bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tumor growth following genetic modifications of mouse prostate tumor cells
Authors: Aguilar Roberto , Zhang Carl Tinghao , Ma Tengchuan , Feng Chongyu 
Published: January 2023

Abstract

Objectives
1) To determine the growth features of MyC-CaP mouse prostate tumors induced in immunodeficient nude mice and in immunocompetent FVB mice; and 2) To assess growth characteristics of MyC-CaP-p53 null tumor cells in vitro.

Methods
MyC-CaP cells were transfected with enhanced green fluorescent protein (eGFP), and the MyC-CaP-eGFP cells were injected intraperitoneally into naive immunodeficient nude recipients and into naive immunocompetetent FVB recipients. The transplanted tumor cells were subsequently tracked by fluorescent stereo microscopy. MyC-CaP-p53 null tumor cells were generated by inducible CRISPR/Caspase-9 gene deletion targeted to the native mutated p53 tumor suppressor gene.

Results
1) Increased growth of MyC-CaP-eGFP tumors occurred in immunodeficient nude mice compared to growth in immunocompetent FVB mice; 2) decreased metastasis of MyC-CaP-eGFP tumors compared to MyC-CaP tumors occurred following injection into immunocompetent FVB mice; and 3) MyC-CaP-p53 null cells showed reduced proliferation and growth in vitro compared to non-modified MyC-CaP tumor cells.

Conclusions
Three conclusions can be drawn from these studies: 1) Injection of MyC-CaP-eGFP tumor cells into naive immunodeficient nude recipients allowed for adequate examination of initial tumor viability and subsequent tumor growth and metastasis; 2) Decreased MyC-CaP-eGFP tumor growth in immunocompetent FVB mice compared to immunodeficient nude mice indicates that an immune response to the eGFP protein inhibited growth of the MyC-CaP-eGFP tumor; and 3) The growth inhibition following deletion of the mutated p53 tumor suppressor gene in MyC-CaP tumor cells indicates that the mutated allele helps drive the malignancy.
TGF-β signature is a potential biomarker for identifying individualized immunotherapeutic approaches in hepatocellular carcinoma
Authors: Chen Jian , Su Xiaoping 
Published: January 2023

Abstract

Background
Several immune checkpoint inhibitors have recently shown promising results in patients with advanced hepatocellular carcinoma (HCC). However, tumor response rates ranged from 14% to 17%. Thus, there remains a large unmet medical need for novel and effective immunotherapeutic strategies against HCC. Dysregulated signaling in the transforming growth factor (TGF)-β pathway plays a central role in immunomodulation, inflammation, and fibrogenesis in the HCC microenvironment. We therefore used The Cancer Genome Atlas database to analyze potential therapeutic approaches combining TGF-β inhibition and immunotherapies for HCC.

Methods
Transcriptomic analyses were performed in 193 HCC cases. Pathway analyses were performed by correlating 70 published signatures with TGF-β signatures.

Results
1. Unsupervised hierarchical clustering of TGF-β genes in HCC revealed four distinct clusters with unique TGF-β signatures: highly activated (14.5%), activated (45.0%), normal (30.6%), and inactivated (9.9%). 2. Highly activated TGF-β signatures were significantly associated with fibrosis and activated stromal signatures. 3. TGF-β signature subtypes were significantly associated with immune cell infiltration, T cell exhaustion, and checkpoint blockade response subclasses. 4. Potentially targetable genes were associated with specific TGF-β signatures. 5. TGF-β signatures provide potential combination therapeutic options for HCC.

Conclusion
We demonstrated a specific immunosuppressive role of TGF-β in mediating immunotherapy resistance in HCC, highlighting the TGF-β signature as a potential biomarker in individualized immunotherapeutic approaches in HCC.
Conventional type 1 dendritic cells and natural killer cells demonstrate strong correlation to cytotoxic T lymphocyte infiltration in cervical cancer tumors
Authors: Dikshit Anushka , Anderson Courtney M , Zhang Bingqing , Ma Xiao-Jun 
Published: January 2023

Abstract
Professional antigen presenting cells such as dendritic cells (DCs) are vital for priming naive CD8+ T cells and developing a sustainable anti-tumor immune response. Natural killer (NK) cells within the tumor microenvironment (TME) recruit a specific population of DCs called conventional type 1 DCs (cDC1s). However, these cells are low in abundance making their detection in the tissue context challenging.
To interrogate the presence of cDC1 and NK cells in the TME and reveal their spatial relationship to each other we utilized the highly sensitive and specific RNAscope Multiplex Fluorescence in situ hybridization (ISH) assay. NK cells and cDC1 cells were identified by using cell specific marker probes in 4 cervical cancer samples. Similarly, CTLs were visualized to determine if there is a correlation between the presence of cDC1 and NK cells and infiltration of CTLs within the cervical cancer tumors.
Our results revealed a strong correlation between the presence of NK cells, cDC1 cells, and CTLs within 3 out of 4 cervical cancer samples. The NK cells showed expression of the chemokines XCL1 and CCL5, suggesting that the XCR1+/CCR5+ cDC1 cells may have been potentially recruited by these NK cells in the TME. Regions high in cDC1 and NK cells also showed significantly higher levels of CTL recruitment, indicated by the presence of CD8+/IFNG+ T cells. Conversely, 1 of the 4 cervical cancer samples demonstrated relatively lower levels of NK cells which correlated with lower cDC1 cells and in turn lower CTL infiltration.
In conclusion, by utilizing the RNAscope Multiplex ISH assay we identified and visualized the spatial relationship between NK cells, CTLs, and cDC1 cells, highlighting the strength of this technology to spatially interrogate the TME.
Heat shock protein gp96 controls regulatory T cell lineage stability via IL-2/pSTAT5 signaling
Authors: Zhou Lei , Kwon Hyunwoo , Rubinstein Mark P , Li Zihai 
Published: January 2023

Abstract
Gp96 is an essential endoplasmic reticulum chaperone for various proteins including GARP, TLRs, integrins, Wnt co-receptor LRP6 and the platelet glycoprotein Ib/IX/V complex. Our laboratory has previously shown that loss of gp96 in murine Foxp3+ regulatory T cells (Tregs) results in their instability as determined by FOXP3 downregulation, impaired immunosuppressive function and production of effector cytokines (Zhang, et al., J Clin Invest., 2015, 125: 859–869). However, the mechanism underlying Treg dysfunction upon gp96 deletion remains unknown. Using tamoxifen-inducible Treg-specific gp96 gene (encoded by Hsp90b1) knockout (KO) mice (Foxp3-ERT2creHsp90b1f/f), we first phenotypically characterized gp96-deleted Tregs in mouse spleen and peripheral lymph nodes (pLNs). Gp96 deficient Tregs showed decreased frequency of TCF1−CD62L− effector population, but were enriched with short-lived KLRG1+ and Helios+ subsets, in both spleen and pLNs. We also confirmed that the expression of Foxp3 was decreased in splenic gp96-deleted Tregs, which correlated with a reduction of cell surface CD25. Intriguingly, both Foxp3 and surface CD25 levels stayed high in gp96-null Tregs in pLNs. In addition, gp96-null Tregs in mouse spleen, but not in pLNs, demonstrated compromised suppressive function. We further found that IL-2/CD25 signaling, as assessed by STAT5 phosphorylation, was more activated in gp96-null Tregs in pLNs, compared to that in spleen. In vitro IL-2 treatment was able to rescue the attenuated Foxp3 expression in gp96-null Tregs. Our work indicates that IL-2/pSTAT-5 signaling may be a key factor in maintaining Treg Foxp3 expression and its lineage stability upon gp96 deletion.
Protein kinase C-eta is required for Treg-mediated suppression of anti-tumor and viral immunity
Authors: Liu Hsin-Yu , Pedros Christophe , Balancio Ann , Kong Kok-Fai , Altman Amnon 
Published: January 2023

Abstract
We previously reported that protein kinase C-eta plays an important role in the contact-dependent suppressive activity of Tregs via its association with CTLA4, and that PKC-eta-deficient (Prkch−/−) Tregs fail to suppress anti-melanoma tumor immunity. Here we extend this study to a genetically engineered mouse model of HCC driven by CRISPR-Cas9-driven deletion of Pten and p53. In the Pten-p53 HCC model, Prkch Treg-specific conditional knockout (cKO) mice developed a lower tumor incidence, fewer tumors, lower degree of steatosis phenotype, and showed higher intratumoral CD4+ T cells than WT mice. In addition, B cells and resident DCs displayed higher levels of the costimulatory ligand CD86 in dLNs of cKO mice. Increased CD4+ T cells, memory T lymphocytes, and cytokines production were observed in spleen of cKO mice. These results indicate that Treg-expressed PKC-eta is required for Treg-mediated suppression of anti-HCC tumor immunity. To further explore the importance of PKC-eta in Teff cells, the LCMVArm acute infection model, as well as the in vitro activation of murine or human CD8+ T cells were used. We found that purified Prkch−/− mouse CD8+ T cells as well as PRKCH knockdown human CD8+ T cells displayed intact T cell activation in vitro as measured by proliferation and expression of GzmB and IFNg. Interestingly, Treg-specific cKO mice showed improved viral clearance and displayed enhanced expression of GzmB and IFNg by virus-specific CD8+ T cells. Thus, global PKC-eta deletion does not impair overall CD8+ T cell-mediated immunity implying that selective pharmacological PKC-eta inhibition could be safely used in vivo to inhibit undesired contact-dependent suppression by Tregs and, thus, enhance tumor- and viral-specific immunity.
Foxp3+ regulatory T cells suppress T cell immunity in inflamed and non-inflamed tumors by distinct mechanisms
Authors: Delfino Teresita , Wang Yingyun , Muller Soren , Iftikar Hina , Modrusan Zora , Schartner Jill , Rutz Sascha 
Published: January 2023

Abstract
Foxp3+ regulatory T cells (Treg) are essential to prevent autoimmunity, but may also suppress anti-tumor immune responses. The relevance of Treg cell-mediated suppression in inflamed and non-inflamed (immune excluded or immune desert) tumors is not well understood. Here, we used Foxp3-DTR transgenic mice to selectively ablate Treg cells in mice with established tumors. We observed robust and rapid tumor clearance in a number of models, independent of the immune infiltration at baseline. These responses were driven by the induction of infiltration, proliferation, and cytokine production by CD8 T cells in tumor tissues. Importantly, pairing single cell TCR sequencing with transcriptome analysis revealed that CD8 T cells in the tumor acquired a progenitor exhausted phenotype and a clonally diverse TCR repertoire following Treg cell depletion. Pharmacological blockade of lymph node egress demonstrated that local Treg cell depletion and the resulting activation of tumor-infiltrating CD8 T cells was sufficient for tumor rejection in inflamed tumors. In contrast, short-term Treg cell depletion outside of the tumor and the resulting de novo generation of CD8 T cell responses facilitated the rejection of immune desert tumors. These data suggest that Treg cells play a critical role in suppressing both priming of novel anti-tumor CD8 T cell responses as well as limiting effector functionality of infiltrating CD8 T cells within the tumor microenvironment. More importantly, our data suggest that transient systemic Treg cell depletion can convert non-inflamed tumors into inflamed tumors.
Identifying a uniform T cell signature associated with protective anti-tumor immunity and autoimmune systemic sclerosis
Authors: Lust Hannah E , Whitfield Michael , Huang Yina H 
Published: January 2023

Abstract
Precise tug-of-war between immunosuppressive regulatory T (Treg) cells and inflammatory IL-17-producing (Th17) cells ensures appropriate reactivity to pathogenic challenge. While an imbalance towards Tregs leads to suboptimal responses as observed in cancer, unchecked Th17 activity leads to autoimmunity. Though numerous mechanisms have been identified for how Tregs suppress effector cells, productive effector T cell responses are believed to result from passive escape of Treg suppression. Here we report a novel active role for Th17 cells in dampening Tregs to promote effector activity in the context of melanoma. We hypothesize that Th17 cells can physically impede Treg suppression resulting in enrichment of CD8 T cell prevalence and effector function. Using a melanoma mouse model to characterize Th17 effectors responsible for inhibiting Tregs, we found elimination of IL-17-producing cells prevents vaccine-induced anti-melanoma immune responses. This suggests Th17 cells are important for induction of CD8-mediated anti-tumor immunity. Using RNA-Seq analysis, we generated a gene expression (GE) signature to characterize Th17 effectors responsible for inhibiting Tregs in the context of tumor vaccination. Interestingly, increased ratio of Th17 to Treg cells may contribute to pathogenesis of localized and systemic scleroderma. Diminished Treg- and enhanced Th17-associated genes are consistently found in scleroderma patient skin, leading us to hypothesize that scleroderma pathogenesis has a similar Th17 GE signature to that associated with enhanced tumor clearance. Using the tumor-associated Th17 GE signature to examine scleroderma patient samples we determined transcriptional overlap between the Th17 cell subsets.
Regulatory T cell control of systemic tumor-specific immunity and response to checkpoint immunotherapy response in liver metastasis
Authors: Lee James , Mehdizadeh Sadaf , Bridge Jennifer Anne , Young Arabella , Mufazalov Ilgiz A. , Daud Adil , Bluestone Jeffrey A 
Published: January 2023

Abstract
A significant number of metastatic melanoma patients have disease progression despite checkpoint inhibitor therapy. We lack salvage strategies for this population and a mechanistic understanding of checkpoint inhibitor refractory disease. Previously, we reported that patients with liver metastasis have worse response rate and survival than those without liver metastasis despite treatment with checkpoint immunotherapy, but how liver metastasis can influence the antitumor immune response in this context is unclear. We hypothesize that tumor antigen-specific tolerance can be induced in the context of liver metastasis, resulting in therapy resistance.
In this study, we developed a syngeneic two-site tumor model in which tumor cells are injected simultaneously into a subcutaneous (SQ) site and into the liver or other organ sites of B6 mice to study the effects of the intrahepatic tumor on antitumor immunity at a distant site. We found that the presence of tumor or tumor antigen within the liver reduced systemic antigen-specific effector T cell function. Tetramer and sc-RNAseq studies suggest a liver and antigen-specific immunoregulatory process associated with the activation of regulatory T cells and the remodeling of innate immune cells. Importantly, this systemic dysfunctional immune state was not reversed by anti-PD-1 monotherapy but rather by deleting (anti-CTLA-4) or destabilizing (EZH2 inhibitor) intratumoral Tregs, suggesting a distinct regulatory mechanism contributing to anti-PD-1 resistance that could be overcome to achieve complete tumor regression at all sites.
Our results reveal a novel mechanism of checkpoint inhibitor resistance and suggest a strategy to improve the outcome for patients with liver metastasis.
Sex differences in immune responses to influenza vaccine in healthcare workers during the 2019–2020 influenza season
Authors: Park Han-Sol , Kuo Helen , Dhakal Santosh , Ursin Rebecca , Shaw-Saliba Kathryn , Fenstermacher Katherine , Gearhart Patricia J , Rothman Richard , Pekosz Andrew , Morgan Rosemary , Klein Sabra L 
Published: January 2023

Abstract
Seasonal influenza epidemics affect 5–15% of the world population with an estimated 290,000–650,000 annual respiratory deaths due to influenza. The Center for Disease Control recommends annual influenza vaccination for healthcare workers (HCWs) because not only does their occupation increase their risk of contracting influenza, but HCWs care for people at higher risk for developing influenza-related complications. Studies have shown immune responses, such as seroconversion, differ between male and female vaccine recipients, but few have focused on HCWs and their influenza vaccine responses by sex. To understand sex differences in immune responses to the influenza vaccine in the HCW population, 50 male and 50 female were recruited during the 2019–2020 influenza season as part of the Johns Hopkins Center for Excellence in Influenza Research and Surveillance (JH-CEIRS) annual serological surveys among Johns Hopkins Hospital HCWs that takes part in the annual hospital-wide vaccination campaigns. Participants were recruited just prior to receiving their annual quadrivalent influenza vaccine (QIV), which targets influenza strains A/Brisbane/H1N1, A/Kansas/H3N2, B/Colorado (Victoria lineage), and B/Phuket (Yamagata lineage). Whole blood was collected at 0, 7 and 28 days post-vaccination. Neutralizing antibody assays will be used to measure pre and post antibody titers against the H1N1 and H3N2 vaccine components. We will also analyze vaccine-specific B cell responses from peripheral blood mononuclear cells (PBMCs) at 7 and 28 days post-vaccination via flow-cytometry. This study will provide extensive data on the effects of patient sex on influenza vaccine-induced antibody responses and B cell phenotypes in the HCW.
Molecular mechanisms underlying T cell co-potentiation by anti-CD3 Fab fragments
Authors: Huynh Hien T. , Nelson Alfreda D , Hoffmann Michele , Abergel Megan , Hu Shuang , Alarcon Balbino , Schrum Adam G , Gil Diana 
Published: January 2023

Abstract
Cognate antigen binding to the T cell receptor (TCR) induces a conformational change of the CD3 complex (CD3Δc), which is required but not sufficient to trigger productive signaling from the TCR/CD3. CD3Δc fails to occur when the TCR ligates poorly immunogenic antigens. In the absence of CD3Δc, T cells do not display productive immune responses. CD3Δc involves the exposure of a conserved proline-rich-sequence (PRS) in CD3ɛ cytoplasmic tail that functions as a docking site for cytoplasmic adaptor proteins like Nck, which participate in the CD3 signaling cascade. The opening of the PRS is controlled by a CXXC motif in CD3ɛ located adjacent to the plasma membrane. We previously described it is possible to mimic CD3Δc on T cells when adding Fab fragments capable to bind to the CD3 complex. When combining TCR engagement to weak antigens with binding of Fabs to the CD3 complex that not interfere with antigen binding, T cell co-potentiation occurs. In this instance, T cells become capable to display responses to poorly immunogenic stimulation. As an example of the consequences of T cell co-potentiation in vivo, we studied anti-mouse CD3 Fabs as a novel immunotherapeutic against poorly immunogenic types of cancer. Anti-CD3 Fabs reduced melanoma burden in B6 mice bearing B16-F10 tumor in lungs. Now, we have explored the molecular mechanisms supporting T cell co-potentiation in monoclonal and polyclonal mouse models. In B6 mice expressing null mutations for either the CXXC or the PRS motifs, anti-CD3 Fabs fail to induce CD3Δc, as well as to co-potentiate murine T cells in vitro and in vivo. These data suggest PRS opening by anti-CD3 Fabs may be linked to their capacity to co-potentiate T cells responses to poorly immunogenic antigens.
Querying resources in the tumor microenvironment using bystander memory T cells
Authors: Sarkar Surojit , Sullivan Peter , Yuzefpolskiy Yevgeniy , Kalia Vandana 
Published: January 2023

Abstract
The general paradigm is that the tumor microenvironment is non-conducive to sustaining productive cytotoxic T cell responses due to suppressive and counter-immunity T cell extrinsic factors. It has been shown that the tumor microenvironment can harbor large numbers of bystander memory cells. However, it is not very clear if given the right stimulation these bystander memory cells can be commandeered to utilize the available resources of the tumor microenvironment, get optimally activated and elaborate productive immune responses. This has wide-ranging implications in hijacking the tumor-resident bystander memory cells for efficacious tumor control. In this study we show that the tumor microenvironment is capable of harboring ultra-functional bystander memory cells, which display complete panoply of memory markers. These tumor–resident bystander memory cells are not only capable of elaborating robust ex vivo and transplantable memory T cell functions, but they can also efficiently utilize the resources within the tumor microenvironment to elaborate in situ recall effector functions. We further show that these memory T cells utilize CXCR3 as a key chemokine receptor for trafficking to the tumor sites. Our data identify bystander tumor-resident memory cells as Spartan warriors to mediate enhanced tumor control during adoptive T cell immunotherapy.
Characterization of serological response to dengue and Zika viruses in pregnant women during the Zika outbreak in Brazil
Authors: Driesse Kaitlin , Tsai Wen-Yang , Brites Carlos , Pedroso Celia , Wang Wei-Kung 
Published: January 2023

Abstract
The outbreak of Zika virus (ZIKV) and associated congenital Zika syndrome in dengue virus (DENV)-endemic regions raised questions about the effects of ZIKV infection on DENV immunity and disease outcome and vice versa, which highlighted the need of serological tests that can discriminate between DENV and ZIKV infections. Previously we developed combined DENV and ZIKV non-structural protein 1 (NS1) enzyme-linked immunosorbent assays (ELISAs) to overcome the cross-reactivity of traditional envelope protein-based serological tests (Tsai et al. Clin Infect Dis 2017;65:1829). In this study, we used this assay to screen 120 serum samples collected from asymptomatic parturient women during the ZIKV outbreak in Salvador, Brazil and identified four serostatus groups. We further characterized antibody responses in 8 participants including 2 primary DENV (pDENV), 2 secondary DENV (sDENV), 1 primary ZIKV (pZIKV), and 3 ZIKV with previous DENV (ZIKVwprDENV) infections. Using depletion experiments with inactivated ZIKV and urea tests, we determined the IgG avidity to DENV. Using depletion experiments with inactivated DENV and endpoint ELISA titers, we determined ZIKV type-specific and cross-reactive antibody and found pZIKV had higher proportion of ZIKV type-specific antibody (82%) than ZIKVwprDENV (10-16%). These results were supported by microneutralization tests. Our findings suggest that combined DENV and ZIKV NS1 IgG ELISAs plus depletion experiments can delineate past and present DENV and ZIKV infections and are a potential tool to further our understanding of pathogenesis and epidemiology of DENV and ZIKV in endemic regions.
Research supported by R01 AI11076901, R21AI135292-01A1 from NIAID, and P30GM114737 from NIGMS, NIH.