bluuebunny/crossref_metadata_2025_split huggingface dataset:
Combination of adoptive T cell therapy with an EGFR x EpCAM-specific antibody for the treatment of a murine melanoma model
Authors: Klaus Richard Carl Philipp , Lampert Christopher , Schmollinger Jan C. , Niederfellner Gerhard , Sustmann Claudio , Klein Christian , Endres Stefan , Kobold Sebastian 
Published: January 2023

Abstract

Background
Adoptive T cell therapy is a new powerful treatment of cancer - however limited by T cell infiltration to the tumor. Transduced antigen-specific T cells with a marker antigen (EGFR) not expressed by normal T cells combined with an EGFR x EpCAM bispecific Antibody (biAB) mediate potent anti-tumor effects. To test the applicability of this approach to different T cell antigens and tumor models, we hypothesized that this synergy could be seen in the B16-OVA-melanoma model using ovalbumin-specific T cells (OT-1).

Methods and Results
We generated a B16-OVA-mEpCAM cell line through retroviral transduction. We expanded CD8+ T cells from OT-1 transgenic mice in vitro. C57Bl/6 mice were subcutaneously injected with B16-OVA-EpCAM melanoma cells and were treated with OT-1 T cells or the combination of the biAB with T cells (n=6 per group). Tumor growth was delayed in the combination group (mean tumor size on day 15, 77 mm2 vs 137 mm2). However we found that the treatment success relied entirely on the biAB alone, while the T cells did not contribute to efficacy (mean tumor size on day 18, 8 mm2 in antibody-treated vs 106 mm2 in untreated mice). We hypothesized that antibody dependent cellular cytotoxicity (ADCC) may be the mode of action of the biAB. We thus sublethally irradiated (5 Gy) mice 24 h after subcutaneous injection of B16-OVA-EpCAM tumor cells and found that irradiation completely abrogated the effect of the biAB treatment (mean tumor size on day 29, 146 mm2 in irradiated vs 117 mm2).

Conclusions
Combination of antigen-specific OT-1 T cells and a biAB does not synergize in the B16-OVA-EpCAM model. The biAB was, however, effective in this model, most likely due to ADCC. Other strategies need to be investigated to enhance ACT in this model.
Low antigen dose in adjuvant-based vaccination selectively induces CD4 T cells with enhanced functional avidity and protective efficacy
Authors: Billeskov Rolf , Wang Yichuan , Solaymani-Mohammadi Shahram , Frey Blake , Kulkarni Shweta , Agger Else Marie , Sui Yongjun , Berzofsky Jay A 
Published: January 2023

Abstract
T cells of high functional avidity sense and respond to low levels of antigen (Ag), and their superior protective efficacy was first described using CD8 T cells cultured in vitro with low levels of Ag resulting in higher avidity and anti-tumor/viral efficacy compared to low avidity T cells cultured with high Ag concentrations. Selectively inducing high avidity by low Ag concentrations has required in vitro expansion so far, as in vivo attempts with low-dose vaccination gave no response.
We show for the first time that functionally high avidity CD4, but not CD8, T cells are selectively primed in vivo by low concentrations of Ag in the strong crosspriming liposomal cationic adjuvant formulations (CAF) DDA/TDB or DDA/MMG/pIC from SSI. Increased functional avidity observed after low-dose vaccinations correlated with lower surface expression of PD-1 and higher per cell IFN-gamma production compared to low avidity CD4 T cells. We further show that IL-15, known to increase functional avidity of CTLs, is essential for high avidity of vaccine specific CD4 T cells.
Importantly, high avidity CD4 T cells offered superior protection in a murine model of latent infection with Mycobacterium tuberculosis compared to low avidity T cells. Furthermore, adoptive transfers of HIV-IIIB gp140-specific CD4 T cells of high functional avidity resulting from low-dose immunizations in combination with TCR transgenic HIV-IIIB gp120 specific CTLs conferred superior protection against viral (vaccinia-HIV-IIB-gp140) challenge compared to TCR-Tg CTLs alone or in combination with low avidity CD4 T cells induced by high-dose immunization. Thus, inducing high avidity CD4 T cells by low-dose immunization is important in clearing viral and bacterial infections.
Study of specific plasmablasts and plasma cells to lipidic particles in the mouse models of lupus generated by liposomes bearing lipidic particles stabilized with different inducers
Authors: Mora Albany Reséndiz , Pardo Elena Sanchez , Galicia Viridiana Galicia , Barbosa Sandra Sanchez , Wong Carlos , Baeza Isabel , Baeza Carlos Wong 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is an inflammatory, multisystem, chronic an autoimmune disease of unknown etiology. In our research group, we developed a mouse model of lupus by the administration of liposomes bearing lipid particles stabilized by chlorpromazine, promazine or manganese in BALB/c mice. In this model, we have detected IgG antibodies against lipid particles, so we are studying the immune mechanisms of how IgG antibodies are generated against a lipid antigen. It is noteworthy that the production mechanisms of anti-lipid IgG antibodies are practically unknown and have not been studied in autoimmune diseases such as SLE. In previous works, we showed that B cells mainly respond through germinal center to generate IgG antibodies against lipid particles. Moreover, against protein antigens after germinal center reaction, B cells differentiate into secondary lymphoid organs to plasmablasts that are maintained in cell cycle, can be divided and are in circulation. Eventually these plasmablasts can differentiate into plasma cells that are no longer in the cell cycle and are set in a niche (mainly in bone marrow or less in secondary lymphoid organs) where they begin to secrete antibodies. In this work, we propose to study the mechanisms of plasmablasts and plasma cells, specific to a lipid antigen, that leads to the production of IgG antibodies. This is very important because the IgG autoantibodies are the main causes of autoimmunity and particularly the anti-lipid particles antibodies, which have been found in patients with SLE, leprosy and preeclampsia, are those that trigger the lupus like disease in the mouse.
Comparative analyses of PAD expression and activity in myeloid cell lines
Authors: Rumble Julie M , Fackelman Elli M , Mobley James L 
Published: January 2023

Abstract
Protein arginine deiminase (PAD) enzymes and the arginine to citrulline reaction that they catalyze have increased in prominence in recent years, as dysregulation is associated with multiple inflammatory and autoimmune diseases. Under normal circumstances, PAD4 is required for the generation of neutrophil extracellular traps (NETs) by neutrophils. PAD4 citrullinates histone H3, initiating chromatin decondensation and subsequent extrusion of nuclear DNA to form a net-like structure. These NETs are important in innate immune responses to infection, as loss of components of this process can lead to the inability to eradicate usually harmless pathogens. We have developed tools to better understand the complete picture of PAD2 and PAD4-catalyzed citrullination and its effects in various systems. HL-60 and THP-1 cells differentially expressed PAD2 and PAD4, depending on differentiation and activation stimuli, as shown by ELISA. The activity of these PAD enzymes was also highly dependent on the differentiation and activation status of these cells. The PAD4 target histone H3 was found to be citrullinated only upon activation of differentiated cells, although PAD4 was expressed in differentiated cells which had not been activated. These data help to elucidate appropriate model systems for researchers looking to investigate PAD activation and downstream targets in immune cells.
The role of Ndfip1 in macrophage-mediated bacterial clearance
Authors: Field Natania S , Moser Emily K , Chandramouleeswaran Prasanna , Subudhi Ipsita , Vasquez Nikoff , Oliver Paula M 
Published: January 2023

Abstract
Macrophages are essential components of the innate immune response to bacteria. While these cells must mount an inflammatory response to harmful pathogens, they require tight regulation to prevent host damage. One way in which macrophages attenuate inflammatory signaling cascades is through ubiquitination. This post-translational modification can target a protein substrate for downstream signaling or for degradation. E3 ubiquitin ligases are required to facilitate the formation of these chains. Nedd4-family interacting protein 1 (Ndfip1) activates multiple E3 ubiquitin ligases, including Itch, which is crucial for attenuating cytokine production by T cells, and Nedd 4-2, which is required for iron regulation by neurons. While Itch has been shown to limit macrophage activity, it is unclear how Ndfip1 functions in macrophages, and how it affects macrophage antibacterial responses.
We generated mice lacking Ndfip1 in myeloid cells (Ndfip1fl/fl LysM Cre). Unlike mice lacking Ndfip1 in T cells, these mice have no evidence of inflammation at baseline. Upon i.n. infection with Klebsiella pneumoniae, Ndfip1 fl/fl LysM Cre mice show reduced bacterial loads three days post infection, despite normal innate cell recruitment. This result could be recapitulated when mice were infected intraperitoneally. Peritoneal macrophages lacking Ndfip1 have unchanged cytokine responses when stimulated with K. pneumonia, suggesting Ndfip1 attenuates macrophage activity through a mechanism other than limiting cytokine production. This project may reveal new mechanisms used by myeloid cells in eliminating bacteria, providing insight into the regulatory mechanisms that prevent excess inflammation.
The dual roles of membrane and soluble forms of gC1qR and C1q in the breast cancer microenvironment.
Authors: Kandov Evelyn , Pednekar Lina , Hosszu Kinga K , Nanda Japbani , Galanakis Dennis K , Preissner Klaus T , Kishore Uday , Peerschke Ellinor I.B , Ghebrehiwet Berhane 
Published: January 2023

Abstract
There is robust experimental data, which show that C1q and gC1qR play a pro- as well as anti-proliferative role in cancer. This opposing role in turn depends on whether each molecule is expressed on the cell surface or pericellular milieu. The aim of the present studies was therefore to more closely examine the functions of soluble and membrane forms of gC1qR and C1q using the SkBr3 cell line as a model for breast cancer. Our results show that: (1) In addition to gC1qR and cC1qR, SkBr3 cells express membrane C1q, and these molecules appear to be co-localized with each other as well as with fibrinogen (FGN) and vitronectin (VN). (2) Co-culture of SkBr3 cells with either purified C1q or with pAb directed against the gC1qR site on the A-chain of gC1q (ghA) resulted in a time-dependent inhibition of cell growth. (3) Co-culture of SkBr3 cells with individual C1q head modules-–ghA, ghB and ghC–showed significant inhibition, with the inhibitory hierarchy being ghA&gt;ghC &gt;ghB. (4) While SkBr3 cells do not secrete C1q, they are able to secrete gC1qR, and the latter in turn is able to negate the anti-proliferative effect of exogenously added C1q supporting the postulate that gC1qR secreted into the tumor cell microenvironment serves as an important molecular checkpoint. Collectively, the data show that both membrane-associated gC1qR and C1q on their own are pro-proliferative. Whereas the proliferative function of gC1qR can be abrogated by blockade of its C1q binding site, the proliferative function of C1q can be abrogated by blockade of its gh domains with specific antibodies indicating therefore that the pro-proliferative function of C1q resides in the its gh domains, with the ghA domain playing a central role.
Endosomal RNA-sensing toll-like receptor stimulated T cells play a unique role in autoimmune manifestations in murine model of DNA accrual.
Authors: Pawaria Sudesh , Marshak-Rothstein Ann 
Published: January 2023

Abstract
DNaseII is a lysosomal DNase that plays a critical role in immune homeostasis. DNaseII deficiency in mice causes embryonic lethality, due to severe anemia associated with excessive production of type I interferon (IFN), and can be rescued by IFN receptor knockout (IFNaR). The DNaseII/IFNaR double knockout (DKO) mice develop a late-onset inflammatory arthritis, dependent on the cytosolic DNA sensor STING (Stimulator of Interferon Genes). They also develop early-onset clinical manifestations of systemic autoimmunity including disrupted hematopoiesis and B cell lymphopoiesis, splenomegaly (due to increased accumulation of Ter119+ cells), and the production of anti-nuclear antibodies (ANA), through a mechanism that is STING-independent, Unc93B1-dependent and likely to result from endosomal RNA-sensing TLRs. Bone marrow (BM) chimera studies have shown that the DNaseII deficiency is essential in both radioresistant host cells and hematopoietic donor cells for all disease manifestations. Development of arthritis requires the presence of STING in both host and donor. The TLR-dependent defects require TLR expression in only the hematopoietic compartment but still need DNaseII−/− recipients. Remarkably transplantation of DKO spleen fragments or injection of DKO spleen cells alone into DNaseII-sufficient Rag−/− mice leads to splenomegaly and disrupted hematopoiesis and lymphopoiesis. Moreover DKO T cells are sufficient to transfer these TLR-associated defects to the Rag−/− mice. As activated T cells are known to be radioresistant, T cells activated in a DKO environment may account for the radioresistant host component required to precipitate the TLR-dependent disease manifestations.
High dimensional analysis of immune signatures in melanoma patients
Authors: Krieg Carsten , Nowicka Malgorzata , Guglietta Silvia , Schindler Sabrina , Hartmann Felix J. , Weber Lukas M. , Dummer Reinhard , Robinson Mark D. , Levesque Mitchell P. , Becher Burkhard 
Published: January 2023

Abstract
Checkpoint inhibitor therapy (CIT) has revolutionized cancer therapy. Notably, inhibition of programmed cell death protein 1 (PD-1) showed benefit in the treatment of metastatic melanoma and has even been promoted to first line therapy in advanced melanoma. Despite an increase in overall survival a minority of patients profits from CIT with anti-PD-1. Therefore, immune components or markers predicting responsiveness are urgently needed. Using phenotypic and functional markers we used high-dimensional single cell cytometry by time of flight (CyTOF), to measure blood samples from anti-PD-1 treated patients before and during therapy. CyTOF uses metal-tagged instead of fluorochrome-tagged antibodies, thus eliminating spectral overlap and, in combination with barcoding, enables the measurement of more than 30 parameters on a single cell in multiple samples in parallel. Due to the complexity of the data set and in order to reduce analytical bias we adapted a custom bioinformatic workflow. Using this approach we find immune signatures which allow us to separate responders from non-responders and might provide insights into the biology of immune therapy.
Cancer derived peptide of vacuolar ATPase ‘a2’ isoform plays immunosuppressive role by recruiting myeloid derived suppressor like cells
Authors: Sahoo Manoranjan , Kulshrestha Arpita , Katara Gajendra K , Beaman Kenneth D 
Published: January 2023

Abstract
Cancer cells express vacuolar ATPase (V-ATPase), a multi-subunit proton pump on the plasma membrane and intracellularly on vesicular membranes. The V-ATPases are responsible for acidification of intracellular vesicles and in certain activated cells are expressed on the surface where they appear to acidify the extracellular environment. The N-terminal domain of the a2 isoform of V-ATPase (a2NTD) is cleaved and secreted in exosomes. a2NTD has immunomodulatory effects on monocytes, macrophages and neutrophils. The changes elicited by a2NTD play an important role during pregnancy and cancer/tumor progression. To further investigate whether a2NTD has an effect on recruitment or stimulation of MDSCs in vivo, we injected a2NTD into the peritoneum of mice and analyzed CD11b+ myeloid cell populations. Here we observed that, intra-peritoneal (i.p.) administration of recombinant a2NTD protein results in recruitment of significantly higher number of CD11b+Ly6G+Ly6C+ MDSC like cells to the peritoneal cavity. We show that these cells are different from neutrophils by their expression of Ly6C on their surface. However, the total percent of CD11b+ cells or CD11b+F4/80+ macrophages remained unchanged. In vitro results demonstrate that addition of recombinant a2NTD protein to primary bone marrow derived macrophages (BMMs) from C57BL/6 mice leads to secretion of anti-inflammatory cytokine IL-10. Collectively, these results demonstrate that a2NTD administration leads to an immunosuppressive environment that might facilitate tumor progression. Further study is needed to understand the mechanism by which a2NTD modulates the MDSC like cells and creates an immunosuppressive pro-tumorigenic environment.
HDAC8 inhibition modulates liver tumor microenvironment: Rationale for combined epigenetic and immunotherapy
Authors: Cheng Alfred Sze , Feng Yu , Zhou Jingying , Yang Weiqin , Sun Hangyong , Chen Zhiwei , Goodwin Tyler , Liu Rihe , Huang Leaf 
Published: January 2023

Abstract
The tumor microenvironment plays an instrumental role in cancer development and treatment response/resistance. Accumulating evidence is underscoring the fundamental importance of epigenetic regulation in tumor immune evasion. Epigenetic modification agents represent a ‘double-edged sword’, as they can exert divergent effects on cancer cells and the milieu of immune and stromal cells. Our integrative epigenomics analysis has elucidated previously unexplored functions of histone deacetylase 8 (HDAC8) in promoting β-catenin-dependent hepatocarcinogenesis through interacting with another critical chromatin regulator enhancer of zeste homology 2 (EZH2). Given the strong oncogenicity of HDAC8, we further investigated the therapeutic potential of a potent and highly-selective HDAC8-specific inhibitor PCI-34051. Using a hepatocellular carcinoma (HCC) orthotopic model in immunocompetent mice, we demonstrated that HDAC8 inhibition exerted a strong anti-tumorigenic effect (Ctrl vs. PCI: 6.32 vs. 0.33*107 ROI intensity; p&lt;0.01), which was comparable with programmed death-ligand 1 (PD-L1) blockade. Intriguingly, PCI-34051 significantly increased tumor-infiltrating multi-functional CD8+ T cells (~3-fold) and specifically reduced regulatory T cells (~2-fold). However, the level of PD-1+CD8+ T cells also increased, implicating potential T cell exhaustion. Our data suggest that selective chromatin modifications by HDAC8 and EZH2 alter the tumor immune surveillance program. By understanding the impact of epigenetic control on the liver tumor microenvironment, rational combinatorial epigenetic and immune checkpoint targeting has the potential to fully unleash T cell responses against HCC.
Immunotherapy against experimental melanoma is improved by beta-adrenergic blockade
Authors: Kokolus Kathleen M , Drabick Joseph J , Repasky Elizabeth A , Schell Todd D 
Published: January 2023

Abstract
Despite recent advances in immunotherapy which have expanded cancer treatment regimens to target a patient’s own immune cells, a significant proportion of patients fail to achieve long-lasting clinical benefit. The stress and immune responses are closely integrated yet few cancer treatment strategies account for this relationship. Activation of stress signaling releases norepinephrine which binds β-adrenergic receptors (βARs) on immune cells leading to regulation of immune functions. βAR signaling is reduced with antagonists (β-blockers) and these drugs, often prescribed for hypertension, are associated with improved cancer outcomes. We determined how βAR blockade impacts the efficacy of two immunotherapies used to treat melanoma: IL-2 and anti-(α)PD-1 which promote T cell proliferation and function, respectively. Mice with established melanomas were treated with β-blockers and immunotherapy (αPD-1, IL-2 or αPD-1/IL-2). Tumor growth was monitored and immune cell accumulation within tumors and lymphoid tissues was evaluated. Blockade of βAR signaling alone had no impact on tumor growth. Attenuation of tumor growth by each immune-based therapy was enhanced with administration of β-blockers, but this effect was most significant with dual αPD-1/IL-2. Likewise, survival of tumor-bearing mice was significantly extended by addition of β-blockers to αPD-1 or αPD-1/IL-2. Importantly, our results from a parallel retrospective clinical analysis of metastatic melanoma patients support a potential role for β-blockers in enhancing immunotherapy. This novel strategy could be translated to potentially increase the number of melanoma patients experiencing long term benefit from immune-based therapies.
Harnessing the immune response for the treatment of cancer by inducing immunogenic cell death through inhibition of a group II chaperonin
Authors: Khaled Annette R , Showalter Anne , Limaye Arati , Bassiouni Rania , Khaled Amr S , Oyer Jeremiah , Igarashi Robert , Copik Alicja 
Published: January 2023

Abstract
Immunotherapy is a promising treatment for cancer but lacks broad application. One way to enhance immunotherapy is by inducing immunogenic cell death (ICD). ICD is characterized by a unique molecular signature, involving the release of molecules that sensitize cancer cells to killing by the immune system. However, most ICD-inducing agents are not effective at clinically relevant doses. We developed a novel cytotoxic peptide, CT20p, that is delivered to cells using nanoparticles (NPs), which causes regression of tumors in mice at nanomolar concentrations. CT20p-treated cancer cells upregulated markers characteristic of ICD, such as calreticulin (Crt) and HSP70. The intracellular target of CT20p is a group II chaperonin called chaperonin containing-TCP-1 (CCT) that is essential for the folding of actin, tubulin and other proteins like STAT3. The CCT complex is composed of eight subunits (CCT1-8). That CT20p cytotoxicity is driven by CCT was proven by inhibiting or overexpressing CCT2 in susceptible or resistant cells, respectively. Disruption of CCT activity by CT20p caused endoplasmic reticulum (ER) stress that initiated intracellular pathways, such as PERK and IRE-1, generating danger signals associated with ICD. As a result, increased phagocytosis of dying cancer cells and enhanced NK cell killing was observed. Importantly, normal epithelial cells, macrophages or NK cells were unaffected. Using the CRISPR-CAS9 system, the genes for CCT were edited. Loss of CCT2 altered the levels of other CCT subunits as well as client proteins such as STAT3, while increasing membrane-exposed Crt. These studies revealed that inhibiting CCT induces ICD to stimulate immune functions essential for generating a robust anti-cancer immune response.