bluuebunny/crossref_metadata_2025_split huggingface dataset:
SP17 as new immunotherapy target and biomarker for triple-negative breast cancer (VAC5P.1121)
Authors: Chiriva-Internati Maurizio , Mirandola Leonardo , Cobos Everardo , Dahlbeck Scott , Nguyen Diane , Figueroa Jose , Layeequr Rahman Rakhshanda 
Published: January 2023

Abstract
Breast cancer (BC) is a main health concern for women worldwide. Immunotherapies directed against specific tumor-associated antigens enhancing cytotoxic T lymphocyte (CTL)-mediated antitumor responses, have not been equally effective against all BCs: targeted, specific anti-hormonal therapies are now available against the estrogen (ER) and progesterone (PR)-positive BC, as well as HER-2/neu receptor. Triple negative breast cancer (TNBC) displays a negative immunohistologic pattern for ER, PR, and HER-2/neu. As neither of the hormone receptors nor the HER-2 are expressed in TNBC, the hormonal and molecular therapies used in other subtypes are not effective. Current efforts are focused on identifying molecular targets for TNBC. The CTA, SP17, is aberrantly expressed in an array of neoplasms. In this study, we investigated SP17 expression and immunogenicity in triple-negative breast cancer cell lines and tissues, and we tested the efficacy SP17 to stimulate a specific CTL-mediated antitumor response. We showed, for the first time, that SP17 was expressed in TNBC cell lines and primary breast tumor samples. Furthermore, we identified specific anti-SP17 antibodies in TNBC patients’ sera and we were able to generate SP17-specific, HLA class I-restricted, cytotoxic T lymphocytes capable of efficiently killing TNBC cells in vitro. Hence, SP17 may function as a novel biomarker, as well as vaccine target in this disease.
Simultaneous monitoring of in vivo humoral and cellular immune responses in the avian model (VAC5P.1123)
Authors: Erf Gisela , Alaamri Olfat , Jang Hyeonmin , Falcon Daniel , Byrne Kristen , Aguilar Zoraida , Dienglewicz Robert 
Published: January 2023

Abstract
Using the dermis (pulp) of growing feathers (GF) as a test-tissue together with blood sampling, we monitored cellular and humoral immune responses in chickens immunized with 3 different formulations of mouse IgG test-antigen (T-Ag). Test-antigen (26 μg) was injected i.m. in 2 groups of chickens either alone, mixed with Alum adjuvant or conjugated to iron-oxide nanoparticles (10 nm). Group 1 was immunized at 7 weeks and Group 2 at 7 and 11 weeks (n = 18 chickens/group). Antibody production (IgM, IgG) to T-Ag was monitored for 4 weeks following both the primary and secondary i.m. immunizations. To examine the local effector response to T-Ag, T-Ag was injected into 20 GF per bird (10 μL/GF) on Day 10 (Group 1) or Day 5 (Group 2) post-primary or secondary immunization, respectively, and GF collected 0.25, 1, 2, 3, 4, 5, & 7 days post-injection. Analysis of GF pulp cell suspensions by flow cytometry revealed temporal, qualitative and quantitative differences (P &lt; 0.05) in leukocyte-infiltration between immunization treatments and primary/memory effector-responses. Cytokine expression analysis (qRT-PCR) is in progress. ELISA also revealed temporal, qualitative, and quantitative differences (P &lt; 0.05) in the humoral responses to T-Ag following the different primary/secondary i.m. immunizations. Minimally invasive, non-terminal procedures such as sampling of injected GF and blood provides unique insight into cellular and humoral immune activities in the same individual over time.
The inhibition of CD1d-mediated antigen presentation upon an HSV-1 infection is VP22-dependent (VIR1P.1134)
Authors: Liu Jianyun , Gallo Richard , Duffy Carol , Brutkiewicz Randy 
Published: January 2023

Abstract
CD1d-restricted T (NKT) cells are important for controlling a herpes simplex virus (HSV) infection. One of the mechanisms of immune evasion by HSV is to down-regulate CD1d-mediated activation of NKT cells. VP22 is an HSV-1-encoded protein responsible for reorganizing the host cell’s cytoskeletal network and viral spreading. We have shown that modification of the cytoskeleton can alter CD1d-mediated antigen presentation. In this study, we found an HSV-1 lacking VP22 (ΔUL49) was impaired in its ability to inhibit CD1d-mediated antigen presentation compared to wild-type (WT) virus; this was reversed by a repair virus (UL49R) in CD1d-expressing cells. Of importance, the effects of VP22 were shown to be exerted on the T322/S323 dyad, critical residues for the virus to reduced antigen presentation. We further demonstrated that CD1d recycling was inhibited by infection with WT and UL49R, but not ΔUL49 virus. Rather than inhibiting antigen presentation by CD1d, ectopic expression of VP22 in CD1d-expressing cells complemented the VP22-deficient virus in inhibiting antigen presentation. Therefore, VP22 is required (but not sufficient) for the inhibition of CD1d-mediated antigen presentation by an HSV-1 infection.
Viral cyclin control of infection and associated disease (VIR1P.1135)
Authors: Niemeyer Brian , Oko Lauren , Van Dyk Linda 
Published: January 2023

Abstract
Gammaherpesviruses (gHV), including Epstein-Barr virus and Kaposi’s sarcoma associated herpesvirus, are lymphotropic viruses, distinctive in their ability to exploit the lymphocytes as their lifelong reservoir. gHV infection is ubiquitous, and can result multiple pathologies in immune-compromised individuals. Murine gHV68 is homologous to the human viruses, and serves as a small animal model. Previously, our lab demonstrated that a conserved virally encoded homolog of host cyclins, (v-cyclin), is critical for specific aspects of gHV68 infection. A virus lacking the v-cyclin is is dramatically defective in reactivation from latency, viral persistence, and progression of disease. Using an enzymatically marked virus, we now identify infected cells during latent infection with v-cyclin sufficient or deficient viruses. We found that cells infected with or without the v-cyclin are not significantly different in infected B cell subset distribution. However, we found the number of cells with detectable levels of gene expression was significantly depressed in the absence of the v-cyclin. These data suggest that the v-cyclin supports active gene expression during latency, likely corresponding to increased reactivation capacity. This system parallels the complex gene expression programs of Epstein Barr virus, and may permit us to distinguish the specific features of infected cells poised for reactivation and virus propagation, along with their relative roles in virus induced pathology.
BNIP3-mediated mitophagy promotes the generation of natural killer cell memory (VIR1P.1139)
Authors: O'Sullivan Timothy , Sun Joseph 
Published: January 2023

Abstract
Natural killer (NK) cells are innate lymphocytes that possess traits of adaptive immunity such as clonal expansion, contraction, and generation of long-lived “memory” cells, processes poorly understood at the molecular level. Here we find that as proliferating NK cells accumulate dysfunctional mitochondria during viral infection, a protective mitophagy pathway is induced during the contraction phase to promote their survival in a reactive oxygen species (ROS)-dependent manner. Inhibition of mammalian target of rapamycin (mTOR) or activation of AMPK during the contraction to memory phase of the antiviral response increased autophagic activity and enhanced memory NK cell numbers through an Atg3-dependent mechanism. Furthermore, we demonstrate a temporally regulated role for mitophagy-inducing proteins BNIP3 and BNIP3L in the generation of robust NK cell memory. Thus, our study reveals the functional importance of mitophagy during the dynamic response of these cytolytic innate lymphocytes against infection.
Dysregulation of the B cell compartment correlates with development of pulmonary arterial hypertension in a non-human primate model of HIV-infection (VIR1P.1142)
Authors: Kling Heather , Tarantelli Rebecca , Norris Karen 
Published: January 2023

Abstract
HIV-infected persons are at increased risk for co-morbidities associated with advanced age, such as pulmonary arterial hypertension (PAH), though underlying mechanisms are not understood. HIV infection is also associated with B cell dysregulation, which may contribute to tissue damage and premature onset of disease. We have previously shown that simian immunodeficiency virus (SIV) infection of macaques results in increased pulmonary arterial pressures and pulmonary vascular remodeling. In the current study, we examine whether B cell activation/exhaustion is associated with PAH in SIV infection. Rhesus macaques (n=10) were mucosally infected with SIVΔB670. Peripheral blood (PB) and bronchoalveolar lavage fluid (BALF) cells were collected prior to and monthly following SIV infection, stained and analyzed by flow cytometry. Right ventricular systolic pressures (RVSP) were assessed by right-heart catheterization at 3-4 month intervals. Percent of CD95+ B cells was increased in PB and BALF (p&lt;0.0001) following SIV infection and correlated with RVSP (PB, p=0.0317; BALF, p=0.0007). Additionally, elevated frequency of exhausted B cells (CD27-CD10-CD21lo) correlated with RVSP (p=0.043). CD95 expression was strongly correlated with age-associated B cells (ABC; CD21-CD11c+) (p&lt;0.0001), the frequency of which also correlated with RVSP (p=0.017). Collectively, these results support the hypothesis that chronic B cell activation in HIV infection is associated with PAH development.
Heart rate variability in humans infected with the cytomegalovirus (VIR1P.1145)
Authors: Koenig Julian , Jarczok Marc , Rector Jerrald , Fischer Joachim , Sternberg Esther , Thayer Julian 
Published: January 2023

Abstract
Humans infected with the cytomegalovirus (CMV+) have been shown to have a higher risk of cardio-vascular-disease (CVD) and mortality. Previous research supports a neuroimmunomodulatory reflex via vagally mediated t-cells in the splenic nerve end. Vagally-mediated heart rate variability (vmHRV) is a independent predictor of CVD mortality and associated with inflammatory processes. Existing studies investigated the association of CMV infection and HRV only in small samples of CMV infected infants or CMV-infected fetuses. The aim of the present study was to explore potential differences on vmHRV in individuals infected with CMV compared to their non-infected counterparts. 24-hour recordings of HRV were obtained by ECG holter monitoring and the high-frequency (HF) power was derived as index of vagal activity. CMV activity was determined by an enzyme-linked immunosorbent assay on plasma from fasted blood samples. Data from a total of 485 participants (10% females; mean age 46±10 years,) was available for analysis. Participants diagnosed as CMV+ (37%, n = 181) showed significantly (t(1.98) = 0.024, p = .024) lower vmHRV (HF 236±16.5 ms2) compared to individuals without CMV (CMV-, n = 304, HF 284±15.9 ms2).
The role of CD40 in LP-BM5 infection of mixed glial cells (VIR1P.1158)
Authors: Grlickova-Duzevik Eliza , Malon Jennifer , Cao Ling 
Published: January 2023

Abstract
LP-BM5 retrovirus infection is an established murine model of HIV infection in humans due to that susceptible strains of mice (such as B6) develop immunodeficiency syndrome. Previously, we have shown that infected B6 mice developed peripheral neuropathy and CD40 knockout (KO) mice displayed less mechanical hypersensitivity (behavioral sign of peripheral neuropathy) post-LP-BM5 infection. The spinal cord viral RNA levels were significantly lower in CD40 KO mice compared to wild type (WT) B6 mice, which is correlated with decreased lumbar spinal cord cytokines, IL-1-beta, IL-6 and IFN-gamma. It is known that LP-BM5 can infect glial cells directly. We hypothesized that glial cells in CD40 KO mice were less susceptible to LP-BM5 infection. To test this hypothesis, primary spinal cord mixed glial cells from both WT and CD40 KO mice were infected with various doses of LP-BM5. Consistent with our in vivo findings, levels of viral RNA are significantly lower in mixed glial cells from CD40 KO mice than that from WT mice. Further, levels of cytokines, IL-1-beta, IL-6, TNF-alpha, IFN-gamma, and IL-12p40 were measured in the supernatants. LP-BM5 only induced minimal increase of these cytokines and there no significant differences between WT and CD40 KO mice were observed. Our data indicate that glial cells lacking CD40 are more resistant to LP-BM5 infection and this resistance does not seem to be due to the decreased production of proinflammatory cytokines by glial cells.
Temporal transcription analysis of innate immune response to respiratory syncytial virus infection (VIR2P.1164)
Authors: Gao Yajing , Song Ran , Dozmorov Igor , Viswanathan Kasthuribai , Levitz Ruth , Kahn Jeffrey , Wakeland Edward 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) is a leading cause of severe respiratory disease in infants. Little is known about the cell-intrinsic innate immune responses of RSV infected epithelial cells or responding immune cells. We studied two clinical-isolates from the RSV-B subgroup: NH1125 that induces high cytokine responses (measured by CCL5 and IL-6); and NH1067, which induces low cytokine responses. We used RNA sequencing technologies to profile the transcriptome of non-immune and immune-lineage cells following infection with these two strains. The innate immune response of A549, a human epithelial cell line, was distinct from that of monocyte-derived macrophages and dendritic cells at 2, 4, 8, and 24 hours following infection. These responses were compared at a high multiplicity of infection (MOI=5) for a homogeneous response and low MOI (MOI=0.3), which more closely resembles in vivo conditions, to determine the extent of communication between infected and un-infected cells. Our analyses revealed 6 classes of genes marking the early- and late-stage cellular responses. Pathway analyses identified miRNA let-7f, EIF2 signaling and mitochondrial dysfunction signaling as early activated pathways and pattern-recognition receptor and Interferon pathway at later time points. A cross-comparison also reveals that RSV gene expression and viral replication is greater in A549 cells than macrophages, consistent with a more potent cell-intrinsic innate defense in myeloid lineage cells.
Extra-cellular Matrix protein 1 (ECM1) Ameliorates Experimental Autoimmune Encephalomyelitis (EAE) through Inhibition of MMP9 Activity
Authors: Sun Bing , Su Pan 
Published: January 2023

Abstract
Multiple sclerosis (MS) is the most common inflammatory demyelinating disease of the central nervous system (CNS). Experimental autoimmune encephalomyelitis (EAE), an animal model representing human multiple sclerosis (MS), is mainly mediated by Th1 and Th17 cells. Matrix metalloproteinase-9 (MMP9), a zinc bound protease, plays an important role in the invasion or infiltration of pathogenic Th cells into CNS during MS/EAE. In current study, we find that ECM1 could inhibit MMP9 activity through protein/protein interaction in vitro and ameliorate MOG-induced EAE via blocking MMP9-mediated invasion/infiltration of pathogenic Th cells into CNS in vivo. Giving ECM1 protein treatment on the late periods of pathogenic T cell migration in EAE model, the EAE disease is significantly attenuated. This treatment functions by reduction of infiltrating pathogenic Th cells into CNS, but had no impact on Th1/Th17 cell development in the peripheral lymphoid tissues. Our results suggest that ECM1 is potential an inhibitor to treat EAE disease.
The innate immune receptor NOD2 is protective against autoimmune-induced arthritis in SKG mice
Authors: Napier Ruth J , Vance Emily , Snow Paige , Lee Ellen J , Sakaguchi Shimon , Rosenzweig Holly L 
Published: January 2023

Abstract
Rheumatoid arthritis remains a major public health concern, estimated to affect 1.5 million adults in the U.S. However, the etiology and underlying immunological mechanisms of this disease remain largely speculative. SKG mice harbor a spontaneous mutation in ZAP-70 and develop arthritogenic T cells, leading to chronic, self-sustained joint destruction. Arthritis in SKG mice replicates a current theoretical paradigm that environmental factors, including microbes, activate preexisting autoreactive T cells in genetically predisposed individuals to cause disease. Currently, the mechanisms by which innate immune receptors augment T cell activation as well as disease onset and progression are not understood. Here we sought to characterize the role of the intracellular pattern recognition molecule, NOD2, in disease pathology. Using NIR-spectroscopy and clinical scoring methods we determined that SKG mice lacking NOD2 develop markedly worse arthritis when triggered by fungal associated molecules, including curdlan and zymosan, as compared to SKG/NOD2+/+ mice. Intracellular cytokine staining showed no difference between Th17 and Th1 cytokine production by CD4+ and CD8+ T cell splenocytes between genotypes. Importantly, neutralization of IL-17A reduced disease severity in SKG/NOD2−/− mice to levels of disease exhibited by SKG/NOD2+/+, suggesting IL-17A as the prominent worsening factor in SKG/NOD2−/− mice. Our data suggest that NOD2 serves a protective role against autoimmune arthritis, in part due to regulation of IL-17A. Thus, understanding how microbial recognition pathways like NOD2 factor into the induction and pathology of arthritis is key to the development of effective a therapy.
IL-23 compensates for the absence of IL-1 signaling on CD4 T cells in EAE model
Authors: Mufazalov Ilgiz , Schelmbauer Carsten , Regen Tommy , Kuschmann Janina , Wanke Florian , Gabriel Laureen , Wolk Michaela , Hauptmann Judith , Muller Werner , Pinteaux Emmanuel , Kurschus Florian , Waisman Ari 
Published: January 2023

Abstract
Interleukin-1 (IL-1) is implicated in numerous pathologies, including multiple sclerosis and its animal model experimental autoimmune encephalomyelitis (EAE), which is believed to be a T cell-mediated disease. However, the lack of specific tools so far limited our understanding of the precise mechanisms of IL-1 action. We therefore generated a mouse strain allowing the conditional deletion of the only known signaling receptor for IL-1, IL-1 receptor type 1 (IL-1R1). Crossing of these mice to CMV-Cre or CD4-Cre transgenic mice resulted in IL-1R1 deletion globally or specifically in αβ T cells, respectively. When we induced EAE in these mice using a standard protocol, we found that Pertussis toxin administration leads to IL-1R1 dependent recruitment of myeloid cells to the draining lymph nodes and subsequent IL-1β production. This myeloid-derived IL-1β did not vitally contribute to the generation and plasticity of Th17 cells in general, but rather promoted expansion of a GM-CSF+ Th17 cell subset, thereby enhancing its encephalitogenic potential. Mice lacking IL-1R1 specifically in T cells, in contrast to complete IL-1R1 deficient animals, were not resistant to active EAE induction but rather developed a mild form of paralysis. CNS-infiltrating CD4 T cells included GM-CSF+ Th17 cells yet in reduced frequencies and numbers compared to wild type control animals. Moreover, the pathogenicity of IL-1R1 deficient T cells was fully restored by IL-23 polarization in vitro prior to transferring of sufficient numbers of these IL-17A+ cells into Rag1−/− recipient mice. Therefore, our data emphasize an important but dispensable function of IL-1 signaling in T cell mediated neuropathology.
Mercury modulates splenic immune cell proportion in mice
Authors: Jin Guang-Bi 
Published: January 2023

Abstract
Mercury is one of the widely spread environmental pollutants, inducing autoimmunity characterized by a T cell-dependent polyclonal activation of B cells. However, its mechanisms remain unclear. In this study, we investigated the role of mercury on the immune cell proportion. Eight-week old BALB/c mice were injected (s.c.) three times a week with HgCl2 (1 mg/kg body weight) throughout the duration of the experiment. Control mice received a sterile saline in the same manner. At 2, 3, 4, 5, 10 and 12 weeks after first HgCl2 injection, single splenic cells were stained with fluorochrome-conjugated mAbs for flow cytometer analysis. Our results showed that mercury treatment decreased the percentage of Treg (CD4+CD25+FoxP3+) in the spleen at time-points of 3–10 weeks, while increase in the percentage of B cells (CD19+CD3−) at 4–12 weeks. Mercury treatment did not affect the proportion of dendritic cells (DCs) in the spleen, including myeloid DCs (CD11chighMHCII+B220−) and plasmacytoid DCs (CD11clowmPDCA1+B220+). The dose and frequency of our HgCl2 injection did not affect body weight in the BALB/c mice. However, mercury transiently increased spleen weight at 2 and 3 weeks. Our findings suggest that mercury reduces Treg in the spleen, which may contribute to mercury-mediated autoimmunity.
Funding: Supported by the Grant from National Natural Science Foundation of China (81460503), and Research Funds from Jilin Provincial Science & Technology Department (20150101207ZC), Education Department of Jilin Province (JIJIAOKEHEZI2015No.3), Health and Family Planning Commission of Jilin Province (2014Z089).
The Ena/VASP family of cytoskeletal proteins selectively regulates activated T cell trafficking by promoting the diapedesis step of transendothelial migration.
Authors: Jacobelli Jordan , Estin Miriam L. , Traxinger Brianna , Fisher Marlie H. , Friedman Rachel S. 
Published: January 2023

Abstract
Vasodilator-stimulated phosphoprotein (VASP) and Ena-VASP-like (EVL) are cytoskeletal effector proteins that have been implicated in T cell actin cytoskeleton remodeling in response to T cell receptor signaling. However, the role of these proteins in T cell motility, transendothelial migration, and in vivo trafficking remained unknown. We investigated the role of EVL and VASP in T cell trafficking and found that these cytoskeletal effectors are selectively required for activated but not naïve T cell trafficking to the lymph nodes and spleen. Moreover, in a model of Multiple Sclerosis, EVL/VASP deficiency severely impaired activated T cell trafficking to the inflamed central nervous system (CNS) of mice with experimental autoimmune encephalomyelitis (EAE). Deletion of EVL and VASP resulted in impaired Very Late Antigen-4 (VLA-4) expression and function, but unexpectedly did not cause alterations in shear-resistant adhesion and motility of T cells on primary brain microvascular endothelial cells in vitro. Instead, deletion of EVL and VASP caused a significant impairment in the ability of T cells to initiate and complete the diapedesis step of transendothelial migration. Furthermore, upon blockade of VLA-4, T cell diapedesis became equivalent between control and EVL/VASP-deficient T cells. Together, these data suggest that that EVL and VASP selectively regulate activated T cell trafficking by specifically mediating the diapedesis step of transendothelial migration in a VLA-4-dependent manner.