bluuebunny/crossref_metadata_2025_split huggingface dataset:
Fomitopsis cajanderi extracts have differential tumoricidal actions on human MCF7 and U-937 cancer cells and inhibit TNF secretion by U-937 monocytes
Authors: Wenner Cynthia A , Froehlich Olivia R , Sitkoff Anna 
Published: February 2023

Abstract
This study aimed to determine if hot ethanol and aqueous extracts from the Fomitopsis cajanderi mushroom species are cytotoxic to human cancer cell lines and modulate TNF secretion by human monocytes. The hot ethanol extract was made via a 2 hour Soxhlet extraction of F. cajanderi fruiting body in ethanol using a cold finger condenser. The hot aqueous extract was made via a 2 hour decoction in water at 100°C. Human MCF7 mammary adenocarcinoma and U-937 histiocytic lymphoma cells were treated with increasing concentrations of extract or solvent controls and were assessed for cytotoxicity by XTT colorimetric assay. Effects of the aqueous extract on TNF secretion by PMA-differentiated, LPS-stimulated U-937 cells were determined by TNF ELISA. Glycosyl content and linkage analysis of the aqueous extract were assessed by the Complex Carbohydrate Research Center (Athens, Georgia, USA). The hot ethanol extract induced concentration-dependent cytotoxicity of MCF7 and U-937 cells, with stronger U-937 tumoricidal effects. The hot aqueous extract was not cytotoxic to MCF7 cells, but had cytotoxic effects on undifferentiated U-937 cells at high concentrations. The aqueous extract, found to contain beta glucans, inhibited TNF secretion by LPS-stimulated U-937 cells in a concentration-dependent manner. Hot ethanol and aqueous F. cajanderi extracts showed differential tumoricidal actions on human cancer cells. The ethanol extract was cytotoxic to the human cancer cells tested. The aqueous extract contained beta glucans reported to be immunomodulatory, and inhibited U-937 TNF secretion. These results warrant further research of F. cajanderi extracts as novel adjunctive anti-cancer and anti-inflammatory agents.
Clearance of established oropharyngeal and genital HPV tumors by mucosal vaccination via induction of multiple cytotoxic effector populations
Authors: Hegde Venkatesh L , Sierra Gloria , Nookala Sita , Yanamandra Ananta , O’Hara Madison , Sastry Jagannadha K 
Published: February 2023

Abstract
High-risk type human papillomaviruses (HPV) are the underlying cause of &gt;5% of all cancers including cervical and Head & Neck cancers in which virus-encoded E6/E7 oncoproteins serve as tumor-specific antigens. Despite the success of prophylactic HPV vaccine, therapeutic strategies targeting the E6/E7 have been ineffective in established HPV+ tumors due mainly to inadequate antigen-specific immunity. Therefore, enhancing the therapeutic potential HPV vaccines to achieve curative efficacy is an unmet clinical need. We present here preclinical evidence for efficient clearance of immunocompetent HPV tumors at the vaginal (TC-1) and oral (mEER) mucosal tissues by intranasal vaccine using HPV16 E6 and E7 short peptides along with the combination of clinically relevant adjuvants QS-21 and CpG-ODN. This vaccine exhibited high therapeutic efficacy with &gt;80% mice with oral or vaginal tumors showing regression as well as significant extended survival advantage over untreated mice or mice treated with vaccine containing single adjuvants. The analysis of immune correlates by multicolor flow cytometry showed robust increases in the overall as well as antigen-specific (E7 tetramer+) cytotoxic CD8 T cells expressing Granzyme B and/or IFNg in both intra-tumoral and systemic compartments in vaccine-treated mice. Furthermore, the intranasal vaccine containing the combination of adjuvants induced significant frequencies of a unique subpopulation of NK cells expressing CD11c, granzyme B and IFNg, termed NKDC. Our results suggest that mucosal HPV E6/E7 peptide vaccine formulation containing QS-21+CpG-ODN adjuvants may provide an effective therapeutic strategy for the treatment of patients with HPV+ oropharyngeal and cervical cancers.
The comparison of two SARS-CoV-2 spike protein antigens in TLR-adjuvanted subunit vaccines.
Authors: Partlow Haley A , Davison Clara J. , Lathrop Stephanie K , Evans Jay T. 
Published: February 2023

Abstract
Adjuvanted subunit vaccines offer improved safety and efficacy over whole inactivated or live attenuated vaccines by incorporating recombinant proteins with highly selective innate immune activators. Selection of an antigen in subunit vaccines can alter immune responses, making it an important consideration. For severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the spike (S)-protein aids in receptor recognition and membrane fusion. Previous studies of SARS-CoV-2 and the closely related SARS-CoV-1 define the receptor binding domain (RBD) portion of the S-protein as the target of most neutralizing antibodies (nAbs). Here the recombinant full spike trimer (FST) and the isolated recombinant RBD are compared as antigens in an adjuvanted subunit vaccine. Induction of Th1-biased CD4+ and CD8+ cellular responses are believed to be important for durable antiviral immunity. Therefore, we used synthetic agonists of the toll-like receptor (TLR)4 or TLR7/8 as vaccine adjuvants for their ability to promote Th1 cellular immunity. We found that a TLR4 agonist was effective at adjuvanting both RBD and FST, while a TLR7/8 ligand provided superior adaptive immune responses with FST. In addition, increased induction of nAbs were demonstrated when vaccinating mice with FST. Thus, we find that early selection and optimization of adjuvant and antigen combinations is critical for the induction of humoral (including nABs) and cell-mediated immunity to SARS-CoV-2 antigens.
Combinations of alum and synthetic toll-like receptor agonists as adjuvants for CoVID-19 vaccines
Authors: Davison Clara J , Partlow Haley A , Lathrop Stephanie K , Siram Karthik , Abdelwahab Walid , Burkhart David J , Evans Jay 
Published: February 2023

Abstract
Aluminum-based adjuvants (“alum”) are the most widely-used adjuvants for human vaccines, with an extensive history of safety and efficacy. Alum is known to elicit high antibody titers by driving a Th2-based immune response, which may not be optimal for many viral infections, such as SARS-CoV-2. Therefore, combining the strong safety and efficacy attributes of alum with a Th1-polarizing adjuvant could improve immunity against viral antigens. Here we test the combination of alum with synthetic TLR4- and TLR7/8 -ligands in vaccines against SARS-CoV-2. The combination of alum and TLR agonists resulted in efficient adjuvantation of both humoral and cell-mediated immunity. Alum adsorption studies showed low association between the small, positively-charged receptor binding domain (RBD) antigen and aluminum hydroxide (Alhydrogel) and high adsorption to negatively charged aluminum phosphate (Adju-phos). However, Adju-phos and Alhydrogel both enhanced immunity to the SARS-CoV-2 RBD antigen, suggesting antigen adsorption may not be required for the immune enhancing effects of alum. These data demonstrated that adjuvants combining alum with a TLR agonist result in an improved anti-viral immune response. However, the ability of alum to adsorb to an antigen may not predict its ability to effectively adjuvant.
STAT4 influences the expression of IL-10 in Th17 cells
Authors: Anderson Ashlyn E , Shin Boyoung , Bryars Morgan , Maynard Craig , Harrington Laurie 
Published: February 2023

Abstract
Autoimmune diseases such as Multiple Sclerosis (MS), Rheumatoid Arthritis (RA) and Crohn’s Disease (CD) affect more than twenty million people in the United States. While the exact etiology of most autoimmune diseases is unknown, single nucleotide polymorphisms in the STAT4 locus are associated with an elevated risk of developing various autoimmune diseases. Specifically, in a mouse model of MS, experimental autoimmune encephalomyelitis (EAE), STAT4 is essential for the development of neuroinflammation. Interestingly, STAT4 is a transcription factor known to be important for Th1 cell differentiation, however we and others have demonstrated that many Th1 associated molecules such as IL-12, IFNγ and Tbet are dispensable for EAE. These data lead us to postulate that STAT4 functions in another effector CD4 T cell, Th17 cells, to drive neuroinflammation. Using an adoptive transfer model, we find that STAT4 is required in a Th17-mediated EAE. Transcriptional profiling of the adoptively transferred Th17 cells reveals that STAT4 influences the expression of over 200 genes, one of them being the immunosuppressive cytokine IL-10. Strikingly, we find an indirect relationship between STAT4 and IL-10 in Th17 cells as well as T regulatory 1 (Tr1) cells. We hypothesize that STAT4 regulates the balance between Th17 and Tr1 cells in the context of autoimmune inflammation. Together, these data illustrate a novel role for STAT4 in controlling the functional and plastic properties of Th17 cells, and this may provide a promising therapeutic target for patients affected by Th17 driven autoimmune diseases.
Single cell profiling identifies IgMMDA-LDL-producing human B cells and a novel role for CD24
Authors: Pattarabanjird Tanyaporn , Nguyen Anh Tram , McSkimming Chantel , Dinh Huy , Marshall Melissa A , Ghosheh Yanal , Gulati Rishab , Durant Chistopher , Vallejo Jenifer , Saigusa Ryosuke , Drago Fabrizio , Taylor Angela M , Tsimikas Sotirios , Miller Yury , Ley Klaus , Hedrick Catherine C. , McNamara Coleen A 
Published: February 2023

Abstract

Objectives:
IgMs that inactivate oxidation specific epitopes (IgMOSE) on phospholipids such as low density lipoprotein (LDL) were shown to confer athero-protection. We developed a 24 antibodies mass cytometry panel (CyTOF) to identify the human B cell subtype producing IgM to malondialdehyde modified LDL (IgMMDA-LDL), a predominant IgMOSE in humans. We also utilized humanized mice model to characterize a role of CD24 mediated IgM production.

Methods:
Peripheral blood mononuclear cells from healthy donors and subjects with coronary artery disease (CAD) undergoing quantitative coronary angiography were used to perform high dimensional analysis using CyTOF, RNAseq and Abseq as well as adoptive transfer into humanized mice model.

Results:
CD20+CD27+IgM+ cells (B27+IgM+) spontaneously produced IgM and produced IgMMDA-LDL in response to MDA stimulation when injected into humanized mice; an effect significantly augmented in B27+IgM+ cells with high expression of CD24 (B27+IgM+CD24hi). CD24 expression also enhanced splenic and bone marrow trafficking of B27+IgM+ cells. Blocking CD24 with a mAb reduced CCR6 expression, increased CCL20-induced CCR6 internalization and impaired migration to the spleen leading to lower IgM production. Lastly, single cell protein and transcriptome sequencing of PBMCs from 60 CAD subjects revealed enhanced CCR6 and IgM signaling in B27+IgM+CD24hi cells in subjects with low compared to high CAD severity.

Conclusions:
Identification of IgMMDA-LDL-producing cells in humans has the potential to allow for the development of therapeutics aimed at cellular targeting that may allow for enhancing production of IgM to the many OSE produced during inflammatory states such as atherosclerosis.
The role of DCAF1-GSTP1-ROS axis in regulating T cell senescence in immunological ageing and tumors
Authors: Guo Zengli , WAN Yisong 
Published: February 2023

Abstract
Immunological aging is characterized by chronic nonspecific inflammation, compromised vaccination and declined immune competence, contributing many age-associated diseases. As the central regulator of immune system, whether and how regulatory T (Treg) cells are regulated during ageing are not fully understood due to a lack of specific markers. By a combination of cellular, molecular and bioinformatic approaches, we discovered that Treg cells senesce more severely than conventional T (Tconv) cells during ageing. Treg cells from aged mice were less efficient than young Treg cells to suppress Tconv cell function in an inflammatory-bowel-disease model and to prevent Tconv cell ageing in the irradiation-induced ageing model. Further study revealed that DCAF1 (DDB1 and CUL4 associated factor 1) was downregulated in aged Treg cells and was critical to restrain Treg cell ageing via glutathione S-transferase P (GSTP1) regulated reactive-oxygen-species (ROS). Functionally, interfering with GSTP1 and ROS pathways reinvigorated the proliferation and function of aged Treg cells. Thus, the DCAF1-GSTP1 regulated ROS contributes to Treg cell senescence, which led to systemic immunological ageing.
In tumor microenvironment, ROS produced by Treg cells and myeloid-derived suppressive cells (MDSCs) is one of the major inhibitory molecules suppressing anti-tumor immunity. We found that there was significant increase of senescent T cells in tumors in a B16 melanoma model, featured with increased p16 expression, elevated senescence associated β-gal activity and increased KLRG1 expression. Further investigating the role of DCAF1-GSTP1-ROS axis in tumor infiltrating T cells will provide novel mechanistic insights to cancer immunotherapy.
Corepressors in CD4+ T cell development and homeostasis
Authors: David Natalie A , Lee Robin D , Owen David L. , Farrar Michael A. 
Published: February 2023

Abstract
Corepressors NCOR1 and NCOR2/SMRT serve as scaffolds for the recruitment of histone deacetylases to transcription factor-DNA complexes. These closely related proteins are functionally non-redundant. Though they were first identified by their interactions with nuclear receptors, NCOR1/2 also interact with non-nuclear receptor transcription factors, including several involved in T cell development and polarization. Previous research identified that NCOR1 promotes survival of double positive thymocytes and protects them from negative selection. We have built upon this work by examining the effect of combined NCOR1/2 deletion in developing thymocytes. NCOR1/2 deletion using a CD4-Cre model results in an 8-fold decrease in the frequency of CD4 single positive T cells, compared to a 2-fold decrease and an insignificant decrease for NCOR1 and NCOR2 deletion alone, respectively. NCOR1/2-deficient T cells demonstrate exquisite sensitivity to TCR stimulation and polarize to Th subsets at different frequencies compared to wild-type CD4+ T cells. Despite a profound loss of CD4 single positive T cells and thymic regulatory T cells (Tregs), the frequency of both Treg progenitor populations remains unchanged. In contrast, the frequency of splenic Tregs is not reduced in NCOR1/2-deficient mice. However, there is a striking increase in activation of conventional T cells, suggesting that T cell tolerance is perturbed. Using an OX40-Cre deletion model which deletes NCOR1/2 in dendritic cells (DCs), activated T cells, and Tregs, we observe a decreased CD4+/CD8+ ratio and high incidence of ulcerative dermatitis. We are currently examining how Treg or DC specific deletion of NCOR1/2 affects T cell development, homeostasis, and tolerance.
Building a Mini Biomedical Ethics Curricular Thread in an Immunology Course in Undergraduate Medical Curriculum
Authors: Zhao Yuan , Olaiya Oluwaseun 
Published: February 2023

Abstract
Education in biomedical ethics has become an essential part of undergraduate medical curriculum. Integrating this topic longitudinally in the curriculum ensures early educational exposure and provides opportunities for continued advancement of students reasoning and behavior. The objective of this study is to develop a mini ethics curricular thread in the immunology course and to raise student awareness of ethical issues associated with disorders of the immune system. In our project, three ethics learning modules are designed and built in a six-week immune system course in the second semester of undergraduate medical curriculum. A mixed teaching approach using multiple teaching tools will be employed. The first module uses a movie about a well-known patient with severe combined immunodeficiency disease. The second module is delivered through a team-based learning case on human immunodeficiency virus. The third module uses small and large group discussions on blood transfusion and bone marrow transplantation. Quizzes and reflective writings are embedded in each module to assess student knowledge and attitudes. Student surveys will provide information on challenges and opportunities in teaching ethics within a biomedical science course. The qualitative analysis of student reflections will be presented in the poster. We believe this curricular design has the potential to help students build connections between science, human disease, and ethics. It will also shed light on how to integrate ethics in biomedical science courses to cultivate professionalism in the early stages of their medical training. Future direction includes the evaluation of the longitudinal impact of this training on students clinical practice.
Investigating the age and sex associated mucosal immune landscape of urinary bladder
Authors: Hamade Ali , Li Deyang , Tyryshkin Kathrin , Shakfa Noor E. , Chenard Stephen , Siemens Robert , Koti Madhuri 
Published: February 2023

Abstract
The incidence of BC is four times higher in males than females, however, females tend to present with a more aggressive disease, a poorer response to immunotherapy, and suffer worse clinical outcomes. Our recent novel findings demonstrated sexual dimorphism in the tumor immune microenvironment of non-muscle invasive bladder cancer, which is primarily treated with local immunotherapy. Investigating sexual dimorphism in the clinical journey of BC beyond the static local immune landscapes remains a challenge. With similarities in mucosal immune physiology between humans and mice, and the recently reported age-associated pathogenic stimuli independent inflammation observed female murine bladders, we conducted a comprehensive evaluation of the sex and age-related immune alterations in healthy murine bladders.
Bulk-RNA sequencing and multiplex immunofluorescence based spatial immune profiling of normal murine bladders from male and female mice of the ages 3, 6, 9, 12, 15, and 18 months were performed. Preliminary findings from the transcriptomic analysis showed a highly altered immune landscape that exhibited sex differences in aged bladders. Spatial profiling of using markers specific to macrophages (CD163, CD11b), T lymphocytes (CD3, CD8), B lymphocytes (Pax5), activated dendritic cells (CD208), high endothelial venules (PNAd), inflammatory cells (Ly6G) and the PD-L1 immune checkpoint, showed sex and age associated differences. Older female mice had a higher density of tertiary lymphoid structures/lymphoid aggregates compared to both young female and male equivalents. Findings from this study will allow the appropriate modeling in pre-clinical studies evaluating immunotherapeutic agents for NMIBC treatment.
CD8 T cell differentiation in cancer is comprised of two distinct phases
Authors: Prokhnevska Nataliya , Cardenas Maria , Valanparambil Rajesh , Jansen Caroline , Master Viraj , Sanda Martin , Kissick Haydn 
Published: February 2023

Abstract
A crucial part of the immune response to tumors are CD8 T cells, with CD8 infiltration predicting disease progression in many cancers. Recent work has shown two subsets of CD8 T cells that respond to tumors, one a stem-like CD8 T cell (TCF1+) that can give rise to a more cytotoxic terminally differentiated cell (TCF1−). To understand the CD8 T cell response to tumors it is important to study how tumor-specific CD8 T cells activate and differentiate. To study this we have made a prostate cancer model which expresses the LCMV glycoprotein (GP) and acts as a tumor-specific antigen. This model allows us to transfer LCMV GP specific P14 CD8 T cells into TRAMPC1-GP bearing mice to study how tumor-specific CD8 T cells activate. These studies have shown that tumor-specific CD8 T cells are activated in tumor draining lymph nodes (TDLN), where they retain an activated undifferentiated phenotype, upregulating CD44, PD1, while maintaining TCF1. These tumor-specific CD8 T cells only differentiate (TCF1−) once they have migrated into the tumor. This model can also be seen in human prostate cancer, with CD8 T cells in TDLNs retaining an activated undifferentiated phenotype (PD1+CD45RA-TCF1+). To determine if these cells are related to the CD8s within the tumor we have shown TCR overlap between the activated CD8s T cells in human prostate TDLNs and the CD8 T cell subsets within the tumor. These data suggest a two-step differentiation process for tumor-specific CD8 T cells where they are activated in TDLNs and differentiate further only in the tumor. This model of two distinct phases of CD8 T cell differentiation adds to the basic understanding of the immune response to cancer.
Gene expression analysis of sarcoma-infiltrating NK cells identifies signatures correlating with favorable prognosis and potential new targets for augmenting NK function
Authors: Judge Sean J , Dunai Cordelia , Razmara Aryana , Cruz Sylvia M , Canter Robert J 
Published: February 2023

Abstract
Success of current T-cell based immunotherapies in soft tissue sarcomas (STS) is limited while pre-clinical and clinical studies have shown evidence of NK activity in STS. To investigate the effects of the tumor microenvironment (TME) on tumor-infiltrating NK cell gene expression, we isolated circulating and tumor-infiltrating NK and CD3 T cells from sarcoma patients undergoing surgery (N=4) and performed RNA sequencing analysis. Comparing Differential Gene Expression (DGE) of flow-sorted intra-tumoral NK cells to circulating NK cells, we identified upregulation of genes involved in mitogen signaling inhibition (DUSP4) and metabolic function (SMPD3, SLC7A5), but not of genes typically associated with cytotoxic function (e.g. IFNG, GZMB). In contrast, intra-tumoral T cells showed significant upregulation of both activating (CD137) and inhibitory genes (TIM-3) compared to circulating T cells. We then queried the TCGA STS dataset and stratified cases into “NK Activating TME” and “NK Suppressing TME” based on the CIBERSORT NK signature analysis program (based on high or low proportion of activated NK cells, respectively). Patients with NK Activating TME had significantly improved survival compared to NK Suppressive TME (log-rank p = 0.03). DGE identified unique tumor-associated gene ontology between NK activated versus suppressive subsets with upregulation of the retinoic acid pathway. Unlike T cells which demonstrate significant DGE between circulating and tumor-infiltrating subsets, NK cells appear to have similar gene expression between blood and tumor. Therefore, although NK cells appear to be prognostic in STS, their anti-tumor effects may be dictated by systemic rather than local factors.