bluuebunny/crossref_metadata_2025_split huggingface dataset:
Quantitative bioimaging of WRvFire and of IHDJ-Luc vaccinia virus dissemination in mice and the effects of prophylactic and therapeutic treatments with IgG and antiviral. (45.18)
Authors: Kapnick Senta , Zaitseva Marina , Scott John , Meseda Clement , Nielsen Henriette , Merchlinsky Michael , Weir Jerry , Golding Hana 
Published: January 2023

Abstract
Whole body bioluminescence imaging was employed to test the effects of IgG-based therapies and Cidofovir on lethality and virus dissemination in BALB/c mice challenged intranasally with 10^5 pfu of recombinant vaccinia viruses (VV) expressing luciferase, WRvFire or IHDJ-Luc. Both viruses induced lethality within 6-8 days, with slightly different kinetics of replication in internal organs. Using statistical analysis of 80 animals/group we determined that replication of IHDJ-Luc (increased Extracellular Enveloped Virion compared to WR) rose faster and peaked earlier than WRvFire (predominantly Intracellular Mature Virion) in the liver and spleen. In the nasal cavity, WRvFire persisted at significantly higher levels than IHDJ-Luc. Vaccinia immunoglobulin for intravenous injection (VIGIV) administered at day -2 protected 100% of animals from WRvFire and IHDJ-Luc infections. Sym002 (26 anti-vaccinia human mAbs) completely protected mice from both challenges at 300-fold lower dose than VIGIV. A single treatment of Cidofovir (100 mg/kg) at day 1, 2, or 3 p.i. fully protected mice from WRvFire challenge. Complete protection conferred by treatments correlated with significant reductions in bioluminescence in nasal cavity or lungs and in both the liver and spleen between days 1-5. Our data demonstrated that despite some differences observed in the kinetics of VV dissemination, infections with these viruses were similarly susceptible to prophylactic and therapeutic treatments.
Development of a sensitive serodiagnostic assay for human Lyme disease based on immunodominant epitopes of an in vivo expressed microbial surface antigen (45.19)
Authors: Coleman Adam , Pal Utpal 
Published: January 2023

Abstract
Lyme disease (LD) is a common bacterial infection transmitted by the ticks belonging to Ixodes scapularis complex. The diagnosis of LD is complicated by several factors, including unnoticed exposure and variation in patient presentation. Serology is currently the primary laboratory means of diagnosis in suspected LD cases, and the development of new diagnostics is an important goal of LD research. Central to this effort is the identification and characterization of new immunodominant antigens to serve as serodiagnostic markers. Borrelia burgdorferi, the causative agent of LD, encodes BBK07, an outer membrane lipoprotein thought to be an important antigen expressed during infection. We show that BBK07 gene expression is selectively induced during mammalian infection when compared to B. burgdorferi grown in vitro or in ticks. We show that BBK07 is surface-exposed on the bacterium, and that an antibody response to BBK07 is detectable throughout the murine infection. We use recombinant BBK07 protein to dissect the early (IgM) and late (IgG) immune response over the course of infection. A synthetic overlapping peptide library of BBK07 was used to identify the most immunogenic epitopes of the antigen using a human LD serum panel. We used a combination of immunodominant peptides to develop a specific and sensitive assay for the diagnosis of LD. These data suggest possible development of a BBK07 peptide-based serodiagnostic kit for the detection of human LD.
Differentiation, distribution and γδ T cell-driven regulation of IL-22-producing T cells in tuberculosis (45.21)
Authors: Yao Shuyu , Huang Dan , Chen Crystal , Halliday Lisa , Zeng Gucheng , Wang Richard , Chen Zheng 
Published: January 2023

Abstract
Differentiation, distribution and immune regulation of human IL-22-producing T cells in infections remain unknown.We demonstrated that M. tuberculosis infection resulted in apparent increases in numbers of T cells capable of producing IL-22 de novo without in vitro Ag stimulation, and drove distribution of these cells more dramatically in lungs than in blood and lymphoid tissues. Consistently, IL-22-producing T cells were visualized in situ in lung tuberculosis (TB) granulomas by confocal microscopy and immunohistochemistry, indicating that mature IL-22-producing T cells were present in TB granuloma. Surprisingly, phosphoantigen HMBPP activation of Vγ2Vδ2 T cells down-regulated the capability of T cells to produce IL-22 de novo. Up-regulation of IFNγ-producing Vγ2Vδ2 T effector cells after HMBPP stimulation coincided with the down-regulated capacity of these T cells to produce IL-22 de novo. Importantly, anti-IFNγ neutralizing Ab treatment reversed the HMBPP-mediated down-regulation effect on IL-22-producing T cells, suggesting that Vγ2Vδ2 T-cell-driven IFNγ-networking function was the mechanism underlying the HMBPP-mediated down-regulation of the capability of T cells to produce IL-22. These novel findings raise the possibility to ultimately investigate function of IL-22 producing T cells and to target Vγ2Vδ2 T cells for balancing potentially-hyper-activating IL-22-producing T cells in severe TB.
Modulation of phosphatidylinositol 3-kinase (PI3K) as a tool to enhance anti-polysaccharide antibody response in the aged (45.23)
Authors: Fallah Mosoka , Chelvarajan R. , Bondada Subbarao 
Published: January 2023

Abstract
The elderly are at risk to increased infection with Streptococcus pneumoniae, gram positive bacteria. This has been shown to be due to a reduced production of pro-inflammatory cytokines (IL-6, IL-12, TNF-α) by splenic macrophages (SM), as well as a decrease in the B-cell class-switching to IgG3 and IgA, isotypes that are critical for the opsonization and clearance of the pathogen. Here we showed that PI3K expression and activity are increased in aged compared to young SM. Phosphorylation of AKT and glycogen synthase kinase-3 (GSK-3), downstream targets of PI3K, was also more in the aged than young SM and B-cells. The PI3K inhibitor, LY294002, enhanced TLR-2 and TLR- 4 dependent pro-inflammatory cytokine secretion by SM from the aged. Moreover, low concentrations of the PI3K inhibitor enhanced TLR-4 induced class-switching to IgG3 and IgA. Interestingly, inhibition of GSK-3 also enhanced TLR-4 induced IgG3 and IgA secretion as well as B-cell proliferation. Hence, a pharmacological modulation of the PI3K-AKT-GSK pathway could be a potential vaccine enhancement strategy against infection with S. pneumoniae in the elderly. (Supported in part by NIH Grants)
Stable expression of a foreign protein by a live rubella viral vector (45.25)
Authors: Berkower Ira , Virnik Konstantin , Ni Yisheng , Prutzman Kirk , Spadaccini Angelo 
Published: January 2023

Abstract
Live attenuated rubella virus has a number of desirable features for a live viral vector. It has a good safety profile, so it can immunize while growing exponentially in the host. It immunizes at a low dose, and one dose protects (against rubella) for life. It grows well in rhesus macaques, which allow a viral challenge. It has no DNA intermediate, cannot integrate, and usually does not persist in the host. A full length infectious cDNA clone of rubella was available for modification, but prior efforts to express foreign genes were frustrated by genetic instability and size limitations on the insert. A deletion of 507 bp in nonstructural gene P150 was permissive, so we asked whether this deletion could make room for insertion of zoanthus GFP (792 bp). By using the deletion/insertion strategy, zGFP was stably expressed for at least 12 passages. In the zGFP hybrid, rubella structural proteins were expressed at normal levels, while the vector grew to high titer (4 X 10^6/ml). Normal P150 function was detected by P150-GFP labeling of functional replication centers in living cells. Rubella may be useful for expressing vaccine antigens as large as most viral proteins, and it is stable enough for vaccine production followed by growth in vivo. This approach may extend the range and utility of rubella to include immunization against other viruses for which attenuation is not currently feasible.
SLAM activation induces ERK and CREB phosphorylation that leads to IFN-γ production during active tuberculosis (TB) (45.27)
Authors: Pasquinelli Virginia , Alvarez Ivana , Jurado Javier , Aspera Romina , Fernandez Do Porto Dario , Samten Buka , Barnes Peter , Garcia Veronica 
Published: January 2023

Abstract
IFN-γ is crucial for protection against M. tuberculosis (Mtb). Previously we reported that signaling through SLAM on Mtb-stimulated cells induced CREB activation increasing IFN-γ. Here, we deeply investigated the potential relationship between SLAM-CREB and IFN-γ. After Mtb-stimulation, most of the IFN-γ+ cells co-expressed SLAM and pCREB (48,68 ± 11,18), whereas a smaller percentage of T cells expressed one of the two molecules. These results demonstrate that the induction of IFN-γ mediated by SLAM in TB depends, at least in part, on CREB activation. It was reported that SLAM signaling would induce the activation of the protein kinase Erk. To analyze the signaling pathways induced by SLAM that contribute to CREB activation and IFN-γ secretion in TB, we studied Erk phosphorylation after SLAM activation. The addition of PD90059 (an Erk inhibitor) to Mtb stimulated cells inhibited the percentage of IFN-γ+pCREB+ T cells, indicating that Erk could be involved on CREB activation. Moreover, the addition of PD90059 to Mtb±α-SLAM stimulated cells significantly decreased IFN-γ production (% inhibition: 71,6 ± 7,4). Finally, α-SLAM induced Erk activation in Mtb stimulated cells. These results demonstrate that IFN-γ production induced by SLAM signaling on Mtb stimulated cells is, at least in part, mediated by Erk and CREB phosphorylation. These data contribute new information about the molecular basis operating during SLAM activation that leads to IFN-γ production in human TB.
Heat shock protein Gp96-Ig-peptide complexes based vaccine induce predominant immune responses at mucosal sites (46.13)
Authors: Strbo Natasa , Vaccari Monica , Pahwa Savita , Kolber Michael , Fisher Eva , Gonzalez Louis , Franchini Genoveffa , Podack Eckhard 
Published: January 2023

Abstract
We have developed a vaccine design that uses the unique ability of the endoplasmic reticulum (ER) chaperone, heat shock protein gp96, to bind antigenic peptides and deliver them to antigen-presenting cells. Secreted form of gp96, gp96-Ig, was generated by replacing the ER retention sequence KDEL with the IgG1-Fc domain. To generate a cellular SIV vaccine, HEK-293 cells were transfected with gp96-Ig and the cDNAs encoding the SIV antigens gag, env and retanef. Irradiated, transfected 293 cells that secrete 1, 5 or 50 micrograms gp96-Ig-SIV complexes in 24h, were injected intraperitoneally (IP) in Mamu-A*01+ Rhesus Macaques at 0, 4 and 25 weeks. To determine whether gp96-Ig vaccination induces mucosal immunity, we used the 3T3-OVA-gp96-Ig/OT-1 model. Our data indicate that the best route of vaccination for induction of mucosal CD8-CTL immunity is the IP route. Gp96-Ig immunization increases frequency of CD11chighMHC classIIhighCD103+ cells in peritoneal cavity and efficiently induces CCR9 on responding T cells. In nonhuman primate model, the frequency of SIV-specific cells in the rectal mucosa reached 30-50% after the third immunization. We conclude that gp96-Ig vaccination startegy induces antigen-specific effector memory CD8T cells migrating to the intestinal mucosa and thus could be extremely useful for improving protection against a range of mucosal pathogens.
Effect of Bacillus-based direct-fed microbials on macrophage functions and Eimeria maxima infection in broiler chickens (46.18)
Authors: Lillehoj H. , Lee Kyungwoo , Guangxing Li , Lee Sung Hyen , Jang Seung Ik , Babu Uma , Lillehoj Erik , Siragusa Gregory 
Published: January 2023

Abstract
The present experiment was conducted to study the effect of dietary Bacillus-based direct-fed microbials (DFMs) on macrophage functions and resistance against experimental coccidiosis in broiler chickens. Birds were fed diets containing one of eight Bacillus subtilis strains (designated Bs2084, LSSAO1, 3AP4, Bs18, 15AP4, 22CP1, Bs27, and Bs278) or one multiple-strain DFM product (AVICORR™) for 21 days, and then the chickens were uninfected or orally infected with 5000 Eimeria maxima (EM) oocysts. At day 21 (before EM challenge), strong phagocytosis ability using BSA-coated fluorescent latex beads and GFP-labeled salmonella was seen in animal given Bs18, 15AP4, Bs27, or Bs278 compared with the no DFM control group. At day 27 (6 days post infection), EM-induced reduction of body weight gain was significantly decreased by adding 15AP4 or Bs27 into broiler diets compared with no DFM/EM inoculated control. Serum Nitric oxide levels were increased in infected animals fed with Bs27 compared with the no DFM/EM inoculated control. The levels of Eimeria-specific antibody were increased in infected chickens fed with LSSAO1 compared with the no DFM/EM inoculated control. Finally, all experimental diets increased concanavalin A-induced splenocyte mitogenesis in infected broilers compared with the no DFM/EM inoculated control. In summary, dietary DFMs reduced the clinical signs of experimental avian coccidiosis and increased various parameters of immunity in broiler chickens.
Enterotoxigenic E. coli CFA/I Fimbriae Abate Collagen-Induced Arthritis (CIA) via Regulatory CD39+ T Cells and IL-35 (47.1)
Authors: Kochetkova Irina , Thornburg Theresa , Callis Gayle , Crist Kathryn , Pascual David 
Published: January 2023

Abstract
Colonization factor antigen I (CFA/I) expressed by Salmonella vaccine vectors can prevent experimental autoimmune encephalomyelitis and CIA attributed to suppression of Th1 and Th17 cells by regulatory and immune deviation mechanisms. We hypothesize that soluble CFA/I fimbriae may confer similar protection against CIA as Salmonella-CFA/I. Single dose, mucosal CFA/I application 14 days post-CIA induction abated clinical development of arthritis similar to Salmonella-CFA/I. IFN-γ, IL-6, and IL-17 responses by CD4+ T cells were suppressed in protected mice via the induction of IL-10 and regulatory FoxP3+ CD4+ T cells. Such regulatory T cells were both CD25- and CD25+, each conferring protection upon adoptive transfer to CIA mice. Further evaluation revealed the protective CD25- CD4+ T cells also expressed CD39, ectonucleoside triphosphate diphosphohydrolase, an enzyme responsible for extracellular ATP hyrolysis. Flow cytometry analysis revealed that CFA/I fimbriae nearly doubled the CD39+ CD4+ T cell subset, ~50% of which were CD25-negative. Moreover, mucosal CFA/I delivery enhanced by 2-fold IL-35+ FoxP3+ CD4+ regulatory T cells. CD39+ CD4+ T cells accounted for most (~70%) of TGF-β production by CD4+ T cells. Upon restimulation, these CFA/I -induced CD4+ T cells did not produce IFN-γ or IL-17, but IL-10 levels were enhanced. In conclusion, CFA/I fimbriae alone can augment regulatory T cells for protection against autoimmune diseases. Work supported by P01 AT-04986.
Th17/Treg immunoregulatory balance in human periapical lesions (47.5)
Authors: Rodrigues Denise , Marçal Juliana Romanelli , Samuel Renata , Fernandes Danielle , Araujo Marcelo , Napimoga Marcelo , Lima-Pereira Sanivia , Napimoga Juliana , Alves Polyanna , Mattar Rinaldo , Rodrigues Virmondes 
Published: January 2023

Abstract
Cysts and granulomas are chronic periapical lesions mediated by a set of inflammatory mediators that develop in order to contain a periapical infection. This study analyzed the inflammatory infiltrate, mast cells and in situ expression of cytokines, chemokines and FoxP3 in human periapical granulomas and cysts compared to controls. Fifty-five lesions (25 cysts, 25 granulomas and 5 controls) were analyzed. The type of inflammatory infiltrate was evaluated by HE staining and the presence of mast cells was analyzed by toluidine blue staining. Indirect immunohistochemistry was used to evaluate the expression of cytokines, chemokines and FoxP3. The inflammatory infiltrate mainly consisted of mononuclear cells. In cysts, mononuclear infiltrates were significantly more frequent than mixed (polymorphonuclear/mononuclear) infiltrates (p=0.04). Mixed inflammatory infiltrates were significantly more frequent in patients presenting fistulae (p=0.0001). The number of mast cells was significantly higher in granulomas than in cystic lesions (p=0.02). A significant difference in the expression of IL-17 (p=0.001) and TGF-β (p=0.003) was observed between cysts and granulomas and the control group. Significantly higher IL-17 levels were also observed in cases of patients with fistulae (p=0.03). Mast cells, TGF-β and IL-17 probably participate in the modulation of the inflammatory process in established periapical lesions, especially in granulomas.
Biodegradable dissolving microneedle arrays effectively deliver antigens and adjuvants to skin DCs for the induction of antigen specific immune responses. (48.12)
Authors: Erdos Geza , Donahue Cara , Williams Mellissa , Ozdoganlar Burak , Falo Louis 
Published: January 2023

Abstract
The skin contains a high density of antigen presenting cells making it an attractive target for vaccine delivery. However, the remarkable barrier function of the skin continues to present a formidable obstacle to topical vaccine delivery. To overcome this barrier we designed carboxy-methyl cellulose microneedle arrays (MNAs) to topically deliver antigens and adjuvants into the skin. MNAs were designed and fabricated to enable advantages in penetration, delivery, and diversity of biocompatibility over existing skin delivery technologies. Initially we fabricated MNAs with integrated tracer antigens. Following MNA immunization labeled antigens were identified in dendritic cells in human and murine skin, and in the draining lymph nodes of immunized animals. To evaluate immunogenicity, we incorporated the model antigen OVA in combination with various adjuvants into MNAs. MNA-mediated delivery of low dose antigen/adjuvant induced potent and durable CTL responses compared to those generated by traditional intradermal immunization. Further, MNA immunization utilizing B16 tumor cell lysates as a source of antigen induced B16 specific DTH reactions and inhibited tumor growth in a B16 tumor model. Taken together, these results demonstrate the feasibility of the use of CMC microneedle arrays for cutaneous immunization and suggest potentially broad applicability for the co-delivery of structurally diverse antigens and adjuvants.
Human decidual tissue contains disarmed CD8+ Effector-Memory T cells (48.2)
Authors: Tilburgs Tamara , Schonkeren Dorrith , Eikmans Michael , Strominger Jack , Roelen Dave , Scherjon Sicco , Claas Frans 
Published: January 2023

Abstract
During pregnancy maternal lymphocytes at the fetal-maternal interface play a key role in the immune acceptance of the allogeneic fetus. Recently, CD4+CD25bright regulatory T cells have shown to be concentrated in decidual tissue where they are able to suppress fetus-specific immune responses. Decidual CD8+ T cells form the largest fraction of T cells at the fetal-maternal interface but limited data is present on the characteristics of these cells. In this study we examined decidual and peripheral CD8+ T cells for CD45RA, CCR7, CD28 and CD27 expression using nine-colour flowcytometry. Our data demonstrate that decidual CD8+ T cells mainly consist of differentiated CD45RA-CCR7- effector-memory (EM) cells while unprimed CD45RA+CCR7+ naïve cells are absent. Unlike peripheral blood EM T cells, the decidual EM T cells do not express perforin and have a reduced expression of granzyme B, which was confirmed by immunohistochemistry of decidual tissue sections. Interestingly, quantitative PCR shows an increased perforin and granzyme B mRNA content in decidual EM T cells in comparison to peripheral blood. The presence of high levels of perforin and granzyme B mRNA in decidual EM T cells suggests that decidual CD8+ T cells pursue alternative means of EM cell differentiation that may include a blockade of perforin and granzyme B mRNA translation into functional proteins. Regulation of decidual CD8+ T cell differentiation may play a crucial role in maternal immune tolerance to the fetus.
Both systemic and mucosal LCMV immunization generate robust viral specific IgG in mucosal secretions, but elicit poor LCMV-specific IgA (48.3)
Authors: Zeng Junwei , Rajini Bheemreddy , Onami Thandi 
Published: January 2023

Abstract
Immunoglobulins in secretions play a critical role in protection at mucosal surfaces. We examined the generation of viral specific IgG and IgA in plasma and mucosal secretions of mice following systemic or mucosal immunization with LCMV, a widely used experimental model of viral infection. While there are early differences in humoral responses depending on the route of immunization, we show that both routes generate comparably robust viral specific IgG in plasma, vaginal, lung, and nasal secretions of immune mice. In contrast, LCMV elicited poor viral specific IgA responses. Mice that were infected i.n. showed elevated viral specific IgA in nasal and lung washes compared to i.p. infected mice, however, LCMV-specific IgG overwhelmingly contributed to the humoral response in all mucosal secretions examined. Thus similar to HIV-1, and several other mucosally encountered microbial infections, these data suggest that LCMV infection fails to induce vigorous viral specific IgA responses.
Long term overexpression of IL-10 increases recruitment of fibrocyte into the lung and induces fibrosis (48.5)
Authors: Sun Lei , Louie Marisa , Vannella Kevin , Moore Bethany , Shanley Thomas 
Published: January 2023

Abstract
Pulmonary fibrosis is characterized by the progressive deposition of extracellular matrix components such as collagen and fibronectin by fibroblasts and myofibroblasts. Three potential sources of fibroblasts and myofibroblasts include: expansion of resident lung fibroblasts, epithelial-mesenchymal transition, or recruitment and differentiation of circulating mesenchymal precursors (fibrocytes). IL-10 is a well-known immunosuppressive cytokine and in some studies has an inhibitory effect on fibrosis, however, this is less well studied. In our study, long-term overexpression of IL-10 (IL-10 OE) induced fibrosis. The total cell number from bronchoalveolar lavage (BAL) increased ten-fold in the IL-10 OE mice, with significant infiltration of T, B lymphocytes and collagen-producing cells. Fibrocytes from collagenase digested lung were identified by flow cytometry with dual staining of CD45 and collagen 1. IL-10 OE increased whole lung fibrocyte numbers by two-fold. Q-PCR analysis on an array of chemokine/chemokine receptor genes showed that only CCL2 and CCL2 receptor (CCR2) were highly upregulated in IL-10 OE mice, suggesting that IL-10 induced fibrocyte recruitment is CCL2/CCR2 specific. We also examined M2 macrophage activation and found significant numbers of M2 macrophages in both BAL and whole lung tissue from the IL-10 OE mice. Taken together, long term overexpression of IL-10 may induce fibrosis, in part, by fibrocyte recruitment and M2 macrophage activation.