bluuebunny/crossref_metadata_2025_split huggingface dataset:
Interleukin (IL)-15 superagonist mediates immunotoxicity by inducing hyperproliferation of activated NK cells and production of IFNγ
Authors: Guo Yin , Luan Liming , Rabacal Whitney , Bohannon Julia , Fensterheim Benjamin , Hernandez Antonio , Sherwood Edward 
Published: January 2023

Abstract
Interleukin (IL)-15 is currently undergoing clinical trials to assess its anti-tumor efficacy. IL-15 is essential for the generation, function and survival of natural killer (NK), natural killer T (NKT) and memory phenotype (MP) CD8+T cells. The combination of IL-15 with its soluble IL-15 receptor α (IL-15Rα) generates a complex termed IL-15 superagonist (IL-15 SA) that exhibits more profound biological activities on NK, NKT and MP CD8+T cells than IL-15 alone. Thus, IL-15 SA is a potentially more attractive therapeutic agent for use against advanced tumors. However, the safety of IL-15 SA administration in vivo is not clearly elucidated. This study was designed to evaluate toxic consequences of IL-15 SA in mice. Administration of 2 μg IL-15 SA for 4 days caused a time- and dose-dependent immune-toxicity, characterized by hypothermia, weight loss, liver damage and death. At 2ug, IL-15 SA expanded NK cells in spleens and livers compared to vehicle and significantly increased NK cell expression of CD69, NKG2D and NKp46 as well as production of IFNγ, granzyme B and perforin. IL-15 SA also prompted NK cell maturity with a preferential expansion of the pro-inflammatory (CD27+ CD11b+) NK subset. Mice depleted of NK cells by anti-asialoGM1or anti-NK1.1 were resistant to IL-15 SA-induced toxicity, whereas adoptive transfer of wild type NK cells to Rag2−/−γc−/− mice re-established IL-15 SA toxicity. While IL-15 SA expanded NKT and mCD8+ T cells, mice depleted of NKT or CD8+ T cells remained sensitive to IL-15 SA-mediated toxicity. In addition, IFNγ KO, but not TNF-α KO or perforin KO, mice were resistant to IL-15 SA-induced toxicity. In conclusion, high-dose IL-15 SA causes systemic immunotoxicity via induction of NK cell proliferation and activation.
FIP200 targets Ago2 and mitochondria in naive T cells to control tumor immunity
Authors: Xia Houjun , Wang Wei , Kryczek Ilona , Wei Shuang , Crespo Joel L , Guan Junlin , Zou Weiping 
Published: January 2023

Abstract
Naive T cells are poorly studied in cancer patients. Here, we report that naïve T cells are susceptible to apoptosis and lost in human and mouse cancer. Accompanied with this feature is a selective loss of autophagy component, FIP200 in naïve T cells in tumor bearing hosts. FIP200-deficient T cells functionally resemble their counterparts in tumor bearing hosts and mediate poor anti-tumor immunity in vivo. Mechanistically, loss of FIP200 disables the balance between pro- and anti-apoptotic Bcl-2 family members via elevating argonaute (Ago) 2 ubiquitination and subsequently eliminating microRNA1198-5p expression and consequently lifting microRNA1198-5p-mediated repression on pro-apoptotic gene Bak1 expression. In vivo rebalance of the Bcl-2 family genes via enforcing Bcl-2 expression partially rescues FIP200-deficient T cell apoptosis. In parallel, specific FIP200 deficiency causes potent mitochondria activation and reactive oxygen species (ROS) release in T cells. In vivo ROS blockade prevents FIP200-deficient T cell apoptosis. Thus, tumor may trigger naïve T cell apoptosis via targeting its FIP200 to escape anti-tumor immunity.
First in-depth investigation of multiple inhibitory molecule expression by bona fide tumor-reactive T cells in ovarian cancer.
Authors: Chacon Jessica A , Yamamoto Tori , Garrabrant Thomas , Kari Gabor , Best Andrew , Ye Qunrui , Powell Daniel J 
Published: January 2023

Abstract
Ovarian cancer causes more deaths than any other cancer of the female reproductive system. However, novel therapies, including immunotherapy are now being utilized to treat this disease. In addition, efforts have been made to identify and evaluate the subsets of cells within the tumor environment that correspond with patient overall survival. The presence of intraepithelial CD3+ T cells and CD8+ T cells have been correlated with ovarian cancer patient survival and our lab has recently identified CD137 as a biomarker for naturally-occurring tumor-reactive T cells in melanoma and ovarian cancers. However, a thorough elucidation of the immune landscape of endogenous bulk and tumor-reactive T-cell has not been conducted. In our study, we used an in-depth multi-parameter flow cytometry panel to determine the expression of multiple inhibitory molecules, Th1 cytokines, and the effector phenotype in the bulk and tumor-reactive subsets in human ovarian cancer. We observed that the CD4+ and CD8+ T cells found within the tumor environment express inhibitory receptors, very limited Th1 cytokine producing cells and a decreased expression of Granzyme B within the CD8+ subset. The tumor-reactive subset expressed inhibitory molecules, but was still able to elicit an effector phenotype and comprise Th1 cytokine producing cells.
In conclusion, using multi-parameter flow cytometry enabled us to investigate the expression of inhibitory molecules and determine the effector phenotype in the bulk TIL and tumor-reactive CD137+ subsets.
“I”-branched Carbohydrates Negatively Regulate Galectin-3-binding and Melanoma Malignancy
Authors: Sweeney Jenna Geddes , Liang Jennifer , Giovannone Nicholas , Schaffer Lana , Head Steven R. , Antonopoulos Aristotelis , Haslam Stuart M. , Widlund Hans R. , Dimitroff Charles J 
Published: January 2023

Abstract
Malignant transformation is often associated with aberrant glycosylation of cell surface proteins. Commonly observed changes in glycan structures include increased presence of sialic acid, altered expression of mucins, and abnormal branching of N-glycans. Increasing evidence supports the idea that the presence of certain glycans correlate with cancer progression by affecting tumor cell invasiveness, ability to bind pro-tumorigenic galectins, and by promoting metastasis to distant organs. Taking this relationship into consideration, we conducted MALDI-TOF mass spectrometry on cell surface glycans as well as glycomic gene profiling from normal human epidermal melanocytes (NHEM) and human metastatic melanoma (MM) cells. We found that a defining feature between NHEMs and MMs was the relative level of “I”-branched and “i”-linear poly-N-acetyllactosamines on N-glycans. While NHEM predominantly expressed “I”-branched structures, MMs expressed “i”-linear modified N-glycans. Furthermore, glycomic gene profiling and confirmatory RT-qPCR data showed that the “I”-branching β1,6 N-acetylglucosaminyltransferase 2, GCNT2, is decreased in MM compared with NHEMs. With regards to its role in malignancy, we found that GCNT2 functioned as a negative regulator of galectin-3 (Gal-3) binding. GCNT2 overexpression in human MM cells inhibited Gal-3 mediated ERK1/2 phosphorylation and significantly reduced primary tumor growth. While GCNT2 knockdown in human MM cells promoted Gal-3 mediated ERK1/2 phosphorylation and significantly enhanced primary tumor growth. These findings highlight new glycobiological insights into melanoma development, implicating GCNT2 as a negative regulator of Gal-3 binding and melanoma malignancy.
Unsaturated fatty acids suppress anti-tumor T cell function.
Authors: Cui Guoliang , Kaech Susan M 
Published: January 2023

Abstract
Remarkable progress in cancer immunotherapy has revealed the power of harnessing immunity to combat cancer. Anti-tumor T cells suppress tumor development by secreting cytotoxic cytokines and activating other immune cells. However, the tumoricidal function of tumor-infiltrating T cells is often suppressed and inefficient to combat tumors. Thus one of the key questions in the field of cancer immunology is to address what causes the immunosuppressive nature of the tumor microenvironment. Using a melanoma mouse model, we found that 1) fatty acids accumulated in the tumor microenvironment, 2) unsaturated fatty acids suppressed T cell effector function through CD36/PPAR pathway and 3) blocking FAs transportation into T cells enhanced T cell cytotoxicity and delayed tumor growth. This study has identified unsaturated fatty acid species as immunosuppressive factors in tumor, which may lead to the development of novel forms of therapy that stimulates anti-tumor immunity by manipulating fatty acid transportation and metabolism.
Detection of HIV Env-mediated B cell receptor aggregation on engineered B cells by flow cytometry.
Authors: Gallaher Brandon L , Stamatatos Leonidas 
Published: January 2023

Abstract
Approximately 20% of individuals infected with HIV-1 produce broadly neutralizing antibodies (bNAbs) against the virus. Understanding how bNAb linked B cell receptors (BCRs) are initially stimulated by the HIV-1 envelope (Env) in the context of natural HIV infection is a major focus of HIV research. While germline-reverted BCR forms of bNAbs do not efficiently recognize Env, recombinant Env proteins have been recently designed to engage and activate some germline BCRs. To better understand how such modified Envs engage BCRs and to further optimize the activation of B cells, we (a) created a stable Ramos cell line that expresses the germline BCR form of a potent and broadly neutralizing antibody, VRC01 and (b) developed a fluorescence-based assay that allows for the monitoring of Env-mediated BCR aggregation. B cells were incubated with different multimeric forms of Env (valency ranging from 1–24) and determined the extent of BCR aggregation and compared it with the extent of B cell activation (measured by Ca2+flux assays). Significant increases in BCR aggregation and B cell-activation were detected in response to heptameric and 24-meric forms of Env. In contrast, lower valency multimeric forms of Env activated B cells inefficiently. No aggregation or calcium flux was detected in response to monomeric Env. A positive correlation was observed between the magnitudes of calcium flux and BCR aggregation. Thus, either BCR aggregation or Ca2+ flux assays can be used to monitor the activation of B cells by recombinant Env proteins that are being evaluated as possible immunogens to elicit bNAbs. Our results are relevant to optimizing the design and formulation of recombinant Env proteins for the elicitation of broadly neutralizing HIV-1 antibodies.
Immune correlates for VEEV replicon vaccine protection against Ebolavirus
Authors: Stonier Spencer W , James Rebekah M , Josleyn Nicole M , Bakken Russell R , Dye John M 
Published: January 2023

Abstract
The progress of filovirus vaccines has advanced greatly due in part to the urgent need to develop countermeasures for the unprecedented Ebolavirus (EBOV) outbreak in West Africa. While a variety of vaccination approaches have been moved into clinical trials, the understanding of what constitutes a protective response after vaccination is still under investigation. Our approach to addressing immune correlates has involved the use of various mouse models to first test whether or not protection is afforded in the absence of a cell population. To this end, wild type (wt), β2m−/− and μMT−/− mouse models were used to assess the protective efficacy of vaccination with a Venezuelan Equine Encephalitis Virus replicon bearing the EBOV glycoprotein (GP). Vaccinated β2m−/− mice, despite lacking CD8 T cells, enjoyed full protection against mouse-adapted (ma) EBOV exposure. Vaccinated μMT−/− mice, on the other hand, succumbed to maEBOV exposure, similar to unvaccinated control mice. A flow cytometry analysis of T cell responses in Wt, β2­m−/− and μMT−/− mice revealed all mice mounted CD4 T cell responses, whereas only wt mice generated CD8 T cell responses. Antibody responses to EBOV GP were predictably lacking in μMT−/− mice after vaccination, but were present at high titers in wt and β2m−/− mice. We were able to achieve protection in vaccinated μMT−/−, but not unvaccinated μMT−/− mice, given serum from vaccinated wt mice before and after maEBOV exposure. These data indicate that T cell responses in this model can contribute to protection when given antibody help. Ultimately, antibody responses appear to be the best protective correlate in this model, though we also provide evidence that T cell responses can be essential in certain conditions.
Meeting the threshold of multifunctional CD8+ TEM by γ-irradiated sporozoite vaccination, essential to ensue sterile protection against Plasmodia liver stage infection
Authors: Patel Hardikkumar Bipinchandra , Yadav Naveen , Parmar Rajesh , Dalai Sarat K 
Published: January 2023

Abstract
People in malaria endemic area develop immunity after several years of exposure to parasite, children particularly take longer to develop protective immunity. Interestingly, the protection is lot more consistent among children living in high transmission area compared to those of low transmission area, suggesting that protection may depend on the frequency of exposure and/or a load of parasite exposure. Thus, we hypothesize children experiencing multiple exposures might develop critical frequencies of anti-parasite lymphocytes ensuring the protection to Plasmodia. CD8+ T cells play a key role in mediating protection against Plasmodia liver-stage infection. In present study, we took whole sporozoite (spz) vaccination strategy and immunized C57Bl/6 mice with multiple doses of γ-spz at different time intervals. We found continuous immunization with 3 and 4 doses of γ-spz, 2 weeks apart, provides ~50% and 100% sterile protection, respectively. However, mice immunized with 3 doses of γ-spz with an interval of 90 days during 2nd – 3rd dose fail to be protected. Upon measuring the CD8+ T cell responses, we found that higher frequencies of liver-stage specific multifunctional CD8+ T cells (CD44hiKLRG-1intCD107+IFN-γ+) were accumulated in mice immunized with 4 doses of γ-spz (180–410 cells/106; Avg. 284), whereas the same among mice receiving 3 doses showed lower frequencies (50–125 cells/106; Avg. 75) before challenge, and mixed frequencies (126–551 cells/106; Avg. 340) following challenge, suggesting that protection against Plasmodia infection could be ensured either by pre-existing multifunctional memory CD8+ T cells before the infection or mobilizing the required frequencies of the same to liver during the course of infection.
ABVIC: an improved serological test to resolve indeterminate diagnoses for herpes simplex virus 2 (HSV-2)
Authors: Sowa Gavin M , Geltz Joshua , Kempa Annie , Bernert Rene , Warren Terri , Halford William P 
Published: January 2023

Abstract
Approximately 50 million Americans are infected with herpes simplex virus type 2 (HSV-2) and may, at any time, transmit infectious virus to sexual partners. Current HSV-2 serological antibody tests are imperfect; 5–6% of those who initiate HSV-2 ELISA IgG screening receive low-positive results, known to include about 50% false-positives compared to HSV Western Blot, the current gold standard test. Even when low-positive samples are run by Western, ~20% obtain indeterminate results. We report a new, flow-cytometry-based method that measures serum antibody-binding to virus-infected cells (ABVIC). Unlike the glycoprotein G-based HerpeSelect assay, ABVIC tests for antibodies to ~30 HSV-2 antigens and appears to be ~100-fold more sensitive than the gG-2-based antibody capture ELISA. We report that when serum was collected from n=17 patients who had previously received “HSV-2 indeterminate” results by Western Blot, the ABVIC test offered a clear answer in every case. Sixteen of 17 were HSV-2 seronegative by ABVIC and only 1 of 17 was weakly HSV-2 seropositive. Most of the “Indeterminate” serum samples exhibited high background, which produces weak reactivity in the HerpeSelect and Western blot assays, but which does not confound the internally controlled ABVIC assay. We conclude that the highly quantitative, flow cytometry-based ABVIC test may be used to resolve the vast majority of “HSV-2 Indeterminate” results that have misled thousands of patients to the erroneous conclusion that they carry HSV-2 and pose a transmission risk to others.
Pathogenic conversion of Foxp3+ T cells into Th1 and Th17 cells in Stromal Keratitis
Authors: Varanasi Siva Karthik , Bhela Siddheshvar , Rouse Barry T 
Published: January 2023

Abstract
Ocular infection with herpes simplex virus 1 (HSV-1) can result in a chronic immuno-inflammatory lesion that is a significant cause of human blindness. IFN-γ producing CD4 + T cells are generally considered the main orchestrators, and lesions are more severe if the regulatory T cell (Treg) response is compromised. Tregs have been shown to lose FoxP3 (Treg lineage factor) and adopt alternate lineage fates in a changing cytokine environment. Moreover, little is known about the stability of Treg cells in an ongoing inflammatory reaction such as is the case of SK. In this study-using fate mapping mice, we were able to demonstrate that the population of ex-Tregs increased at the site of infection that is the cornea and also at the secondary lymphoid organs after HSV-1 infection. In vivo studies showed that these ex-Tregs acquired the Th1 and Th17 phenotype, which may play a destructive role in stromal keratitis. We also demonstrate in vivo that CD25lo subset of Tregs is less suppressive and more prone to conversion than CD25hi Tregs. Additionally, in-vitro studies suggested that Tregs in the presence of IL-12 converted into ex-Tregs, however this plasticity was prevented when Tregs were generated in the presence of DNMT inhibitor (Azacytidine). Thus, plasticity of Tregs might represent a problem during SK, which needs to be controlled with appropriate therapeutic procedures. Overall, our results indicate that Tregs lose FoxP3 expression in ocular inflammatory setting and drugs that reverse Treg plasticity could be useful to restore Treg function and to inhibit ocular immunopathology.
PD-1 inhibition results in reduced dendritic cell (DC) activation of HIV-1-specific de novo CTL and enhancement of DC-induced CTL memory recall responses
Authors: Garcia-Bates Tatiana M , Kim Eun , Gambotto Andrea , Rinaldo Charles R , Mailliard Robbie B 
Published: January 2023

Abstract
Combination strategies consisting of DC vaccines and suppression of immunoregulatory pathways such as PD-1 blockade have been advocated to elicit optimal CTL responses in chronic diseases including cancer and HIV infection. We have shown that inactivated HIV-1 virus loaded, high IL-12 producing, mature, type-1 polarized DC (DC1) can activate CTL from naïve CD8+ T cell precursors that more effectively kill HIV-1-infected cells than CTL derived from memory CD8+ T cells. Here we tested DC1 transfected with an adenoviral (Ad) vector encoding anti-PD-1 antibody (DC.αPD1) enhancement of both primary as well as memory CTL responses to HIV-1. Ad-αPD1-transduced DC1 (DC.αPD1) secreted high levels of functional anti-PD1 Ab without affecting their phenotype or IL-12p70-producing capacity. We next compared HIV Gag peptide epitope-loaded DC.αPD1 to DC1 that were transfected with an empty vector for their ability to activate either purified, autologous naïve or memory HIV-1-specific CD8+ T cells from chronic HIV-1-infected participants of the Multicenter AIDS Cohort Study. When compared to the control DC1, antigen-loaded DC.αPD1 enhanced the overall magnitude of HIV-1-specific CTL responses induced, as determined by the expansion of HIV-1-peptide responsive CD107a and IFNγ expressing T cells. In contrast, the overall number HIV-1 antigen-reactive CTL derived from the naïve CD8+ T cell fraction sharply decreased when using the DC.αPD1-based approach. These results suggest previously unrecognized, opposing roles of the inhibitory receptor PD-1 in DC1-induced primary versus memory recall CD8+ T cell responses to HIV-1.
A large scale screening of HLA-class II-binding peptides from HBs and HBc peptide libraries
Authors: Miyadera Hiroko , Okabe Yuki , Chen Cindy , Tokunaga Katsushi , Mizokami Masashi 
Published: January 2023

Abstract

Objectives
Variations in the human leukocyte antigens (HLA)-DP are associated strongly with susceptibility/protection to chronic hepatitis B virus (HBV) infection. It has been reported previously that HLA-DPB1*05:01 and *09:01, the major HLA-DP alleles in East Asians, are associated with susceptibility, while HLA-DPB1*04:01 and *04:02, the major alleles in Europeans, confer protection against chronic hepatitis B (Kamatani, et al. Nat Genet (2009); Nishida, et al. PLoS One (2012)). To identify the peptides that are potentially involved in immunological responses against HBV infection, we screened the peptide libraries of the HBV surface antigen (HBs) and core antigen (HBc) through the cell-surface HLA expression assay.

Methods
The six major HLA-DP haplotypes that are associated with susceptibility to or protection against chronic hepatitis B were used in this study. Peptide libraries were designed for the entire HBs and HBc antigens, based on the consensus sequence of the genotype C. To identify the HLA-DP-binding region, we expressed HLA-DP in fusion with each peptide, using fibroblast cell line as an expression host.

Result
The level of HLA-peptide interaction was estimated by measuring the level of cell-surface expression of each HLA-peptide fusion construct. Out of &gt;70 peptides that cover the entire HBs and HBc antigens, we found several regions that bound strongly to certain HLA-DP allele products. In this presentation we show an overview of the methodology and discuss potential mechanisms that might underlie association of HLA-DP with chronic hepatitis B.
Dysregulated CD8 T cells resemble Tfh-like cells and promote B cell activation during autoimmunity.
Authors: Hoyer Katrina K , Valentine Kristen , Davini Dan , Sindi Suzanne 
Published: January 2023

Abstract
T follicular helper (Tfh) cells provide essential help for the development of antigen specific antibodies during normal and autoreactive immune responses. IL-2-deficient (KO) mice suffer from excessive lymphoproliferation and spontaneous, systemic autoimmune disease. Depletion of CD4 T cells in IL-2-KO mice eliminates the generation of RBC-specific antibodies, augmenting survival. Surprisingly, depletion of CD8 T cells in IL-2-KO mice also reduces disease and prolongs survival. Several groups have demonstrated a protective role for CD8 T cells in inflammation and autoimmunity, while others have suggested a pathogenic role. However the mechanisms of CD8 T cell influence on autoimmune disease are largely unknown. We have identified a new CD8 Tfh-like population in an autoimmune setting. RNA-sequencing of day 12 IL-2-KO CD8 T cells revealed an increase in the expression of genes associated with the CD4 Tfh lineage (Bcl-6, CXCR5, PD-1, ICOS and IL-21) compared with wild-type CD8 T cells. Flow cytometry confirmed the presence of expanded CD4 Tfh and CD8 Tfh-like cells, and the production of IL-21 in IL-2-KO mice. Germinal center tissue staining revealed localization of both CD4 and CD8 T cells to the germinal center during autoimmunity. In vitro, IL-2-KO CD8 T cells induced proliferation and activation of B cells. Our results indicate that autoreactive CD8 T cells contribute to the autoimmune disease in part through dysregulated follicular-like differentiation and function. NIH funding R00HL090706.