bluuebunny/crossref_metadata_2025_split huggingface dataset:
Liver metastasis restrains immunotherapy efficacy via macrophage-mediated T cell elimination
Authors: Kryczek Ilona , Liu Jali , Green Michael , Chinnaiyan Arul , Cieslik Marcin , Zou Weiping 
Published: January 2023

Abstract
Metastasis is the primary cause of cancer mortality, and cancer frequently metastasizes to the liver. It is not clear whether liver immune tolerance mechanisms contribute to cancer outcomes. We report that liver metastases diminish immunotherapy efficacy systemically in patients and preclinical models. Patients with liver metastases derive limited benefit from immunotherapy independent of other established biomarkers of response. In multiple mouse models, we show that liver metastases siphon activated CD8+ T cells from systemic circulation. Within the liver, activated antigen-specific Fas+CD8+ T cells undergo apoptosis following their interaction with FasL+CD11b+F4/80+ monocyte-derived macrophages. Consequently, liver metastases create a systemic immune desert in preclinical models. Similarly, patients with liver metastases have reduced peripheral T cell numbers and diminished tumoral T cell diversity and function. In preclinical models, liver-directed radiotherapy eliminates immunosuppressive hepatic macrophages, increases hepatic T cell survival and reduces hepatic siphoning of T cells. Thus, liver metastases co-opt host peripheral tolerance mechanisms to cause acquired immunotherapy resistance through CD8+ T cell deletion, and the combination of liver-directed radiotherapy and immunotherapy could promote systemic antitumor immunity.
This work was supported in part by research grants from the NIH/NCI grants for WZ (CA248430, CA123088, CA099985, CA193136, and CA152470); AC (1UM1HG006508); TSL (U01CA216449); IEN (CA233487); FW (S10OD020053); MAM (CA240515) and the NIH through the University of Michigan Rogel Cancer Center Support Grant (P30CA46592).
The B cell adapter for PI3K (BCAP) promotes M2 macrophage phenotype and safeguards tumors from immune surveillance
Authors: Saha Irene , Pasare Chandrashekhar 
Published: January 2023

Abstract
The tumor microenvironment is a complex milieu consisting of different populations of infiltrating immune cell populations. Macrophages and DCs within the tumor microenvironment are vital in priming naïve T cells within tumors and tumor draining lymph nodes and collectively guide responses to various immunotherapies. Macrophages can exist in both pro-inflammatory (M1) state and produce IL-6, IL-12, TNF-α and ROS to promote anti-tumor responses, and reparative (M2) state that support angiogenesis and secrete IL-10 and TGF-β favoring tumor growth. We have previously identified BCAP as an adapter that links TLRs to PI3K/Akt activation and negatively regulates inflammatory responses. Recent work has shown a critical role of BCAP in promoting the transition of M1 macrophages to M2 phenotype. Since tumor associated macrophages (TAMs) in their M2 state have tumor promoting roles, we wanted to investigate if deficiency of BCAP could drive anti-tumor responses. BCAP null mice with implanted MC38 tumors had significantly reduced tumor burden and higher infiltration of leukocytes. This included higher presence of CD4+ and CD8+ T cells as well as macrophages and inflammatory monocytes in the tumor microenvironment. We are currently investigating macrophage and T cell intrinsic role of BCAP on tumor growth. Finally, to understand the molecular events leading to differential tumor progression in BCAP deficient condition, we will interrogate the transcriptional profile of tumor infiltrating myeloid and lymphoid cells at different stages of tumor growth and examine the potential role of MyD88 and PI3K/Akt signaling in myeloid cells in influencing BCAP dependent tumor growth.
Supported by grant from NIH (R01 AI113125)
Enhanced anti-tumor immunity in ST8Sia6 knockout mice
Authors: Friedman David J , Kizerwetter Monika , Belmonte Paul , Rajcula Matthew , Theodore Keith , Lee Hyun Se Kim , Shapiro Michael J , Dong Haidong , Shapiro Virginia Smith 
Published: January 2023

Abstract
Inhibitory receptors have a critical role in the regulation of immunity. Siglecs are a family of primarily inhibitory receptors expressed by immune cells, which recognize specific sialic acid modifications on cell surface glycans. Sialyltransferases are the enzymes that catalyzing the addition of sialic acids onto glycoproteins and glycolipids. Siglec-E binds to α2,8-linked disialic acids, generated by the enzyme ST8Sia6 on O-linked glycoproteins and functions to inhibit activation of innate immune cells. Many tumors have increased sialic acid incorporation. Overexpression of the sialyltransferase ST8Sia6 on tumors led to altered macrophage polarization and increased tumor growth through engagement of ligands generated by ST8Sia6 with Siglec-E. Here, we examine the role of ST8Sia6 on immune cells in regulating anti-tumor immunity. The loss of ST8Sia6 in tumor bearing mice reduced tumor growth, increased survival, and altered the immune response as compared to ST8Sia6 sufficient tumor bearing mice. Using MC38, B16, and B16-OVA flank tumor mouse models, tumor growth was decreased in ST8Sia6 KO mice as compared to C57Bl/6 mice. Within the tumor microenvironment of tumors present in ST8Sia6 KO mice compared to tumors present in C57Bl/6 mice, macrophages and dendritic cells altered their phenotype towards enhanced immune activation. Tregs from tumors in ST8Sia6 KO mice also displayed an increase in RORγt and a reduction in TNFRII, suggesting a possible shift to a less suppressive Treg. These alterations in the immune response to tumors were due to the loss of ST8Sia6 in macrophages and dendritic cells and not T-cells. Thus, ST8Sia6 generates ligands for Siglecs that dampen anti-tumor immunity.
Supported by grants from NIH (R01CA243545, 2T32AI007425) and Mayo Clinic Center of Biomedical Discovery
Tumor-associated macrophages found at early tumor sites exhibit altered metabolism and contribute to tumor progression
Authors: Kurt Robert A , Chang Musea , Ellmaker Katherine , Esposito Abigail , Pa Sophea , Roberts Meghan 
Published: January 2023

Abstract
Previously, we reported that macrophages at early tumor sites, before the tumors are even palpable, have a mixed phenotype with production of both pro- and anti-inflammatory cytokines, decreased production of reactive oxygen species, and increased dependence on oxidative phosphorylation. Here we show that depleting these tumor associated macrophages (TAM) impacted subsequent growth of the EMT6 tumor and metastasis of the 4T1 tumor supporting the contention that macrophages found at early tumor sites contribute to later stages of tumor progression. Since these TAM could not be clearly defined as M1 or M2 we set out to characterize them further focusing on their metabolism. We found that TAM present at early sites of 168, EMT6 and 4T1 tumors had elevated expression of pkm2, glut1 and ampk, and transwell assays revealed that these tumors could induce expression of pkm2, glut1 and ampk in bone marrow derived macrophages. Since tumor-derived lactate is known to induce alterations in white blood cells we investigated lactate production by these tumors and whether lactate could induce expression of these same genes. The data revealed that the 168, EMT6 and 4T1 tumors produced significant amounts of lactate and that lactate induced expression of pkm2, glut1 and ampk in bone marrow derived macrophages. Finally, we found that targeting lactate dehydrogenase or eIF2 at early tumor sites impacted growth of the EMT6 tumor, and targeting AMPK or GCN2 impacted metastasis of the 4T1 tumor. Collectively, these data support the contention that alterations in macrophages are already present at the early stages of tumor growth, before a tumor is even palpable, and that targeting metabolism at these early tumor sites can impact later stages of tumor progression.
Activation of IRE1 RNase in dendritic cells curtails antitumor adaptive immunity in melanoma.
Authors: Osorio Fabiola , Flores Felipe , Fernandez Dominique 
Published: January 2023

Abstract
The unfolded protein response (UPR) sensor IRE1 (inositol-requiring enzyme 1 alpha) and its target, the transcription factor XBP1s (X-box binding protein 1, spliced) critically regulates the function of dendritic cell (DC) subtypes. However, the contribution of the IRE1/XBP1s axis in DC subsets to the initiation of antitumor immunity is not entirely understood. In this work, using reporter mice we found that DCs, in particular type 1 conventional DCs (cDC1s), are major targets of IRE1 RNase activity in melanoma tumors. Deletion of XBP1s in DCs resulted in augmented tumor growth, impaired effector T cell responses and marked accumulation of TIM-3+ CD8+ T cells with a terminal exhausted profile. Transcriptomic studies further revealed that XBP1s deletion in tumor cDC1s induced the loss of major proteostasis core genes and elicited mRNA degradation via regulated IRE1-dependent decay (RIDD), which accounted for the dysregulated T cell immunity in melanoma. Thus, our work demonstrates that a strict regulatory circuit involving IRE1 RNase and XBP1s in DCs ensures optimal antitumor T cell immunity in melanoma models.
Supported by HHMI international research scholar grant # 55008744 FONDECYT (National Fund for Scientific and Technological Development) #1200793 CONICYT-PFCHA/DoctoradoNacional/2017-21170366
CD95/Fas-ligation and resting NK or IL-2 activated NK (LAK) attack limit NK-92 efficacy.
Authors: Navarrete-Galvan Lydia T , Guglielmo Michael , Amaya Judith Cruz , Merica Rebecca , Smith-Gagen Julie , Lombardi Vincent C , Hudig Dorothy 
Published: January 2023

Abstract
Adoptively transferred NK-92 cells and genetically modified variants must undergo irradiation to prevent proliferation, since NK-92 cells are themselves tumors. This malignant quality also makes NK-92 cells susceptible to attack by patients’ NK cells. Here, we investigated the effect of gamma-irradiation and/or Fas-ligation on NK-92 killing against K562 tumor cells, as well as the effect of irradiation on the susceptibility of NK-92 cells to attack by primary NK cells. To evaluate killing, we used a 51Cr-release assay with multiple effector to target (E:T) K562 tumor cell ratios at 4- and 8-hour time points, 10 Gy irradiation and IgM antibody ligation of Fas. (K562 cells lack detectable Fas). NK-92 cells, maintained in 1000 U/ml of recombinant IL-2, retained high cytotoxic activity immediately after irradiation, but lost &gt;50% activity one day after 10 Gy irradiation. NK-92 cells were highly resistant to death initiated by CD95-ligation, whether irradiated or not. However, the surviving NK-92 cells suffered loss of cytotoxicity following Fas-ligation. This loss of cytotoxicity was more pronounced when Fas-ligation and irradiation were combined. Irradiation also made NK-92 cells more sensitive to attack by resting NK cells and IL-2 activated NK (LAK) cells. These findings demonstrate that Fas-ligation and irradiation are detrimental to NK-92 functionality and support the recommendation that genetic removal of CD95/Fas may increase cytotoxicity by NK-92 cells and potentially, other cell lines used for adoptive immunotherapies.
This research was supported in part by a UNR Foundation Award, an NIH P30 GM110767 for the Cytometry Center, a Nevada Undergraduate Research Award, and by the Nevada INBRE program (NIH GM103440) for research scholarship and supplies that helped support the project.
Characterization of the Tumor Immune Microenvironment in Soft Tissue Sarcoma Patients Undergoing Surgery
Authors: Cruz Sylvia M , Judge Sean J , Darrow Morgan A , Perry Lauren M , Farley Lauren E , Iranpur Khurshid R , Dunai Cordelia , Chen Shuai , Thorpe Steven W , Canter Robert J 
Published: January 2023

Abstract
Tumor infiltrating lymphocytes (TILs) have been shown to predict survival in soft tissue sarcomas (STS), but the specific contribution of natural killer (NK) and CD8+ T cells to outcomes is undefined. Therefore, we sought to characterize the extent of NK and CD8+ T cell infiltration in STS.
Prospectively, we evaluated 15 patients using fresh tumor from surgery for flow cytometric analysis. Retrospectively, we evaluated archived tumor tissue from 90 STS patients by immunohistochemistry (IHC) for CD3, CD8, CD45RO, NKp46, TIGIT, MHC-I, and p53. We analyzed metastasis-free survival (MFS) and overall survival (OS) by Kaplan-Meier method and log-rank test.
By flow cytometry, we observed significant variability in CD45+ leukocytes in the STS TME (mean 29±24% of total live cells) with low percentages of tumor-infiltrating CD3-CD56+ NK cells (1.7±1.9% of total live cells and 5.3±3.0% of live CD45+ cells) and CD8+ T cells (1.6±1.6% of total live cells and 29.6±30.5% of live CD45+ cells). By IHC, NK and T cell infiltrates were low (median H score 0, range 0–66.5 and 2.7, range 0–110, respectively). We confirmed a positive correlation between CD8+ T cell infiltration and significantly improved OS (P&lt;0.05) and a trend for improved MFS. We also observed a trend for improved OS among patients with higher NKp46 scores (P=0.07). MHC-I expression positively correlated with both T and NK cell infiltration (P&lt;0.05), whereas TIGIT expression positively correlated with T cell infiltration (P&lt;0.05), but not NK infiltration.
Infiltration of NK and CD8+ T cells is overall low in STS patients undergoing surgery but associated with superior OS. Further characterization of the immune infiltrate in STS may yield better biomarkers of prognosis and immune targeting.
Supported by the following grants: NIH/NCI 1R03CA252793 NIH/NCI T32 CA251007-01
Tumor resident memory CD8 T cell formation and concomitant tumor immunity is CD40L dependent and CD4 independent.
Authors: Gough Michael J , Kramer Gwen , Bambina Shelly , Alice Alejandro , Blair Tiffany , Duhen Thomas , Duhen Rebekka , Crittenden Marka R 
Published: January 2023

Abstract
The implantation of cancer cell lines into immunocompetent mice acts as a vaccination event, resulting in T cell immunity that impacts both tumor progression and the response to subsequent therapy. We have previously shown that blocking immunity at tumor implantation using anti-CD40L antibodies blocks development of an intratumoral antigen-specific CD8 T cell population with tissue resident memory (Trm) phenotypes, and also limits responsiveness to radiation therapy and immunotherapy combinations. Using pancreatic tumors derived from Pdx1-Cre and Pdx1-Cre /R26LSL-LSIY mice crossed with KrasLSL-G12D/+ Tp53LSL-R172H/+ mice, we explored the role of CD4 T cells in providing CD40-CD40L help to generate Trm and conventional T central memory CD8 T cells (Tcm) cells in the tumor and systemically. Using depleting antibodies, we identified that Trm cells in the tumor could form independently of CD4 T cells but were lost following CD40L blockade. By contrast, systemically circulating Tcm cells that could re-expand after antigen-specific challenge were dependent on both CD4 T cells and CD40-CD40L interactions. Notably, concomitant immunity generated by tumor implantation that can prevent tumor growth on the opposite flank could also occur independently of CD4 T cells, but was lost on CD40L blockade. Systemic vaccination with Listeria monocytogenes expressing SIY efficiently induced the formation of SIY-specific Tcm cells but did not impact tumor implantation. These data suggest that tumor implantation generates Trm cells independently of CD4 T cells, and that these cells, not Tcm cells, sustain concomitant immunity. These data may impact immune therapies designed to control metastatic seeding in patients.
Supported by grants from NIH (R01CA182311, R01CA244142, R01CA208644).
TGF-β-dependent Lymphoid Tissue Residency of Stem-like T cells Limits the Response to Tumor Vaccine
Authors: Zhang Nu , Li Guo , Wang Liwen , Srinivasan Saranya , Ma Chaoyu , Liao Wei , Liu Yong , Mishra Shruti , Zhang Xin , Qiu Yuanzheng , Lu Qianjin 
Published: January 2023

Abstract
Stem-like CD8+ T cells represent the key subset responding to multiple tumor immunotherapies, including tumor vaccination. However, the signals that control the differentiation of stem-like T cells are not entirely known. Most previous investigations on stem-like T cells are focused on tumor infiltrating T cells (TIL). The behavior of stem-like T cells in other tissues remains to be elucidated. Tissue-resident memory T cells (TRM) are often defined as a non-circulating T cell population residing in non-lymphoid tissues. TILs carrying TRM features are associated with better tumor control. Here, we found that stem-like CD8+ T cells differentiated into TRMs in a TGF-β and tumor antigen dependent manner almost exclusively in tumor draining lymph node (TDLN). TDLN-resident stem-like T cells were negatively associated with the response to tumor vaccine. In other words, after tumor vaccine, TDLN stem-like T cells transiently lost TRM features, differentiated into migratory effectors and exerted tumor control.
This work is supported by NIH grants AI125701 and AI139721, Cancer Research Institute CLIP program and American Cancer Society grant RSG-18-222-01-LIB to N.Z. Data generated in the Flow Cytometry Shared Resource Facility were supported by the University of Texas Health Science Center at San Antonio (UTHSCSA), NIH/NCI grant P30 CA054174-20 (Clinical and Translational Research Center [CTRC] at UTHSCSA), and UL1 TR001120 (Clinical and Translational Science Award).
Pre-assembled universal SNAP-CARs show potent anti-tumor function
Authors: Ruffo Elisa , Kvorjak Michael , Adams Eric , Lohmueller Jason 
Published: January 2023

Abstract
Universal chimeric antigen receptors (CARs) are engineered T cell receptors that instead of directly binding to an antigen, recognize one or more adaptor molecules that bind to a target antigen. Universal CARs are of high clinical interest due to their tunable activity by antibody adaptor dose and the capabilities of targeting multiple antigens on the same cancer or different cancers by combining with different adaptors. We previously developed a unique universal CAR, “SNAP-CAR,” that carries out a chemical reaction to covalently fuse to antibody adaptors with a benzylguanine tag. Our previous experiments showed potent SNAP-CAR function with co-administered antibody adaptor. We now expand on these findings evaluating the function of the SNAP-CAR when pre-assembled with the adaptor prior to therapy. Primary human SNAP-CAR T cells were pre-incubated with different amounts of antibody adaptor, showing a titratable level of receptor assembly. When challenged with antigen positive target cells, SNAP-CAR T cells showed potent activation and killing activity that correlated with the amount of preloaded adaptor in a dose-dependent manner. The activity of pre-assembled SNAP-CAR T cells was then tested in vivo in a human tumor xenograft NSG mouse model targeting the HER2 antigen. A single dose of preloaded SNAP-CAR T cells showed significant tumor regression that could be further enhanced by additional injections of the preloaded SNAP-CAR T cells. Administration of adaptor along with preloaded SNAP-CAR T cells further improved tumor regression. In conclusion, preloading of SNAP-CAR T cells prior to therapeutic administration can deliver a potent pulse of anti-tumor activity with the capability of further programming by additional adaptor.
Supported by NIH: R01GM142007-01A1
Sex hormones more than sex chromosomal complement predict sex differences in influenza vaccine-induced immunity and protection in mice
Authors: Seddu Kumba , Dhakal Santosh , Ganesan Abhinaya , Jacobsen Henning , Klein Sabra L 
Published: January 2023

Abstract
Following influenza vaccination, adult female mice have greater quantity and quality of antiviral antibodies than males, which is a correlate of protection against live influenza virus challenge. Whether sex steroid hormone concentrations, sex chromosome complement, or both underlie greater immunity and protection following vaccination has not been determined. To dissect the relative contribution of steroid hormones and chromosome complement, we used the Four Core Genotype (FCG) mouse model on a C57BL/6 background to evaluate how gonadal sex (i.e., testes or ovaries) and sex chromosome complement (i.e., XX or XY) can independently or together contribute to sex differences in influenza immunity. Adult FCG mice were vaccinated intramuscularly with inactivated mouse-adapted A/California/04/09 (ma2009 H1N1). Blood samples were collected at several time points post vaccination to measure total IgG and neutralizing antibodies. All mice were challenged intranasally with live ma2009 H1N1 drift variant virus and used to either measure lung virus replication kinetics or disease pathogenesis. Gonadal females (i.e., XXF and XYF), regardless of chromosome complement, produced greater antibody responses and had less pulmonary virus replication than gonadal males (i.e., XYM and XXM). Gonadal sex and sex chromosomal complement intersected to impact disease severity following challenge, in which among gonadal females, XX females experienced less morbidity than XY females. In contrast, there was no effect of chromosome complement among gonadal males. Our data suggests that sex steroid hormones more than sex chromosome complement underlie sex differences in influenza vaccine-induced immunity and protection.
Financial support provided by NIH/OWRH/NIA U54AG062333
Improvement of the aging-related reduced efficacy of the H1N1 pandemic influenza vaccine by the complex of poly-γ-glutamic acid and alum
Authors: Kim Jaemoo , Jihyun Yang , Haryoung Poo 
Published: January 2023

Abstract
Highly contagious respiratory diseases caused by viral infections, like influenza are a constantly emerging threat, particularly among the elderly. Vaccination is one of the most effective strategy for preventing influenza, but the aged-related reduced efficacy of the influenza vaccine remains an unresolved problem. Here, we demonstrate that the aging-related reduced efficacy of the influenza vaccine can be improved by the complex of poly-γ-glutamic acid (γ-PGA) and alum called PGA/Alum, a new adjuvant capable of inducing innate and adaptive immune responses in aged mice. PGA/Alum induced activation of dendritic cells isolated from spleen of aged mice, with increases of expression of co-stimulatory molecules and production of pro-inflammatory cytokines. Although aged mice were observed decrease of T and B cell population and weaken efficacy of pandemic H1N1 influenza vaccine than young mice, PGA/Alum recovered the vaccine efficacy in aged mice. Notably, the vaccine-PGA/Alum-immunized aged mice increased hemagglutination-inhibition titers, serum-neutralization titers, cytotoxic T lymphocyte activity, and generation of multifunctional CD4+ T lymphocytes. Taken together, these results suggest that PGA/Alum could be a potent vaccine adjuvant that improves the aging-related reduced efficacy of the influenza vaccine, thereby resulting in effective prevention against influenza virus infection in the elderly.
This research was funded by a grant from KRIBB Research Initiative Program (KGM9942112).