bluuebunny/crossref_metadata_2025_split huggingface dataset:
To clone tumor specific T cell receptor in cancer patients after immunotherapy with patient-specific antigens
Authors: Han Yanyan , Long Jiangting , Zhou Xiangjun 
Published: January 2023

Abstract

Background
Adoptive T cell transfer was recently often used to treat cancer patients who have failed in standard therapies. An immunotherapy named MASCT (Multiple Antigen Stimulating Cellular Therapy) was applied to cancer patients, which is composed of patient-specific antigen pulsed dendritic cells (DCs) and autologous tumor specific T lymphocytes.

Methods
Patients were treated by the autologous T cells stimulated with mature DCs pulsed with multiple tumor antigens. The tumor antigen specific immune responses were examined during the treatment, and the antigens were selected accordingly. The tumor specific T cell receptor (TCR) would be further cloned from these patients who have demonstrated both immunological and clinical responses.

Results
A human papilloma virus (HPV) positive cervical cancer patient with bone metastasis on the right sacroiliac joint have repeatedly received treatment of MASCT. Specific responses against tumor antigens were detected by IFNγ ELISPOT assay, such as telomerase, CEA, and HPV18/58. Based on patient’s specific immune response, we adjusted her antigen peptide pool by saving the antigens, which had induced specific responses and removed the peptides that did not. The adjusted antigen pool clearly further boosted the specific responses. Moreover, this patient was evaluated as partial remission after MASCT treatment with patient-specific antigens. Therefore, we are now trying to clone the CEA and telomerase specific TCRs from this patient.

Conclusion
Our study provides a promising method to clone tumor specific TCRs from cancer patients, who have shown enhanced immunological responses as well as clinical benefits after immunotherapy with patient-specific antigens.
Development of LAMP-based vaccine for cashew allergy immunotherapy
Authors: Shen Wei , Marketon Anthony , Su Yan , Macauley Juliete , Heiland Teri 
Published: January 2023

Abstract
Tree nut allergy is the second most common triggers of anaphylaxis behind peanut in children and adults. Proactive therapies to decrease reaction severity are currently not available. Here, we utilized Lysosomal-associated membrane protein-1 (LAMP-1) as a targeting signal to make novel DNA vaccines against three major cashew allergens (Ana O1, Ana O2, Ana O3). Briefly, the chimeric constructs encoding the DNA sequences of Ana O1 or Ana O2 or Ana O3 or multivalent plasmid fused to LAMP-1 were synthesized and administrated to BALB/c mice through a needle-free delivery method (Biojector). By ELISA analysis, it showed that LAMP-Ana O 1/Ana O 2/Ana O 3 immunized mice produced high titers of IgG2a and moderate titers of IgG1 antibodies against each antigen. In contrast, IgE response was reduced in the vaccine immunized mice. Moreover, induction of IFN-γ and reduction of allergen-stimulated Th2 cytokine IL-4 were observed in vitro in splenocyte cells of LAMP-Ana O1/Ana O2/Ana O 3 treated mice after antigen stimulation, suggesting a predominant Th1 type immune response was elicited via DNA immunization. To further explore the therapeutic effect of this vaccine, we pre-sensitized the C3H/HeJ mice by oral gavage of protein of cashew extracts, and then immunized the mice with either single LAMP-Ana O plasmids or multivalent plasmid. LAMP-Ana O 1/Ana O 2/Ana O 3 treated mice exhibited strong IgG2a response and decreased IgE level, while control LAMP-vector treated mice demonstrated low IgG2a titer and higher IgE level accompanied by elevation of plasma histamine and IL-4 producing basophils in the blood. These results indicate that immunotherapy with LAMP-Ana O1/Ana O 2/Ana O 3 is effective, and may be a potential approach for treating cashew allergy.
Global dysregulation of gene expression in macrophages from different chicken haplotypes correlates with disease resistance
Authors: Drechsler Yvonne , Irizarry Kris , Griggs Lisa , Kopulos Renee , Keeler Calvin , Boettger Cynthia , Carr Thomas , Clark Jory , Bryden Randall , Downs Eileen , Collisson Ellen 
Published: January 2023

Abstract
Disease resistance and susceptibility has been associated with the major histocompatibility complex (MHC) in several species. This study aimed to better characterize the molecular basis for dramatically different nitric oxide production and associated immune function between the B2 and the B19 haplotype chicken macrophages in response to IFNγ stimulation, previously shown by our laboratory. A next generation RNA sequencing approach was employed to analyze gene expression from purified macrophages from each haplotype group (B2 vs. B19) across 9 time points spanning seven days. Ultimately, our data shows global dysregulation of gene expression in B19 haplotypes correlating with increased disease susceptibility of this haplotype. Our results demonstrate that transcription factors (TF) exhibit divergent expression between B2 and B19 haplotype cells both prior to and after stimulation. Our results also reveal that miRNAs exhibit divergent expression between B2 and B19 haplotype cells both, prior to, and after stimulation with IFNγ. In contrast to TFs, miRNAs can modulate gene product levels post-transcriptionally. We also identify ubiquitin-related enzymes and factors that exhibit divergent expression between B2 and B19 haplotype cells. Some of our results indicate that snoRNAs exhibit divergent expression between B2 and B19 haplotype cells, which is noteworthy because snoRNAs can modulate genes epigenetically. Taken together our results suggest that the global dysregulation of gene expression observed in B19 haplotypes can at least in part be attributed to differential epigenetic regulation.
Establishment and characterization of a porcine lymphoma cell line
Authors: Rahe Michael C , Borgerding Anthony J , Wiseman Barry , Cenatti Leonardo , Martin Daniel , Murtaugh Michael P 
Published: January 2023

Abstract
The lack of well characterized, established domestic porcine cell lines hinders advancement of porcine cellular immunology understanding in disease resistance and anti-viral immunity. Numerous cases of multicentric lymphoma were diagnosed in pigs at the time of slaughter. Affected organs were harvested and submitted for single cell isolation and analysis. Culture of disaggregated single cell suspensions in RPMI 1640 with weekly passage resulted in clusters of dividing cells in about 2% of attempted isolations. In one pig, cells grew in approximately 50% of wells. Cell lines were established by limiting dilution repeated 3 times from splenic and subiliac lymph node lymphomas. Initial flow cytometry analysis showed a population of CD3+, CD79a+, CD21+, CD4−, and CD8− cells which have grown and been maintained in culture for more than 7 months and more than 10 subcultures. To further characterize the nature of this paradoxical homogenous population of CD3+ and CD79a+ cells, transcriptome analysis is being carried out to validate initial cytometric findings and identify key gene pathways. This new porcine lymphoma cell line will be a valuable resource for more in-depth cellular investigations into the porcine immune system, as well as providing a potential tool for the growth of lymphotropic viruses of pigs and humans.
A novel role for Osteopontin in facilitating West Nile virus neuroinvasion
Authors: Paul Amber M , Acharya Dhiraj , Le Linda , Stokic Dobrivoje S , Leis A Arturo , Bai Fengwei 
Published: January 2023

Abstract
West Nile virus (WNV) is a positive sense, single-stranded RNA flavivirus that can cause human neuroinvasive diseases, including encephalitis and meningitis. The mechanisms by which WNV enters the central nervous system (CNS) and the host-factors that are involved in WNV neuroinvasion are not completely understood. In this study, we found increased production of soluble osteopontin (OPN) in human plasma following WNV infection compared to uninfected controls, indicating a role for OPN in WNV pathogenesis. In a mouse model, after challenge with WNV (2000 plaque forming units, PFU), OPN deficient (Opn−/−) mice exhibited a higher survival percentage (80%) than wild-type (WT) mice (20%). However, both groups had comparable levels of virus in circulating blood and peripheral organs. Interestingly, viral burden in the brains of Opn−/− mice were significantly lower than WT mice, which was coupled to reduced polymorphonuclear neutrophil (PMN) infiltration. Soluble OPN has been previously described as a chemokine for PMN recruitment, and intracerebral supplement of recombinant OPN (50 ng/mouse) followed by an intraperitoneal challenge of WNV (2000 PFU), resulted in increased PMN infiltration and WNV load within the CNS. Furthermore, we also confirmed that infected-PMNs carried WNV into the CNS. Together, these data suggest OPN plays a negative role in recruiting WNV-infected PMNs into the CNS, thus mediating WNV neuroinvasion.
Porcine reproductive and respiratory syndrome virus non-structural protein Nsp2TF down-regulates Swine Leukocyte Antigen class I (SLA class I) expression
Authors: Cao Qian , Subramaniam Sakthivel , Cao Dianjun , Meng Xiang-Jin 
Published: January 2023

Abstract
Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is arguably the most economically-important global swine pathogen. The virus typically induces sub-optimal immune responses especially a gradual increase of cellular immune-mediated (CMI) responses in the host. In this study, we demonstrated that PRRSV reduced SLA-I surface expression on porcine alveolar macrophages, monocyte-derived dendritic cells and a porcine kidney epithelial cell line, which may partly explain for the impaired CMI responses observed against PRRSV in pigs. To identify the PRRSV proteins responsible for SLA-I down-regulation, we screened all 22 PRRSV proteins by transient transfection assays. Our results indicated that Nsp1α, Nsp2TF and GP3 significantly down-regulated SLA-I surface expression of which Nsp2TF showed the greatest effect. To further validate whether Nsp2TF directly contributes to the PRRSV-induced SLA-I down-regulation, a panel of mutant viruses were generated with abolished Nsp2TF expression by disrupting −2 ribosomal frameshifting elements essential for the Nsp2TF protein synthesis as well as by introducing stop codons in TF domain of the protein. The infections with these mutant PRRSV strains fully restored the SLA-I surface expression in the infected cells. Additionally we demonstrated that the last 68 amino acids of TF domain are critical for this function. Our study for the first time uncovered an immune-modulatory function of PRRSV Nsp2TF in down-regulating SLA-I expression. Further studies are warranted to generate Nsp2TF-knockout vaccine strains and investigate their efficacies in inducing better CMI responses against the virus in pigs.
Regulatory role of neutrophils in the inflammatory responses to paramyxovirus infection in mice
Authors: Cheemarla Nagarjuna R , Banos-Lara Rocio , Guerrero-Plata Maria Antonieta 
Published: January 2023

Abstract
Neutrophils are the most abundant leukocytes (50% to 70%) in humans and are the first immune cell population recruited to the sites of infection. They are known to act in the first line of innate immune defense against invading pathogens, and more recently, to playing a crucial role in orchestrating adaptive immune responses. However, the role of neutrophils in the control of adaptive immune response during respiratory viral infections is still largely unknown. In order to elucidate the role of neutrophils in respiratory antiviral defense, we employed an experimental mouse model of human metapneumovirus (hMPV) infection. HMPV is a member of the Paramyxoviridae family, and a leading respiratory pathogen causing severe symptoms such as bronchiolitis and pneumonia in the young, elderly and immunocompromised patients. Our findings demonstrate that neutrophils are rapidly recruited in high numbers to the lungs of infected mice (&gt;80%) during the acute phase of infection. Specific depletion of neutrophil in vivo using a monoclonal antibody and simultaneous infection with hMPV exhibited significantly higher levels of inflammatory cytokines, pulmonary inflammation and severe clinical disease (as indicated by an enhanced body weight loss and clinical illness) compared to hMPV-infected competent mice. Interestingly, the lack of neutrophils altered the T cell responses in the lung, particularly the γδ-T-cell subset. Finally, we also observed that the recruitment of neutrophils to the lung was controlled by the expression of the hMPV attachment protein. These novel findings demonstrate that neutrophils regulate T cell responses and are critically important for the control of the hMPV-induced inflammatory responses in the respiratory tract.
Insight into the role of immunosenescence during varicella zoster virus infection (shingles) in the aging cell model
Authors: Kim Ji-Ae , Park Seul-Ki , Shin Ok Sarah 
Published: January 2023

Abstract
Varicella zoster virus (VZV) is the etiological agent of shingles, a painful skin rash that affects a significant proportion of the elderly population. In the present study, we used two aging cell models, Hutchinson-Gilford progeria syndrome (HGPS) fibroblasts and stress or replicative senescence-induced normal human dermal fibroblasts (NHDFs), to investigate age-associated susceptibility to VZV infection. VZV infectivity titers were significantly associated with donor age in HGPS fibroblasts and senescence induction in NHDFs. High throughput RNA-sequencing (RNA-seq) analysis was performed to assess global and dynamic changes in the host transcriptomes of VZV-infected aging cells. Analysis of differentially expressed genes (DEGs) indicated that VZV infection in aged HGPS fibroblasts resembled that in senescent NHDFs, particularly in terms of genes associated with pattern recognition receptors in virus sensing network, providing novel insights into the mechanisms of senescence associated susceptibility to VZV infection. Additionally, we identified stimulator of interferon genes (STING) as a potential VZV sensing receptor. Knockdown of STING expression resulted in increased viral replication in primary fibroblasts, whereas STING overexpression led to suppression of VZV plaque formation. In conclusion, our findings highlight the important role of immunosenescence following VZV infection and provide significant insights into the mechanisms underlying cellular sensing of VZV infection and the induction of immune responses in aged skin cells.
Pathobiology of West Nile Virus infection in the Collaborative Cross mouse model
Authors: Ireton Renee C. , McMillan Aimee , Thomas Sunil , Treuting Piper , Voss Kathleen , Hendrick Duncan , Wilkins Courtney , Green Rich , Mooney Michael , McWeeney Shannon , Gale Michael J 
Published: January 2023

Abstract
West Nile Virus (WNV) is an emerging neuroinvasive flavivirus that causes serious encephalitis disease that is lethal in 6% of human cases. The immunopathology of WNV infection has been well-studied in C57BL/6J mice, but the model lacks the diversity in susceptibility and outcomes that are observed in humans, thus limiting its usefulness as a model to understand the host genetics and overall breadth of WNV infection outcomes and immune response. We are using the Collaborative Cross (CC) mouse model to evaluate the pathobiology and immune regulation that underlies WNV disease progression and neuroinvasion. Our studies reveal that the CC model captures the diversity in susceptibility and outcomes of WNV observed in humans. We generated a standardized histology scoring system to evaluate viral localization and immunopathology of WNV infection in tissue including brain isolates from CC lines. Our studies show a diverse array of histological phenotypes that link with the susceptibility and disease outcomes of the CC mice with WNV infection. Time course studies also showed variation in the temporal progression of viral induced-pathology and immunopathogenesis among different CC lines. Our findings show that the CC model of WNV is a relevant and powerful mouse model for mechanistic studies of WNV infection and neuropathology. Application of the CC model to genetic mapping of quantitative trait loci that impart immune regulation of WNV infection will be presented.
IL-36 cytokines regulate antiviral immunity
Authors: Jensen Liselotte E , Milora Katelynn A 
Published: January 2023

Abstract
Viruses may succeed as pathogens by deploying immune evasion mechanisms aimed at limiting detection by the host. If the host has developed counter strategies to ensure viral clearance is less clear. HSV-1 blocks inflammatory signaling from infected cells by preventing extracellular release of IL-1β. We found that HSV-1 infected keratinocytes retain the ability to signal danger to the surrounding tissue through secretion of IL-1α. This inflammatory signal appeared essential in vivo for early recruitment of immune cells to infected sites. Furthermore, mice deficient of the common receptor for IL-1α and IL-1β were more susceptible to lethal outcome of HSV-1 infection. Hence, IL-1α may have evolved to counteract an HSV-1 immune evasion strategy targeting IL-1β. We here extend these studies to the novel IL-1 family members IL-36α, IL-36β and IL-36γ, which signal via their common receptor IL-36R. We previously showed that poly(I:C), a double stranded RNA analogue often used to simulate viral infections, promoted expression and release of IL-36γ through a cell death mechanism called pyroptosis, suggesting a role for IL-36γ in immunity against viruses. Surprisingly, using the flank skin HSV-1 infection model, IL-36γ knockout (KO) mice developed disease indistinguishable from that observed in wild type mice. However, IL-36β KO mice progressed to more severe disease as reflected by larger skin lesions and increased mortality. Interestingly, inflammation induced by the anti-viral drug imiquimod was dependent upon IL-36α. Hence, the immune system appears to have several layers of redundancies that ensure a productive immune response.
Interferon-gamma inducible GTPases disrupt viral replication complex upon their targeting by the LC3 conjugation system of autophagy
Authors: Hwang Seungmin , Biering Scott , Choi Jayoung , Park Sungwoo , Williams Lelia , Dominici Erin 
Published: January 2023

Abstract
Viruses with a single strand (+) sense RNA genome replicate within vacuole-like structures in cytoplasm, called replication complex (RC). RC is made through reorganization of cellular organelle membranes, and it provides a favorable microenvironment for the viruses to replicate. How the host immune system counteracts viral RCs has been obscure. Recently, we found that interferon-gamma (IFNγ) inhibits the replication of murine norovirus (MNV) at the stage of RC formation. Intriguingly, this anti-viral activity of IFNγ depends on a protein complex involved in cellular autophagy. Autophagy is an evolutionarily conserved pathway that sequesters cytoplasmic materials in membrane-bound autophagosomes and delivers them to lysosome for degradation. To form an autophagosome, the microtubule-associated-protein-1-light-chain-3 (LC3) conjugation system (Atg7, Atg3, Atg12-Atg5-Atg16L1) is essential. Surprisingly, only the LC3 conjugation system of autophagy, but not lysosomal degradation through autophagy, was required for IFNγ to inhibit MNV RC formation. Through a comparative mechanism study, we found that the LC3 conjugation system was required to target IFNγ inducible GTPases - immunity related GTPases (IRGs) and guanylate binding proteins (GBPs) - to the RC. Both IRGs and GBPs are known to be targeted to the membrane of vacuoles containing bacteria, protozoa, or fungus. The targeted membranes are vesiculated and vacuoles get ruptured, leading to the death of exposed pathogens. Similarly, the GTPases disrupt the membrane structures of the RC and thus the replication of MNV. These data suggest that IFNγ utilizes the effectors against vacuolar pathogens for the control of viruses replicating in cytosolic membranous structures.
Bias-free imaging of antigen degradation using bioorthogonal antigens
Authors: Pawlak Joanna B. , Hos Brett , Megantari Otty A , Ossendorp Ferry A , van Kasteren Sander I 
Published: January 2023

Abstract
Degradation of antigen into short MHC-loadable polypeptides by antigen presenting cells presents a Heisenbergian conundrum: reporter strategies for imaging these processes affect the processes itself to such an extent that the information obtained from reporter antigen strategies may not be representative of the process itself.
For example fusion constructs of antigens are only imagable until they are degraded or separated from the antigen, which is essential if a peptide from the antigen is to be loaded on an MHC and reach the surface for sampling by T-cells. This results in an imaging bias to only those parts of the pathway in which reporters remain intact. Further complications can arise from the fact that reporter-constructs can alter the rate of proteolysis and routing by virtue of changing overall physiochemical properties of the antigens.
I will present our recent developments of ‘bioorthgonal antigens’, a bias free antigen labelling strategy that allows the imaging of antigen routing with minimal interference in the pathway. These antigens carrying amino acids with tiny chemical modifications (2 or 3 atoms in size) in their side-chains that are stable to proteolysis and other conditions found in the cell, can be selectively visualised using click chemistry. This allows, for example the localization of degrading antigen inside an APC or the determination of the rate of epitope appearance on the cell surface without the need for peptide-MHC-specific antibodies. This offers the first opportunity to have a detectable group that does not affect processing and presentation
Type I interferon signaling acts directly on transitional T2 B cells to promote RNP-Ag tolerance loss in BXD2 mice
Authors: Hamilton Jennie A , Wu Qi , Yang PingAr , Luo Bao , Li Jun , Liu Shanrun , Hsu Hui-Chen , Mountz John D 
Published: January 2023

Abstract
In lupus, autoreactive B cells fail to be removed at the transitional B cell stage. Lupus prone BXD2 mice exhibit transitional B cell tolerance loss and production of autoAb against RNA binding proteins (RNP). The purpose of this study was to determine direct effects of IFNα on developing transitional B cells. Analysis of IFNαR expression in B6 and BXD2 mice revealed the highest expression levels on transitional B cells with lower levels on mature FO, MZ, and GC B cells. Interestingly, both in vitro and in vivo, peripheral B cell responsiveness to IFNα started at the IgMhiCD23lo T1 stage and was significantly upregulated at the T2 stage, as measured by CD69 induction (76±5% in B6 vs 89±3% in BXD2 T2). In vivo treatment of B6 and BXD2 mice with IFNα promoted B cell upregulation of CD69 and CD86 on T2 B cells. Using an antigen (Ag) tetramer approach for isolation and characterization of La and RNP (La13-27 and snRNP357-373) autoreactive B cells, we found that IFNα stimulation increased maturation of transitional RNP autoreactive B cells and expansion of the autoantigen engaged IgMlo T3 compartment in BXD2 mice. Consistent with abnormal signaling and activation in vivo, transitional B cells from spleens of BXD2 mice exhibited elevated basal phospho-tyrosine and BCR induced activation, beginning at the T2 stage. Single cell analysis of transitional T2 B cells from BXD2 mice revealed a heterogeneity of gene expression within this population and a negative correlation between the expression of IFNαR gene and Id3, an important transcription factor for B-cell anergy. Our results suggest that B-cell tolerance loss in BXD2 mice occurs at the transitional stage, and IFNα exhibits the potential to break tolerance to RNP in transitional B cells.