bluuebunny/crossref_metadata_2025_split huggingface dataset:
Targeting CARP-1 in breast cancer to decrease tumor growth and enhance immunogenicity
Authors: Shissler Susannah Claire , Drechsler Jennifer M , Derakhshandeh Roshanak , Rishi Arun K , Webb Tonya J 
Published: January 2023

Abstract
Natural killer T (NKT) cells are important in antitumor immunity, but these cells are significantly reduced in breast cancer (BC) patients. The tumor-associated factors that lead to the reduction in NKT cells are unclear; however, it is known that during malignancy BCs upregulate pro-survival proteins in order to proliferate and evade immune detection. Cell Cycle and Apoptosis Regulatory Protein (CARP)-1, a perinuclear phosphoprotein that regulates cell growth and apoptosis, is overexpressed in chemotherapy-treated cancers and several BC cell lines, such as MCF-7. CARP functional mimetics (CFMs) are small molecule inhibitors that interfere with the interaction of CARP-1 and the anaphase promoting complex, causing cell cycle arrest and apoptosis. Thus, we hypothesize that targeting CARP-1 through the use of CFMs will induce apoptosis and restore anti-tumor immune responses. In this study, BC cell lines were treated with CFM-4&5 and proliferation, cytotoxicity, and immunogenicity were assessed. It was found that pretreatment of human MCF-7 and murine E0771 BC lines with 20μM CFM-4&5 resulted in 80–100% cell death within 48–72h. In order to assess immunogenicity in the absence of cytotoxicity, BC cell lines were pretreated with 10μm for 24h, washed and cultured in fresh medium. Conditioned medium was collected and used to treat CD1d-expressing cells for 4h prior to coculture with NKT cell hybridomas. To test the effects of CFMs on immune cell activation, peripheral blood mononuclear cells (PBMC) from healthy and BC patients were treated with CFM-4&5 in the presence of stimuli and immuno-activity was assessed by ELISA. Collectively, these data implicate a novel role for targeting CARP-1 for the treatment of BC.
E3 ubiquitin ligase NKLAM is important for bacterial killing in a murine model of streptococcal pneumonia
Authors: Lawrence Donald W. , Kornbluth Jacki 
Published: January 2023

Abstract
Natural killer lytic associated molecule (NKLAM) is an important component of the innate immune system. Previous in vitro studies using NKLAM-KO macrophages have demonstrated that NKLAM positively affects the transcriptional activity of STAT1, a transcription factor crucial for proper immune function. Therefore, we hypothesized that NKLAM-KO mice would be unable to mount an effective immune response when exposed to Streptococcus pneumoniae. Using a nasal inhalation infection model we found that the lungs of NKLAM-KO mice had a significantly higher bacterial load than WT mice. This observation was associated with increased lung phosphatase activity and decreased STAT1 phosphorylation. STAT1 regulates a large number of genes, including iNOS and MCP-1, which are important for successful pathogen eradication. In our model, we observed that NKLAM-KO mice expressed less iNOS in their lungs at 24 and 48 hr post-infection in comparison to WT mice. Additionally, infected NKLAM-KO lungs contained less TNF-alpha, IL-12, IFN-gamma and MCP-1 than lungs from WT mice. NKLAM-KO mice had significantly less neutrophil lung infiltration than WT mice, an effect that is likely due to the decreased expression of chemokines including MCP-1. In summary, these novel findings demonstrate the importance of NKLAM in innate immunity and begin to define a role for NKLAM in leukocyte trafficking to the lung upon S. pneumoniae infection.
Autophagy induced by human neutrophil exovesicles controls intracellular growth of Mycobacterium tuberculosis H37Rv in autologous macrophages
Authors: Estrada-Garcia Iris , Alvarez-Jimenez Violeta , Garcia-Martinez Mariano , Vazquez-Flores Luis , Leyva-Paredes Kahiry , Garcia-Perez Blanca , Serafin-Lopez Jeanet , Chacon-Salinas Rommel , Wong-Baeza Isabel , Castañeda-Casimiro Jessica , Ruiz-Sanchez Bibiana Patricia , Estrada-Parra Sergio 
Published: January 2023

Abstract
We have previously shown that human neutrophils (NEU) stimulated with live Mycobacterium tuberculosis H37Rv (Mtb) release exovesicles, named EXO-Mtb. When autologous macrophages were incubated with EXO-Mtb, they increased the expression of CD86 and MHC II and produced inflammatory cytokines. Here we demonstrate that EXO-Mtb can also reduce the intracellular load of Mtb in autologous macrophages. Since autophagy has been shown to control mycobacterial growth, LC3II expression was measured on infected macrophages after incubation with EXO-Mtb (4, 6 and 24h). Control EXO preparations included those released spontaneously from NEU, or after stimulation with PMA or fMLP. Our results showed that of all EXO preparations used, EXO-Mtb induced the largest amounts of LC3II protein, which correlates with a significant reduction of colony forming units (CFU). This phenomenon could be reverted with the addition of Wortmannin, an autophagy inhibitor. It is important to remark that there was not a 100% control of intracellular mycobacterial growth. Further experiments should be done to better understand the role of ectosomes in NEU-macrophage communication through exovesicles.
Adiponectin inhibits eosinophil recruitment and is protective in invasive aspergillosis
Authors: Templeton Steven , Amarsaikhan Nansalmaa 
Published: January 2023

Abstract
Aspergillus fumigatus is a fungal pathogen associated with allergy, asthma, and invasive infection in susceptible individuals. Our previous results suggested that eosinophils may be a novel therapeutic target in invasive aspergillosis (IA) patients that do not respond to conventional pharmacotherapy. Eosinophil recruitment is induced by inhalation of the fungal cell wall polysaccharide chitin. However, the mechanisms of immune regulation that drive this recruitment are not well understood. We observed decreased levels of the metabolic-regulating cytokine adiponectin in the lungs and serum of mice after chitin aspiration, suggesting that adiponectin may be involved in the regulation of type 2 lung immune responses. Co-aspiration of adiponectin with chitin significantly reduced eosinophil accumulation in the lungs, and airway eosinophil recruitment in adiponectin-deficient mice was increased with chitin aspiration. In addition, chitin aspiration resulted in decreased surface expression of the adiponectin receptor AdipoR1 on eosinophils. Most significantly, airway accumulation of eosinophils, disease severity, and fungal burden were increased in adiponectin knockout mice with IA. This may be due to a direct inhibition of eosinophil recruitment, as recombinant adiponectin inhibited chemotaxis of bone marrow-derived eosinophils in vitro. These results suggest that adiponectin inhibits potentially detrimental lung eosinophil recruitment in response to chitin-containing pathogens. Increased understanding of the relationship between metabolic and immune regulation may ultimately result in novel therapies for obesity and allergic and infectious disease.
NK cells require both inhibitory and activation Ly49 receptors to mediate MHC I Dk-dependent MCMV resistance.
Authors: Gamache Awndre Elijah , Nash William , Teoh Jeffrey , Gillespie Alyssa , Brown Michael G 
Published: January 2023

Abstract
The MHC class I molecule, Dk, is a key genetic factor of murine cytomegalovirus (MCMV) resistance. NK cells expressing the C57L-derived inhibitory receptor Ly49G2 (G2) mediate Dk-dependent MCMV control, however, a precise in vivo role for activation receptors has not been delineated. We studied a role for activation receptors in Dk-disparate B6.NKCc57l congenic mice by injecting neutralizing mAbs prior to infection. We found that only Ly49R (R) blockade specifically, and completely, abrogated host resistance. R+ NK cells had a differentiated phenotype (CD27−, CD11b+, KLRG1+, DNAM1−) and displayed higher granzyme B during MCMV infection, in comparison to R− NK cells. R+ G2+ NK cells responding to MCMV also showed higher proliferative potential and cytokine responsiveness compared to R+ G2− NK cells. To further interrogate the role of these NK cell subsets in vivo, we exploited differences in expression of Nkrp1 (NK1.1) and Ly49 receptor alleles in our NKC congenic mouse strains. Endogenous NK cells in H-2 matched host B6.Dk mice were either selectively depleted with mAb PK136 (NK1.1) or neutralized with mAb 3D10 (a-Ly49H), rendering them susceptible to MCMV infection, prior to adoptive transfer of donor B6.NKCc57l NK cells. We found that responding donor NK cells were mostly R+ G2+ and actively expanding in contrast to R+ G2− NK cells. Ongoing adoptive transfer experiments with sorted NK cell populations suggest MCMV control coincides with donor R+ G2+ NK cells. We are continuing to investigate how inhibitory Ly49 receptors modify NK cell responsiveness via activation Ly49 receptors. We infer from these data that inhibitory and activation Ly49 receptors work in tandem to deliver specific NK cell-dependent virus control.
Targeting stem cell-derived viral antigen-specific T lymphocytes for HBV immunotherapy
Authors: Haque Mohammad Shamsul 
Published: January 2023

Abstract
Adoptive cell transfer (ACT) of hepatitis B virus (HBV)-specific CD8+ cytotoxic T lymphocytes (CTLs) is a promising treatment for chronic HBV infection. Naive or central memory T cell-derived effector CTLs known as “highly reactive” cells are optimal populations for cell-based therapies. However, such ACT is often not feasible due to difficulties in obtaining sufficient numbers of CTLs from patients. Pluripotent stem cells (PSCs) have the capacity to be reprogrammed into functional viral antigen (Ag)-specific CTLs, which have a potential to be used for ACT-based immunotherapy. However, the optimal approach to differentiate viral Ag -specific CTLs from PSCs remains unclear. In this study, we generated HBV-specific CTLs from induced PSCs (iPSCs), i.e., iPSC-CTLs, and utilized a mouse model of chronic HBV infection to test ACT of iPSC-CTLs for HBV immunotherapy. Murine iPSCs that genetically modified with HBV-specific T cell receptor (TCR) were co-cultured with OP9 cell line expressing Notch ligands Delta-like 1 and 4 (OP9-DL1/DL4 cell) for a week and adoptively transferred into recipient C67BL/6 mice for in vivo development. The combination of Notch signaling and incorporation of HBV-specific TCR genes in iPSCs drove the development of HBV-specific iPSC-CTLs. In addition, In a mouse model of chronic HBV infection in which the HBV viral genome is delivered into the mouse liver through hydrodynamic injection, HBV-specific iPSC-CTLs infiltrated into liver tissue, and inhibited HBV replication after ACT. These results confirm that ACT of viral Ag-specific iPSC-CTLs can substantially suppress HBV replication, and indicate that stem cell-derived highly reactive viral Ag-specific CTLs may be used for HBV immunotherapy.
Loss of CCR5 signaling results in early NK cell and neutrophil recruitment into the liver and increased hepatic inflammation in a mouse model of acute hepatitis B
Authors: Stevens Kathleen , Thio Chloe , Osburn William 
Published: January 2023

Abstract
Human genetic studies demonstrate that a non-functional CCR5 receptor sequence variant is associated with enhanced control of acute hepatitis B virus (HBV) infection. To better understand the role of CCR5 in immune responses to HBV, we employed a mouse model of acute HBV in which Ccr5 +/+(WT) or Ccr5 −/−(KO) C57BL/6 mice were intravenously infected with adenovirus containing an overlapping HBV1.3 construct (AdHBV) or vector control (AdShuttle). Mice (N=4–6/group) were euthanized at days 2, 3, 7, 10, and 14 post injection. Increased plasma ALT levels were observed in both Ccr5 WT and KO AdHBV mice at day 7, but levels were significantly higher in the Ccr5 KO compared to WT mice by day 14 (554 vs 274 U/L, p &lt; 0.05). Plasma ALT levels were not significantly different between Ccr5 WT and KO AdShuttle mice. Analysis of intrahepatic leukocytes by flow cytometry revealed significantly increased intrahepatic CD11b+ NK cells (26.3 vs 6.9, % of live leukocytes, p&lt; 0.01) and neutrophils (7.3 vs 1.3, % of live leukocytes, p&lt; 0.05) at day 3 in Ccr5 KO compared to WT AdHBV mice. By day 7, these differences in cell populations were no longer present. At Day 14, significantly higher levels of intrahepatic Ly6c+ macrophages were detected in Ccr5 KO compared to WT AdHBV mice (17.0 vs 6.4, % of live leukocytes, p&lt; 0.01). These differences in intrahepatic leukocytes were not observed in AdShuttle mice. Loss of Ccr5 function did not significantly alter intrahepatic CD4+ and CD8+ populations at the time points investigated. Taken together, these findings suggest that loss of Ccr5 signaling increases intrahepatic innate cells and inflammation during acute HBV infection, providing evidence for a role of CCR5 in regulation of innate immune responses to acute HBV infection.
Role of CD28 superagonist in preventing HSK lesions in mice
Authors: Jaggi Ujjaldeep , Rouse Barry T 
Published: January 2023

Abstract
Ocular infection with herpes simplex virus (HSV) can result in a chronic immune-inflammatory lesion that is a significant cause of human blindness. A key to controlling stromal keratitis (SK) lesion severity is to identify cellular and molecular events responsible for tissue damage. Regulatory T cells (Tregs) are known for preventing autoimmune diseases like systemic lupus erythematosus (SLE), multiple sclerosis (MS), Type 1 diabetes (T1D), rheumatoid arthritis (RA) and Inflammatory bowel disease (IBD). Furthermore, Tregs play an important role in reducing the severity of SK lesions by inhibiting the immune responses which cause immunopathology. In T cell activation and clonal expansion, the costimulatory receptor CD28 and its closest structural relative, the de-activating receptor CTLA-4 (CD152) are of central importance. Both bind to CD80 and CD86 expressed by APC. Interestingly, there is a class of “superagonistic” CD28 specific mAb which are able to preferentially expand and activate Tregs. Previous studies have shown that treating mice with a CD28 superagonistic antibody (CD28SA) ameliorated the acute signs of arthritis and completely prevented the development of chronic destructive arthritis in a mouse model. In the present study, we report that a single injection with CD28SA mAb given at day 3 post infection, reduced the severity of SK lesions. We found increased numbers of Tregs in mice treated with CD28SA. Additional mechanisms involved will be further evaluated during the course of the study. Thus, our studies demonstrate the usefulness of CD28SA as a therapeutic agent to control virus induced immunopathology.
Late-rising CD4 T cells resolve viral persistence.
Authors: Benedict Chris A , McDonald Bryan , Seumois Gregory , El Morabiti Rachid , Ghosh Raima , Fu Zheng , Adler Barbara , Vijayanand Pandurangan , Gupta Ankan 
Published: January 2023

Abstract
Robust CD4 T cell responses help to control chronic infection and restrict disease, but their specific requirements for resolving viral persistence remain poorly defined. During mouse cytomegalovirus (MCMV) infection, CD4 T cells recognizing an epitope derived from the viral M09 protein expand at late times of infection and display a unique phenotype compared to their conventional counterparts, including high IFNγ-production and low expression of the activated CD43 isoform. Ablating these late-rising CD4 T cells by mutating the MCMV M09 genomic epitope, or inducing them through vaccination, revealed their critical role in resolving persistent replication by overcoming IL-10 mediated immune suppression. Together, these data show that unique subsets of late-rising CD4 T cells are qualitatively superior in resolving chronic infection when compared to those that expand earlier, results that should be considered for vaccine development.
IL-33 promotes antiviral CD8+ T cells through the mTORC1 pathway and limits liver injury via recruiting immunosuppressive neutrophils
Authors: Liang Yuejin , Yi Panpan , Jie Zuliang , Yuan Denley Denley Ming Kee , Soong Lynn , Cong Yingzi , Sun Jiaren 
Published: January 2023

Abstract
Viral hepatitis is a major public health problem worldwide. The multifunctional IL-33 is critical for the activity of antiviral cytotoxic T lymphocytes (CTLs) in persistent viral infection; however, the signaling mechanism defining how IL-33 promotes immune cell activation is unclear. In addition, IL-33 can also recruit neutrophils, suggesting that it may regulate liver injury through modulating myeloid cell functions. In this study, we revealed an essential role for endogenous IL-33 regarding antiviral T cell responses in lymphocytic choriomeningitis virus (LCMV) infection. Deficiency of CTL effector activity in IL-33 knockout (KO) mice was restored by recombinant IL-33 treatment. Mechanistically, IL-33 induced CD8+ T cell activation and proliferation through the PI3K/mTORC1 singling pathway. Interestingly, IL-33 treatment also induced significant neutrophil accumulation, but decreased CD8+ T cell numbers, in infected liver. By depleting neutrophils in vivo, we demonstrated that neutrophils played an immunomodulatory role and attenuated liver injury by inhibiting infiltration and the activity of CTLs at the contraction stage of immune response. IL-33-induced immunosuppressive neutrophils expressed higher levels of arginase-1 through type 2 innate lymphoid cells (ILC2)/IL-13/STAT6 axis. This study, therefore, demonstrates a dual role of IL-33 in promoting antiviral CD8+ T cell response at the initial stage, but inducing immunomodulatory neutrophils through induction of ILC2 to limit liver injury during the contraction stage of immune response.
The cytotoxic formulation, INT230-6, induces protective T cell immunity
Authors: Bloom Anja , Bender Lewis H , Walters Ian B , Clark Katharine T , Terabe Masaki , Berzofsky Jay A 
Published: January 2023

Abstract
The benefits of anticancer agents extend beyond direct tumor cell lysis. One major aspect of cell death is the potential to release antigen and induce a long lasting immune response. In this study, a novel tissue and cell diffusive cytotoxic formulation, INT230-6, was administered intratumorally over 5 sequential days into subcutaneous 300mm3 murine Colon26 tumors. Treatment resulted in regression from baseline of 100% of the tumors and up to 80% complete response (CR) with protection from re-challenge. We then analyzed the role of T cells in the protection induced by INT230-6. After CD8 or CD8/CD4 double-depletion at treatment onset no animals achieved a CR, indicating a critical role of T-cells in complete tumor regression. Mice with CRs challenged by subcutaneous re-inoculation of the Colon26 cells also lost protection following CD4/CD8 double-depletion prior to the re-challenge. This result indicates that immunological T cell memory was induced by INT230-6. Colon26 tumors express the endogenous retroviral protein gp70 containing the AH-1 CD8 T cell epitope. AH-1-specific CD8 T cells were detected in tumors of mice without treatment. However, these cells highly expressed PD-1, and did not lead to tumor regression. Hence, INT230-6 given locally to treat tumors appears to induce tumor-specific protective T cell immunity that may overcome suppressive signals or exhaustion. INT230-6 is currently in clinical testing as monotherapy and will be tested in combination with anti-PD1 compounds.
Efficacy of combining radiotherapy and immunotherapy for metastatic colorectal cancer and the modulation of tumor microenvironment
Authors: Wu Chia-Jen , Tsai Yi-Ting , Chang Ching-Cheng , Lee I-Jung , Wu Ping-Yi , Tao Mi-Hua 
Published: January 2023

Abstract
Immune checkpoint blockade antibodies specific to programed death-1/ligand-1 (PD-1/PD-L1) shows durable clinical responses in various cancers but benefit only to a small portion of patients. The failure of immunotherapy is likely due to the complex network of immunosuppressive mechanisms present in advanced tumors. Radiation (RT) modifies tumor microenvironment and causes tumor antigen release, and thus augment efficacy of immunotherapy. Interleukin 12 (IL-12) and granulocyte macrophage colony-stimulator factor (GM-CSF) have shown potent antitumor activities through activating innate and adaptive immunities, as well as reprogramming myeloid cells to a phenotype that favors immune activation. The aim of this project is to compare the therapeutic efficacy of two types of combination therapy: RT combined with IL-12/GM-CSF (RT/12GM) versus RT combined with anti-PD-L1 antibody (RT/PD-L1), and to dissect the molecular mechanisms of the antitumor responses. Our data showed that RT/12GM led to tumor regression in a majority of mice with large established tumors and suppressed liver metastasis, while RT/PD-L1 could only suppress primary tumors but not metastatic tumor. Mechanistic analysis showed that RT/12GM was superior to RT/PD-L1 in inducing accumulation of CD8+ T cells in tumor and in augmenting both NK and CD8+ T cells cytotoxicity in tumor and liver. Interestingly, RT/12GM also induced infiltration of tumor-associated neutrophils (TAN) and impaired their immunosuppressive functions. Depletion experiments showed that both CD8+ T cell and TAN were required for tumor regression in RT/12GM. Together, our results demonstrate that RT/12GM represent a powerful alternative therapy against both primary and metastatic tumors.
Development of a curative therapeutic vaccine (TheraVac) for the treatment of large established tumors
Authors: Yang De , Nie Yingjie , Trivett Anna L , Han Zhen , Chen Xin , Oppenheim Joost J 
Published: January 2023

Abstract
Harnessing immune system to treat cancer requires simultaneous generation of tumor-specific CTLs and curtailment of tumor immunosuppressive environment. Here, we developed an immunotherapeutic regimen capable of eliminating large established mouse tumors using HMGN1, a DC-activating TLR4 agonist capable of inducing anti-tumor immunity. Intratumoral delivery of HMGN1 with low dose of Cytoxan that acts as a functional checkpoint inhibitor cured mice bearing small (diameter ~ 0.5 cm), but not large (diameter ~ 1.0 cm) CT26 tumors. Screening for activators capable of synergizing with HMGN1 in activating DC identified R848. Intratumoral delivery of HMGN1 and R848 plus Cytoxan eradicated large established CT26 tumors. The resultant tumor-free mice were resistant to subsequent challenge with CT26, indicating the generation of CT26-specific protective immunity. This immunotherapeutic regimen caused homing of tumor-infiltrating DC to draining lymph nodes and increased infiltration of T cells into tumor tissues. Cytoxan in this regimen could be replaced by anti-CTLA4) or anti-PD-L1. Importantly, this immunotherapeutic regimen was also curative for large established mouse Renca and EG7 tumors. Thus, we have developed a curative therapeutic vaccination regimen dubbed ‘TheraVac’ consisting of HMGN1 and R848 plus a checkpoint inhibitor, that can, without using exogenous tumor-associated antigen(s), eliminate various large tumors and induce tumor-specific immunity.