bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human T cell responses to IL-6 and IL-27 are inhibited during cancer and chronic inflammation (P2019)
Authors: Miyahira Andrea , Simons Diana , Murad John , Lee Peter 
Published: January 2023

Abstract
In the complex interplay between an evolving tumor and its host, the functionality of the immune system is a key determinant in patient outcome. Optimal immune responses eliminate tumors while immune dysfunction allows disease progression. Cytokines play key roles in many immune functions known to be abrogated in cancer patients. We have previously shown that interferon responses are impaired in cancer patient lymphocytes. We hypothesized that additional alterations in cytokine signaling pathways occur in cancer patients and contribute to immune dysfunction. We used phospho-flow cytometry to assess if STAT phosphorylation in response to additional cytokines is impaired in peripheral blood immune cells from patients with various carcinomas or psoriasis, a chronic inflammatory skin condition. Compared with healthy controls, T cells from cancer and psoriasis patients exhibited significantly defective STAT1 and STAT3 responses to the gp130 cytokines IL-6 and IL-27, and correlating decreased expression of the gp130 and IL-6Rα receptors. Chronic exposure of healthy PBMCs to IL-6 led to down-regulated receptor expression and hyporesponsiveness to IL-6 and IL-27. Our studies indicate that that chronic inflammation which occurs in the settings of cancer and psoriasis contributes to T cell IL-6 and IL-27 hypo-responsiveness via exposure to well known systemically elevated IL-6 levels. We are exploring the functional consequences of these defects on T cell function in cancer.
Protective effects of orally administered fucoidan mix on immunosuppression by an experimental cancer chemotherapy (P2045)
Authors: Miyazaki Yoshiyuki , Nakamizo Masaji , Shibasaki Tetsuya , Kirino Tomomi , Saito Yuzu , Kawahara Kazuhiro , Otsuka Ken , Tachibana Hirofumi , Yamada Koji , Tachikawa Daisuke 
Published: January 2023

Abstract
Fucoidan are sulfated polysaccharides derived from brown seaweeds that mainly consist of L-fucose. Peoples in Asian countries eat seaweeds willingly as healthy foods. In previous studies, we have demonstrated that fucoidan augment anti-tumor immunity in an experimental mouse model and in healthy human subjects. However, availability of fucoidan to protect immune system from hazardous side effects of cancer chemotherapy has not been verified enough. Therefore, we performed a mouse model experiment in which immunosuppression was induced by subcutaneous 5-fluorouracil (5-FU) injection (1 mg/mouse/day, 5 times on alternate days) and evaluated protective effects of orally administered fucoidan mix (40 mg/mouse/day). The immunosuppression by 5-FU s.c. was confirmed with the loss in spleen weight of 5-FU-injected mice. NK cell activity was lower in 5-FU-injected mice than sham-treated control mice, but the decline was recovered by the fucoidan mix feeding. The intake of fucoidan mix significantly alleviated the reduction in interferon (IFN)-gamma production of splenocytes from 5-FU-injected mice. Furthermore, flow cytometric analyses revealed that specific population such as Th1 and CD11b+ cells were supported by the fucoidan mix feeding even in the 5-FU-injected mice. These results suggested that oral administration of fucoidan mix is effective to maintain tumor immunity during immunosuppressive cancer chemotherapy.
Deficiency in IL-4Rα signaling is associated with impaired tumor immunity (P2057)
Authors: Manzar Gohar , Kim Eun-Mi , Zavazava Nicholas 
Published: January 2023

Abstract
We recently made a seminal observation that Ym1, a chitinase-like lectin, activates NK cells, thereby promoting robust anti-tumor responses in vivo. This activation requires dendritic cells. We next sought to characterize the significance of endogenously expressed Ym1 in tumor immunity. In previous studies, Ym1 has been described as a marker for alternatively activated macrophages. Furthermore, the Th2-associated cytokines IL-4 and IL-13 were shown to induce Ym1 expression in macrophages. From these data, we hypothesized that IL-4Rα-/- mice are deficient in Ym1 and therefore more susceptible to tumor metastasis and growth. Indeed, IL-4Rα-/- mice exhibited reduced serum levels of Ym1. When WT or IL-4Rα-/- mice were i.v. challenged with the allogeneic murine melanoma cell line B16-F1, we observed a 12-fold increase in tumor burden in the lungs of IL-4Rα-/- mice compared to WT mice. Next, we generated bone marrow chimeras in which IL-4Rα-/- mice were adoptively transferred with either WT or IL-4Rα-/- bone marrow cells. Subsequently, the mice were challenged with B16-F1 cells and tumor burden evaluated after two weeks. Mice that received WT bone marrow exhibited significantly fewer tumor nodules in the lungs compared to control mice. In addition, their serum levels of Ym1 had been restored by the introduction of WT bone marrow cells. Altogether, these data indicate that IL-4 is important for tumor immunity via the induction of Ym1.
Immunogenicity assessment of in silico-selected T cell epitopes for a burkholderia biodefense vaccine (P4282)
Authors: Liu Rui , Desrosiers Joe , Martin Barbara , Moise Lenny , Groot Annie 
Published: January 2023

Abstract
Background: Burkholderia pseudomallei (BPM) and Burkholderia mallei (BM), the causative agents of Melioidosis and Glanders respectively, are Category B pathogens. Burkholderia cepacia (BC) causes severe disease in Cystic Fibrosis patients. No vaccines are available for these pathogens. We are developing a single Burkholderia vaccine that is effective for all three pathogens. Previously, we used informatics tools and HLA binding assays to select 70 promiscuous Class II T epitopes that are highly conserved in BPM, BM and/or BC. Here, an epitope-based vaccine is produced and its immunogenicity is evaluated in a mouse model. Methods: Epitopes were concatenated to form 3 multi-epitope gene constructs (PB1, PB2 and PB3) to produce DNA and protein vaccines. The VaxCAD algorithm ordered epitopes to minimize potential immunogenicity at epitope junctions. HLA DR3 transgenic mice were immunized by a DNA-prime/peptide- or protein-boost strategy. Immunogenicity was determined by IFNγ production to individual peptides by ELISpot assay. Results and Conclusion: Following DNA/peptide immunization, significant IFNγ production (p&lt;0.01) was found in PB3- but not PB1-and PB2-immunized mice. Notably, only PB3 concatameric protein successfully expressed. DNA/PB3 protein immunization studies are ongoing. We describe the step-wise process of developing genome-derived vaccines and identify solutions to the problem of engineering epitope-rich constructs for a broadly reactive Burkholderia vaccine.
A novel multi-stage subunit vaccine against paratuberculosis induces significant immunity and reduces bacterial burden in tissues (P4304)
Authors: Thakur Aneesh , Aagaard Claus , Riber Ulla , Skovgaard Kerstin , Andersen Peter , Jungersen Gregers 
Published: January 2023

Abstract
Effective control of paratuberculosis is hindered by lack of a vaccine preventing infection, transmission and without diagnostic interference with tuberculosis. We have developed a novel multi-stage recombinant subunit vaccine in which a fusion of four early expressed MAP antigens is combined with a MAP protein expressed in latent infection (FET11 vaccine). FET11 vaccine proteins were formulated with CAF01 adjuvant and injected to MAP challenged calves at two different ages. 28 calves divided into two FET11 vaccine groups, a commercial vaccine and a control group were used in the study and followed for a year. The FET11 vaccine induced a significant T cell response against constituent vaccine proteins characterized by a high percentage of CD4+ T cells and participation of polyfunctional CD4+ T cells. Of the two different age groups, late FET11 vaccination conferred protective immunity characterized by a significant containment of bacterial burden in gut tissues compared to non-vaccinated animals. There was no cross-reaction with bovine tuberculosis in vaccinated animals. This novel multi-stage vaccine has the potential to become a marker vaccine for paratuberculosis.
Antigen-specific B cells responding to pneumococcal polysaccharide vaccination in newly diagnosed HIV infected individuals. (P4264)
Authors: Leggat David , Khaskhely Noor , Iyer Anita , Westerink M.A. Julie 
Published: January 2023

Abstract
The peripheral blood of newly diagnosed HIV infected individuals is subjected to high uncontrolled viral titers. The decimation of both T and B cells thus leaves these individuals highly susceptible to opportunistic infections, including Streptococcus pneumoniae, despite the availability of vaccines. We have previously shown using directly labeled fluorescent pneumococcal polysaccharides that the majority of pneumococcal polysaccharide-specific B cells in healthy individuals were IgM+ memory B cells. Using this method, we identified specific deficiencies in B cell populations responding to pneumococcal polysaccharide vaccination in HIV infected individuals. Namely, a significant decrease in pneumococcal polysaccharide-specific IgM+ memory B cells compared to healthy individuals. Consequently, HIV infected individuals showed reduced antibody response and opsonophagocytic titers compared to healthy individuals. Furthermore, we correlated changes in pneumococcal polysaccharide-specific B cell populations post-vaccination with individual CD4 T cell counts. We observed a decreasing trend in pneumococcal polysaccharide-specific IgM+ memory B cells with lower CD4 T cell counts. These data indicate that the loss of pneumococcal polysaccharide-specific IgM+ memory B cells in HIV infected individuals correlates with CD4 counts and results in poor responsiveness to pneumococcal vaccination.
Recombinant Ehrlichia P29 protein, but not P13 protein, induces a protective immune response in a mouse model of ehrlichiosis (P4280)
Authors: Thirumalapura Nagaraja , Crocquet-Valdes Patricia , Saito Tais , Dadfarnia Tahereh , DiCicco Beau , Walker David 
Published: January 2023

Abstract
Ehrlichioses are emerging tick-borne bacterial diseases of humans and animals. The diseases are caused by obligately intracelluar bacteria belonging to the genus Ehrlichia. Currently, no vaccine is available against any ehrlichial diseases. Several immunoreactive proteins of ehrlichiae identified using immune sera from human patients and animals that include the major outer membrane proteins, ankyrin-repeat proteins and tandem repeat proteins (TRPs). In the present study, we evaluated E. muris P13 and P29 proteins, which are orthologs of E. chaffeensis TRP32 and TRP47, respectively, as subunit vaccines in a mouse model of ehrlichiosis. Immunization of mice with recombinant P29, but not P13, induced significant protection against a challenge infection. The protection induced by recombinant P29 was associated with the induction of a strong IgG antibody response. In contrast to development of IgG antibodies to P13 in all E. muris-infected mice, fewer animals infected with E. muris had IgG antibodies to P29. We hypothesize that polyclonal activation of B cells by CD4+ Th1 cells and subsequent development of hypergamaglobluinemia during persistent E. muris infection impairs formation of antibodies to antigens, such as P29, with low B cell precursor frequencies. In conclusion, our study indicated that Ehrlichia P29, but not P13, induces protective immune response, and induction of antigen-specific antibody responses impaired during persistent ehrlichial infections.
MicroRNAs induced during Mycobacterium leprae infection and their effects on immune response (P3019)
Authors: Mehta Manali , Liu Philip , Modlin Robert 
Published: January 2023

Abstract
MicroRNAs have been shown to be crucial in regulation of immune cell development and cancer progression. Recently, it has become increasingly clear that their involvement in regulation of immune responses to infection is also important. Here we use Mycobacterium leprae, the causative agent of leprosy, as a model to determine the potential roles of microRNAs in infectious disease. The immune response to M. leprae has been well-characterized as presenting as a spectrum, ranging from a contained infection characterized by a Th1 response in tuberculoid (T-lep) patients to a disseminated infection reflecting a Th2 response in lepromatous (L-lep) patients. We performed mRNA and microRNA microarray analysis on 6 T-lep and 4 L-lep lesions and subsequently integrated this data using Ingenuity. Preliminary gene network analysis of differentially expressed microRNAs in L-lep versus T-lep yielded multiple networks potentially targeted, including antigen presentation. A supervised analysis of Th1 and Th2 gene targeting indicates that microRNAs more highly expressed in L-lep lesions, such as miR-21 and miR-146a, may preferentially target proinflammatory genes required for a Th1 immune response. Further analysis to determine the functional effects of these microRNAs and their involvement in skewing towards a Th2 immune response to infection could explain the bias in disease outcome.
Activated CD8+ T cells exhibit blunted effector functions in early HIV-1 disease (P3035)
Authors: Crawford Timothy , Hecht Frederick , Ndhlovu Lishomwa , Barbour Jason 
Published: January 2023

Abstract
Elevated, persistent CD8+ T cell activation is a hallmark of HIV-1 infection. CD8+ T cell activation is measured by expression of CD38 and HLA-DR antigens on CD8+ T cells from the blood. How CD8+ T cell activation state may define impairments to CD8+ T cell function remains unclear. ERK1/2 signaling is critical for multiple T cell functions, and interruption of this pathway due to immune activation and inflammation may blunt CD8+ T cell function. We previously demonstrated that a major proportion of activated (CD38+HLA-DR+) CD8+ T cells from recently HIV-1-infected antiretroviral-treatment-naïve adults are refractory to phosphorylation of the ERK1/2 kinases (p-ERK1/2-refractory). Here we hypothesized that p-ERK1/2-refractory CD8+ T cells would exhibit reduced effector function compared to CD8+ p-ERK1/2-responsive cells. We combined single-cell phospho-kinase flow cytometry with intracellular cytokine staining into a single assay (PFICS), to examine IFN-γ and perforin responses in CD8+ T cells by ERK1/2 signaling ability. On a per cell basis, p-ERK1/2-refractory cells produced less IFN-γ in response to polyclonal or HIV-1 Gag peptide stimulation (% IFN-γ+ cells was 5.5-fold lower, p=0.006 for polyclonal and 1.5-fold lower, p=0.048 for Gag) and expressed reduced levels of perforin (2.1 and 1.6-fold lower perforin MFI). Blunted CD8+ T effector functions, secondary to ERK1/2 signaling deficits concentrated within activated CD8+ T cells, may contribute to HIV immunodeficiency.
Higher expression of several interferon-stimulated genes in HIV-1-infected females after adjusting for the level of viral replication (P3039)
Authors: Chang J. Judy , Woods Matt , Lindsay Robert , Doyle Erin , Griesbeck Morgane , Chan Ellen , Robbins Gregory , Bosch Ronald , Altfeld Marcus 
Published: January 2023

Abstract
Background. Clinical studies have shown faster disease progression and stronger immune activation in HIV-1-infected females as compared to males for the same level of HIV-1 replication. This study assesses whether the elevated levels of HIV-1-induced IFN-α production observed in females were associated with higher interferon-stimulated gene expression levels in T cells, and hence suggesting type-I-IFN as a mechanism for the higher HIV-1-associated immune activation observed. Methods. Fluorescence activated cell sorting was used to isolate T cells and dendritic cells from PBMCs of treatment-naïve chronically HIV-1-infected individuals enrolled in ACTG 384. The expression of 98 genes involved in toll-like receptor and type-I-IFN signaling pathways were quantified using Nanostring technology on sorted cell populations. Results. Several interferon-stimulated genes were significantly correlated with HIV-1 viral load and/or CD4+ T cell count. Higher expression levels of a subset of these interferon-stimulated genes were observed in cells derived from females compared to males after adjusting for viral load, and were correlated to higher levels of T cell activation. Conclusion. These data show that higher IFN-α production and resulting higher ex vivo expression of several interferon-stimulated genes in females can contribute to higher levels of immune activation, and subsequently the observed faster HIV-1 disease progression in females for a given level of viral replication.
B cell exhaustion in sickle cell disease patients is associated with inhibited memory generation after influenza vaccination (P3009)
Authors: Szczepanek Steven , Adami Alex , Louis Deirdre , Farrington Narye , Bracken Sonali , Natarajan Prabitha , Secor Eric , Jellison Evan , Thrall Roger , Andemariam Biree 
Published: January 2023

Abstract
Dysregulated immunity is a hallmark of Sickle Cell Disease (SCD), and clinical evidence indicates that SCD patients mount adequate initial antigen-specific antibody responses only to have significant drops in titer levels 1-2 years post-vaccination. We thus hypothesized that SCD patients do not generate sufficient B-cell memory after vaccination. We conducted a translational study (IRB #11-026-1) using flow cytometric analysis of memory B-cells from frozen PBMCs taken from 11 SCD patients and 9 healthy controls before and 4-6 weeks after receiving the influenza vaccine. Percentages of CD19+ B-cells were similar between groups before vaccination, but only SCD patients showed a significant increase in B-cells after vaccination (P=0.04). However, control subjects generated approximately 2-fold more CD27+ memory B-cells after vaccination (P=0.03) while almost no change was observed in the SCD patients. This indicates that SCD patients generate naïve, but little to no memory B-cells after vaccination, thereby explaining the diminishing antibody titers observed by clinicians. Further analysis demonstrated that SCD patients have a significantly higher proportion of FcRL4+ exhausted B-cells at baseline when compared to controls (33% more; P=0.02). Given that FcRL4 down-regulates BCR signaling and is associated with reduced memory B-cell generation in people with chronic infections, we propose that chronic inflammation in SCD leads to B-cell exhaustion through BCR signal dampening.
Pro-inflammatory response of human bronchial epithelial cells following Burkholderia cenocepacia infection: focus on IL-1β. (P1265)
Authors: Gillette Devyn , Shah Prexy , Cremer Thomas , Gavrilin Mikhail , Besecker Beth , Sarkar Anasuya , Knoell Daren , Cormet-Boyaka Estelle , Wewers Mark , Butchar Jonathan , Tridandapani Susheela 
Published: January 2023

Abstract
Burkholderia cenocepacia is the causative agent of the fatal lung disease, Cepacia syndrome. Epithelial cells are the first cells challenged in airway infections, yet, their pro-inflammatory response to B. cenocepacia remains under investigated. Here, we aimed to determine pro-inflammatory mechanisms initiated by airway epithelial cells following B. cenocepacia infection. We first examined signaling pathways activated post B.cenocepacia infection, and found that phosphorylation of Erk, Akt, Iκk, and NF-κB occurred. Cytokine profiles showed that while epithelial cells secreted TNFα, IL-6, and IL-8, they did not secrete IL-1β. Strikingly, qRT-PCR showed significantly elevated mRNA levels of IL1b. Examination of IL-1β within cell lysates revealed pro-IL-1β was present at significant amounts, but the mature form failed to be secreted. This inability to secrete IL-1β was not specific to B. cenocepacia, as LPS stimulation showed similiar results. Caspase-1 is required for pro-IL-1β processing and mature IL-1β release. Interestingly, we found that caspase-1 was expressed at minimal levels in both mRNA and protein forms within epithelial cells. Transfection experiments showed that restoration of caspase-1 expression in epithelial cells permits IL-1β secretion. Our data show, for the first time, that deficiency of fundamental inflammasome components, particularly caspase-1, may be responsible for the lack of IL-1β secretion by infected epithelial cells.
Glutathione reductase is essential for host defense against bacterial infection (P1239)
Authors: Liu Yusen , Yan Jing , Ralston Melissa , Meng Xiaomei , Bongiovanni Kathleen , Jones Amanda , Nelin Leif , Rogers Lynette , Smith Charles 
Published: January 2023

Abstract
Glutathione reductase (Gsr) catalyzes the reduction of glutathione disulfide to glutathione. We have recently shown that Gsr is essential for host defense against Escherichia coli in a mouse model of sepsis. While we have demonstrated that Gsr is required for the oxidative burst and the development of NETs, the role of Gsr in other phagocytic functions remains unclear. It is also unclear whether Gsr-null mice exhibit host defense defects against Gram-positive bacteria. Here we characterized the effects of Gsr deficiency on the innate immune responses to group B Streptococcus and LPS. Like E. coli, group B Streptococcus also resulted in a substantially more robust cytokine response, a markedly higher morbidity and mortality, and greater bacterial burden in Gsr-null mice than in wildtype mice. Interestingly, Gsr-deficient mice did not exhibit a greater sensitivity to LPS than did wildtype mice. Analysis of Gsr-null neutrophils revealed an impaired phagocytosis. In response to thioglycollate stimulation, Gsr-null mice mobilized far fewer phagocytes, including neutrophils, macrophages, and eosinophils, into their peritoneal cavities than did wildtype mice. The defective phagocyte mobilization is associated with heavy oxidation and aggregation of ascitic proteins, particularly albumin. Our results indicate that the oxidative defense mechanism mediated by Gsr is required for an effective innate immune response against bacteria, likely by preventing phagocyte dysfunction due to oxidative damage.
Mice lacking tissue factor in myeloid cells are susceptible to Mycobacterium tuberculosis infection (P1243)
Authors: Venkatasubramanian Sambasivan , Paidipally Padmaja , Barnes Peter , Mackman Nigel , Vankayalapati Ramakrishna 
Published: January 2023

Abstract
Tissue Factor (TF) is a trans membrane glycoprotein important for initiation of the coagulation cascade. Recent observations indicate that TF is expressed in cancer, metabolic diseases and infections. In the current study, we determined the role of TF in M. tuberculosis (M. tb) infection using mice lacking the TF gene in myeloid cells (Cre+). One month after infection with the M. tb virulent laboratory strain, H37Rv, the lung bacterial burden was higher in Cre+ than Cre- mice (3.7 ± 0.2 X 106 vs 1.9 ± 0.1 X 106 CFU, p=0.01). M.tb infected Cre+ mice had more CD4+ and CD8+ lymphocytes and expressed 33 and 38 fold less of IFN-γ and IL-17 mRNA in the lungs, compared to Cre- mice, as measured by real time PCR. In contrast, lung IL-10 mRNA levels were higher in M. tb-infected Cre+ mice than Cre- mice (16.04 ± 3.3 vs 4.4 ± 1 fold, p=0.03). Immunohistochemical analysis indicated that M. tb-infected Cre+ mice had more foamy macrophages in the lungs than Cre- mice. M. tbH37Rv grew more rapidly in peritoneal exudate macrophages of Cre+ mice, compared to Cre- macrophages (6.1 ± 1 X 106 vs 1.5 ± 0.6 X 106 CFU, p=0.02). Further studies indicated that M. tb-infected Cre+ macrophages undergo less mitochondrial apoptosis than Cre- macrophages (16.2 ± 2.1vs 27.9 ± 1.4 %, p=0.01). Studies are underway to determine the mechanism/s involved in increased IL-10 production, reduced mitochondrial apoptosis and increased bacterial burden of M tb infected Cre+ mice.