bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sensitivity and specificity of an indirect ELISA using Hawaii 31 kDa Angiostrongylus cantonensis antigen in experimentally-infected rats (Rattus norvegicus)
Authors: Jarvi Susan Irene , Steel Argon , Jacob John 
Published: August 2023

Abstract
The parasitic nematode Angiostrongylus cantonensis is the leading cause of neuroangiostrongyliasis, is an emerging tropical disease, and is the leading cause of eosinophilic meningitis worldwide. Diagnosis is generally presumptive and can be easily confused with other central nervous system disorders. Several studies have demonstrated the promise of immunodiagnosis, most frequently in the form of ELISA assays. Taking advantage of plasma samples from previous rat studies, we developed an indirect ELISA assay based on the 31 kDa antigen isolated from A. cantonensis collected in Hawai’i. Using IgG, our assay showed 100% sensitivity, and specificity in diagnosing A. cantonensis infection in lab-reared rats fed 50 live larvae six weeks previously. We saw diminishing sensitivity to IgM, IgA, and IgE, respectively, and no antibody response in rats fed killed A. cantonensis larvae. Our experiments validate the use of indirect ELISA with 31 kDa antigen as an effective immunodiagnostic assay for rats, and our use of different antibody isotypes provided an opportunity to investigate in more detail the humoral immune response in lab-reared rats to A. cantonensis infection.
Daniel K. Inouye College of Pharmacy, Hawaii State Legislature
Fasciola hepatica Fh15 promote survival in a mouse septic shock model and downregulates inflammatory cytokines
Authors: Armina-Rodriguez Albersy , Ocasio-Malavé Carlimar , Méndez-Torres Loyda B , Valdés-Fernández Bianca , Espino Ana M 
Published: August 2023

Abstract
Parasitic helminths and helminth-derived molecules, possess an inherent anti-inflammatory capacity. Fasciola hepatica has been reported to decrease in vivo particular Th1 immune responses caused by concurrent bacterial infections. Studies have shown that some Fasciola hepatica’s excretory-secretory products has the capacity to reduce phagocytic activity, affect cell recruitment, and induce activation of alternative activated macrophages. We hypothesized that recombinant F. hepatica FABP (Fh15) may be an excellent target for the development of new anti-inflammatory drugs against sepsis. For this, we developed a mouse model of septic shock to determine the capacity of Fh15 to suppress the cytokine storm and prevent the mortality rate of mice exposed to lethal doses of LPS. BALB/c mice were allotted into two experimental groups of 5 animals each, which received an IP injection with 12mg/kg body wt. of LPS from E. coli 0111: B4. One hour after LPS injection one group received a single IP injection with 100 μg Fh15 and the other group received two injections of 100 μg Fh15 two hours apart. Control groups received IP injection with LPS, Fh15 or PBS alone. Animals were monitored for mortality for 72 hrs. Blood samples were collected from all animals at baseline and by the time they were about to die or survived the LPS lethal challenge. As expected, all animals from LPS-control group succumbed between 12 to 24h following the LPS challenge. In contrast, between 40 to 100% of animals that received the Fh15 treatment survived to the LPS challenge. This notable increase in the survival rate was found associated to significant suppression in the production of pro-inflammatory cytokines such as IL-12, TNF-a and IFN-g, which were measured in serum by BioPlex.
Sialophorin is an essential host element for vaccine immunity against pulmonary fungal infections
Authors: Nanjappa Som G , Sharma Jaishree , Deepe George S , Mudalagiriyappa Srinivasu 
Published: August 2023

Abstract
The global incidence of fungal infections is alarming due to the increasing immunocompromised population. Vaccine measures are necessary to prevent fungal diseases. Identifying a protective host element as a functional phenotypic marker is immensely valuable as a target to enhance vaccine responses, measure vaccine efficacy, and assess disease outcomes. We identified a host element, O-glycosylated sialophorin, preferentially associated with antifungal memory Tc17 (IL-17A+) cells. Using a mouse model of fungal vaccine immunity during CD4+ T-cell deficiency, we found that sialophorin was essential to bolster vaccine-induced CD8+ T-cell responses and immunity to pulmonary infection. Sialophorin-sufficient CD8+ T cells were superior in responses than the deficient cells under a similar inflammatory micromilieu. Supplementation of sialophorin mAb that binds to O-glycosylated form potentiated the CD8+ T cell responses to fungal antigens, and an inhibitory mAb that binds to non-O-glycosylated form selectively reduced Tc1 (IFNγ+) but not Tc17 cell responses. Sialophorin-mediated immunity was independent of T cell trafficking into the lung. Finally, using fungal antigens vaccination, we show that O-glycosylated sialophorin is a marker of vaccine-potency and immunity. Our study revealed that sialophorin is an essential host element to bolster vaccine responses and serves as a potential biomarker of fungal immunity.
NIAID-NIH 5R01AI153522
Updating the definition of V(D)J recombination signal sequences with high-throughput recombination assays
Authors: Hoolehan Walker Aloysius , Rodgers Karla K 
Published: August 2023

Abstract
V(D)J recombination initiates the production of a diverse antigen receptor repertoire in developing B and T cells. Antigen receptor diversity is required for adaptive immunity in all jawed vertebrates. Recombination activating proteins, RAG1 and RAG2 (RAG1/2), catalyze V(D)J recombination by cleaving adjacent to recombination signal sequences (RSSs) that flank antigen receptor gene segments. Previous studies defined the consensus RSS as containing conserved heptamer and nonamer sequences separated by a less conserved 12 or 23 base-pair spacer sequence. However, many RSSs deviate from the consensus sequence, and the molecular mechanism for semi-selective V(D)J recombination specificity is unknown. Here, we developed a cell-based, high-throughput V(D)J recombination assay to evaluate RAG1/2 activity on thousands of RSSs in parallel. Specifically, we focused our study on the RSS heptamer and adjoining spacer region because it undergoes extensive conformational changes during RAG-mediated DNA cleavage. While the consensus heptamer sequence (CACAGTG) was marginally preferred, RAG1/2 was highly active on a wide range of non-consensus sequences. RAG1/2 generally selected purine/pyrimidine motifs instead of specific bases. Molecular dynamics simulations of RSSs showed that DNA sequences preferred by RAG1/2 had dynamic structural features that may accommodate heptamer unwinding in the RAG1/2 active site. Our results suggest RAG1/2 specificity for RSS heptamers is primarily dictated by dynamic DNA properties dependent on purine/pyrimidine pattern. Further investigation of RAG1/2 specificity will help elucidate the genetic instructions guiding V(D)J recombination to antigen receptor gene segments.
National Institutes of Health (AI128137, AI15635); Oklahoma Center for Advancement in Science and Technology (OCAST) [HR21-142]; Presbyterian Health Foundation of Oklahoma City; National Institute of Allergy and Infectious Diseases Immunology training grant [T32 AI7633-16]
IL-9 responsive macrophages utilize Arginase 1 to enhance lung tumor growth
Authors: Cannon Anthony Michael , Kaplan Mark H 
Published: August 2023

Abstract
In cancer, the immunosuppressive function of myeloid cells that support tumor progression is controlled by secreted factors in the tumor microenvironment. In the tumor microenvironment, arginine and arginine-derived metabolites have been demonstrated to be crucial factors in tumor development. Within macrophages, arginine metabolism influences the polarization of macrophages and therefore, tumor growth by eliciting an anti-tumoral or pro-tumoral phenotype. Interleukin 9 (IL-9) is a pleiotropic cytokine that signals through the IL-9 receptor (IL-9R) and can function as a positive or negative regulator in tumor immunity. Recently, our lab has demonstrated that IL-9 signaling promotes tumor progression in the lung by expanding pulmonary interstitial macrophage populations and inducing Arginase 1 (ARG1), an enzyme associated with pro-tumoral macrophage function, activity. However, the mechanism by which IL-9R/ARG1+ interstitial macrophages promote tumor development remains unknown. Here, using a B16F10 model of metastatic lung cancer, we demonstrate that knockdown of ARG1 using macrophage targeting Arg1 siRNA containing nanoparticles results in altered pulmonary T-cell and macrophage proportions. Moreover, attenuation of IL-9 signaling and ARG1 expression in macrophages impacts arginine and arginine-derived metabolite concentration in lung tissue and BAL fluid which correlates with decreases in CD4+ T Cell exhaustion marker expression and a concomitant reduction in PDL1 expression on pulmonary interstitial macrophages in tumor-bearing mice. Thus, our work suggests that the IL-9R/ARG1 axis in macrophages is sufficient to alter arginine in the tumor microenvironment and promote tumor growth.
Stem-like CD4 T differentiation controls CD8 T cell immunity or tolerance in cancer
Authors: Cardenas Maria A , Prokhnevska Nataliya , Jansen Caroline S , Sobierajska Ewelina , Master Viraj , Kissick Haydn T 
Published: August 2023

Abstract
In this work we wanted to understand how CD4 T cell differentiation shapes the CD8 T cell response in cancer. We identified a population of tumor specific CD4 T cells that maintain an activated, undifferentiated phenotype throughout the cancer response. These CD4 T cells were the dominant population within tumor draining lymph nodes (TDLNs), characterized by high expression of TCF1 and little to no expression of other lineage defining transcription factors. Tumor specific TCF1+ CD4s exhibited functional stem-like properties, as they self-renewed and differentiated almost exclusively into Tregs in tumor bearing mice. These cells, however, were not limited to Treg differentiation, as sorted TCF1+ CD4s from a 5-week tumor response expanded and gave rise to TH1 and TFH effectors in response to LCMV Armstrong infection. Interestingly, we also found a similar population of activated PD1+ TCF1+ CD4s with stem-like capabilities infiltrating various human tumors, indicating this phenotype of CD4s is comparable in human cancer and mouse models. Importantly, the choice of lineage differentiation of TCF1+ CD4s regulated the quality of the CD8 T cell anti-tumor response. When Tregs were selectively depleted, TCF1+ CD4s differentiated into TH1s, which enhanced tumor specific CD8 T cell effector differentiation in both tumor and TDLNs, resulting in reduced tumor burden. Using a bone marrow chimeric CRISPR-Cas9 approach, we found that IFNg from TH1s was intrinsically required for CD8 stem to effector differentiation within TDLNs, promoting an improved anti-tumor response. Overall, this work shows that CD4 differentiation dictates tolerance or immunity in cancer by regulating the phenotype and magnitude of the effector CD8 T cell response.
Varied T cell repertoire bearing intact TCR/CD3 complexes required to observe maximal therapeutic anti-tumor effects of CD3 Fab fragments when binding to T cells
Authors: Mahadevappa Chandrashekara Puthanapura , Huynh Hien , Okpasuo Onekachi , Radhakrishnan Vijay , Cheatham Christa , Vu Nhan , Schrum Adam G , Gil Diana 
Published: August 2023

Abstract
A cognate antigen engagement of TCR induces a conformation change in the CD3 complex (CD3Δc) that is required for productive T cell activation. CD3Δc fails when TCRs interact with poorly immunogenic antigens, like many tumor-associated-antigens (TAAs), which do not active T cells. Anti-CD3 monovalent Fab fragments (Mono-Fabs) bound to the CD3 complex mimic CD3Δc, producing T cell “co-potentiation” when associated with TCRs bound to weak antigens that enhances T cell activation, neither stimulating non-engaged T cells, nor interrupting T cell responses to strong antigens. Previously, we showed T cell co-potentiation can be exploited as a novel immunotherapeutic principle by reducing melanoma burden in B6 mice treated with anti-CD3 Mono-Fabs. TCR signaling initiation requires CD3Δc for the exposure of a conserved sequence in the cytosolic domain of CD3ɛ rich in prolines (PRS) that enables the recruitment of adaptor protein Nck. The mutation of cysteines in the CXXC motif of CD3ɛ extracellular domain inhibits the transmission of TCR signaling across transmembrane that defects CD3Δc and PRS accessibility. Here, using the B6 syngeneic melanoma cell lines Yumm 1.7 and Yummer1.7, which differ in the number of somatic mutations they carry, in B6 mouse strains either wild or mutated in either the CXXC or the PRS motifs in CD3ɛ, we have established the need of tumor cells to carry potential tumor neoantigens, and a more varied repertoire of T cells responding to the tumor with intact TCR/CD3 complexes to observe full anti-tumor effects by anti-CD3 Mono-Fab treatments.
MU Start-up funds
NIH, NCI, U01 CA244314
NIH, NIAID, R01 AI097187
CNPY2-orchestrated unfolded protein responses play redundant roles in regulating anti-tumor immunity of CD8+ T cells
Authors: Wang Yi , Li Anqi , Song No-joon , Shannon Ariana E , Hong Feng , Searle Brian C , Li Zihai 
Published: August 2023

Abstract
Tumors evade immune surveillance by creating a hostile environment that impairs the functions of immune cells. Persistent perturbations including chronic antigen stimulation and metabolic challenges cause endoplasmic reticulum (ER) stress and unfolded protein responses (UPR) in T cells. CNPY2 is one of the key initiators of UPR in the ER, but its roles in T cell activation and persistence have not been reported. Herein, we found that the expressions of CNPY2 and other molecules in UPR pathways were upregulated in both activated and exhausted T cells, indicating UPR activation. However, genetic deletion of Cnpy2 in CD8+T cells did not result in changes in viability, proliferation, mitochondrial functions, and protein translation rate under acute or chronic antigen stimulation in vitro. Furthermore, no differences were seen between wild-type and CD8-specific Cnpy2 knockout mice in protection against multiple syngeneic tumor models including colon (MC38 and CT26), bladder (MB49), melanoma (YUMM1.7 and B16-F10) and breast tumors (4T1). Although CD8+ T cells from tumors were moderately more activated in the absence of CNPY2, no apparent impact was observed on CD8+ infiltration and effector differentiation. Finally, by mRNA profiling and proteomic analysis, we found that the downstream UPR signaling pathways remained intact in T cells despite loss of CNPY2. In summary, we concluded that persistent TCR stimulation and the tumor microenvironment induce CNPY2 expression and UPR in CD8+ T cells. However, ablating CNPY2 alone failed to overcome UPR-mediated T cell dysfunction, suggesting that targeting multiple initiators of UPRs in CD8+ T cells need to be considered to improve cancer immunotherapy.
Impaired IL-17 and cytotolytic responses to influenza vaccine antigens in aging nonhuman primates
Authors: Rout Namita , Walker Edith , Bodogai Monica , Mattison Julie , Jazwinski Stanislaw Michal , Biragyn Arya 
Published: August 2023

Abstract
Despite widespread flu vaccination, influenza remains a major cause of morbidity and mortality in the elderly. The early immune response, including CD4+ helper T cells, and CD8+ CTLs, plays a critical role in efficient control of virus and initiation of adaptive immunity. However, the correlates of protection in the elderly are not well understood. Here, we sought to investigate early cellular responses to re-exposure with seasonal flu vaccine antigens in a cohort of pre-vaccinated young and old rhesus macaques (human age equivalent of 30–45 yrs, and 55–85 yrs). In vitro T cell responses were examined following overnight stimulation of PBMCs with flu vaccine antigens by flow cytometry for Th1-type and Th17-type cytokine responses, and cytolytic functions. Notably, older macaques showed significantly impaired IL-17 production and CD107a upregulation in response to vaccine antigens, along with greater HLA-DR+/PD-1+CD8 T cells. Further, they produced significantly higher TNF-α than young macaques. These results suggest that circulating flu vaccine-specific T cells in older macaques are deficient in mucosal barrier-protective and cytotolytic functions and possess a dominant proinflammatory function. These findings suggest that re-exposure to flu vaccine antigens in older individuals likely induces a proinflammatory response that is impaired in cytolytic capacity and the ability to repair lung mucosal damage during influenza infection, thereby resulting in more severe disease outcomes. Further studies on in vivo modulation of proinflammatory T cells while enhancing IL-17 functions during vaccination could support the development of more effective influenza vaccine strategies for older adults.
The adaptation model of immunity: tumor cell adaptation to tumor-reactive immune responses
Authors: Manjili Masoud H 
Published: August 2023

Abstract
Tumor cells are genetically unstable carrying continuously changing mutations which are the basis for neoantigen vaccines. Pre-existing immune responses against tumor-associated self-antigens as well as against semi-nonself neoantigens have been reported in cancer patients. These data are against the self-nonself (SNS) model of immunity, proposing that tumor cells expressing self-antigens induce tolerance. The adaptation model proposes that expression of many adaptation receptors (AdRs) linked to anti-apoptotic pathways in tumor cells, one of which is reported to be B7-H1, is responsible for the survival of tumor cells in the presence of anti-tumor immune responses. To this end, anti-tumor efficacy of immune checkpoint inhibitors is mainly because of the blockade of survival pathway in tumor cells downstream of B7-H1. Limited efficacy of these therapies is because several AdRs may be expressed by tumor cells which compensate for the blockade of B7-H1. In fact, B7-H1 is mischaracterized as PD-L1 ligand while it is a bi-directional receptor that relays survival signal both as trans, when engaged with PD-1 on T cells, and as cis, when both B7-H1 and PD-1 or B7.1 are expressed on DCs or tumor cells. AdRs could also exist as a dual receptor system consisting of those linked to anti-apoptotic pathways, i.e., TNFR2, and countered by those linked to pro-apoptotic pathways, i.e. TNFR1. The outcomes of tumor-reactive T cell responses would depend on the presence/absence of AdRs or balance between an AdR and its counter-receptor on tumor cells. Therefore, current immunotherapies should focus on the discovery and inhibition of tumor-associated AdRs, or knocking out the nominal AdLs in tumor-specific T cells.
This work was supported by the Office of the Assistant Secretary of Defense for Health Affairs through the Breast Cancer Research Program under Award No. W81XWH2210793. Opinions, interpretations, conclusions, and recommendations are those of the authors and are not necessarily endorsed by the U.S. Department of Defense. This work was also supported by multi-investigator pilot funding from the VCU Massey Cancer Centre, supported in part with funding from NIH/NCI Cancer Centre support grants P30 CA016059.
Development of an Ultrahigh-plex Antibody Panel for Spatial Phenotyping of Murine Cancer Models
Authors: Nikulina Nadezhda , Fisher Jeremy , Loebelenz Jean , Jensen Samuel , LeungLee Paulina , Braubach Oliver 
Published: August 2023

Abstract
Single cell spatial phenotyping of protein biomarkers is an established tool for cancer researchers aiming to understand the biology of the tumor microenvironment. This technology has been developed around human tissues and there remains a paucity of applications available to study murine cancer models. Yet, mouse models are widely used for immuno-oncology (IO) research, justifying the development of spatial phenotyping applications for use in mice. We developed a &gt;30-plex mouse IO antibody panel by selecting biomarkers for tumor, immune cell lineages, immune activation and checkpoints, as well as proliferation and structural markers. Purified antibodies, previously IHC validated in mouse models using rigorous protocols developed by Cell Signaling Technology (CST), were then conjugated to PhenoCycler barcodes, using a standardized conjugation protocol, and confirmed to show positive staining. We then performed ultrahigh-plex imaging using this IO antibody panel on the PhenoCycler-Fusion Platform, using murine lymphoid organs and/or tumor tissues. The resulting data were analyzed for cellular phenotypic compositions as well as the presence of cellular neighborhoods. The work presented here demonstrates the feasibility of conducting ultrahigh-plex spatial phenotyping in FFPE mouse tissues. The tools that were developed as part of this study can readily be deployed for discovery and translational research and will engender future comparative mouse and human spatial phenotyping studies that can further our understanding of cancer biology.
Indolent B-cell splenic lymphomas have T cells with an exhausted phenotype
Authors: Herne Christine Ann , Bruno Jennifer E , Baran Andrea M , Peterson Derick R , Mosmann Tim R , Quataert Sally A , Evans Andrew G , Zent Clive S , Chu Charles C 
Published: August 2023

Abstract
T cell dysregulation has been observed in many tumors, but has not been well-characterized in rare B-cell splenic lymphomas. We began defining the T cell populations in the tumor microenvironments of four human indolent B cell splenic lymphomas: splenic marginal zone lymphoma (SMZL), hairy cell leukemia (HCL), hairy cell leukemia variant (HCLv), and splenic diffuse red pulp small B-cell lymphoma (SDRPL). After Institutional Research Subjects Review Board approval, we studied de-identified cryopreserved bulk splenic tumor suspension cell isolates from fresh (less than 4 h) splenectomy tissue and matched intact formalin-fixed paraffin embedded (FFPE) tissue sections from 17 splenic lymphoma patients and 4 trauma patients (controls). Initial immunohistochemical staining analysis of FFPE tissue sections suggests that the distribution of intratumoral T cells is markedly different among B cell splenic lymphomas. In HCL, residual T cell zones appear retained, albeit diminished, while HCLv and SDRPL have markedly reduced T cells throughout with non-distinct zonation, and SMZL has prominent peritumoral collections of T cells at the interface of the neoplastic white pulp. High-parameter (32-color) fluorescence spectral flow cytometry (Cytek Aurora) analysis suggests that splenic lymphomas exhibit CD4+ and CD8+ T cells with higher proportions of transitional memory, regulatory, and exhausted phenotypes than controls. T cell activation may be enhanced in HCLv. HCLv and SMZL may have decreased naïve T cells. Finally, splenic lymphomas had higher proportions of T cells with an exhausted phenotype compared to trauma samples. In summary, our data suggests T cells are dysregulated in B-cell splenic lymphomas.
American Association of Immunologists, Hairy Cell Leukemia Foundation / Sass Foundation for Medical Research, and generous donations by Elizabeth Aaron.