bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of effector B and T lymphocytes and cytokine production in the protective host response to vaccination with heat killed bacilli against Burkholderia mallei (43.14)
Authors: Whitlock Greg C. , Rowland Caroline , Lukaszewski Roman A. , Judy Barbara M. , Paessler Slobodan , Torres Alfredo G. , Estes D. Mark 
Published: January 2023

Abstract
Burkholderia mallei, the etiologic agent of glanders disease, is a gram-negative, intracellular bacterium for which no vaccine is available. Consequently, we are interested in defining protective host immune responses to B. mallei infection with the goal of developing candidate vaccines. Here, we report on initial cell/cytokine depletion studies to investigate the possible role these effectors may play in response to vaccination with heat killed bacilli in a susceptible BALB/c mouse model of infection. While protection with heat killed bacilli does not result in sterilizing immunity, protection is afforded against an otherwise lethal infection and provides insight into potential host protective mechanisms. Our results demonstrate that mice treated with either anti-B220, TNF-α or IFN-γ monoclonal antibodies exhibited decreased survival rates, indicating a key role for vaccine induced antibody and early cytokine production in protection from lethal challenge infection by the intraperitoneal route.
PD-1 plays a downmodulatory role in a murine immunodeficiency syndrome (43.26)
Authors: Green Kathy A , Okazaki Taku , Honjo Tasuku , Green William R. 
Published: January 2023

Abstract
An immunodeficiency syndrome is induced by infection of C57BL/6 (B6) with the murine leukemia virus LP-BM5. Other labs have shown that this syndrome requires CD4 T and B cell interactions. We have found it to be dependent on: CD154/CD40 and CD40 associated TRAF 6 and nondependent on CD80 and CD86. Gene chip analysis on spleen cells from LP-BM5 infected B6 mice revealed that the PD-1 and IL-10 genes are highly expressed after LP-BM5 infection. PD-1, a transmembrane cellular protein, was originally identified in a T cell line undergoing cell death and its ligation is generally considered to result in negative regulation of immune cell interactions and responses. We examined the role of PD-1 in LP-BM5 induced immunodeficiency via the use of B6.PD-1 k.o. mice and found that they develop substantially more severe disease than w.t. mice, as a function of; viral dose delivered in vivo and kinetics. PD-1 ligands, PD-L1, and PD-L2 have been identified, and flow cytometry shows that PD-L1 is preferentially upregulated during in vivo LP-BM5 infection. In in vivo antibody blocking studies, after LP-BM5 infection, anti-PD-L1, but not anti-PD-L2 treatment led to exaggerated disease. Other labs have reported that PD-L1 ligation of PD-1 leads to increased IL-10 production. We have found IL-10 k.o. mice are more susceptible to LP-BM5 induced immunodeficiency than w.t. mice. Thus, PD-1/PD-L1 and IL-10 pathways both appear to play roles in the moderation of LP-BM5 pathogenesis. Adoptive transfer experiments, using combinations of w.t., vs PD-1 k.o., CD4 T vs B cells to reconstitute LP-BM5 infected Rag 1 k.o. mice, suggest that B-cell expressed PD-1 is critical for the moderation of LP-BM5 induced pathogenesis.
Vaccinia Virus-specific CD4+ T cell Responses Target a Set of Antigens Largely Distinct from those Targeted by CD8+ T cell Responses (43.34)
Authors: Moutaftsi Magdalini , Bui Huynh-Hoa , Peters Bjoern , Sidney John , Salek-Ardakani Shahram , Oseroff Carla , Pasquetto Valerie , Crotty Shane , Croft Michael , Grey Howard , Sette Alessandro 
Published: January 2023

Abstract
Vaccinia virus (VACV) is a potent and effective vaccine, inducing long-lasting protective cellular immune responses. Recent studies defined the antigens recognized in mice and humans by CD8+ T cell responses. However, little is known about the specificities of CD4+ T cell responses, which makes it problematic to study and monitor cellular immune responses to VACV infections.
Herein, we report for the first time the identification of VACV-specific CD4+ T cell epitopes. By screening a large set of 15-mer peptides (2146 in total) in C57BL/6 mice, we identified 14 IAb-restricted T cell epitopes. These epitopes account for about 30% of the total anti-VACV CD4+ T cell response and are derived from 13 different structural and viral genome regulation proteins. Surprisingly, no CD4+ T cell epitopes were identified that target virulence factors. While early antigens are recognized, late antigens predominate as CD4+ T cell targets. These results are in sharp contrast to what was previously found in CD8+ T cells responses, where early antigens including virulence factors are prominently recognized.
Taken together, these results highlight fundamental differences in immunodominance of CD4+ and CD8+ T cell responses to a complex pathogen and have potential implications for the development of subset vaccines.
Funded by the NIH: Contract # HHSN266200400024C and RO1 grant # RO1-AI-56268. Kirin Publication Number: 837
Cervicovaginal epithelial cell cytokine production in response to microorganisms associated with bacterial vaginosis (43.40)
Authors: Trama Jason , Libby Erika , Mordechai Eli , Adelson Martin 
Published: January 2023

Abstract
Bacterial vaginosis (BV) is the most common vaginal disorder among women of reproductive age. However, the pathogenesis of BV is poorly understood. Although direct causality has not been demonstrated, local epithelial cells responding to microorganisms are proposed to be the source of elevated IL-1β, IL-6 and IL-8 in the vaginal washes of women with BV. Here, we determine the inflammatory cytokine responses of immortalized cervical and vaginal epithelial cells (CVEC) when co-cultured with lactobacilli and BV-associated microorganisms including Atopobium vaginae, an anaerobic, gram-positive bacterium common in women with heterogeneous vaginal flora and more specific for BV than Gardnerella vaginalis. Dose-dependent induction of IL-8 was seen when CVEC were co-cultured with A. vaginae, but not when CVEC were co-cultured with lactobacilli or G. vaginalis at MOIs of up to 100. The induction of IL-8 was similar in endocervical, ectocervical and vaginal epithelial cells. The release of IL-8 reached a maximum by 4 hours, was sustained for 24 hours and was abrogated by heat treatment of A. vaginae. Examination of IL-1β, IL-6, IL-8, IL-10 and TNFα secretion by bead array showed induction of IL-6 and IL-8 by A. vaginae. These results suggest that A. vaginae and CVEC are contributing factors to the cytokine production and the inflammatory sequelae associated with BV.
Supported by Medical Diagnostic Laboratories LLC.
Nippostrongylus brasiliensis infection modulates Mycobacterium tuberculosis induced Th1 response (43.45)
Authors: bhatt kamlesh , Liu Zhugong , Gause W C , Salgame P 
Published: January 2023

Abstract
Modulation of Th1 response generated against pathogens like M tuberculosis (Mtb) by helminths is an important, area of investigation, because of the prevalence of coinfections in TB endemic regions. In this study we have designed an approach to investigate how the Th1 response generated by Mtb is modulated by helminthic infection. We have used the ovalbumin – specific TCR transgenic adoptive transfer system, wherein the ability of Mtb-pulsed dendritic cells to initiate a Th1 response was studied on the background of a helminth infection. BALB/c mice were infected with N. brasiliensis, and groups of uninfected and infected mice were injected with CFSE labeled OVA-specific TCR transgenic T cells followed by intratracheal instillation of Mtb and ovalbumin peptide-pulsed bone marrow-derived dendritic cells. Analysis of the labeled transgenic T cells harvested from the draining lymph nodes indicated proliferation of T cells from both groups of mice. However, we observed significantly reduced level of IFNγ and a reciprocal increase in IL-4 in T cells derived form the helminth-infected mice. These findings indicate that helminth infection does not abrogate the priming ability of Mtb-pulsed DCs, but interferes with their ability to promote a Th1 response. The significance of the findings lies in the potential of helminth infections to modulate resistance to tuberculosis, and also to interfere with the efficacy of tuberculosis vaccines.
Type I interferon and IL-12 induce phenotypic variegation of human CD4+ T cells (43.7)
Authors: Davis Ann M , Matthews Loderick A , Ramos Hilario J , Farrar J David 
Published: January 2023

Abstract
Antigen presenting cells secrete distinct patterns of innate cytokines upon activation via Toll-like receptors (TLRs). These innate cytokines shape adaptive immune responses to pathogens. For example, interleukin 12 (IL-12) is produced in response to bacterial recognition via TLRs 2, 4, 5, and 6. IL-12 induces CD4+ T helper 1 (Th1) cell development through the activation of signal transducer and activator of transcription 4 (STAT4), leading to the secretion of interferon gamma (IFN-γ). Conversely, during viral infection, signaling through TLRs 3, 7, and 9 results in the secretion of both IL-12 and type I interferon (IFN-α/β). The effects of combined IL-12 and IFN-α/β stimulation on T helper development are not well-studied. We have demonstrated that activation of naïve CD4+ T cells in the presence of IFN-α/β alone fails to induce Th1 development. However, a combination of IL-12 and IFN-α/β induces phenotypic variegation to generate two populations of T helper cells. An effector population secretes both IFN-γ and IFN-α/β, and these cells are efficient in eliminating viral infections in vitro. A second subset displays memory properties and may be involved in long-term viral immunity. Furthermore, these two subsets can be distinguished by expression of chemokine receptors. These data have important implications for the understanding of viral infections as well as for the development of novel vaccines and therapies.
Inhibition of dendritic cell IL-12 production by Giardia lamblia (44.1)
Authors: Kamda Joel D. T. , Singer Steven M. 
Published: January 2023

Abstract
Unlike most other enteric infections, Giardia lamblia infection does not typically induce mucosal inflammation. We therefore studied the response of bone marrow-derived dendritic cells (BMDCs) to Giardia extracts. G. lamblia is only a weak activator of BMDCs. Compared to LPS, co-incubation of G. lamblia extract results in only moderate up-regulation of the co-stimulatory molecules CD80 and CD86. In addition, G. lamblia extract causes minimal induction of IL-6 and TNF and no detectable IL-10 or IL-12. Interestingly, pre-exposure of BMDCs to G. lamblia extract inhibits the secretion of the pro-inflammatory cytokine IL-12 by LPS activated BMDCs. IL-12 suppression by G. lamblia is apparently dependent on its activation of the phosphotidylinositol 3-Kinase (PI3-K) pathway, as it is abrogated by pretreatment of BMDCs with the specific PI3-Kinase inhibitor, wortmannin. These results suggest that Giardia actively prevents inflammation, and suggests a distinct mechanism of immune regulation by this intestinal parasite.
The pattern recognition molecule mindin promotes intranasal clearance of influenza viruses. (44.10)
Authors: Jia Wei , He You-Wen 
Published: January 2023

Abstract
The innate immune response is essential for host defense against microbial pathogen infections and is mediated by pattern recognition molecules recognizing pathogen associated molecular patterns (PAMPs). Our previous work has demonstrated that the extracellular matrix protein mindin functions as a pattern recognition molecule for bacterial pathogens. Here we examined the role of mindin in influenza virus infection. We found that intranasal infection of mindin-deficient mice by influenza viruse resulted in dramatically increased virus titers in the lung and intranasal cavity of mutant mice. In contrast, lungs from intratracheally infected mindin-deficient mice contained similar influenza virus titers. We showed that mindin interacted with influenza virus particles directly and that mindin-deficient macrophages exhibited impaired activation after influenza virus infection in vitro. Furthermore, intranasal administration of recombinant mindin significantly enhanced the clearance of influenza virus in wildtype mice. Together, these results demonstrate that mindin plays an essential role in host innate immune response to influenza virus infection and suggest that mindin may be used as an immune enhancing agent in influenza infection.
1. This work was supported by NIH grants AI054658 and AI061364.
Role of dectin-1 in the host defense to infection with Cryptococcus neoformans (44.17)
Authors: Miyazato Akiko , Nakamura Kiwamu , Saijo Shinobu , Iwakura Yoichiro , Kaku Mitsuo , Kawakami Kazuyoshi 
Published: January 2023

Abstract
pathogen-associated molecular patterns (PAMP) are recognized by Toll-like receptors (TLR) and C-type lectin recptors including Dectin-1. Previously, we indicated that neither TLR2 nor TLR4 was involved in the host defence to infection with Cryptococcus neoformans, an opportunistic fungal pathogen in AIDS patients (FEMS Immunol. Med. Microbiol. 47: 148–154, 2006). In the current study, we examined the role of Dectin-1, a receptor for β-glucan, in this response. Dectin-1-deficient mice were resistant to intratracheal and intravenous infection with C. neoformans at a comparable level to wild-type mice. IFN-γ production in lung an serum was not largely different between these mice. There was not significant difference in the synthesis of IL-12p40 and TNF-α by bone marrow-derived dendritic cells upon stimulation with this fungal pathogen. Taken together, these results demonstrated that Dectin-1 did not play a major role in the host protective responses to C. neoformans infection.
This work was supported in part by a Grant-in -Aid for Science Research (C) (KAKENHI18590413) from the Ministry of Education, Culture, Sports, Science and Technology and by Tohoku University 21st COE program ′CRESCENDO′
LPS deacylation by a host enzyme shortens endotoxin tolerance (44.20)
Authors: Lu Mingfang , Munford Robert 
Published: January 2023

Abstract
Prior exposure to bacterial lipopolysaccharide (LPS) alters animals’ responses to a second LPS exposure. Although this reprogramming (also called endotoxin tolerance) may prevent harmful inflammatory reactions, it also may decrease resistance to subsequent infection. How animals recover from tolerance is not known. Acyloxyacyl hydrolase (AOAH) is a host lipase that inactivates LPS by cleaving the secondary fatty acyl chains from lipid A. We found that 21 days after an initial dose of LPS, long after the responses of normal mice had returned to baseline, Aoah−/− mice had diminished serum IL-6 and TNF responses to LPS challenge. Ten days after receiving LPS i.p., macrophages explanted from Aoah−/− mice and stimulated ex vivo with LPS produced less IL-6 and TNFα than did macrophages from LPS-primed control mice. Whereas the LPS that was associated with peritoneal cells of Aoah+/+ mice was completely deacylated by 10 days, LPS remained undeacylated in Aoah−/− peritoneal cells. LPS-primed Aoah−/− mice were unable to control bacterial growth and therefore were more susceptible to lethal E. coli infection. LPS inactivation by AOAH shortens the duration of tolerance and its associated immunosuppression. A better understanding of how tolerance is terminated may help ameliorate post-infection immunosuppression in humans.
Supported by NIAID grant AI18188.
Role for platelet toll-like receptor 4 (TLR4) in the formation of Neutrophil Extracellular Traps (NETs) in sepsis (44.22)
Authors: Ma Adrienne , Clark Stephen , Tavener Samantha , Patel Kamala , Chakrabarti Subhadeep , Sinclair Gary , Keys Elizabeth , Allen-Vercoe Emma , DeVinney Rebekah , Doig Christopher , Green Francis , Kubes Paul 
Published: January 2023

Abstract
It has been known for many years that LPS, neutrophils and platelets all participate in the pathogenesis of severe sepsis, but the inter-relationship between these players is completely unknown. Using flow chambers in vitro we suggest a novel innate immune response leading to enhanced trapping of bacteria in blood vessels. The mechanism involved platelet TLR4 detecting LPS in blood and inducing a unique response, specifically platelet binding to adherent neutrophils, but not platelet aggregation or P-selectin expression. Subsequently, the platelets stimulated very robust neutrophil activation leading to formation of NETs. These NETs retained their integrity under flow conditions and functioned to ensnare bacteria within the vasculature. Plasma from severely septic patients also induced TLR4-dependent platelet-neutrophil interactions leading to the production of NETs. We propose that this novel bacterial trapping mechanism would only occur under extreme conditions such as severe sepsis and platelet TLR4 (not leukocyte TLR4) functioned as the threshold switch for this innate immune response to occur. With the advent of antibiotics perhaps reducing the need for NET formation, we would propose that inhibiting platelet activation with TLR4 inhibitors may inhibit NET formation and reduce inadvertent tissue injury.
This work was funded by: CIHR and AHFMR
CCL2 antisense therapy improves the resistance of burn mice to Enterococcus faecalis translocation (44.23)
Authors: Shigematsu Kenji , Tsuda Yasuhiro , Kobayashi Makiko , Herndon David N , Suzuki Fujio 
Published: January 2023

Abstract
M2Mϕ are inhibitors of the Mϕ conversion from resident Mϕ to M1Mϕ, an antibacterial effector cell in innate immunities. CCL2 has been described as an inducer of M2Mϕ. To manipulate the inhibitory function of M2Mϕ on the generation of M1Mϕ, in this study we performed CCL2 antisense therapy in burn mice. CCL2 antisense ODN was administered s.c. to mice 2 hrs after burn injury (3rd degree, 25% TBSA burn). One day after burn injury, Mϕ were prepared from mesenteric lymph nodes of these mice, and cultured in transwells with a 10x higher number of resident Mϕ (1 x 106 cells/ml, lower chamber) in media supplemented with CpG DNA (10 μg/ml). To detect the M1Mϕ generation, the culture fluids obtained were assayed for CCL3 (a typical biomarker for M1Mϕ). In some experiments, decontaminated burn mice were infected orally with a lethal dose of E. faecalis (107 CFU/ml), and treated s.c. with CCL2 antisense ODN once daily for 5 days. In the results, M1Mϕ were not generated from resident Mϕ transwell-cultured with M2Mϕ even when they were stimulated with CpG DNA. However, M1Mϕ were generated when the same transwell-cultures were performed with resident Mϕ and Mϕ from burn mice that were previously treated with CCL2 antisense ODN. After E. faecalis infection, 75% of burn mice died, while 80% of burn mice subjected to CCL2 antisense gene therapy survived. These results indicate that M1Mϕ are inducible in burn mice subjected to CCL2 antisense therapy, and the resistance of burn mice to E. faecalis translocation is improved after this antisense gene therapy.
This study was supported by SHC NA #8840.
Inhibitory effect of glycyrrhizin on the expression of c-fos by an immunosuppressive subset of murine neutrophils (PMN-II) (44.30)
Authors: Kobayashi Makiko , Yoshida Tsuyoshi , Herndon David N , Suzuki Fujio 
Published: January 2023

Abstract
The root of Glycyrrhiza glabra (licorice) and its principle component (glycyrrhizin) have revealed various immunomodulatory properties that fight off attacking pathogens. In our previous studies, glycyrrhizin was shown to prevent infectious complications developed in mice with systemic inflammatory response syndrome (SIRS) through the inhibition of CCL2 production by PMN-II. c-fos (one of the nuclear transcription factors) is known as a CCL2 gene promoter. Therefore, in the present study, c-fos mRNA expression in PMN-II treated with or without glycyrrhizin was analyzed. For these studies, SIRS was induced by a flame burn injury (3rd degree, 25% TBSA burns). PMN-II were isolated from the peripheral blood of mice 18 hrs after burn injury by Ficoll-Hypague and dextran sedimentations, followed by the depletion of contaminating monocytes using magnetic beads coated with anti-F4/80 mAb. In the presence of Staphylococcus aureus antigen (0.0075%), PMN-II (1 x 106 cells/ml) were cultured with various doses of glycyrrhizin for 12 hrs, and the culture fluids obtained were assayed for CCL2. Also, the obtained cells were analyzed for c-fos mRNA by RT-PCR. In the results, 3.7 ng/ml of CCL2 was detected in the culture fluids of PMN-II, while PMN-II treated with glycyrrhizin did not produce CCL2. c-fos mRNA was expressed in PMN-II, but not in PMN from normal mice. When PMN-II were treated with 10 μg/ml of glycyrrhizin, c-fos mRNA expression was greatly decreased. These results suggest that c-fos expression in PMN-II may be controlled by glycyrrhizin when CCL2 production by these PMN is inhibited by the compound.
This study was supported by SHC NA #8840.
TLR3 and TLR9 mRNA expression is impaired in macrophages from severely burned mice (44.31)
Authors: Kobayashi Makiko , Yoshida Tsuyoshi , Herndon David N , Suzuki Fujio 
Published: January 2023

Abstract
Classically activated Mϕ (CAMϕ) are not generated in mice with severe burn injuries even when they are stimulated with a mixture of poly I:C and CpG DNA (well-known inducers for CAMϕ). To investigate why the Mϕ conversion from burn mouse-derived Mϕ to CAMϕ was not induced after stimulation with a mixture of poly I:C and CpG DNA, the expression of receptors for poly I:C (TLR3) and CpG DNA (TLR9) in burn mouse-derived Mϕ were examined. BALB/c mice exposed to a flame burn injury (3rd degree, 25% TBSA burns) were used for these studies. Mϕ were prepared from the peritoneal cavities of mice 6, 24, 48, 120 and 168 hrs after burn injury using magnetic beads coated with anti-F4/80 mAb. Peritoneal Mϕ from unstimulated normal mice were utilized as a control. Then, Mϕ preparations obtained were analyzed for TLR3 and TLR9 mRNA expression using RT-PCR. In the results, TLR3 and TLR9 mRNAs were shown to be expressed by Mϕ from normal mice. However, TLR3 mRNA expression was not demonstrated in Mϕ from mice 6 to 168 hrs after burn injury. TLR9 mRNA expression was strongly impaired in Mϕ from mice 6 hrs after burn injury, and then it gradually recovered until 120 hrs after burn injury. These results indicate that burn mouse-derived Mϕ are shown to not be a carrier of TLR3 and TLR9. The unresponsiveness of burn mouse-derived Mϕ to the CAMϕ inducers may be related to the impaired expression of TLR3 and TLR9 that are receptors for poly I:C and CpG DNA, respectively.
This study was supported by SHC NA #8550.
Loss of TLR7 receptor expression in HCV viral infection as a potential mechanism to evade the innate immune system (45.13)
Authors: Chang Serena , Szabo Gyongyi 
Published: January 2023

Abstract
Hepatitis C Virus (HCV), a single stranded RNA virus that causes chronic inflammation in the liver, has evolved ways to interfere with the innate and adaptive immune system for survival. Recent studies show that Toll-like Receptor (TLR) 7 agonists decrease HCV expression in patients and infected cell lines, through a Type 1 IFN-mediated pathway. As it is unknown whether HCV interferes with TLR7 or its pathway, we investigated changes in TLR7 expression during HCV infection. Compared to 75% of the positively stained TLR7 expressing controls, human hepatoma cells infected with the replicating JFH-1 HCV clone, showed only 25% TLR7 expression by flow cytometry. Cells expressing full-length or sub genomic HCV replicons followed the same pattern as JFH-1 with 11% and 18% TLR7 expression compared to the wild-type expressing 56% and 42%. Micro array analysis of HCV infected livers showed a decrease in TLR7 mRNA expression suggesting in vivo correlation. TLR7-mediated signaling involves activation of IRF7 and IRF3 for type I IFN induction. IRF7 nuclear protein levels in JFH-1 cells were equivalent to control cells, as opposed to a drastic reduction in IRF3 levels and IP-10, an IFN-inducible gene, in JFH-1 cells vs. control cells. These results demonstrate an attenuation of TLR7 expression in Hepatitis C Virus infection that may represent a novel mechanism to prevent sufficient innate immune responses in HCV elimination.