bluuebunny/crossref_metadata_2025_split huggingface dataset:
Phosphatidylserine targeting antibody enhances anti-tumor activity of adoptive cell therapies in a mouse melanoma model
Authors: Schad Sara , Hirschhorn-Cymerman Daniel , Budhu Sadna , Zhong Hong , Yang Xia , Merghoub Taha , Wolchok Jedd D 
Published: January 2023

Abstract
Adoptive cell therapy has emerged as a viable strategy to treat cancer. T cells that recognize tumor antigens can be reinvigorated ex-vivo or autologous T cells can be genetically modified to express anti-tumor T cell receptors (TCRs) or chimeric antigen receptors (CARs). However, once re-infused into patients, these tumor specific T cells are subjected to immunosuppressive signals within the tumor. A critical immune checkpoint within tumors is phosphatidylserine (PS), a phospholipid that is exposed on apoptotic cells and tumor cells. Innate cells exposed to PS secrete suppressive cytokines that can significantly impair the function of tumor specific T cells. Antibodies that target PS can reactivate anti-tumor immunity by reducing the number of MDSCs in tumors and promoting the maturation of functional APCs. Our lab has shown that the mouse chimeric version of PS Targeting monoclonal antibody Bavituximab (1N11), in combination with transgenic CD4+ T cells that recognize melanoma antigen Trp1, can regress advanced melanoma tumors in mice. Here, we demonstrate a 2nd generation CAR T cell, that binds Trp1 on the surface of B16 melanoma, in combination with 1N11 can improve anti-tumor activity and survival in B16 tumor bearing mice. Flow cytometry analysis of immune responses in the tumor of mice treated with tumor specific T cells and 1N11 shows a decrease in M2 macrophages and FoxP3+ regulatory T cells. These findings highlight that diminishing suppressive mechanisms locally with PS targeting can enhance the efficacy of transgenic TCR and CAR T cells to improve the outcome in patients with advanced-stage melanoma. Our studies may inform the design of clinical trials combining PS Targeting antibodies with CAR T cell therapy in solid tumors.
T-cells modified with CD70 as an alternative cellular vaccine for antitumor immunity
Authors: Cho Hyun-Il , Lee Sang-Eun , Shin A-Ri , Sohn Hyun-Jung , Kim Tai-Gyu 
Published: January 2023

Abstract
Successful tumor eradication primarily depends on generation and maintenance of a large population of tumor-reactive CD8 T-cells. Dendritic cells (DCs) have applied to clinics as potent antitumor therapeutic agents. However, high cost and difficulty in obtaining sufficient amounts for clinical use are the crucial drawbacks of DC-based vaccines. Here, antigenic peptide-loaded T-cells transfected with retrovirus encoding costimulatory ligands CD70, CD80, OX40L, or 4-1BBL were assessed for antigen-specific CD8 T-cell responses, and evaluated antitumor effects along with immunization of a mixture of synthetic peptides, poly-IC and anti-CD40 antibodies (TriVax). T-cells expressing CD70 (CD70-T) exhibited similar level of stimulatory functionality and therapeutic efficacy as DCs. Moreover, CD70-T prime followed by TriVax booster heterologous vaccination elicited therapeutic antitumor effect against B16 melanoma where mediated by CD8 T-cells but not CD4 T-cells or NK cells. The combination with PD-L1 blockade led to potent therapeutic efficacy which exhibited increased tumor infiltrating CD8 T-cells. CD70-T pulsed with multi-antigenic peptide generated multiple antigen-specific polyvalent CD8 T-cells that were capable of inhibiting tumor growth effectively. Moreover, CD70-T vaccination resulted in higher expansion and migration of adoptively transferred T-cells into tumor sites, and elicits enhanced therapeutic effects with peptide-based booster immunization. These results imply that T-cells endowed with CD70 enable the design of effective vaccination strategies against solid cancer, which may overcome current limitations of DC-based vaccines.
Lysosomal Protease Creation of Chimeric Epitopes for Diabetogenic CD4 T cells via Transpeptidation
Authors: Reed Brendan Kearney , Crawford Frances , Hill Ryan , Jin Niyun , Krovi Sai Harsha , White Janice , Marrack Philippa , Hansen Kirk , Kappler John 
Published: January 2023

Abstract
The identification of the peptide epitopes presented by major histocompatibility complex class II molecules (MHCII) that drive the CD4 T cell component of autoimmune diseases has presented a formidable challenge over several decades. In type-1 diabetes (T1D) recent insight into this problem has come from the realization that several of the important epitopes are not directly processed from a protein source, but rather pieced together by fusion of different peptide fragments to create new chimeric epitopes. We have proposed that this fusion is performed by a reverse proteolysis reaction called transpeptidation, occurring during the catabolic turnover of pancreatic proteins when secretory granules fuse with lysosomes. Here we demonstrate that highly antigenic chimeric epitopes for diabetogenic CD4 T cells are produced by digestion of the appropriate fragments of the diabetogenic granule proteins with the lysosomal protease, cathepsin L (CatL). This pathway has implications for how self-tolerance can be broken in T1D and other autoimmune diseases.
Lipid dysregulation and antigen presentation during intestinal inflammation
Authors: Russo Matthew V , Tordoff Jesse , Chiang Shian-Huey , Berasi Stephen , Buhlmann Janet 
Published: January 2023

Abstract
Inflammatory bowel diseases are associated with a loss of epithelial barrier integrity that can lead to an imbalance in gut homeostasis and severe inflammation. Studies in mice have shown that CD1d, a lipid antigen-presenting molecule, and natural killer T (NKT) cells play important roles in forming and maintaining homeostasis in the intestines. However, it is currently unknown what specific lipids are presented to NKT cells during homeostasis or inflammation in the gut. We hypothesize that during inflammation, CD1d ligands are replaced with pathogenic lipids that alter the activation of NKT cells and contribute to inflammation. Mice were administered dextran sodium sulfate (DSS) in their drinking water for 7 days to induce colitis symptoms, then switched to normal water for an additional 7 days to recover. Lipid species were measured in colons from day 7 inflamed mice and day 14 recovered mice. Compared to control mice, the lipid composition of inflamed colon was significantly altered. Day 7 enriched ceramide and hexosylceramide species remained elevated at the recovery time-point. These lipids stimulated inflammatory cytokine production in lamina propria cultures in a CD1d-dependent manner. To determine if these lipids were being generated endogenously, we measured the gene expression of several key enzymes in the hexosylceramide synthesis pathway. Expression of Degs1 and Ugcg were up-regulated in lamina propria cells during colitis, suggesting that immune cells could be responsible for the altered lipid composition we observed in DSS treated mice. Overall, we have demonstrated that colitis-induced lipid alterations in lamina propria cells of the gut can contribute to intestinal inflammation through CD1d antigen presentation.
The Outer Face of the IgV Domain of Butyrophilin 3A1 is Critical for the Sensing of Prenyl Pyrophosphates by Human Vγ2Vδ2 T cells: A New Paradigm for γδ T Cell Recognition
Authors: Morita Craig T , Rhodes David A. , Al-Janabi Mohanad Hameed Nada , Tanaka Yoshimasa , Sakuraba Shun , Wang Hong 
Published: January 2023

Abstract
γδ T cells expressing Vγ2Vδ2 TCR play critical roles in human immunity to microbes and tumors. These cells are uniquely able to use their TCRs for pattern recognition to monitor phosphorylated metabolites in isoprenoid biosynthesis in a process requiring butyrophilin (BTN) 3A1. Metabolites are sensed by binding to the intracellular B30.2 domain and requires the juxtamembrane coiled coil region. The role played by the extracellular domains (ECD) is unclear. To assess the ECD, surface IgV and IgC residues were mutagenized to alanine. Mutant BTN3A1 cDNAs were expressed in 293 T embryonic kidney cells lacking BTN3A1, -A2, and -A3, and the transfected cells pulsed with zoledronic acid and used to stimulate Vγ2Vδ2 T cells. Mutation of 13 residues scattered on the IgC domain had no effect on stimulation. Mutation of 22 residues scattered on the inner, side, and outer faces of the IgV domain also had no effect. However, 4 residues localized to a region on the outer face of the IgV domain either abrogated or diminished stimulation. Others have identified 2 additional residues in this region and mapped recognition to the Vγ2 HV4 loop paralleling BTNL3 and Btnl6 recognition by other γδ TCR. BTN3A2 and BTN3A3 can complement mutant BTN3A1 proteins suggesting that they form heterodimers or can alter their conformation. Modeling of the potential binding interface identifies BTN3 residues that can form ionic or hydrogen bonds to HV4. Given that our previous studies have shown the requirement for all CDRs of Vγ2Vδ2 TCR, a putative second ligand for the conventional binding side is also required. Taken together, we propose that Vγ2Vδ2 T cell stimulation is dependent on the simultaneous binding of BTN3A1 to HV4 and a second ligand to the conventional Vγ2Vδ2 binding site.
TLR7/8 agonist accelerates lupus development with heart disease in lupus-prone mice
Authors: Elshikha Ahmed Samir , Teng Xiangyu , Li Wei , Brown Josephine Michelle , Abboud Georges , Choi Seung-Chul , Kanda Nathalie , Zou Xueyang , Morel Laurence 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is a chronic autoimmune disease characterized by loss of self-tolerance to nuclear antigens leading to the production of pathogenic autoantibodies, immune complex deposition and multi-organ damage. Although the prevalence of heart disease is higher than 50% in SLE patients, its etiology remains unclear but both genetic and environment factors are likely involved. Here, we compared the effect of TLR7/8 agonist Resiquimod (R848) on lupus and cardiac lesion development in lupus-prone B6.Sle1.Sle2.Sle3 (TC) and congenic normal mice B6 mice. Two weeks of R848 treatment increased the production of antinuclear antibodies and induced antiphospholipid autoantibodies lined to vascular pathology in TC mice. Correspondently, R848 expanded the frequency of Tfh cells and plasma cells in TC mice as compared to R848 treated B6 mice. More importantly, unlike the near normal appearance in hearts of R848-treated B6 mice, R848 induced severe cardiac pathology in TC mice with infiltration of CD45+ cells, edema, hemorrhage, enlarged endothelial cells and myocyte damage. Our results indicate that heart disease develops by combining genetic predisposition to lupus together with TLR7/8 activation. This novel model may lead to the identification of autoimmune mechanisms responsible for lupus-associated heart disease.
Novel B cell-dependent multiple sclerosis model using extracellular domains of myelin proteolipid protein
Authors: Boyden Alexander W. , Brate Ashley A. , Karandikar Nitin J. 
Published: January 2023

Abstract
Therapeutic success of B cell-targeting approaches in multiple sclerosis (MS) has intensified research into the role B cells play in pathogenesis and regulation of disease. Historical dissection of B cell function in the MS mouse model experimental autoimmune encephalomyelitis (EAE) has largely been confined to disease induced with either the immunodominant epitope of myelin oligodendrocyte glycoprotein (MOG35–55) or full length recombinant human MOG protein (hrMOG), the latter representing the only available B cell-dependent EAE model. We have demonstrated that myelin proteolipid protein (PLP178–191)-specific CD8 T cells possess striking EAE-suppression capacity. However, little is known regarding the role of B cells in these contexts. Interestingly, unlike MOG35–55, where genetic absence or depletion of B cells yields a more severe disease course, PLP178–191 yields identical EAE progression in both WT and μMT mice, suggesting suboptimal B cell engagement. Given the reports that hrMOG drives a B cell-dependent demyelinating disease versus its MOG35–55 counterpart, we hypothesized that a longer PLP antigen consisting of more epitopes may better engage B cells and provide an additional model for the field. To this end, we designed a novel peptide encompassing the extracellular domains (ECD) of PLP and compared its immunogenicity and disease-inducing capacity with PLP178–191 in μMT and WT mice. We demonstrate here that PLPECD-immunized B cell-deficient mice failed to exhibit EAE disease compared to WT counterparts. PLPECD also induced EAE in B cell-sufficient mice incapable of secreting antibodies, suggesting a predominant antigen presentation role. These results establish a novel, efficient B cell-dependent EAE model.
Novel shared antibody specificities in anti-Ro/ La antibody negative Sjögren’s Syndrome
Authors: Longobardi Sherri , Georgescu Constantin , Lawrence Christina , Moya Charmaine , Wren Jonathan , James Judith A , Sivils Kathy L , Farris A. Darise 
Published: January 2023

Abstract

INTRODUCTION
Sjögren’s syndrome (SS) is a rheumatic autoimmune disease characterized by focal lymphocytic infiltrates in the lacrimal and salivary glands, severe dry mouth and eyes, pain and debilitation. Diagnosis requires autoantibodies to ubiquitous Ro antigens or a lip biopsy positive for focal lymphocytic infiltrates. Here we used human proteome arrays to identify novel antibodies in plasma from Ro positive and Ro/La antibody negative SS patients compared with healthy controls.

METHODS
Anti-Ro positive (n=15) and anti-Ro negative (n=15) cases meeting 2016 ACR/EULAR classification criteria for SS were age, race, and sex matched with each other and healthy controls (n=15). Plasma IgG binding to human proteome arrays containing &gt;19,500 recombinant human proteins representing &gt;80% of the human proteome (HuProt v3.2 arrays, CDI Laboratories) was assessed. Data were normalized by the Robust Linear Model using the PAA Bioconductor Package in R and log intensity values for each protein generated. Thresholds of mean + 4SD were established using the controls. Antigens bound by IgG more frequently in cases compared to controls (p&lt;0.05, one-tailed Fisher’s exact test) were considered significant.

RESULTS
IgG from Ro positive SS cases significantly bound 18 proteins, including the canonical SS antigens Ro60 and Ro52. IgG from Ro negative SS cases significantly bound 4 proteins compared to controls, 3 of which were shared with the Ro positive group. Binding to any one of 4 novel proteins identified 73% of the Ro negative SS cases.

CONCLUSION
A total of 17 novel antigen specificities were identified in SS, with 4 antigens being bound by plasma IgG from Ro/La negative SS cases. These antigens may be useful for diagnosing SS without a lip biopsy.
Expression of ANA patterns by Indirect Fluorescent Assay in a local Hong Kong population from private sector
Authors: Lau Kwun Hung , Hung Marco Hing-Lam , Kwan Po-Yi , Lam Tsoi-Hin , Kai Cheuk-Him , Lai Ho-Hin , Yau Peter Kin-Nam 
Published: January 2023

Abstract

Objective
Our aim was to investigate the association among age, sex and Antinuclear Antibody (ANA) patterns expressed in Hong Kong population for improving the autoimmune diagnosis by using ANA HEp-2 slides under fluorescent microscope.

Methods
A total of 65 blood samples from both male and female were collected and examined under fluorescent microscope. Haemolysed samples were excluded. In this research, a HEp-2 complete Kits with the methodology of indirect fluorescent antibody (IFA) was utilized for the detection and semi - quantification of antinuclear antibodies to distinguish positive ANA patterns classified in International Consensus on ANA Patterns (ICAP) for human body.

Results
In this experiment, we obtained that the number of positive samples was 6 (9%), the number of suspected (weak) positive samples was 4 (6%) and the number of negative samples was 55 (85%). In 6 positive samples, they were all females aged between 59 – 89. 5 in 6 positive samples in the result were over age 69 (83%). The positive ANA patterns included AC2 – Speckled, AC3 – Centromere, AC6 – Discrete Nuclear Dots, AC7 – Discrete Nuclear Dots, and AC8 – Nucleolar patterns separately. No AC-29 - DNA topoisomerase I was examined and resulted. We found that Hong Kong have a higher distribution of ANA positive frequency for female (9.23%) than male.

Conclusion
Our data indicated that ANA patterns were found in female aged above 69 is more likely to suffer from autoimmune disease related to ANA, providing an important foundation for considering and studying the trend in the elderly who are easier to have an ANA related autoimmune disease in Hong Kong.
The Use of Interactive Educational Tools to Engage Students in Microbiology and Immunology Classroom
Authors: Simmons Jazmyne Cymone Durrah , Fadhil Shams , Edquiban Marianne , Olk Sandra , Fazal Nadeem 
Published: January 2023

Abstract
Microbiology and Immunology is a required 4 credit hour course for pharmacy students to progress in Chicago State University College of Pharmacy program. Twenty lectures are offered to first-year pharmacy students to grasp fundamentals of Immunology. We surveyed this course for five weeks and observed the classroom lectures, student’s participation, and the learning materials that were provided to students to use in this course. We noticed the major hindrance in learning was lack of student engagement in the classroom. Our survey indicated 77.6% of the students could not focus during lectures for more than 10 minutes. In the classroom environment, traditional PowerPoint lectures were given, students were quizzed during the lecture to keep them attentive, but still there was an obvious lack in lively participation and learning process. The objective of this study was to use interactive educational tools to help engage students. Our hypothesis was that interactive educational tools will improve student’s engagement in the classroom and possibly enhanced student learning. We experimented with various interactive educational tools, such as Moodle (Millionaire game, Quiz, Adaptive Quiz), Facebook livestream, and a private YouTube channel to keep students engaged in the classroom. We observed that simply using various diverse educational tools during the lectures drew student attention and participation rather than sticking to only one method or tool of teaching. The results showed that students were not bored and seemed to enjoy learning and lively contributed to their own learning process. In conclusion, the use of interactive educational tools improved both student engagement in the class as well as boosted the morale of the Instructor.
Conditional abrogation of Ddx3x reveals its essential role in lymphocyte development and partial compensation by Y chromosome
Authors: Krishnamurthy Durga , Seelamneni Harsha , Ochayon David , Harley John B. , Waggoner Stephen N. 
Published: January 2023

Abstract
Ddx3x on the X-chromosome encodes a DEAD-box RNA helicase with putative roles in RNA metabolism, cell cycle progression, apoptosis, and viral immunity. We previously found that hematopoietic-specific hemizygosity of Ddx3x studies caused a marked reduction in the frequencies of natural killer (NK) and B cells in peripheral lymphoid organs of male mice (Liu et al. bioRxiv 2018). No homozygous Ddx3x-deficient female pups were ever obtained, suggesting that hematopoietic loss of this gene contributes to the embryonic lethality observed in germline Ddx3x knockout mice. To assess cell-lineage intrinsic roles for Ddx3x in B-cell and NK-cell development, we bred Ddx3x-floxed mice with Mb1-Cre and Ncr1-iCre mice, respectively. While hemizygous male Ddx3x-deficient mice display mild reductions in B-cell (Mb1-Cre) and NK-cell (Ncr1-iCre) frequencies, homozygous Ddx3x-deficient female mice from each were almost completely devoid of the relevant cell lineages. B-cell loss was evident from the pro-B cells stage, with B cells surviving beyond this stage showing evidence of having escaped deletion of Ddx3x. Ncr1-iCre-mediated deletion of Ddx3x completely abrogated development of both NK cells and other Ncr1-expressing innate lymphoid cells. Thus, DDX3 plays an essential role in the lymphocyte development, where Ddx3y may partially compensate for loss of Ddx3x in these lineages.
Enhanced development of functional human innate immune cells in a novel mouse FLT3null NSG mouse strain expressing human FLT3L
Authors: Vaidya Shantashri , St. Louis Pamela , Burzenski Lisa , Greiner Dale L , Brehm Michael A , Shultz Leonard D 
Published: January 2023

Abstract
Humanized mice are being applied widely to study human immune system homeostasis, function, and as a testing platform for therapeutics. A significant limitation for many humanized mouse models is the lack of mature human innate immune cells with full functionality. Previous studies have demonstrated that delivery of human FLT3L into immunodeficient mice that lack mouse FLT3, promotes the development of human innate immune subsets following engraftment with human HSC. Here we describe a NOD-scid IL2rgnull (NSG) mouse that lacks the expression of mouse FLT3 and expresses human FLT3L transgenically (NSG-mFLT3null Tg(HuFLT3L) or NSG-FLT3L. In these studies, NSG-FLT3L and NSG mice were engrafted with human UCB CD34+ HSC and compared for human immune system development and function. HSC-engrafted NSG-FLT3L and NSG mice show similar levels of total human CD45+ cells in blood over the course of 18 weeks post-engraftment. However, HSC-engrafted NSG-FLT3L mice show significantly increased levels of human immune cells reconstituting the gut mucosa. HSC-engrafted NSG-FLT3L mice show significantly higher levels of human CD141+ and CD1c+ DC subsets, CD123+ pDC, CD14+ monocytes, CD56+ NK cells and CD3+ T cells in the blood as compared to NSG mice. CD56+ NK cells and CD3+ T cells from HSC-engrafted NSG-FLT3L mice express granzyme A and granzyme B, indicating cytotoxic activity. Following treatment of HSC-engrafted NSG-FLT3L mice with LPS, significant levels of human cytokines were detected in serum samples, confirming innate immune system function. Overall these results demonstrate that the NSG-FLT3L model supports enhanced development of functional, human innate immune cells and is a novel tool to study human immunobiology.