bluuebunny/crossref_metadata_2025_split huggingface dataset:
Textpresso Site Specific Recombinases: a text mining tool to aid in identifying Cre expressing mice and conditional alleles (42.23)
Authors: Condie Brian G , Urbanski William M 
Published: January 2023

Abstract
The use of site-specific recombinases such as Cre and Flp in mouse genetics has risen dramatically. The number of mouse strains designed to express Cre recombinase activity in specific tissues and the number of mouse strains carrying conditional "floxed" alleles has increased explosively. As a result, the scientific literature describing these valuable research resources has burgeoned to more than 6000 peer reviewed publications. The large size and rapid growth of this literature makes it very easy to miss key papers that contain critical information relevant to your research. To assist investigators in identifying appropriate Cre recombinase expressing mice, conditional floxed alleles and other research resources based on site specific recombinase technology we have generated a new online resource based on the widely used Textpresso text mining software (http://www.textpresso.org). Our text mining tool, Textpresso Site Specific Recombinases (Textpresso SSR) allows researchers to search a database containing the full text of over 6000 peer-reviewed publications. This database contains most of the publications that describe or use Cre expressing mice as well as papers describing mouse strains carrying conditional mutant alleles. The database can be searched using keywords or combinations of keywords and categories. For each publication that matches the search terms Textpresso SSR displays the abstract as well as the individual sentences that match the query. Textpresso SSR also provides a unique tool for the in depth text mining of publications that describe the results of tissue specific or temporal specific mouse gene knockouts. Textpresso SSR can be accessed at http://ssrc.genetics.uga.edu.
This work was supported by the University of Georgia.
Delivery to Macrophages and Efficacy of Orally Administered RNAi Compounds (42.27)
Authors: Kamens Joanne , Khvorova Anastasia , Pavco Pamela , Rodgers Amanda , Ford Glenna , Bulock Karen , Smith-Anzures Bernice , Lapierre Jennifer , Lam Jessica , Libertine Lyn , Flannery-Rossi Kehlen , Metterville Jake , Byrne Michael , Czech Michael , Ostroff Gary 
Published: January 2023

Abstract
Data will be presented on a method for RNAi compound delivery utilizing RNA encapsulated in beta-glucan shells. When orally administered to mice, these encapsulated nucleic acids traverse the intestinal epithelia by an M cell transcytosis mechanism and are then endocytosed by macrophages. Macrophages containing RNAi compounds migrate to sites of inflammation, and the targeted RNA is specifically inhibited by RNA interference. This formulation presents some unique advantages for therapeutic RNAi including oral mode of administration, low dose (ug/kg/day) and specific delivery to immune cells possibly affording safety advantages for some targets. This novel mode of RNA administration can be used to down-regulate target genes involved in multiple inflammatory disease indications.
Consistency and sensitivity of immune response measurements among multiple laboratories using a standardized ELISpot assay kit (42.28)
Authors: Patke Deepa S , Dryga Sergey A , Marking Christine , Jeffrey Lisa , Manak Mark 
Published: January 2023

Abstract
ELISpot assays measure antigen specific cell mediated immunity, and are useful indicators of vaccine and therapeutic efficacy. Methods that yield consistent and reproducible results are required to compare data in longitudinal studies or among laboratories. Here, we describe the performance of a validated ELISpot assay using PBMC controls and standardized antigenic stimulants among laboratories participating in an External Quality Assurance (EQA) Program. Responses from four PBMC controls (high, medium, low and negative responders) were compared in an IFN-γ ELISpot assay using SeraCare (SC) kits and individual laboratory specific assay protocol and kits in an ELISpot EQA. SC kit performance was also compared to competitor kits in previous rounds of an EQA, each with 13 participants. SC kits showed high sensitivity in detection of low responders (Spot forming cells [SFC] range 10.9 - 49.2/well) compared with in house kit results (SFC range 0.4 - 67.2). Competitor kits yielded larger numbers of Too Numerous To Count results with high responders and greater variability in comparison to SC kits. SC kits yielded a broad linear dose response curve (10-1000 SFC/well) as a result of smaller spot size and improved resolution. SC kits and standardized assay protocols used by multiple laboratories allowed improved sensitivity of detection of low level antigenic responses, higher consistency and a broad linear range.
Reduced inflammation and altered cytokine expression in neonates compared to adults during paramyxoviral infection (43.13)
Authors: Bhattacharya Somashubhra , Janowski Ann , Shornick Laurie P 
Published: January 2023

Abstract
Neonates are especially susceptible to respiratory viral infection. However, there is little known about the neonatal immune response to respiratory virus. We compared the adult and neonatal responses to Sendai Virus (SeV). 6-8 week old C57BL/6 adults and 2-day-old C57BL/6 pups were infected with 500 pfu SeV/g body weight. Adults exhibited transient weight loss, which peaked at day 7 post-infection. Neonatal mice were in a rapid growth period and did not lose body weight during the course of infection. However, neonatal mice infected with SeV showed a slowing of growth after day 8 compared to neonates inoculated with UV-inactivated virus. Viral titer, measured by plaque forming assay, showed similar viral loads and rates of viral clearance in the adults and neonates. Adults exhibited significant neutrophilic inflammation in the lung that peaked at day 7 post-infection. Surprisingly, neonatal lungs had significantly less inflammation in the lung despite comparable levels of virus. ELISA of lung homogenates showed higher levels of IL-1b in the adults. In contrast, neonates had higher levels of TGF-b consistent with reduced inflammation in the neonatal lungs. These results suggest that neonates may be protected from lung inflammation, despite having significant viral load during primary paramyxoviral infection. Further studies will address the ability of the neonates to mount a secondary challenge to SeV. This research was funded by Saint Louis University.
Protection or immunopathology upon heterologous virus infection: A decision of memory cells (43.15)
Authors: Wlodarczyk Myriam F , Kraft Anke , Chen Hong , Selin Liisa K 
Published: January 2023

Abstract
Respiratory viral infection can cause subclinical to severe lung disease. As hosts are continuously exposed to environmental pathogens, a history of past infection might alter the immune response to a subsequent infection. This concept termed heterologous immunity has been largely overlooked. Prior immunity to influenza A (FLU) was shown to have a profound impact on acute infection with LCMV increasing virus titers and immunopathology, a phenomenon that we have called deranged immunity. We observed that LCMV infected FLU-immune mice displayed an alteration of the kinetics of their immune response with increased and more activated CD8 T cells. This is consistent with the activation of cross-reactive memory T cells. We showed that the FLU-induced memory CD8 and/or CD4 T cells played a role in enhancing lung immunopathology, and virus titers. We also identified a cross-reactive CD8 T cell response between the two viruses that maybe playing a role in modulating disease. Consistent with this result, the Vbeta TCR repertoire of some FLU epitope-specific memory CD8 T cells was altered upon LCMV infection. Enhanced IFNg produced by these crossreactive memory cells seemed to play a role in mediating the enhanced immunopathology. These finding have important implications for understanding variability in infectious disease susceptibility and in designing more efficient vaccines against respiratory infectious diseases.
NK1.1+ cells are required for neutrophil recruitment to sites of early viral infection (43.4)
Authors: Hosiawa-Meagher Karoline A. , Clingan Jon , Matloubian Mehrdad 
Published: January 2023

Abstract
LCMV replication is controlled by type I and II interferon (IFN) responses that drive antigen specific T cell expansion and function. It is unclear whether innate immune cells contribute to the initiation of T cell responses. We focused on two innate cell populations, natural killer (NK) cells and CD11b+ cells (Ly6G+ Ly6C+ neutrophils and Ly6G- Ly6C+ monocytes), hypothesizing that sensing of LCMV infection in the spleen drives recruitment of innate cell populations to sites of early infection. C57BL/6-WT or IFN-αR1, IFN-γR, MyD88 or MAVS deficient mice were infected with 2x106 PFU LCMV-Armstrong. Mice were sacrificed at 14, 24, 72 hrs post infection (pi) at which time neutrophil and NK cell recruitment to LCMV infected cells in the spleen was determined by immunofluorescence staining. In WT mice, neutrophils and NK cells were recruited to sites of infection as early as 14 hrs, but dissipated by 72 hrs pi. Depletion of NK1.1+ cells prior to infection abrogated neutrophil recruitment at 24 hrs pi. Furthermore, recruitment of neutrophils to sites of infection was fully dependent on MyD88 and partially dependent on MAVS, but was independent of type I or II IFN receptors. To conclude, our data demonstrate that neutrophil recruitment to sites of early LCMV infection is completely dependent on NK cells and MyD88 signaling, partially dependent on MAVS signaling, and independent of IFN responses.
Supported by Arthritis Foundation
Adaptive Immune Features of Natural Killer Cells (44.1)
Authors: Sun Joseph C , Beilke Joshua N , Lanier Lewis L 
Published: January 2023

Abstract
In an adaptive immune response, naïve T cells proliferate during infection and generate long-lived memory cells that undergo secondary expansion following re-encounter with the same pathogen. Although Natural Killer cells traditionally have been classified as cells of the innate immune system, they share many similarities with cytotoxic T lymphocytes. In a mouse model of cytomegalovirus (MCMV) infection, we demonstrate that, like T cells, NK cells bearing the virus-specific Ly49H receptor proliferate 100-fold in the spleen and 1000-fold in the liver following infection. Following a contraction phase, Ly49H+ NK cells reside in lymphoid and non-lymphoid organs for several months. These self-renewing "memory" NK cells rapidly degranulate and produce cytokines upon reactivation. Adoptive transfer of these NK cells into naïve animals followed by viral challenge results in a robust secondary expansion and protective immunity. These findings reveal novel properties of NK cells previously attributed only to cells of the adaptive immune system.
Attenuation of lymphocytic choriomeningitis virus relaxes interferon-dependence of adaptive immune responses and of virus control (44.12)
Authors: Bergthaler Andreas , Flatz Lukas , Steinborn Ralf , Lutz Hans , Suter Mark , Pinschewer Daniel D 
Published: January 2023

Abstract
Interferons (IFNs) are essential for CD8+ T cell responses against lymphocytic choriomeningitis virus (LCMV) and for humoral responses against vesicular stomatitis virus (VSV), and they have a profound impact on control of these infections. Recently, we have developed LCM viruses carrying the VSV envelope gene (rLCMV/VSVG) as live-attenuated arenavirus vaccines. Here, we assessed how innate and adaptive immunity contribute to control of rLCMV/VSVG.
Even in mice lacking IFN type I and II receptors, rLCMV/VSVG was efficiently controlled and induced neutralizing antibodies at titers similar or even higher than in wildtype mice. Similarly, significantly higher frequencies of specific CD8+ T cells were detected which remained protective for more than 400 days. Mice lacking both IFN type I receptor and RAG1 (AR) failed to control the virus and became viremic while RAG1 single-deficient mice exhibited circulating viral RNA but not overt viremia. Type II IFN was found to have an effect on rLCMV/VSVG control only on an AR background, while NK cells did not seem to contribute.
Thus, viral attenuation for vaccine use was found to considerably relax the IFN-dependence of virus control and of antiviral adaptive immune responses. This provides a better understanding of the attenuated behavior of these vaccine candidates and highlights the varying impact of IFN in LCMV infection.
MHC H-2k prompts Ly49G2+ NK cell responsiveness to murine cytomegalovirus and acquisition of virus-specific effector T cells (44.16)
Authors: Stadnisky Michael David , Xie Xuefang , Bullock Timothy N. , Brown Michael G. 
Published: January 2023

Abstract
An MHC H-2k locus is critical in NK cell-mediated resistance to murine CMV (MCMV) in MA/My mice. Recently we found that congenic mouse strains C57L.M-H2k(R7) and C57L.M-H2k(R2) which express different MHC class I D alleles, also exhibit remarkable variation in MCMV resistance. Interestingly, MCMV resistance (Cmvr) in R7 mice corresponds with selective expansion and activation of inhibitory Ly49G2 receptor bearing NK cells after virus infection. Here we examined a potential role for Ly49G2+ NK cells in MCMV resistance, maintenance of splenic dendritic cell subsets and establishment of virus-specific CD8+ T cells. We show that Ly49G2+ NK cells acquire functional competence most efficiently in Dk-expressing R7 animals. In addition, we have observed a loss of splenic CD8α+ and CD11b+ dendritic cells shortly after MCMV infection in susceptible (Cmvs) R2 mice, whereas R7 animals maintain these subsets. We further report that efficient MHC H-2k NK-mediated virus immunity led to a robust and significant expansion of MCMV-specific CD8 T cells by day 6 in R7 mice in contrast with R2 or C57L (Cmvs) animals, which failed to mount efficient CD8 T cell immune responses. We conclude that MHC H-2k enhances virus specific T cell immunity in R7 congenic mice, in addition to establishing functionally competent Ly49G2+ NK cells, which can rapidly respond to MCMV infection. Supported by NIH R01 AI050072.
Opposing Effects Of IL-17/Th17 Response In Modifying Host Resistance Against Respiratory Chlamydia Infection Via Divergent Immune Mechanisms (44.22)
Authors: Wang Jun , Zhou Xiaohui , Chen Rose , Ramirez Elizabeth Acosta , Beer Jennifer , Moore Jessica M , Brown Tyler , Kolls Jay K , Stadnyk Andrew 
Published: January 2023

Abstract
The role of IL-17/Th17 response in host resistance against Chlamydia trachomatis (Ct) infection is investigated in C57BL/6 and C3H/HeN mice. We showed that C3H/HeN mice developed a skewed Th17/Th1 immune profile which led to a massive accumulation of neutrophils in the lung, impaired bacterial clearance, severe body weight loss and high mortality rate. In comparison, a relatively moderate IL-17/Th17 response was developed in C57BL/6 mice. By using recombinant adenovirus vector-mediated IL-17 transgene delivery approach, we have found that IL-17/Th17 response has both protective and detrimental roles in host resistance against Ct. On one hand, IL-17 protects host from Ct infection via stimulating pulmonary DC function for establishing a protective immunity. On the other hand, IL-17 stimulates neutrophils which counteract the protective effect of IL-17 via promoting intracellular growth of Ct. Of importance, the final outcome of two opposing effects manifests in a time-dependent and dose-dependent manner. Over expressing IL-17 at later stages of Ct infection in the airway could convert C57BL/6 mice into Ct-susceptible phenotype, indicating that an exacerbated Th17 response is responsible for an enhanced Ct susceptibility. This is the first report that a multi-facet role of IL-17 is documented in modifying host immune responses via divergent immune mechanisms in a single disease model.
T cell senescence results in persistent simian varicella virus infection in aged rhesus macaques; a model for herpes zoster (44.4)
Authors: Messaoudi Ilhem , Barron Alexander , Wellish Mary , Li Helen , Wong Scott W , Gilden Donald H , Mahalingam Ravi 
Published: January 2023

Abstract
Herpes zoster (HZ, shingles) is caused by the reactivation of latent varicella zoster virus (VZV) and causes significant morbidity and sometimes mortality in the elderly. Several clinical studies suggest that the waning of T cell immunity with age is the underlying cause of VZV reactivation. However, the exact defects in T cell immunity remain poorly understood due to the fact that VZV infection of laboratory animals, including nonhuman primates, does not result in disease. We have developed a novel animal model wherein rhesus macaques are inoculated with the closely related simian varicella virus (SVV). SVV infection of rhesus macaques reproduces hallmarks of VZV infection in humans: 1) generalized varicella; 2) the development of cellular and humoral immunity; and 3) the establishment of latency with limited transcriptional activity in sensory ganglia. This is the only model where acute infection and latency can be studied in the same animal. Furthermore, whereas adult rhesus macaques can resolve acute SVV infection, aged animals remain persistently viremic. The SVV-specific antibody response generated by elderly animals was comparable to that of adult animals. On the other hand, the SVV-specific T cell response in aged animals was delayed and reduced in magnitude. These data suggest that, similar to clinical observations about VZV, the inability of aged animals to control SVV viremia is due to a reduced T cell response.
The role of the TRAF2/3 binding site in LMP1 and CD40 signaling (44.6)
Authors: Graham John P , Bishop Gail A 
Published: January 2023

Abstract
The Epstein-Barr virus protein, LMP1, is a mimic of the cellular receptor CD40. However, LMP1 signals to B lymphocytes in an amplified and sustained manner compared to CD40. LMP1 contributes to the development of B lymphoma in HIV patients or in immunosuppressed transplant recipients. LMP1 binds the signaling adaptor TRAF2 with lower affinity than CD40, and TRAF2 is needed for inducing CD40-mediated degradation of TRAFs 2 and 3. LMP1 doesn't induce TRAF degradation, and employs TRAF3 as a positive mediator of cell signaling, whereas CD40 signals are inhibited by TRAF3. We thus tested the hypothesis that relative affinity for TRAF2 and/or, distinct sequence differences in the TRAF2/3 binding sites (TBS) of CD40 vs. LMP1, controls the disparate ways in which CD40 and LMP1 use TRAFs 2 and 3, and their distinct signaling characteristics. We examined CD40 and LMP1 mutants in which the TBS sequences were swapped, testing TRAF binding and degradation, and induction of B cell activation. Results revealed that TRAF binding affinity and TBS sequence dictate some, but not all CD40 vs. LMP1 signaling properties. Examination of TRAF binding, degradation, cytokine production, IgM secretion, and the activation of c-Jun kinase and NF-kB revealed that some events are dictated by TBS sequences, others partially regulated, and still others appear independent of the TBS sequence. NIH grant CA099997
STAT1 and STAT2 are required for control of RSV replication in vivo (44.7)
Authors: Cline Troy , Jewell Nancy , Mertz Sara , Kotenko Sergei , Schindler Christian , Durbin Russell , Durbin Joan 
Published: January 2023

Abstract
IFN-α/β are thought to be a critical component of the antiviral immune response, but susceptibility to virus infections is not always impacted by the absence of the IFN-α/β receptor. Using a mouse model of respiratory syncytial virus (RSV) infection we observed that mice lacking both the IFN-α/β and the IFN-γ receptors (IFN-αβ γR-/-) controlled virus replication as well as wild-type mice, whereas STAT1-/- or STAT2-/- mice did not. RSV infection of STAT1-/- or STAT2-/- mice resulted in a 100-fold increase in lung virus titers. We also demonstrated extensive virus spread in the nasal cavity of STAT1-/- and STAT2-/-, but not wild-type or IFN-α/β R-/- mice. The recently discovered type III, orλ, IFNs have also been shown to exert an antiviral effect via activation of the STAT1/STAT2 containing transcription factor ISGF3 which is formed in response to either IFN-α/β or IFN-λ signaling. We found that while RSV infection of wild-type mice induced production of both type I and type III IFNs, we were unable to detect IFN-λ expression or ISGF3 activation following infection of IFN-α/β R-/- mice. These data demonstrate that both STAT1 and STAT2 are necessary for inhibition of RSV replication in vivo, and suggest the existence of a novel pathway leading to their activation in the context of RSV infection.
This research was supported by NIH grants R01 AI047226 and R01 DC00648
Novel roles for T-bet in CD8 differentiation during chronic viral infections (44.8)
Authors: Kao Charlly C. , Intlekofer Andrew I. , Reiner Steven L. , Wherry E. John 
Published: January 2023

Abstract
The T-box transcription factor, T-bet, was shown recently to play an important role in memory CD8 T-cell differentiation. During an acute infection, higher levels of T-bet induced the formation of shorter-lived effectors, while lower levels induced long-lived memory CD8s. However, the role of T-bet, or any transcription factor, in regulating CD8 differentiation during chronic infections has not been examined. We infected mice with the chronic Clone-13 strain of lymphocytic choriomeningitis virus (LCMV), whose antigen-specific CD8s become highly dysfunctional (i.e. exhausted). Surprisingly, T-bet levels in these exhausted CD8s exposed to a chronic inflammatory environment are lower than ag-specific cells generated during infection with an acute (Armstrong) strain. Its expression correlated inversely with that of the inhibitory surface receptor, PD-1, which is known to contribute to T cell exhaustion. T-bet deficient mice infected with Clone-13 had higher levels of PD-1; in addition, they exhibited a severe defect in viral control and lost their antigen-specific CD8s after 2-3 weeks. Finally, we show that T-bet binds to the PD-1 promoter in CD8s from LCMV-infected hosts. Thus, during a chronic viral infection, T-bet is critical for the maintenance of antigen-specific CD8 T-cells, and it may regulate T-cell exhaustion, at least in part, by acting directly on the PD-1 locus.
Enhanced T helper 1 (Th1) responses with Fluzone® adjuvanted with a synthetic TLR4 agonist in an emulsion (45.15)
Authors: Baldwin Susan L. , Shaverdian Narek , Goto Yasuyuki , Duthie Malcolm S. , Evers Tara , Mompoint Farah , Vedvick Tom S. , Bertholet Sylvie , Coler Rhea N. , Reed Steven G. 
Published: January 2023

Abstract
Impairments in anti-influenza Th1 responses are associated with the significantly greater risk of influenza related mortality in the elderly population. Developing appropriate adjuvants for seasonal influenza vaccines could therefore improve protection against influenza in the elderly. In this study we compare the activity of three different adjuvants, an oil-in-water emulsion and a synthetic lipid A adjuvant formulated with or without an emulsion, to enhance the immunogenicity of the Fluzone influenza vaccine. Fluzone combined with lipid A plus an emulsion was the most effective vaccine tested in mice, which induced greater vaccine-specific IgG2a and IgG titers, enhanced HI titers and the production of Th1-mediated cytokine responses (IFN-γ and IL-2) to each of the Fluzone components. In contrast, while the Fluzone vaccine plus the emulsion induced good antibody responses and HI titers to all of the vaccine components following a boost, a Th2-mediated cytokine response (IL-5) and IL-10 was induced. We demonstrate that in addition to antibody responses, cytokine responses may also be needed for evaluation and characterization of new adjuvants formulations for influenza vaccines.
This work was supported in part by Grant #42387 from the Bill & Melinda Gates Foundation