bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expansion of influenza M1-specific memory cells cross-reactive with Epstein-Barr Virus (EBV) lytic epitopes correlates with severity of EBV-induced infectious mononucleosis (130.14)
Authors: Aslan Nuray , Watkin Levi , Pishtari Jonila , Xu Huiqing , Sullivan John L , Luzuriaga Katherine 
Published: January 2023

Abstract
Cross-reactive memory T cells specific to previously encountered influenza A contribute to the lymphoproliferation characteristics of EBV-associated infectious mononucleosis (IM). We showed that cross-reactive CD8 T cell responses commonly occur during EBV infection. Strong cross-reactive T cell responses specific to influenza M1 (Flu-M1) and EBV-specific lytic epitope EBV-BRLF-1 have been observed in 10/10 HLA-A2+ IM patients. We also detected cross-reactive T cell responses that recognized both Flu-M1 and lytic EBV-BMLF-1 epitopes in 14/26 HLA-A2+ IM patients. In addition, 8/19 IM patients demonstrated some intra-viral cross-reactivity between EBV BRLF-1 and BMLF-1 specific epitopes. Cross-reactive T cell responses between Flu-M1 and EBV-BRLF-1/EBV-BMLF-1 epitopes were observed in all IM patients but at different time points after infection. Our analyses showed that Flu-M1 and EBV-BMLF-1 used few Vbeta TCR families that were stable overtime. However, EBV-BRLF-1-specific Vbeta TCR repertoire used multiple Vbeta families that varied between individuals and varied during the time course of IM in the same patient. Disease severity of IM directly correlated with percentage of Flu-M1-specific CD8 T cells but did not correlate with EBV-BMLF-1 or BRLF-1-specific responses.
Human T cell immunity to influenza: magnitude, breadth, and antigenic variation (130.17)
Authors: Subbramanian Ramu A. , Brady Rebecca , Basha Saleem , Shata Tarek , Bernstein David I 
Published: January 2023

Abstract
Unraveling the nature of T cell immunity to influenza is currently an active area of research. Several lines of evidence now suggest that cell mediated immunity plays a significant role in controlling influenza disease in humans. While influenza hemagglutinin (HA)-specific humoral responses have been studied in considerable detail, our understanding of the coexistent cellular responses elicited by HA in humans is woefully limited. Utilizing quantitative approaches including ELISPOT and multiparametric flow cytometry we determine the magnitude of cell mediated responses to HA, the lineage of the responding T cells, and their frequency of prevalence in a tightly-defined cohort of healthy subjects. We demonstrate that the magnitude of HA-specific T cell memory populations are maintained at a low, finite frequency and vary significantly among individuals. HA-specific humoral- and T cell immunity do not correlate with each other. Importantly, the contribution of the T cell responses elicited by the variant and conserved regions of HA varies significantly among high- and low-responders. The ramifications of these observations to our understanding of influenza immunity in humans and development of effective vaccines against this pathogen will be discussed.
Effect of plant extracts on epithelial cell response to influenza infection (130.6)
Authors: Warren Kristi J , Naik Shibani , Kohut Marian 
Published: January 2023

Abstract
Plant extracts derived from Echinacea and Prunella have been used to treat and/or prevent various ailments including respiratory infection. In this study we examined whether plant extracts obtained from Echinacea purpurea (E.pur), E. angustifolia (E.ang), E. pallida (E.pal) or Prunella (Prun) altered cytokine and chemokine response to influenza infection of human bronchial epithelial cells (A549). A549 cells were treated with the plant extracts for 2 hours and subsequently exposed to virus (~ 40 HAU of Influenza A/PR/8/34) or media alone. Cell supernatants were collected 24 hrs later and assayed for cytokines/chemokines with a multiplex assay. The results showed that two extracts (E. ang & E. pal) increased IL-6, IL-8, G-CSF, IP-10 and RANTES, in cell cultures of media alone. In cultures with influenza virus, IL-1β, IL-15 and MIP1β were increased by E. ang, whereas IL-6, IL-8, IP-10 & RANTES were increased by all four plant extracts. These findings suggest that the plant extracts may have direct effects on epithelial tissue in addition to the known effects on monocytes and macrophages. It is possible that the early induction of these cytokines/chemokines promotes an earlier or more potent innate immune response.
Identification of a novel antigen presenting cell population modulating anti-influenza type-2 immunity (131.3)
Authors: Yoo Jae-Kwang , Galligan Carole , Burke Daniel , Virtanen Carl , Fish Eleanor N 
Published: January 2023

Abstract
Anti-influenza type 2 (T2) immunity contributes to both immunoprotection and immunopathology, yet the underlying mechanisms modulating T2 immunity remain ill-defined. Here, we describe a novel murine antigen (Ag) presenting cell (APC) population, designated virus activated plasmacytoid (VAP) cells: mPDCA1+CD11c-TcR-β-B220-CD19-CD38+CD44intCD45+Gr1+. VAP cells are bone marrow derived leukocytes. Phenotypic, morphologic and molecular characterization distinguished VAP cells from other immune cell populations. Pulmonary influenza A virus infection activates circulating VAP cells to enter the lungs and produce interleukin (IL)-25, a strong inducer of T2 pulmonary inflammation. Subsequently, VAP cells capture viral Ag in the lungs and migrate to the draining lymph node (DLN) and spleen, with delayed kinetics relative to DCs, where they show APC activity. In the DLN, influenza virus activated VAP cells produce IL-4 and induce T helper (TH) 2 effector cell polarization by modulating GATA-3 expression. Finally, adoptive transfer of influenza virus activated VAP cells enhanced TH2 effector T cell responses in influenza virus infected recipient mice and subsequently, anti-influenza T2 humoral responses including IgG1 and G2b were significantly increased. Collectively, our results highlight the importance of VAP cells as novel immuno-modulators of T2 immunity against influenza A virus infection in mouse.
These studies were supported by a Canadian Institutes of Health Research grant MOP-15094 to E.N.F.
J.K.Y. was supported by a Connaught Scholarship and an Ontario Graduate Scholarship in Science and Technology.
The Pseudomonas aeruginosa toxin Cif reduces MHC class I antigen presentation and viral immunity (131.4)
Authors: Bomberger Jennifer , Ely Kenneth , Bangia Naveen , O'Toole George , Green William , Enelow Richard , Stanton Bruce 
Published: January 2023

Abstract
Pseudomonas aeruginosa is an opportunistic pathogen critical to the pathogenesis of nosocomial infections, chronic obstructive pulmonary disease, ventilator-associated pneumonia, bronchiectasis and Cystic Fibrosis. Patients chronically infected with P. aeruginosa often experience debilitating viral exacerbations, the cause of which is unknown. Previously, we demonstrated that Cif, a bacterial toxin secreted by P. aeruginosa, promotes the degradation of several ABC transporters involved in innate immunity and the drug bioavailability, including CFTR and Pgp. The current study was conducted to test the hypothesis that Cif enhances the degradation of the transporter associated with antigen processing (TAP) in airway epithelial cells, resulting in reduced class I MHC antigen presentation and therefore, diminished immune responses to viral pathogens. Cif inhibited the deubiquitinating enzyme, ubiquitin specific protease 10, resulting in a time-dependent increase in the polyubiquitination and proteosomal degradation of TAP1. The Cif-mediated decrease in TAP1 expression resulted in a decrease in class I MHC expression at the plasma membrane. Furthermore, Cif decreased MHC class I antigen presentation and cytotoxic T lymphocyte (CTL) recognition of influenza virus antigen, as measured by TNF-α secretion, in a time-dependent manner. These observations suggest that P. aeruginosa, by secreting Cif, inhibits the ability of CTL to eliminate viral infections, severely compromising the immune defenses of the lung.
Human plasmacytoid dendritic cells preferentially conjugate with and fuse with acute or chronically HIV-IIIB infected cells. (131.7)
Authors: Jacobs Evan S , George Thaddeus C , Singh Sukhwinder , Wnek Richard , Fischer Paul , Nordone Shila K , Dean Gregg A , Bocarsly Patricia Fitzgerald 
Published: January 2023

Abstract
Plasmacytoid Dendritic Cells (PDC) produce copious amounts of IFN-a in response to TLR-7 or -9 stimulation and serve as a link between the innate and adaptive immune systems. In HIV infection, there is a gradual depletion of PDC as well as a functional deficiency in remaining circulating PDC. The mechanism of PDC depletion is not yet understood. We have demonstrated using imaging flow cytometry (ImageStream®) that PDC preferentially take up membrane and cytoplasm from HSV and Influenza infected cells in an endocytic process called "nibbling" that results in IFN-α production by the PDC. Using acutely HIVIIIB infected CD4 T cells or chronically HIVIIIB infected H9 T cells, PDC preferentially form conjugates with HIVIIIB infected vs uninfected T cells, but, rather than being nibbled, the majority of these encounters lead to PDC/infected T cell fusions. Acutely infected CD4 T cells but not chronically-infected H9 cells stimulated PDC to produce IFN-α, albeit it lower levels than other TLR7/9 inducers. Fusions were blocked with the addition of the specific fusion inhibitors (T-20 or AMD-3100), but there was no increase in conjugate formation or uptake of material. We hypothesize that this successful subversion of PDC nibbling and IFN-α production by HIV-infected cells contributes to loss of PDC and the deficient IFN-α production seen in HIV-infected patients. Supported by AI26806.
[alpha]-GalCer analogues with branched acyl chain enhance humoral and cellular immune responses in a nasal influenza vaccine (132.11)
Authors: Lee Yoon-Sook , Lee Kyoo-A , Lee Jae-Young , Kang Min-Hee , Kim Sang-Hee , Kang Chang-Yuil 
Published: January 2023

Abstract
α-galactosylceramide (α-GalCer) can act as a safe and effective adjuvant for a nasal vaccine that induces protective immune responses against tumors and viral infections. In this study, α-GalCer analogues were designed to have branched acyl chain by modification of their fatty acyl chain based on the CD1d/glycolipid structures. Two α-GalCer analogues with various branched chain lengths; LDH-II-28 and LDH-II-30 were prepared and evaluated for their efficacy as a nasal influenza vaccine adjuvant. These analogues displayed improved solubility over α-GalCer and effectively stained NKT cells both in mouse and human. They also potently stimulated NKT cells to exhibit different cytokine release profiles from α-GalCer in vitro and in vivo. When profiling serum cytokines in vivo, LDH-II-30 provoked both Th1/Th2 cytokines, while LDH-II-28 induced more Th2-biased cytokine release with diminished IFN-γ production. We found that single immunization of inactivated influenza virus A/PR/8/34 (PR8) together with α-GalCer analogues enhanced PR8-specific humoral and cellular immune responses in both systemic and mucosal compartments. Notably, LDH-II-30 exhibited potent adjuvant effects with significantly higher systemic IgG, mucosal sIgA Ab titers and enhanced CTL generation compared to immunization with inactivated PR8 alone, while addition of LDH-II-28 to inactivated PR8 only marginally increased PR8-specific immune responses. These results suggest that α-GalCer analogues with branched acyl chain could be used as an effective mucosal adjuvant for the induction of influenza vaccine-specific humoral and cellular immune responses.
Characterization of the anti-lethal factor (LF) antibody response in Anthrax Vaccine Adsorbed (AVA) individuals (132.15)
Authors: Crowe Sherry R , Engler Renata J M , Ballard Jimmy D , Harley John B , Farris A Darise , James Judith A 
Published: January 2023

Abstract
The currently licensed anthrax vaccine (AVA) has been designed to elicit primarily an anti-protective antigen response. However, animal studies have clearly demonstrated that antibodies to the lethal factor component of the toxin can provide protection against challenge. This study was designed to identify AVA vaccinated individuals that generate anti-LF responses and to determine if these anti-LF antibodies can mediate protection. Plasma was obtained from one thousand vaccinated individuals and thirty unvaccinated controls. Only 69 individuals (6.9%) had significant levels of anti-LF antibodies. The presence of these antibodies did not correlate with in vitro protection against lethal toxin. To determine if antibodies against specific regions of LF could mediate protection, we used overlapping decapeptides to identify six common antigenic regions within those individuals with high levels of anti-LF antibodies and high toxin neutralizing activity. Epitope-specific antibodies directed against three of these common antigenic regions were isolated and select antibodies were able to protect in both in vitro tissue culture and in vivo mouse toxin challenge models. Findings from these studies have important implications for vaccine design and immunotherapeutic development. NIAID: 5U19AI062629
Memory B cells are required to maintain long-lived plasma cells in mice following viral infections (132.16)
Authors: Langley William A , Mueller Scott N , Ahmed Rafi 
Published: January 2023

Abstract
Antibody responses have been found to last for life following lymphocytic choriomeningitis virus (LCMV) infection in mice and have been estimated to have a half-life of up to 200 years for some pathogens in humans. These antibodies are produced by long-lived plasma cells which have been shown to have a half-life of at least 138 days in mice. However, the role of memory B cells in the maintenance of plasma cell numbers remains unclear. We have been attempting to define this role by infecting hCD20 transgenic mice with viral pathogens, treating them with rituximab (an anti-CD20 antibody), and then following the antibody levels in these mice. Rituximab treatment of hCD20 mice selectively depletes naive and memory B cells while leaving plasma cells untouched. During long-term treatment of LCMV immune mice, there is a slow, steady decline of antibody levels and a decrease in the number of LCMV-specific antibody secreting cells in the spleen and bone marrow. This decrease in antibody levels was also observed with influenza and vesicular stomatitis virus (VSV) infected mice and correlates with decreased hemagglutination inhibition titers in influenza infections and decreased neutralization titers in VSV infections. These results suggest a critical role for memory B cells in the maintenance of long-lived plasma cells following viral infections.
This work was supported by NIH grant AI30048.
Efficacy of HIV-1 and FIV subunit vaccines against FIV (132.3)
Authors: Coleman James K , Pu Ruiyu , Yamamoto Janet K 
Published: January 2023

Abstract
Defining the minimum essential vaccine components required for protective immunity to HIV and FIV remains a major challenge. A recent study reported that recombinant HIV-1 capsid p24 vaccines conferred cross-protection against FIV in cats. Current studies were performed to confirm this protection and identify other protective HIV/FIV proteins. Pathogen-free cats were immunized 3X with HIV p24 (CA), HIV p66 (reverse transcriptase RT), FIV p24, FIV p66, FIV p15 (matrix MA), or combination in adjuvant (plus LPS/IL-12), and were challenged 3 wk later with FIV. Infection was monitored by immunoblot for FIV antibodies, and by FIV RT and PCR tests of lymphoid cells. Cellular immunity was monitored by virus-specific IFN-γ ELISPOT and CFSE-based proliferation of T cells. Protection rates were 2/8 cats for HIV p24, 2/8 HIV p24+p66, 3/7 HIV p66, 1/8 FIV p24, 0/3 FIV p15, 2/7 FIV p24+p66, 5/7 FIV p66, 2/4 HIV p24+FIV p66, and 0/8 adjuvant or PBS. Vaccinated cats had strong IFN-γ and CD4/CD8-CFSE responses to HIV or FIV peptides, but developed no virus neutralizing antibodies. Combined protection rates for current and previous studies are 40/64 cats for HIV p24, 4/25 FIV p24, 5/7 FIV p66, 3/7 HIV p66, 0/3 FIV p15, 0/22 adjuvant, and 0/37 PBS. Thus, cross-protective immunity between HIV-1 and FIV is achievable, and selection of vaccine proteins and epitopes based on structural homology that risk viral fitness is an effective strategy.
Severely burned patient peripheral linage– CD34+ cells are inhibitory on the antimicrobial peptide production by human epidermal keratinocytes (133.10)
Authors: Kobayashi Makiko , Jeschke Marc G , Benjamin Debra , Yoshida Shohei , Kawasaki Takashi , Kotani Takuya , Herndon David N , Suzuki Fujio 
Published: January 2023

Abstract
The importance of epidermal keratinocyte-produced antimicrobial peptides in the control of skin infections has been described. However, these peptides are not produced sufficiently in tissues surrounding the burn area. In this study, linage- CD34+ cells that infiltrated into burn area tissues were shown to be inhibitory on the antimicrobial peptide production by human epidermal keratinocytes (NHEK). Peripheral blood mononuclear cells (PBMC) taken from 5 severely burned patients (3rd degree, more than 40% total body surface area burn) enrolled in these experiments. NHEK (1 x 105 cells/ml, lower chamber) were cultured with 1 x 105 cells/ml of healthy donor PBMC or patient PBMC in a dual-chamber transwell. Forty-eight hrs after cultivation, cells in the lower chamber were assayed for human β-defensin-1 (HBD-1) and cathelicidin (CAP-18) mRNAs by RT-PCR. Without any stimulation, mRNAs for HBD-1 and CAP-18 were consistently expressed by NHEK. mRNA expression for HBD-1 and CAP-18 by NHEK was not influenced when peptide producer cells were transwell-cultured with normal donor PBMC. However, the expression of HBD-1 and CAP-18 mRNAs by NHEK was greatly suppressed when they were transwell-cultured with patient PBMC. Suppressor cells in patient PBMC for the peptide production by NHEK were determined to be linage- CD34+ cells. Linage- CD34+ cells isolated from healthy donor PBMC did not have any suppressor cell activities. These results indicate that linage- CD34+ cells isolated from severely burned patient PBMC are suppressor cells on the antimicrobial peptide production by epidermal keratinocytes. This study was supported by SHC NA #8610.
Commensal microflora induce host defense to E.coli pneumonia through toll-like receptors (133.13)
Authors: Chen Lee-Wei , Hsu Ching-Mei 
Published: January 2023

Abstract
The role of commensal microflora in host response to pneumonia has not been defined. This study is to investigate whether commensal microflora in gut increase the host defense against pneumonia through TLRs signaling.We treated C3H/HeN and C3H/HeJ mice with antibiotics in drinking water for 4 weeks to deplete gut commensal microflora. At week 3, drinking water was supplemented with lipopolysaccharide (LPS); a ligand for TLR4, to trigger TLRs in intestinal tract. E.coli was injected to trachea to induce E.coli pneumonia.
Commensal depletion induced a 15-fold and a 2.5-fold increase in E.coli pneumonia-induced bacterial counts in lung and blood, respectively and a 30% decrease of lung MPO activity. Commensal depletion induced a suppression of TLR4, TNFα&, KC, and CXCR2 expression; and NF-κB activity in intestine and LPS supplement reversed them. LPS supplement increased lung MPO activity and decreased E.coli pneumonia-induced bacterial counts in WT mice but not in C3H/HeJ mice. Commensal depletion induced a decrease of phagocytic activity of alveolar macrophages, reduced NF-κB activity, and increased KC, MIP-2, and IL-1β mRNA expression of lung in WT mice but not in C3H/HeJ mice. In conclusions, commensal microflora are critical in inducing TLR4 expression and NF-κB activation of intestine, increasing the lung immunity, and decreasing the inflammatory signaling in lung after E.coli pneumonia. Given that the stimulatory effect of LPS supplement on lung immunity is abolished in TLR4-deficient mice, we conclude that both commensal microflora and TLR4 ligand increase host defense against pneumonia through toll-like receptors.
Innate immunity to tuberculosis: a forward genetic approach (133.15)
Authors: Tobin David M. , Ray John P. , Moens Cecilia , Ramakrishnan Lalita 
Published: January 2023

Abstract
We have developed a forward genetic screen in the zebrafish to identify host genes that control resistance and susceptibility to tuberculosis. Using facile live imaging techniques to screen mutant larvae for alterations in the pathogenesis of Mycobacterium marinum, we have identified multiple genetic loci that alter myeloid development and functions, including macrophage microbicidal capacity, migration and aggregation. Through a positional cloning approach, we have identified genes affected by three mutations causing hypersusceptibility to M. marinum. One of these mutations lies in the gene encoding leukotriene A4 hydrolase (LTA4H), the biosynthetic enzyme that produces leukotriene B4 (LTB4). In spite of LTB4's role as a potent leukocyte chemoattractant, the mutant is not compromised in the initial migration of macrophages to the site of infection. Rather mutant macrophages are defective in limiting intracellular bacterial growth and are quickly killed by the infection. Chemical inhibitors of this pathway mimic the mutant phenotype. Finally, we show that the protective effect of LTA4H in limiting mycobacterial growth may be attributable to a role in the regulation of TNF, as mutant fish fail to induce TNF transcription in response to infection.
Interleukin-12 is dispensable for functional interferon (IFN)-gamma producing CD8+ cytotoxic T cells in Toxoplasma gondii infection (133.28)
Authors: Daniel Benjamin J , Liu AiJie , Pandeswara Srilakshmi , Ludwig Sara M , Brumlik Michael J , Church Kristina M , Kious Mark J , Curiel Tyler J 
Published: January 2023

Abstract
Interleukin (IL)-12 is a proinflammatory cytokine generally regarded as a master regulator of type 1 T cell mediated immunity against intracellular pathogens, including T. gondii and cancers. A critical measure of IL-12 activity is its ability to induce IFN-γ production by NK and T cells. Using a transgenic T. gondii expressing ovalbumin (OVA), we show here that by flow cytometry analysis T. gondii-OVA infected BL6 IL-12-/- mice generate OVA-pentamer+CD8+ T cells similar to WT and surprisingly, T. gondii-infected IL-12-/- mice produce more IFN-γ +CD8+ T cells than T. gondii-infected WT mice (avg 9.6% in IL-12-/- vs 3.7% in WT). We confirmed that function of OVA-specific CD8+ T cells in IL-12-/- was comparable to BL6 WT using an in vivo OVA+ target lysis assay. As expected, IFN-γ +CD4+ T cells decreased from an avg 29% to 9% in WT mice compared to IL-12-/- mice respectively when infected with T. gondii. These results demonstrate a novel IL-12-independent pathway to CD8+ CTL that may be less dependent of CD4+ T cell help than previously thought.