bluuebunny/crossref_metadata_2025_split huggingface dataset:
NLRP3-dependent IL-1β production by FcγR-mediated Syk kinase signaling confers protective immunity in Francisella tularensis vaccination (VAC7P.966)
Authors: Periasamy Sivakumar , Duffy Ellen , Harton Jonathan 
Published: January 2023

Abstract
Pneumonic tularemia is a fatal disease caused by Francisella tularensis (Ft), a category ‘A’ biothreat agent with no FDA approved vaccine. Previous studies have shown that inactivated-Ft (iFt) complexed with anti-Ft LPS mAb (iFt-mAb) elicits protective immunity against lethal Ft challenge. Other vaccine studies have also shown that IL-1β and NLRP3 play a role in adaptive immunity. The AIM2 inflammasome is critical for innate immunity to Ft, but NLRP3 appears to be unnecessary. We investigated the role of inflammsomes in immune responses to iFt-mAb and found that NLRP3 is essential in this vaccine strategy. An iFt-mAb vaccination completely failed to protect IL-1RKO and NLRP3KO mice against lethal Ft challenge compared to Wt mice. Importantly, Ft-specific antibodies were significantly lower in both KO mice than Wt mice after vaccination and challenge. NLRP3-deficient mBMDM or THP-1 cells produced significantly less IL-1β compared to WT cells after iFt-mAb treatment. To probe the mechanism by which iFt-mAb activates NLRP3 and IL-1β, the role of FcγR and TLR2 signals were explored. Internalization of iFt-mAb and activation of NLRP3 were FcγR-dependent, while NLRP3-dependent IL-1β production required TLR2 and FcγR-signaling through Syk kinase. Our results highlight the role of IL-1β and NLRP3 in successful iFt-mAb vaccination that elicits better antibody and cellular responses. Specifically, FcγR signaling is required for NLRP3 inflammasome activation in this vaccine strategy.
Predicting vaccine efficacy against tuberculosis using immunologic and radiologic markers in the non-human primate model (VAC7P.974)
Authors: DiFazio Robert , Gideon Hannah , Coleman Teresa , Maiello Pauline , Frye Lonnie , Tomko Jaime , O'Malley Melanie , Johnston Paul , Fillmore Daniel , Rodgers Mark , Bigbee Carolyn , Chedrick Chelsea , Lin Philana , Flynn JoAnne 
Published: January 2023

Abstract
The only licensed vaccine (BCG) for tuberculosis (TB) has variable efficacy and does not protect against pulmonary TB. New vaccines are in development and in human clinical trials. However, vaccine trials are long and expensive without useful surrogate markers of protection. A problem thus emerges: what does a protective response to TB look like? This project’s goal is to identify markers of vaccine efficacy that can further be developed as new clinical endpoints. We posit that exploiting differences in vaccine outcome (protected vs. unprotected) will identify novel immunologic markers that correlate with outcome, and serial PET/CT will predict outcome early in infection. Vaccine-induced protection could work at any point in early infection: movement of bacilli from airway to parenchyma; formation of primary lesions; migration of bacteria to LN; and development of secondary lesions. In both our rhesus and cynomolgus macaque models of infection, prime-boost vaccination with BCG followed by viral-expressed or protein antigens protects against TB disease better than BCG alone or no vaccine. However, granuloma formation is not prevented and the vaccines do not reduce initial granuloma numbers. However, total PET signal, as a surrogate for inflammation and disease, in lungs of vaccinated animals is significantly lower at early time points. Future work will develop the predictive markers to elucidate early immune mechanisms that confer protection in the lungs.
Pentameric gH complex is the primary target of neutralizing antibodies generated in cytomegalovirus vaccine immunized rabbit (VAC7P.982)
Authors: Tang Aimin , Freed Daniel , Tang Qi , Li Fengsheng , Meng Weixu , Wang Dai , An Zhiqiang , Fu Tong-Ming 
Published: January 2023

Abstract
Human cytomegalovirus (HCMV) belongs to the herpesvirus family and can cause severe diseases in immune-compromised individuals and birth defects in babies from in utero infection. Understanding the protective immunity is the pre-requisite in developing an effective vaccine against HCMV. Humoral immunity can provide protection against HCMV infection and that such a protection comes from antibodies primarily targeting antigens of pentameric gH complex. To support a vaccine concept based on the pentameric gH complex, we generated and characterized 45 monoclonal antibodies (mAb) from a rabbit immunized with an experimental vaccine virus in which the expression of pentameric gH complex restored. From 45 mAbs, we identified 25 mAbs with neutralizing activities. We demonstrated that binding affinity to virions is not correlated with neutralizing activity and potent neutralizing mAbs (Elite neutralizer) preferentially bind to the pentameric gH complex. Using phylogenetic analysis, we further demonstrated that these antibodies are grouped into 26 distinct B cell lineages and the neutralizing and binding mAbs were largely segregated into different groups. In addition, neutralizing antibodies have longer complementarity-determining region (CDR) 3 than those non-neutralizing mAbs. In conclusion, the pentameric gH complex is the primary target of neutralizing antibodies by vaccination with a HCMV vaccine.
A polyantigenic genotype 1a/1b consensus hepatitis C virus DNA vaccine induces broadly reactive HCV-specific cellular immune responses in both mice and non-human primates (VAC7P.985)
Authors: Latimer Brian , Toporovski Roberta , Yan Jian , Pankhong Panyupa , Khan Amir , Sardesai Niranjan , Jacobson Jeffrey , Weiner David , Kutzler Michele 
Published: January 2023

Abstract
Chronic HCV is a surreptitious disease that can lead to cirrhosis and cancer decades following infection. Each year, 3-4 million individuals are infected. However, there are currently no vaccines available to prevent chronic HCV infection. From patients who acutely resolve (A.R.) infection, a strong and broadly reactive cytotoxic T lymphocyte (CTL) response is associated with viral clearance. With in vivo electroporation (E.P.), DNA vaccines induce strong CTL responses, making them well suited for an HCV vaccine. Our vaccine strategy maximizes recipients' opportunity to induce a broad T cell response with genotype 1a/1b consensus sequences of multiple HCV antigens. Immunogenicity of the HCV vaccine was assessed in mice and non-human primates (NHP). Animals were immunized three times, two weeks apart for mice and six weeks apart for NHP as an intramuscular injection followed by E.P. In mice, interferon-γ (IFNγ) responses spanned all HCV antigens and with total values exceeding 6,000 spot-forming units per million mononuclear cells (SFU) within both the spleen and liver by IFNγ ELIspot. Similarly in NHP, immune responses up to 4,680 SFU per million PBMCs were detectable by IFNγ ELISpot. Flow cytometry revealed HCV-specific CD4+ and CD8+ T cell responses by cytokine staining and significant frequencies of cytolytic CTL compared to baseline. Here, we have demonstrated the magnitude and breadth of HCV-specific immune responses elicited by an HCV DNA vaccine in mice and NHP.
A systems biology approach revealed distinct host responses between ALVAC-SIV and NYVAC-SIV vaccines during an evaluation of immunogenicity in rhesus macaques (VAC7P.992)
Authors: Yang Yajie , Gordon Shari , Doster Melvin , Liyanage Namal , Chang Jean , Williams Chris , Agy Michael , Mclain Randy , Agricola Brian , Law Lynn , Franchini Genoveffa , Palermo Robert , Katze Michael 
Published: January 2023

Abstract
Though only modestly effective, RV144 is the first HIV vaccine clinical trial to show efficacy against heterosexual transmission. It tested ALVAC-HIV (a canarypox vector expressing HIV Gag, Pro and gp120) in the prime-boost combination with AIDSVAX B/E (a HIV bivalent gp120). To understand the immune correlates and inform future trials, we repeated this regimen in a rhesus model and contrasted two different pox vectors. Our study consisted of 24 animals in four groups: two initially receiving ALVAC-SIV or the vaccinia vector NYVAC-SIV (both expressing SIV Gag, Pol and Env) as the prime, with later addition of SIV gp120 and alum boosts; and two control groups receiving empty ALVAC or NYVAC and alum only. Microarray analysis of whole blood samples showed that as early as 6 hours after the initial immunization, the gene expression features of the ALVAC-SIV group were distinct from the animals receiving NYVAC-SIV, with innate immunity pathways activated in an ALVAC-SIV-specific manner. By 24 hours post vaccination, ALVAC-SIV induced more differentially expressed (DE) genes than the other three groups relative to the pre-immunization baseline, and NYVAC-SIV and NYVAC control induced similar numbers of DE genes. We will relate these very early host responses to the differences in immunogenicity observed over the course of immunization. The impact of these early events on the later outcomes will be determined once the protective efficacies of the two vaccines are determined.
Identification of Chlamydia trachomatis proteins recognized by T cells from high-risk women resistant to reinfection and/or protected from ascending infection. (VAC7P.961)
Authors: Darville Toni , Zheng Xiaojing , O'Connell Catherine , Picard Michele , Wiesenfeld Harold , Hillier Sharon , Flechtner Jessica 
Published: January 2023

Abstract
Murine and human data indicate IFNγ-producing Chlamydia-responsive CD4 T cells are key to protection from genital tract disease, and chlamydial proteins have been identified that induce protection in mice. A preventative vaccine against C. trachomatis requires the identification of T cell antigens that induce protection in humans. We analyzed peripheral blood CD4 and CD8 T cell responses from high-risk women defined as “resistant” or “susceptible” based on negative evidence for reinfection or detection of repeated reinfection during a 12-month follow-up period. Autologous APCs preincubated with E. coli clones expressing individual chlamydial ORFs were coincubated with CD4 or CD8 T cells and IFN-γ measured by ELISA. CD4 T cells from resistant women recognized 16 proteins significantly more frequently when compared to susceptible women, and only 2 of 16 were among a subset of immunodominant CD4 proteins suggesting restricted MHC recognition. No proteins recognized by CD8 T cells met immunoprotective criteria. Five CD4 and 5 CD8 antigens were more frequently recognized by women who had cervical infection but no evidence of endometrial infection by PCR. Thus, although protection from reinfection appears dependent on CD4 T cells, both CD4 and CD8 T cells may contribute to disease prevention. The use of samples from persons with evidence of natural immune protection for antigen discovery should advance development of a vaccine against C. trachomatis.
A West Nile virus NS4B P38G mutant induces a differential innate cytokine response in human cell lines. (VAC8P.1005)
Authors: Xie Guorui , Luo Huanle , Mann Brian , Wicker Jason , McBride Jere , Tesh Robert , Barrett Alan , Wang Tian 
Published: January 2023

Abstract
West Nile virus (WNV)-induced neurological disease has become a public health concern. No vaccines have been approved for human use. We have previously shown that an attenuated WNV, the nonstructural(NS)4B-P38G mutant, induced stronger innate cytokine and T cell responses than wild-type WNV. Mice immunized with this mutant were all protected from subsequent lethal wild-type WNV infection. Furthermore, TLR7 and MyD88 expression is required for host protection from NS4B P38G infection. Here, we have examined innate cytokine response in THP-1 cells (human monocytic cells) and THP-1 derived macrophages following with NS4B P38G infection. We have found that NS4B P38G induced stronger type 1 IFN and pro-inflammatory cytokine responses in THP-1 cells than those infected by wild-type WNV. Interestingly, we noted that there was a higher production of pro-inflammatory cytokines, but not IFN-b in NS4B P38G infected THP-1 derived macrophages. Our results also showed an upregulation of TLR4, TLR7 and RIG-I expression in THP-1 cells following NS4B P38G infection, whereas TLR2 expression was significantly increased in mutant infected THP-1 derived macrophages. Similar as our findings in mice, NS4B P38G mutant had a very low replication rate in both types of human cells. Overall, these results suggest that NS4B P38G induced a differential innate cytokine response in human monocytic cells and macrophages, possibly via different pathogen recognition receptor signaling pathways in these cells.
Manipulating TLR pathways activated by oncolytic vaccinia virus elicits a potent immune response against tumors (VAC8P.996)
Authors: Rojas Juan , Sampath Padmavathi , Hou Weizhou , Stephen Thorne 
Published: January 2023

Abstract
The immune response raised by oncolytic Vaccinia Virus after the infection and destruction of cancer cells is believed to be a key factor in the promising recent clinical results seen with these viruses. However, Vaccinia Virus primordially activates TLR2, which is associated with Th2 conversion and strong humoral responses. For cancer therapy, elicitation of cellular responses is thought to be more effective for the direct destruction of tumor cells. In this work, we switched the TLR response that a modified oncolytic Vaccinia Virus elicits. Firstly, we looked to activate the TLR3 pathway after infection with Vaccinia Virus, which is more associated with cellular responses. For this purpose, we constructed an oncolytic Vaccinia Virus that codifies for the TRIF protein, a key adaptor of the TLR3 pathway. The resulting virus increased the level of cytokines, chemokines and DAMPs released after infection, resulting in improved cytotoxicity in cancer cells and significantly enhanced therapeutic effects in mouse tumor models. Concurrently, we demonstrated that enzymatic deglycosylation of the surface of the virus is able to ablate the activation of cell surface TLRs, increasing the level of delivery and early viral gene expression in tumors in vivo. Finally, combination of both strategies demonstrated increased amounts of CTLs targeting the tumor, reduced levels of anti-viral neutralizing antibody and a further increase in antitumor activity in orthotopic mouse models.
A Non-lytic form of listeriolysin o induces human dendritic cells to produce IL-12 and IL-23. (VAC8P.1000)
Authors: Hernández Flores Karina , Thomas Dupont Pablo , Vivanco Cid Hector 
Published: January 2023

Abstract
Introduction. Listeriolysin O (LLO), a pore-forming citolysin is a key virulence factor of Listeria monocytogenes (LM). LLO induces a wide range of biological effects during the infection by LM or by itself as recombinant antigen. Here we studied the effects of a non hemolytic form of LLO on the production of two key cytokines for coordinating innate and adaptive immune responses against intracellular pathogens such as IL-12 and IL-23. Objective. We aimed to test the innate immune response of Human Monocyte-derived Dendritic cells (hDCs) to a non-hemolytic form of LLO named as dtLLO (mutated at C484A, W491A, W492A positions). Materials and methods. hDCs were stimulated with dtLLO ( 1μg/mL), in the presence or absence of 50 μg/mL of polymyxin B; 24 hours after stimulation, sups were tested to measure the concentration of TNF-α, IL-6, IL-10, IL-12 and IL-23 by ELISA technique. Results: dtLLO induced the production of IL-6 (1,660.86 pg/mL), TNF α (2,732.36 pg/mL), IL-12 (2,242.92 pg/mL) IL-10 (930.66 pg/mL) and IL-23 (349.47 pg/mL). Cytotoxic effects were undetected. Conclusion: We have identified a potent innate immune response when dtLLO is used as an exogenous antigen on hDCs. The production of IL-12 and IL-23 by human DCs after the recognition of LLO could be very important for the immune response during the human infection by Listeria. As a potential adjuvant for human vaccination purpose, dtLLO could be use to polarized the immune responses toward a Th1 or Th17 responses.
miR-155 overexpression augments anti-viral CD8+ T cell responses (VIR4P.1007)
Authors: Hope Jennifer , Stairiker Christoper , Gracias Donald , Stelekati Erietta , Carey Alison , Mueller Yvonne , Katsikis Peter 
Published: January 2023

Abstract
MicroRNAs (miRNAs) are highly conserved, small, single-stranded non-coding RNAs that play an essential role in regulating cellular processes. One miRNA in particular, miR-155, has been well characterized in many immune cell subsets. Recently, our lab has demonstrated an essential role for miR-155 in CD8+ T cell responses. Using murine models of influenza virus and Listeria monocytogenes (LM) infection, we have shown that CD8+T cells deficient in miR-155 demonstrate an intrinsic defect in anti-viral and anti-bacterial responses. Currently, we are investigating and characterizing the effect of miR-155 overexpression in CD8+ T cells. Adoptively transferred miR-155-overexpressing OT-I CD8+ T cells demonstrate a greater than 2-fold increase in the absolute number of anti-viral CD8+ T cells in the lungs of infected animals when compared to control OT-I CD8+ T cells at days 8 and 10 post-influenza infection. Interestingly, a significant number of miR-155 overexpressing OT-I CD8+ T cells demonstrate IFNγ- and TNFα-production and LAMP1-expression ex vivo. Overexpression of miR-155 in CD8+ T cells does not affect memory formation. These data suggest that overexpression of miR-155 augments effector CD8+ T cell responses.
CD27high/KLRG1low CD8+ T cells that persist throughout MCMV infection are highly expansive and have the ability to reestablish MCMV immunity (VIR4P.1011)
Authors: Quinn Michael , Turula Holly , Snyder Christopher 
Published: January 2023

Abstract
Cytomegalovirus (CMV) is a herpesvirus that establishes life-long latency in 60-80% of Americans. Constant immune surveillance is necessary to prevent viral reactivation from latency and results in the accumulation of functional CMV-specific CD8+ T cells (CD8s) over time, a process termed memory inflation. As such, CMV reactivations remain a clinical concern for immunosuppressed patients and reconstituting CMV immunity is critical for the long-term prevention of CMV disease. Understanding the maintenance of memory inflation may reveal novel approaches to restore CMV immunity. Previous work has shown that the majority of inflationary CD8s express a terminally-differentiated “effector” (TEFF) phenotype, have a short half-life and appear unable to sustain long-term CMV immunity. Interestingly, inflationary populations also include a minor subset of CD8s that express a “memory” phenotype (TM). Utilizing the murine model, MCMV, our data showed that the minor TM population was responsible for the majority of the inflationary population’s expansive capacity following transfer into naive mice and MCMV challenge. In addition, transferred TM cells produced both TM and TEFF progeny, which inflated and persisted in the recipients. Critically, when TM cells were transferred into chronically-infected mice, they survived and produced TEFF CD8s if host immunity was lost. Thus, TM CD8s appear to be a crucial component for the establishment and maintenance of CMV immunity.
Decreased frequency of regulatory T cells related to liver injury in acute viral hepatitis (VIR5P.1032)
Authors: Lee Jeewon , Choi Yoon Seok , Shin Eui-Cheol 
Published: January 2023

Abstract
FoxP3+ regulatory T cells (Tregs) are essential for maintaining self-tolerance and homeostasis in the immune system. Their role and character are already shown in many disease model, however, still concealed in acute viral hepatitis. In this study, we investigated alteration in the peripheral blood Treg population during acute hepatitis A (AHA) and its implication in the immune-mediated liver injury. Using isolated peripheral blood mononuclear cells (PBMCs) of 62 AHA patients, the decreased frequency and attenuated function of Tregs were observed; those results were inversely correlated with the severity of liver damage, represented by serum alanine aminotransferase (ALT) level. In addition, the frequency and suppressive function of Tregs were recovered at the convalescence phase when serum ALT level was completely normalized. We found a cause of the reduced frequency of Tregs from elevated expression level of Fas receptor, related to apoptosis, on Tregs in AHA patients. Indeed, the Tregs of AHA patients were more susceptible to Fas-mediated apoptosis than those of healthy controls in vitro experiments. In present study, we demonstrate that the frequency and suppressive function of Tregs were attenuated during acute HAV infection, resulting from apoptosis of Tregs induced by Fas-mediated mechanism.
Live attenuated rubella vectors express HIV Env and SIV Gag antigens in the context of an acute infection and elicit high antibody titers, memory B cells, and Gag specific CD8+ T cells (VIR5P.1033)
Authors: Berkower Ira , Virnik Konstantin , Hockenbury Max , Rosati Margherita , Alicea Candido , Valentin Antonio , Pavlakis George , Felber Barbara 
Published: January 2023

Abstract
Live attenuated viral strains are among our most potent vaccines. We have used the live attenuated rubella vaccine strain RA27/3 as a vector to express HIV MPER and SIV Gag antigens. These vectors combine the potency, durability and safety of rubella vaccine with the antigenicity of the inserts. We have recently completed the first successful trial of live rubella vectors in rhesus macaques. Rubella vectors grew robustly in macaques, and the vaccine inserts were strongly immunogenic. Simultaneous immunization with two vectors elicited antibodies to both inserts. Anti-Gag antibodies after immunization reached titers equal to natural SIV infection. Antibodies persisted for at least one year and declined at the same rate as anti-rubella antibodies, which protect for life. Re-exposure to the vector boosted the immune response, indicating the induction of memory B cells. Priming with DNA vaccine and boosting with the vector produced high levels of Gag specific CD8+ T cells. The vector can stably express larger inserts, up to 320 amino acids for SIV Gag proteins, including part of MA and all of CA and P2 proteins. They also express engineered HIV Env outer domains up to 180 amino acids that include the CD4 binding site and bind monoclonal VRC01. Rhesus macaques will provide the ideal animal model for evaluating novel vaccine inserts for immunogenicity, induction of neutralizing antibodies, and protection against SIV or SHIV challenge.
SHP-1 regulates neuroinvasion of neurotropic virus into the CNS (VIR5P.1035)
Authors: Schneider Karin , Massa Paul 
Published: January 2023

Abstract
Viral infections in the central nervous system (CNS) may have devastating clinical consequences to the host. Neurotropic virus infections often begin in peripheral tissues, which then spread to the CNS via peripheral nerves following entry into nerve terminals by receptor-mediated endocytosis and subsequent retrograde axonal transport into the CNS compartment. In our lab, a possible novel role for the protein tyrosine phosphatase, SHP-1, in regulating neuroinvasion via peripheral nerves of a neurotropic virus (Theiler’s Murine Encephalomyelitis Virus, TMEV) is being investigated. Preliminary data using immunohistochemistry of TMEV showed extensive staining of spinal cord neurons after intraperitoneal inoculation in mice with a null mutation in the SHP-1 gene (me/me mice) compared to wild type mice. Further, intramuscular (IM) inoculations of TMEV caused virus infection in spinal cord neurons and subsequent paralysis of SHP-1-deficient mice but not of wild type mice. IM and intraperitoneal injections (IP) of retrograde axonal transport tracers fluorogold and horseradish peroxidase showed increased tracer levels in the spinal cords of SHP-1-deficient mice compared to wild type mice. Therefore, we propose SHP-1 may play a role in regulating viral neuroinvasion by controlling retrograde axonal transport from peripheral sites of infection. In this sense, SHP-1 may play novel roles in antiviral responses including inhibition of virus invasion and spreading in the CNS.