang Guizhong , Cheng Jian , Liu Zhao , Deng Tian , Adeshakin Funmilayo Oladunni , Wan Xiaochun 
Published: January 2023

Abstract
CD317 is frequently overexpressed in various human cancers. Although our previous study had demonstrated its pro-proliferative role in HCC development, the functions of CD317 in tumorigenesis, especially in tumor immune evasion, remain unclear. Herein we report that CD317 protects cancer cells from immune cell-mediated cytotoxicity. CD317 knockdown in cancer cells affects neither cell-cell conjugation nor effector cell activation, but markedly increases the vulnerability of cancer cells by which renders cancer cells more sensitive to YTS- or CAR-modified NK cell-mediated cytotoxicity. This sensitivity alteration cannot be restored by adding recombinant CD317 extracellular domain (317ECD) protein, excluding the possibility that CD317 acts as a traditional immune checkpoint protein that provides inhibitory signaling to effector cells. Mechanically, immunoprotective function of CD317 is mainly dependent on the interaction with RICH2, a known CD317-binding partner that is involved in the cytoskeleton regulation. Knocking down RICH2 significantly rendered CD317 incapable of protecting cancer cells from immune cell-mediated killing, suggesting CD317 facilitates tumor immune evasion by regulating tumor cell cytoskeleton to confront immune killing. Taken together, our findings reveal a novel means of tumor immune evasion and suggest CD317 as an attractive target for improving the efficiency of cancer immunotherapy.
TLR2 Signaling Promoted Tumor Development through Enhancing Inflammation and Regulatory T cells to Suppress Anti-Tumor Immunity
Authors: Shen Chia-Rui , Lee Meng-Hua , Kuo Ming-Ling 
Published: January 2023

Abstract
Inflammation is a relevant physiologic process necessary for immune protection, tissue repair and remodeling. However, chronic inflammation is known to contribute tumor development, including tumor initiation, promotion and progression. Recent studies have shown that up-regulation of Toll-like receptors (TLRs) in tumor microenvironment induced the inflammatory responses. Here, we hypothesized that the TLR2 signaling within tumor microenvironment elicits an inflammatory response and such facilitates tumor development. A murine B16F10 melanoma model was utilized to investigate the role of endogenous TLR2 signaling in tumor development. Anti-tumor immunity and the presence of tumor-associated inflammatory mediators were analyzed by flow cytometry and ELISA in the TLR2 knockout (TLR2 KO) and wild-type (WT) tumor-bearing animals. When compared to the TLR2 deficient mice, WT mice had a significantly aggressive-appearing pathology, such as elicited tumor growth and metastasis. When the splenocytes cultured with tumor-conditional medium, WT produced elevated levels of pro-inflammatory cytokines, whereas TLR2KO or the WT receiving anti-TLR2 antibody treatment blocked the inflamed-like phenomenon and consequently showed a better anti-tumor IFN-γ response. Additionally, tumors from WT mice exhibited comparably higher infiltration levels of CD4+CD25+Foxp3+ regulatory T cells, but much less CD8+ T cells. These results support that cancer cells-derived components through TLR2 signaling generate an inflammatory microenvironment hospitable for tumor growth, and it indicates TLR2 as a potential therapeutic target for cancer immunotherapy.
Immune checkpoint molecule V-set Ig domain-containing 4 (VSIG4) expression is associated with poor prognosis in advanced gastric cancer patients
Authors: Kim So-Woon , Roh Jin , Jung Jiwon , Pak Hyo-Kyung , Lee A-Neum , Park Young-Soo , Park Chan-Sik 
Published: January 2023

Abstract

Background
New therapeutic approaches are needed for the treatment of advanced gastric cancer (AGC). V-set Ig domain-containing 4 (VSIG4) is an immune checkpoint molecule that may have prognostic or predictive utility in the treatment of AGC. The aim of this study was to investigate expression of VSIG4 and its clinical significance in AGC and other major cancers.

Methods
We analyzed VSIG4 expression and its correlation with survival rate in various carcinomas including 1,034 surgically resected AGCs from two academic hospitals. Patients were divided into a discovery (N = 572), internal validation (N= 230), and external validation (N= 232) sets.

Results
VSIG4-positive AGC tumors were significantly associated with overall survival (OS) (hazard ratio (HR) = 3.02, 95% confidence interval (CI) = 2.32–3.94, P&lt;0.001,) and event-free survival (HR = 2.8, 95% CI=2.18–3.72, P &lt;0.001). These findings was successfully validated in the independent cohorts. VSIG4-positive status was also significantly correlated with low intratumoral CD8+ T-cell infiltration (P = 0.036) and high FoxP3+/CD8+ intratumoral T-cell infiltration ratios (P = 0.016). VSIG4 expression in other cancers was not associated with higher 5-year OS rates (colon, P = 0.459, lung, P = 0.275, kidney, P = 0.121, breast, P = 0.147).

Conclusions
VSIG4 is an independent prognostic factor in AGC. Notably, VSIG4 expression was significantly associated with low CD8+ T-cell infiltration and high regulatory T cell/CD8+T cell ratios, implying a cancer tolerating immune environment. Our results may be an important basis for future gastric cancer-specific immunotherapeutic drug development.
A TruCulture Whole Blood Assay to Evaluate Annual Flu Vaccine Recall Responses
Authors: Hwang Shen-An , Eisinger Dominic , LaBrie Samuel 
Published: January 2023

Abstract
Evaluation of vaccine efficacy to protect against future infection remains a challenge. One of the difficulties lies in developing an antigen specific recall assay that can be applied across clinical sites. TruCulture is a whole blood collection and culture system that was designed for ex vivo/in vitro evaluation of immune responses for clinical trials. This abstract demonstrates a TruCulture tube that is highly effective in eliciting recall responses against Influenza A hemagglutinin (HA) proteins after the annual flu vaccine. Whole blood from 10 healthy donors were collected at 2 days pre- and 14 days post-vaccination (Afluria, Sequiris). Blood was collected in TruCulture tubes containing 5mg recombinant HA antigens (Immune Technology Corporation). After incubation at 37°C for 48 hours, the supernatants were collected and analyzed for 13 analytes by OptiMAP and 47 analytes by TruCulture MAP (Myriad RBM). Several analytes (such as IFNγ, IL-1β, IL-3, IL-4, IL-6, IL-10, IL-12, IL-17, and IL-23) demonstrated an increase at 14 days after the flu vaccination compared to samples taken prior to vaccination. This increase is specific to the recombinant HA antigens from flu strains included in the flu vaccine administered. The increase in these analytes were either not observed or at significantly reduced levels when the recall response was conducted using recombinant HA antigens from flu strains not included in the vaccine administered. TruCulture preloaded with HA antigens can be a valuable tool to evaluate human immune responses to the annual flu vaccine and indicates that a similar strategy using antigen-specific TruCulture tubes can be developed for the evaluation of any novel vaccine.
Evaluating Influenza A virus vaccines in a dirty mouse model better mimics the human immune response
Authors: Fiege Jessica K , Block Katharine E , Pierson Mark J , Pape Kathryn A , Jenkins Marc K , Hamilton Sara E , Masopust David , Langlois Ryan A 
Published: January 2023

Abstract
Current seasonal influenza A virus (IAV) vaccines often exhibit reduced efficacy to seasonal strains and offer limited protection to novel pandemics. A number of therapies, including vaccines, that are successful in mice often fail to translate to humans. This could be due to housing animals in specific pathogen free (SPF) conditions. Humans are exposed to a variety of natural host pathogens that SPF mice are protected from. Unlike SPF mice, the immune system of pet store mice more closely recapitulates the immune system of humans. When SPF mice are co-housed with pet store mice harboring natural mouse pathogens, termed dirty mice, these co-housed mice obtain phenotypes observed in the human immune system. In order to determine if dirty mice are an improved mouse model for vaccine development/testing, we sought to study the adaptive immune response to IAV in dirty mice. We infected SPF housed mice or dirty mice with IAV and assessed viral replication, clearance and adaptive immune responses. When we assessed memory responses to IAV in control or dirty mice, we observed altered B and T cell responses in dirty mice. These data demonstrate the importance of studying IAV in the dirty mouse model to facilitate more effective vaccine development and testing before human studies are performed.
The humoral immune response elicited upon immunization with recombinant BCG vaccines protects against hRSV and hMPV in a murine preclinical model
Authors: Galvez Nicolas M. S. , Soto Jorge A. , Bueno Susan M. , Kalergis Alexis M. 
Published: January 2023

Abstract
Human respiratory syncytial virus (hRSV) and human metapneumovirus (hMPV) are among the main etiological agents responsible of acute lower respiratory tract infections in vulnerable populations worldwide. Despite their clinical relevance, the licensing of a safe and effective vaccine has not been achieved. Considering this, our laboratory developed two different recombinant Mycobacterium bovis BCG strains as vaccine candidates. These recombinant BCGs express either the hRSV-Nucleoprotein (rBCG-N) or the hMPV-Phosphoprotein (rBCG-P). Both vaccines prototypes were able to promote cellular protection against their respective virus. However, the humoral response elicited upon immunization with these vaccines has not been characterized. Here we show that, upon immunization and challenge of mice with the respective rBCG and virus, the induced humoral response was able to protect against the viral infection, promoting the secretion of specific antibodies against each virus and several of their proteins, as measured in sera samples by ELISA assays. We also described that immunization and challenge promoted the induction of antibodies with enhanced neutralizing capacities, as seen during in vitro assays. Moreover, we show that sera transferred from immunized and challenged mice into naïve mice is enough to protect from the infection, when these naïve mice were challenged with the viruses. This protection was characterized by a decreased in the viral replication and pulmonary disease. These results support the notion that the use of rBCG vectors could be an effective vaccine platform against these two major respiratory pathogens and places our vaccine candidates as suitable approaches for the treatment against both hRSV and hMPV.
Ursolic acid moderates Th17-mediated immunity in respiratory syncytial virus infections of young cattle
Authors: Slate Jamison Ray , HONG SUYEON , McGill Jodi L 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) is a leading cause of severe lower respiratory tract disease in infants and young children. Bovine RSV is a common cause of respiratory disease in young cattle with high genetic and immunological similarity to human RSV, making it a relevant clinical model. While RSV is highly prevalent in both humans and cattle, the disease pathogenesis remains poorly understood and therapeutic options are limited. Th17-mediated immunity is intrinsically linked to RSV infections and characterized by the production of the inflammatory cytokine IL-17A; however, excessive IL-17 production may contribute to immunopathology by exacerbating neutrophil recruitment and activation in the lungs. Herein, we sought to elucidate the role of Th17-mediated immune responses by inhibiting Th17 cell differentiation with ursolic acid (UA), a small molecule agonist of the RORγt transcription factor. Calves were prophylactically treated with UA before being challenged with aerosolized bovine RSV. Challenged calves were monitored for clinical disease, gross and microscopic lung pathology, and immunologic responses in the periphery and respiratory tract. Local and systemic neutrophils and monocytes were analyzed for phagocytic and oxidative burst functions. Cellular adaptive immune responses were measured in the peripheral blood and airways by antigen recall assays. Our results suggest that UA-treatment may result in reduced RSV-associated immunopathology by shifting the immune response from Th17-mediated immunity to a more productive Th1-mediated immune response.
Glioblastoma infiltration of both tumor- and virus-antigen specific cytotoxic T cells correlates with experimental virotherapy responses
Authors: Nakashima Hiroshi , Alayo Quazim , Chiocca E. Antonio 
Published: January 2023

Abstract
The mode of action for oncolytic viruses (OVs) in cancer treatment is thought to depend on a direct initial cytotoxic effect against infected tumor cells and subsequent activation of immune cell responses directed against the neoplasm. To study both of these effects in a mouse model of glioblastoma (GBM), we employed murine GBM cells engineered to constitutively express the type I Herpes Simplex Virus (HSV1) HSV-1 receptor, nectin-1, to allow for more efficient infection and replication by oncolytic HSV (oHSV). These cells were further engineered with a surrogate tumor antigen to facilitate assays of T cell activity. We utilized MRI-based volumetric to measure GBM responses after injection with the oHSV and bioluminescent imaging (BLI) to determine oHSV replicative kinetics in the injected tumor mass. We found increased infiltration of both surrogate tumor antigen- and oHSV antigen-specific CD8+ T cells within 7 days after oHSV injection. There was no increase in tumor infiltrating CD8+ T cells expressing “exhaustion” markers, yet oHSV infection led to a reduction in PD-1+ CD8+ T cells in injected GBMs and an increase in IFNγ+ CD8+ T cells. There was a significant direct correlation between oHSV-mediated reduction in GBM volume, oHSV intratumoral gene activity and increased infiltration of both viral and tumor antigen-specific CD8+ T cells. These findings imply that both intratumoral viral replication and CD8+ T cell cytotoxicity against both tumor and viral antigens are important in oHSV-mediated GBM therapy.
Phenotypic small-molecule screening identifies JAK inhibitors as pharmacologic modulators of T cell exhaustion
Authors: Marro Brett S. , Zak Jaroslav , Pratumchai Isaraphorn , Lairson Luke , Oldstone Michael B.A. , Teijaro John R 
Published: January 2023

Abstract
Immune checkpoint blockade (ICB) therapy has emerged as viable first-line treatment for cancers such as melanoma. Despite the striking long-term clinical benefits that were previously unattainable, the effectiveness of ICB therapy remains limited in a majority of patients and cancer types. This is due in part to insufficient activation and/or restoration of anti-tumor T cell responses. Thus, understanding the signaling pathways that potentiate T cell exhaustion is essential for developing pharmacologic drugs that reactivate T cells and overcome the limitations of current ICB therapy. To this end, we developed a biologically relevant phenotypic screening system using the LCMV-CL13 infection model coupled with high-throughput flow cytometry to identify small-molecules that resurrect hypofunctional T cells. We discovered 19 hits following a screen of a manually curated collection of 12,000 repurposed drugs. Among the lead compounds identified were janus kinase (JAK) inhibitors including the anti-myelofibrosis drug ruxolitinib. Phenotypic rescue of exhausted T cells in vitro following ruxolitinib treatment resulted in a unique transcriptional signature that diverged from anti-PD-L1 treatment. Lineage-defining genes known to preserve precursor exhausted CD8+ T cells, including transcription factor 7 (Tcf7), were significantly upregulated following exposure to ruxolitinib. Mechanistically, ruxolitinib attenuated the cumulative pSTAT signaling signature within resurrected cells to enhance their survival. Collectively, these results demonstrate a disease-relevant framework for identifying small-molecule modulators of dysfunctional T cells and suggest that JAKs are viable targets for cancer immunotherapy.
T helper differentiation of CAR T cells augments function and target cell killing
Authors: Nelson Andrew Scott , Khan Ibrahim , Kaplan Mark H 
Published: January 2023

Abstract
Chimeric antigen receptor (CAR) T cell therapy involves genetically manipulating T cells to express an antigen receptor specific for tumor protein, resulting in a powerful anti-cancer therapeutic. However, trafficking to tumor sites and the suppressive nature of the tumor microenvironment have prevented effective treatment of solid tumors. Previous work has shown that antigen specific T helper (Th) subsets have variable tumor killing capabilities, however, it is not completely defined how different transcriptional differentiation programs effect the fitness and function of CAR T cells. It was hypothesized that differentiation of T cells transduced with CAR improves cancer cell killing in an antigen specific manner. To test this hypothesis, splenic CD4+ and CD8+ T cells were cultured under T cell activating (T0), Th2 (T2), or Th9 (T9) cell differentiation conditions and transduced with a 2nd generation CD19-specific CAR. Control cells were transduced with CD19-specific CAR lacking the intracellular domain. CAR T cells were co-cultured with CD19+ B cells and target cell killing was measured by flow cytometry. Preliminary results show that CAR T9 cells, but not T2 cells, kill target cells similar to the clinic relevant T0 cells. FACS analysis of T cells after co-culture with target cells revealed antigen-experience promoted production of lineage specific cytokines as well as granzymes A and B. T9 conditions maintained a larger proportion of CD8+ T cells than T2 conditions, suggesting that T9 conditions promote expansion of effector CD8+ T cells. These results suggest that the Th9 transcriptional program promotes target cell killing, transduction efficiency, and generates a better ratio of CD4:CD8 T cells for adoptive cell therapy.
CD4+ T cell help is required for the formation of a cytolytic CD8+ T cell subset that protects against chronic viral infection and cancer
Authors: Zander Ryan A , Schauder David , Xin Gang , Nguyen Christine , Wu Xiaopeng , Zajac Allan J , Cui Weiguo 
Published: January 2023

Abstract
CD8 T cells responding to chronic viral infection can be classified into two major subsets, with a TCF-1+ subset serving as a self-renewing progenitor population that can give rise to a more terminally exhausted PD-1hi subset. However, whether additional heterogeneity exists amongst virus-specific CD8 T cells remains unclear. Here, using single cell RNA-sequencing and flow cytometric approaches, we demonstrate that in addition to the progenitor and exhausted subsets, CD8 T cells responding to persistent LCMV infection also encompass a previously unrecognized T cell subset that exhibits potent cytolytic activity in vitro, and is required for viral control in vivo. This cytotoxic subset is characterized by high expression of killer cell lectin-like receptors, the chemokine receptor CX3CR1, and the transcription factors T-bet and Zeb2. Notably, we further demonstrate a critical role for CD4 help via IL-21 production in facilitating the differentiation of this cytotoxic subset, as depletion of CD4 T cells or deletion of IL-21R signaling in P14 cells (which express a transgenic TCR specific for GP33–41) wholly abrogated the development of CX3CR1+CD8 T cells. Moreover, PD-L1 blockade, an immunotherapy known to partially reverse T cell exhaustion, failed to rescue this differentiation defect of “un-helped” CD8 T cells. Lastly, using a B16 melanoma model, we found that provision of IL-21+CD4 T cells was sufficient to induce over a 2-fold increase in the development of CX3CR1+CD8 TILs, which was associated with enhanced control over tumor growth. Collectively, our findings demonstrate a critical role for CD4 help in the generation of protective CX3CR1+CD8 T cells, and that this differentiation checkpoint may be exploited for immunotherapy.
HSV-2 infects T follicular helper cells to promote HIV reactivation
Authors: Pierce Carl A , Loh Lip Nam , Preston-Hurlburt Paula , Herold Kevan C , Herold Betsy C 
Published: January 2023

Abstract
HSV-2 is one of the most significant factors fueling the global HIV epidemic and coinfection is associated with higher HIV plasma viral loads and increased rates of HIV transmission and HSV-2 recurrences. The mechanisms underlying this syndemic likely reflect local and systemic effects. To explore potential systemic effects, we characterized PBMC from a cohort of HIV-1+ women who were or were not HSV-2 seropositive (HSV-2+) and identified phenotypic changes in CD4 T cells including increased frequencies of activated cells and cells expressing co-receptors for HIV entry in HIV+/HSV-2+ vs. HIV+/HSV-2 seronegative (HSV-2−) women. Surprisingly, IL-32, a proinflammatory cytokine, was lower in CD4 T cells from HSV-2+ women. Addition of IL-32γ to CD4 T cells from HIV+ women blocked HIV reactivation following stimulation with phytohemagglutinin, suggesting that HSV-2 induces phenotypic changes in CD4 T cells to promote HIV reactivation. Subsequent RNAseq analysis of CD4 T cells infected ex vivo with HSV-2 also showed lower levels of IL-32, as well as elevated bcl6, itch, and irf4 transcripts along with decreased ccr7, psgl1, and s1pr1, suggesting a transcriptional landscape consistent with T follicular helper (TFH) cells. Flow cytometry studies using CD4 T cells isolated from PBMC from HIV− or HIV+ donors, as well as from tonsils, showed an increased frequency of TFH cells in HSV-2 infected CD4 T cells, suggesting preferential infection of this subpopulation, which is known to harbor latent HIV. HSV-2 infection of CD4 T cells isolated from HIV+ donors or latently infected T cell lines triggered HIV reactivation. These findings demonstrate that HSV-2 directly infects CD4 T cell subpopulations including TFH cells to promote HIV reactivation.
Concurrent microglial activation in CD8 T cell-mediated CNS vascular permeability during Theiler’s virus infection
Authors: Khadka Roman , Zheng Jiaying , Ayasoufi Katayoun , Jin Fang , Tritz Zachariah Peter , Wu LongJun , Johnson Aaron J 
Published: January 2023

Abstract
Immune-mediated blood brain barrier (BBB) disruption is a prominent feature of various neurological conditions for which an emerging role of CD8 T cells is being realized. Our laboratory has developed a unique model of CD8 T cell-mediated BBB disruption which employs a variation of the Theiler’s murine encephalomyelitis virus infection (TMEV) infection. Upon intravenous administration of VP2121–130 viral peptide during peak anti-viral adaptive response, Db:VP2121–130 epitope specific CD8 T cells induce BBB disruption in a perforin dependent manner. In this model, we addressed the role of microglia in tandem with CD8+ T cells. Using real time two-photon in vivo imaging, we demonstrate that microglia adopt distinct morphological features including enlarged cell body and fewer ramified processes as early as 6 hours post administration of VP2 peptide. Notably, perforin-deficient mice failed to display BBB disruption and had significantly diminished microglial expression of MHC-II and CD68 in this model. This study demonstrates CD8 T cell promote microglia activation through a perforin-dependent process during BBB disruption. Importantly, microglia are activated concurrently with the onset of vascular permeability and could serve as a critical cell type in this process.