bluuebunny/crossref_metadata_2025_split huggingface dataset:
Developing MHV-1 infection as an animal model for Severe Acute Respiratory Syndrome (SARS) (43.30)
Authors: Khanolkar Aaruni , Pewe Lecia , Tifrea Delia , Perlman Stanley , Harty John T. 
Published: January 2023

Abstract
SARS is a life threatening infectious disease that acquired epidemic proportions in late 2002 affecting 8000 people worldwide and claiming nearly 800 lives. It is caused by a novel group 2 coronavirus HCoV-SARS. No suitable small animal model currently exists for this disease. Developing a good animal model is crucial for generating effective anti viral therapies. Preliminary data indicate that intranasal infection of A/J mice with a related coronavirus MHV1 closely mimics the disease observed in humans infected with HCoV-SARS. Following infection these mice experience severe weight loss and the lungs show evidence of patchy mononuclear infiltrates and pneumonitis. Significant mortality is observed beginning around day 6–7 post infection suggesting an adaptive immune component to the pathogenesis of the disease. Interestingly, wild type BALB/c and B6 mice are relatively resistant to disease however, resistance is compromised in BALB/c mice lacking both perforin and IFNγ, and B6 mice deficient in receptors to IFNαβ. Our overall goal is to characterize the phenotype and function of the virus-specific T cell response in susceptible strains of mice to ascertain if a dysregulated T cell response contributes to morbidity and mortality. We believe this study will provide useful information regarding the pathogenesis of SARS and aid translational initiatives aimed at infection control and vaccine design
The T cell response to staphylococcal superantigens is modulated by the bacterial cell wall (43.33)
Authors: Madrenas Joaquin , Chau Thu , McCully Michelle L , Britnell Bill , Haeryfar Mansour , McCormick John K , Cairns Ewa , Heinrichs David E 
Published: January 2023

Abstract
Bacterial superantigens (SAgs) are pyrogenic exotoxins that cause oligoclonal T cell activation leading to toxic shock and death. The selective advantage of SAg expression by bacteria remains unknown. We hypothesized that the T cell response to SAg is modulated by the bacteria. If so, one should see intense T cell responses when SAg are introduced into the body alone, but not when the producing bacteria are also present.
We examined the responses of primary T cells to a panel of pure staphylococcal SAg or in the presence of heat-killed bacteria. We found that the IL-2 response to SAg was inhibited by the presence of bacteria, while the TNF-? and IFN-? responses were not affected or were even increased. Such a modulation of T cell responses to SAg was mediated by TLR2/6 agonists but not by agonists of TLR1/2, TLR-9 or NOD proteins, involved activation of the NF-_B pathway, and was prevented by anti-TLR-2 blocking antibodies. At the cellular level, this modulation resulted from death of antigen-presenting cells (APCs) but not of T cells. Thus, we show that the response to SAg is modulated by TLR2/6 agonists from the bacterial cell wall. Such a modulation involves the induction of APC death that then limits the capacity of SAg to cause massive T cell activation. Our findings provide an explanation for the long-standing question of selective advantage of SAg expression, and point to novel therapeutic strategies for SAg-associated diseases.
Role(s) of Pulmonary Macrophage and Dendritic Cells in SARS-CoV-Induced Host Inflammatory Responses: An in vitro Model. (43.39)
Authors: Yoshikawa Tomoki , Wang Nan , Peters Clarence J , Tseng Chien-Te K 
Published: January 2023

Abstract
SARS is a highly contagious respiratory disease with the lungs as a major target. Pulmonary macrophage (MΦ), airway epithelium, and dendritic cells (DC) are three key cells in host innate defenses against respiratory infections. Pulmonary MΦ are situated at the inner epithelial surface, whereas DC reside abundantly underneath the epithelium. Such strategic anatomic locations of these two effector cells within the airway system make it highly relevant and important to investigate their interrelationship with airway epithelial cells in the course of SARS coronavirus (SARS-CoV) infection. In this study, we established highly polarized human bronchial Calu-3 cell cultures in transwell membrane inserts which allowed us to harvest cytokines produced by the apical or the basolateral surfaces of SARS-CoV-infected Calu-3 cells. Here, we report that inflammatory cytokines released by infected Calu-3 cells differ between the two surfaces and are potent in modulating the intrinsic functions of both MΦ and DC. Specifically, they prompt the production of various inflammatory mediators by both MΦ and DC. However, they also significantly compromised the abilities of DC and MΦ in priming naive T cells and phagocytosis, respectively. These results suggest that the interplays among permissive lung epithelial cells, pulmonary MΦ and DC are critically involved in the regulation of inflammatory responses in the course of acute SARS-CoV infection and pathogenesis.
The L Antigen of the LCM Virus is a Target of Dominant CD8+ Responses (43.47)
Authors: Kotturi Maya , Peters Bjoern , Buendia-Laysa Fernando , Sidney John , Oseroff Carla , Botten Jason , Grey Howard , Buchmeier Michael , Sette Alessandro 
Published: January 2023

Abstract
CD8+ T cell responses control lymphocytic choriomeningitis virus (LCMV) infection in H-2b mice. Although antigen-specific responses against LCMV infection are well studied, we found that up to one third of the CD8+CD44hi response to LCMV in H-2b mice was not accounted for by known epitopes. We screened peptides predicted to bind MHC Class I, and overlapping 15-mer peptides spanning the complete LCMV proteome for their capacity to induce interferon-γ (IFNγ) production from CD8+ T cells derived from LCMV-infected H-2b mice. We identified thirteen novel epitopes. These together with the 7 previously known epitopes account for the total CD8+CD44hi response, diminishing the possible role for bystander T cell activation. Strikingly, 9 out of the 13 new epitopes were derived from the viral L polymerase, which, until now, was not recognized as a target of the cellular response induced by LCMV infection. The L epitopes induced significant levels of in vivo cytotoxicity and conferred protection against LCMV challenge. Interestingly, protection from viral challenge was best correlated with the cytolytic potential of CD8+ T cells, whereas IFNγ production and peptide avidity appear to play a lesser role. Taken together, these findings illustrate that the LCMV-specific CD8+T cell response is more complex than previously appreciated.
Bo-lysin: a potential candidate as a biomarker of protection after vaccination against tuberculosis (43.49)
Authors: Scherer Charles Fernando Capinos , Endsley Janice J. , de Aguiar Juliana B. , Waters Ray , Estes D.Mark 
Published: January 2023

Abstract
Tuberculosis (TB) is still a major health problem worldwide. A Th1 type response with release of IFNγ and cytotoxic granules such as granulysin and perforin, play a major role in the disease. Measurements of protection after TB vaccination include IFNγ measures and T cell proliferation assays, which frequently fail to correlate well with protection. The lack of better vaccines than BCG and good correlates of protection demands that new biomarkers are found. A bovine homologue of granulysin (bo-lysin), cloned by our group, may be a good candidate as a biomarker molecule for TB vaccination. In the present study we examined the kinetics of bo-lysin, compared to perforin, IFNγ and Fas-L in vitro, after different stimulation conditions in different bovine T cell populations. Gene expression profile showed bo-lysin and IFNγ expression increase in an equal manner, with IFNγ showing up as early as 4 hours of stimulation and Bo-lysin at 24 hours, in BCG vaccinated animals. No expression was detected in non-vaccinated animals. Detection of IFNγ and perforin by flow cytometry showed increasing percentages of antigen specific perforin+ or IFNγ+ CD4+ and perforin+ or IFNγ+CD8+ T cells in BCG vaccinated animals, but none were observed in non-vaccinated animals. Results show that granulysin is a good candidate as a biomarker of protection after vaccination against TB.
Project funded by grant #USDA-NRICGP-2005-35204-16039
Biomarkers of CD4+ CTL cell mediated immunity to tuberculosis (43.51)
Authors: Endsley Janice J , Hogg Alison , Shell Lis , McAlauy Martin , Scherer Charles Capinos , Coffey Tracey , Howard Chris , Nonnecke Brian , Waters W. Ray , Estes D. Mark 
Published: January 2023

Abstract
The immune responses mediated by interactions between T-lymphocyte subsets and mycobacteria-infected macrophages are critical for control of tuberculosis . In these studies, the bovine model was used to characterize the cytolytic and mycobactericidal CD4+ T cell response induced by BCG vaccination. Antigenic stimulation of CD4+ T-cells from BCG vaccinated cattle induced expression of perforin and IFNgamma in cells expressing a CD45RA−, CD45RO+, and CD62L+ cell surface phenotype. Antigen specific enhancement of granulysin, IFNgamma, perforin, IL-4, IL-13, and IL-21 mRNA expression was detected and not detected for IL-2, IL-6, IL-10, IL-15, TNFα, FasL, and CD40L. Following antigenic stimulation, CD4+ T cells from BCG vaccinated animals contributed to reduction of intracellular BCG in infected macrophages. These results demonstrate that vaccination with BCG induces a subpopulation of mycobacteria-specific CD4+ T cells that are characterized by the expression of a cell-surface memory phenotype, enhanced expression of mycobactericidal molecules, and anti-mycobacterial activity against intracellular M. bovis.
This work was supported by the NIAID Fellowship for Training in Emerging and Re-emerging Infectious Disease and the Sealy Center for Vaccine Development, UTMB, the Texas-United Kingdom Collaborative Research Initiative, and the Institute for Animal Health by DEFRA and the BBSRC, UK.
M. bovis-BCG elicits memory CD8+ T cell responses to mycobacteria in cattle (43.53)
Authors: Hogg Alison , Worth Andrew , Endsley Janice , Howard C.J. , Beverley P. , Villarreal-Ramos Bernardo 
Published: January 2023

Abstract
Tuberculosis (TB) remains one of the leading causes of death due to infectious disease. Current aims to improve vaccines against TB have focused on enhancing immunity conferred by the attenuated M. bovis BCG (BCG) vaccine. The design of a vaccine to specifically boost the protective features of BCG requires a greater understanding of the immune response induced by BCG vaccination. In this work we show that vaccination with BCG induces a population of CD8+CD45RO+ T cells that proliferate, produce IFN-gamma, up-regulate expression of perforin and granulysin, and lyse BCG-infected macrophages, following antigen specific stimulation. To determine whether a similar response could be detected in vivo, vaccinated animals were re-inoculated intradermally with BCG or PBS eight weeks after primary BCG vaccination. Draining lymph nodes were removed 3 days after the second inoculation for analysis by immunofluorescence and confocal microscopy for the presence of activated CD8+ T cells. Clusters of CD8+CD3+perforin+ cells were readily detectable in lymph nodes from animals re-inoculated with BCG but absent in the animals given PBS in the second inoculum. These results suggest that the memory CD8+ T cells induced by BCG vaccination have the capacity to mount secondary responses to mycobacteria in vivo.
Support for these studies was provided by the BBSRC, DEFRA and the Jenner Institute.
Cellular Immune Responses in a Murine Model of Borrelia burgdorferi and Babesia microti coinfection (43.55)
Authors: Vinasco Javier , Braga Walter , Zegarra-Moro Ofelia , Moro Manuel H 
Published: January 2023

Abstract
Active coinfection of Borrelia burgdorferi and Babesia microti has been suggested to result in an increased disease severity due to changes in immune response patterns. However, the immunological mechanisms involved in this processes are still unknown. The aim of this study is to characterize the cell immune responses during the progression of B. burgdorferi and B. microti coinfection. To address this issue, C3H.HeN and BALB/c mice cohorts were coinfected or singly infected and monitored for 1, 7, 10, 14, 21 and 60 days post-infection. Spleens and lymph nodes from infected and non-infected mice were removed for tissue culture experiments. Mononuclear cells were analyzed by flow cytometry assays using monoclonal antibodies. Interestingly, we found minimum changes among CD4+/CD3+, CD8+/CD3+, gamma-delta and NKT cells in C3H and BALB/c strains. In contrast, B cell populations were significantly augmented in BALB/c than C3H mice after infection. Furthermore, significant increased levels of B cells were also detected in coinfected BALB/c mice at days 14, 21 and 60. Spleen macrophage populations reached a peak at day 7 in both C3H and BALB/c strains for singly infected mice, in contrast with coinfected mice that showed significant decreased population of these cells. The reduction of spleen macrophages in coinfected mice may suggest an impair immune response during B.burgdorferi and B. microti coinfection resulting in an increased disease severity and persistence of Lyme disease. NIH P20RR016443-04.
Chronic Psychogenic Stress (predator exposure) Improved Survival and Altered the Cytokine/Chemokine Profile in Two Sepsis Models (44.11)
Authors: Dugan Amy , Gregerson Karen , Gardner Jason , Neely Alice , Buckley Donna , Noel Greg , Ogle Cora , Horseman Nelson 
Published: January 2023

Abstract
We determined the impact of chronic psychogenic stress (predator exposure) on sepsis-induced mortality and serum cytokine alterations. Male 7-week-old C57Bl/6 mice, individually housed, were exposed to either an unoccupied rat cage (non-exposed) or a rat cage containing 1 male Long Evans rat (predator-exposed), but with no physical contact between the mouse and rat. Exposure sessions lasted 1 hour and occurred on 9 consecutive days. Immediately before the 3rd exposure, non-exposed and predator-exposed mice received either saline, LPS (from E. coli, 20 mg/kg), or live E. coli (1 X 108 CFU) i.p. and were monitored for survival over the remaining 7 days. Predator exposure markedly improved survival rates after either LPS or bacteria injection. Mortality in the non-exposed mice (LPS) was 58%, but only 8% among the predator-exposed mice (p = 0.030). Following E. coli injection, mortality was 67% in the non-exposed mice, but only 17% in predator-exposed mice (p = 0.038). In a separate experiment mice were killed 24 hours after injection and serum cytokines were measured. Predator exposure attenuated the sepsis-induced increase of TNFα while enhancing IL-6. We hypothesize that predator-induced alterations in cytokine profiles during sepsis may be responsible for the observed survival advantage.
This work was funded by the Shriners of North America
Differential secretion of IL-12 and IL-18 based on intracellular localization of Listeria monocytogenes (44.2)
Authors: Meeks Karen D. , Sieve Amy N. , Balch Joshua B. , Berg Rance E. 
Published: January 2023

Abstract
Listeria monocytogenes (LM) is a gram-positive, intracellular bacterium that is commonly used to investigate the immune response to infection. Pro-inflammatory cytokines, such as IL-12 and IL-18, are secreted during LM infection to promote clearance of the bacteria through the induction of IFN-γ. LM secretes listeriolysin O (LLO) which allows the bacterium to invade the cytoplasm. Neither heat-killed LM (HKLM) nor LLO deficient LM (LLO−LM) actively produce LLO. When bone marrow derived macrophages (macs) and dendritic cells (DCs) are infected with LM, HKLM, or LLO−LM and co-cultured with naïve splenocytes, only macs or DCs infected with LM are able to induce IFN-γ secretion. ELISA data demonstrates that IL-12 is secreted from LM, HKLM, or LLO−LM infected macs and DCs, but IL-18 is only secreted from macs and DCs infected with LM. Therefore, IL-12 can be secreted by macs and DCs regardless of cytoplasmic invasion of LM whereas the secretion of IL-18 requires LM to gain access to the cytoplasm. When mice are infected with HKLM, there is no detectable IFN-γ secretion and this can be attributed to a lack of both IL-12 and IL-18. This implies that IL-12 is induced differently in response to HKLM in vivo versus in vitro. Further studies are ongoing to determine which pattern recognition receptors and cell types are responsible for recognizing and secreting IL-12 and IL-18 during in vivo LM infection.
Supported by NIH AI064592.
Listeria induces NKDC to produce IFN-gamma via IL-12, IL-18 and TLR9 ligation (44.24)
Authors: Plitas George , Chaudhry Umer I. , Kingham T. Peter , Stableford Jennifer , DeMatteo Ronald P. 
Published: January 2023

Abstract
Introduction: Natural killer dendritic cells (NKDC, NK1.1+CD11c+) are innate effector cells that secrete IFN-γ when stimulated. Initial control of Listeria monocytogenes (LM) infection depends on IFN-γ. The mechanism of IFN-γ secretion by NKDC in response to LM is unknown.
Methods: NKDC and NK (NK1.1+CD3−) cells were isolated by fluorescence activated cell sorting from the spleens of C57BL/6 mice and cultured with LM strain 10403s at a multiplicity of infection of 1 for 3 days. IL-18 (20ng/ml), IL-12 blocking antibody (10μg/ml), or an inhibitory CpG (iCpG) sequence (50μg/ml) was added. Supernatant IFN-γ was analyzed by cytometric bead array.
Results: LM infection or IL-18 alone did not induce significant IFN-γ production by NK cells or NKDC, however the combination of IL-18 and LM induced substantial amounts of IFN-γ from NKDC (19.3±2.7 ng/ml), and to a lesser extent, NK cells (2.0±0.6 ng/ml). Addition of anti-IL-12 antibody abrogated the IL-18/LM induced IFN-γ production of both NK cells and NKDC (&lt;0.4 ng/ml). The addition of iCpG led to an 85% reduction of IL-18/LM induced IFN-γ production by NKDC and a 20% reduction by NK cells.
Conclusion: IL-18, IL-12, and TLR9 ligation regulate NKDC IFN-γ production in response to LM.
This work was supported by grant DK068346.
Distinct functions of IKKα and IKKβ in the induction of type I IFN and proinflammatory cytokine genes following infection with Theiler’s murine encephalomyelitis virus (TMEV). (44.29)
Authors: So Eui Young , Kim Byung S 
Published: January 2023

Abstract
The initial inflammatory responses and antiviral activity against virus infections are largely mediated by cellular pattern-recognition receptors (PRRs). Among these PRRs, we have recently shown that TLR3 is critical for TMEV-induced cytokine and chemokine production in mouse astrocytes (Glia, 2006). The activation of NF-κB is also involved in these TMEV-induced signaling pathways. While TMEV infection stimulates induction of proinflammatory genes in mouse astrocytes by mechanisms mainly dependent upon TLR3, the steps involved in the association of TLR engagement to NF-κB activation during TMEV infection are unknown. In this study, we showed that the upstream NF-κB activators IKKα and β are directly involved in TMEV-induced innate immune responses. MEFs deficient in IKKα and β resulted in a greatly reduced production of IFN-α/ β, IL-6, and RANTES during TMEV infection compared to normal MEFs. Interestingly, activation of IRF3 following infection with TMEV was impaired in IKKβ-KO MEFs, but not in IKKα-KO MEFs. However, TMEV-induced activation of IRF7 and NF-κB was severely impaired in both IKK-deficient MEFs. Therefore, our study strongly suggests that both IKKα and IKKβ are pivotal components of TLR3-triggered immune responses during TMEV infection. However, the roles of IKKα and IKKβ appear to be distinct in the signaling pathway for these cytokine responses.
(Supported by NS33008 and NS28752).
Sensing and recognition of bacteria by NKT cells (44.3)
Authors: Kronenberg Mitchell , Kinjo Yuki , Tupin Emanual , Nagarajan Niranjana 
Published: January 2023

Abstract
Natural Killer T (NKT) cells are a conserved T lymphocyte subpopulation that participates in host defense against infections. NKT cell express an invariant TCR alpha chain and they recognize glycolipids presented by CD1d. We found that NKT cells also respond to IL-12 and IL-18 produced by APC that have been activated by TLR ligands. As a result, NKT cells rapidly produce IFN gamma that stimulates the innate immune response. By responding to activated APC, NKT cells can amplify the innate immunity to many different microbes.
The invariant TCR expressed by mouse and human NKT cells recognizes glycosphingolipids with alpha linked sugars, which are unique to Sphingomonas bacteria. Mice that lack NKT cells have reduced Sphingomonas clearance. Mouse and human NKT cells with an invariant TCR also recognize glycosyl diacylglycerols from Borrelia burgdorferi, which causes Lyme disease. Interestingly, the response to these compounds was highly dependent on the nature of the aliphatic chains, with the addition of a single unsaturated bond having an enormous influence on antigenic potency. NKT cells are activated during B. burgdorferi infection. These data provide evidence that NKT cells recognize a new category of microbial glycolipids, diacylglycerols, which is more broadly distributed in pathogenic microbes, and they suggest that this TCR-mediated recognition provides protection from microbial pathogens.
CCL2 and IL-10 produced by Gr-1+CD11b+F4/80+ cells as an inhibitor of β-defensin 3 production by epidermal keratinocytes (44.35)
Authors: Jones Vickie C , Shigematsu Kenji , Kobayashi Makiko , Herndon David N , Suzuki Fujio 
Published: January 2023

Abstract
We have previously reported that Gr-1+CD11b+F4/80+ cells demonstrated in tissues surrounding the burn site (TSBS) are inhibitory on the production of antimicrobial peptides by epidermal keratinocytes (EK). In this study, the mechanism by which Gr-1+CD11b+F4/80+ cells inhibit the production of β-defensin 3 (MBD-3) by EK was investigated. Burn mice were BALB/c mice exposed to a gas flame (3rd degree). Twelve hrs after burn injury, Gr-1+CD11b+F4/80+ cells were isolated from their TSBS using a magnetic bead technique. To determine the effect of Gr-1+CD11b+F4/80+ cells on MBD-3 production, LPS-stimulated EK prepared from skin tissues of normal mice (1 x 106 cells/ml, lower chamber) were cultured with Gr-1+CD11b+F4/80+ cells (upper chamber) in dual-chamber transwells. Culture fluids harvested were assayed for MBD-3 by ELISA. After stimulation with LPS (0.1 μg/ml), EK produced 25 μg/ml of MBD-3, and only 0.4 μg/ml of MBD-3 was produced by EK transwell-cultured with Gr-1+CD11b+F4/80+ cells. When the same transwell cultures were performed with media supplemented with a mixture of mAbs for CCL2 and IL-10 (2 μg/ml each), 36 μg/ml of MBD-3 was detected in the culture fluids. In contrast, MBD-3 production by EK stimulated with LPS was inhibited by a mixture of recombinant CCL2 (50 ng/ml) and IL-10 (10 ng/ml). CCL2 and IL-10 were detected in the culture fluids of Gr-1+CD11b+F4/80+ cells. These results indicate that, through the production of CCL2 and IL-10, Gr-1+CD11b+F4/80+ cells appearing in association with burn injuries inhibit the production of MBD-3 from EK.
This study was supported by SHC NA #8610.