bluuebunny/crossref_metadata_2025_split huggingface dataset:
Peripheral T cell-specific role of CDC42 of Rho GTPase family in T cell homeostasis and activation (P1286)
Authors: Guo Fukun , Yang Jun-Qi , Zhang Shuangmin , Zheng Yi 
Published: January 2023

Abstract
We have reported that deletion of CDC42 of Rho GTPase family in T cells at early developmental stage (CD4-CD8-CD44-CD25+) affects T cell homeostasis by the use of CDC42flox/flox/LckCre mice. Here, we generated CDC42-deficient mice with selective expression of the Cre recombinase beginning at a late stage of T cell development (after positive selection of CD4+CD8+) using the distal promoter of the Lck (dLck) gene. As expected, CDC42 deficiency (CDC42flox/flox/dLckCre) did not affect T cell development. However, splenic CD8+, but not CD4+, T cells were significantly reduced in CDC42- deficient mice. Further analysis revealed that there was a marked increase of both proliferation and apoptosis in vivo in naïve CD8+ T cells from CDC42-deficient mice. IL-7/IL-7R signaling is critical for T cell survival. The expression of IL-7R in naïve CDC42-deficient CD8+ T cells was down-regulated. These changes were not significant in naïve CDC42-deficient CD4+ T cells. CDC42 deficiency impaired in vitro activation of both CD4+ and CD8+ T cells. Furthermore, CDC42 deficiency significantly reduced Th1 and Th2 cytokine secretion, but increased Th17 cytokine secretion from CD4+ T cells under non-polarized culture conditions. These results are significantly different from that obtained by using CDC42flox/flox/LckCre mice and suggest a peripheral T cell-specific role of CDC42 in T cell homeostasis and activation.
Blocking IL-1β prevents M2 muscarinic receptor dysfunction in parainfluenza infected guinea pigs. (P6025)
Authors: Rynko Abby , Fryer Allison , Jacoby David 
Published: January 2023

Abstract
RATIONALE: Viral infections cause asthma attacks. Viruses cause loss of function of inhibitory M2 receptors on parasympathetic nerves. Blocking IL-1β prevents M2 receptor dysfunction in antigen-challenged guinea pigs. Here we investigated whether blocking IL-1β would prevent M2 receptor dysfunction is virus infected guinea pigs. METHODS: Guinea pigs were infected with parainfluenza. In some animals, IL-1β was blocked with anakinra (30mg/kg, i.p.) 30min before infection and every 24 hours after infection. Four days after infection, guinea pigs were mechanically ventilated and M2 receptor function was measured as the ability of gallamine to increase vagally stimulated bronchoconstriction. Viral titers were determined by measuring viral RNA in the lungs. White cells in blood and lung lavage were counted. RESULTS: Anakinra prevented M2 receptor dysfunction in infected animals. White cell counts were increased in lavage and decreased in the blood of infected animals. Anakinra did not affect viral titers or white cell counts. CONCLUSIONS: Treatment with an IL-1β receptor antagonist prevents M2 receptor dysfunction in parainfluenza infected guinea pigs.
Characterization of IgE+ B cells during primary and secondary immune responses (P6035)
Authors: Allen Christopher , Yang Zhiyong 
Published: January 2023

Abstract
IgE antibodies may have a protective role in parasite immunity, but they are best known for contributing to the pathology of allergic diseases. Exaggerated IgE responses can have dangerous consequences such as systemic anaphylaxis. However, direct studies of the B cells that express IgE have been hindered by technical challenges. We have generated fluorescent IgE reporter mice and also developed an improved flow cytometry procedure, enabling specific detection of IgE+ B cells in vivo. With these tools, we have analyzed various types of immune responses and determined general features of the genesis and fate of IgE+ B cells. The frequency of IgE+ B cells was typically very low compared with other isotypes such as IgG1. The majority of IgE+ B cells differentiated into short-lived plasma cells. IgE+ B cells transiently participated in germinal centers, and showed limited affinity maturation. Consistent with these findings, IgE+ B cells expressing memory cell markers were virtually undetectable, and no persistent population of IgE+ B cells could be identified several weeks after immunization. However, secondary immunizations generated faster IgE+ plasma cell responses compared with primary immunizations, suggesting that other memory precursors likely give rise to IgE plasma cells in recall responses.
Blockade of JAK1/3 signaling with a selective inhibitor, R256, attenuates Th2 differentiation and development of allergen-induced airway hyperresponsiveness (AHR) and airway inflammation (P6047)
Authors: Ashino Shigeru , Takeda Katsuyuki , Pine Polly , Gelfand Erwin 
Published: January 2023

Abstract
JAK are major downstream regulators of signaling through cytokine receptors, activating STAT. Effective targeting of the Th2-associated JAK-STAT pathway would represent a new therapeutic approach for the treatment of asthma. Here, we studied the effects of a novel selective JAK1/3 inhibitor, R256, on Th1, Th2, and Th17 differentiation in vitro and on the development of ovalubmin-induced AHR and airway inflammation in vivo in mice. R256 prevented only Th2 differentiation in vitro and was associated with the downregulation of STAT5 and STAT6 phosphorylation. In contrast, Th2 cells were unaffected by the inhibitor when added after polarization was complete. In vivo, R256 administered during the allergen sensitization phase prevented the development of AHR, airway eosinophilia, mucus hypersecretion, and Th2 cytokine production without altering Th1 and Th17 cytokine production, indicating that selective blockade of Th2 differentiation was critical. Inhibitor administration after allergen sensitization but during the challenge phase also suppressed AHR, airway eosinophilia, and mucus hypersecretion but no effect on Th2 cytokine levels in BAL could be detected, suggesting effects on downstream cytokine receptor signaling pathways. Thus, R256-mediated inhibition of JAK1/3 activation was effective when administered both prior to and after Th2 polarization to attenuate allergen-induced Th2 dominant airway inflammation and AHR.
Tyrosine phosphatase PTP1B functions to limit cytokine responses and leukocyte trafficking during allergic inflammation (P6057)
Authors: Berdnikovs Sergejs , Abdala Valencia Hiam , Cook-Mills Joan 
Published: January 2023

Abstract
Allergic inflammation is orchestrated by an intricate cellular communication network operated by cytokines. Members of protein tyrosine phosphatase family are potentially critical in balancing cytokine-triggered cellular activation. However, virtually nothing is known about function of specific tyrosine phosphatases in immune-mediated diseases. We interrogated role of protein tyrosine phosphatase PTP1B during allergic inflammation using ovalbumin asthma models in PTP1B KO and WT mice. Flow cytometry and intravital microscopy were used to characterize trafficking of leukocytes to the lung. In this study, we observed exaggerated allergen-driven eosinophilopoiesis and early-induced alterations in leukocyte recruitment to the lungs of KO mice. Intravital microscopy revealed profound differences in kinetics of leukocyte binding to endothelium between challenged WT (in hours) and KO mice (in minutes). In concert with stimulated eosinophilopoiesis and enhanced leukocyte trafficking, there was an increase in eotaxins and a number of Th2 and tissue-derived cytokines in KO mice compared to WT mice. Eosinophils and mast cells cultured from bone marrow of PTP1B KO mice exhibited cytokine-induced hyperproliferation, hyperactivation, as well as an increased chemotaxis and upregulation of markers for migration. In summary, phosphatase PTP1B is a critical endogenous negative regulator of inflammation and a possible target for cytokine-oriented therapies in asthma.
IKKalpha is critical for the maintenance of a normal pool of regulatory T cells and for the expansion of both regulatory and effector T cells (P1095)
Authors: Chen Xin , Willette-Brown Jami , Wu Xueqiang , Howard O.M. Zack , Hu Yinling , Oppenheim Joost 
Published: January 2023

Abstract
Our previous studies show that TNF activates and expands Tregs through TNFR2. It was reported that TNF-TNFR2 interaction results in the activation of alternative NFκB pathway through p100 processing, in IKKα-dependent manner. We therefore hypothesize that IKKα may play an important role in determining the size of Treg pool. To this end, we generated a mouse strain with conditional KO of IKKα in CD4 cells (CD4-IKKα-/-) which showed a marked reduction in the number of Tregs and in the proportion of Tregs in CD4 cells, indicating that IKKα KO had more profound impact on Tregs than Teffs. In vitro, naïve CD4 cells deficient in IKKα could be differentiated as usual into Foxp3-expressing cells by TGFβ, suggesting that reduction of Tregs in CD4-IKKα-/- mice was likely due to a defect in the maintenance of the Treg pool, rather than a malfunction in the induction of Foxp3 expression. The function of Tregs deficient in IKKα was examined by co-transfer to Rag1-/- mice together with naïve CD4 cells. Although WT Tregs was able to inhibit colitis induced by transfer of WT naïve CD4 cells, IKKα-deficient Tregs failed to inhibit colitis which was associated with their incapacity to reconstitute in Rag1-/- mice. Nevertheless, naïve CD4 cells deficient in IKKα also failed to reconstitute Rag1-/- mice. Therefore, our study indicate that IKKα is important for the maintenance of normal Treg pool and is critical for the expansion of Treg cells as well as Teff cells.
A non-coding interspecies conserved motif of the Immune System Released Activating Agent induced potential immunoregularory effects (P1071)
Authors: Bakhiet Moiz , Taha Safa 
Published: January 2023

Abstract
We have recently discovered ISRAA as a novel nervous system-induced factor inducing immune responses in mouse spleen. ISRAA gene was cloned and its sequence was mapped to chromosome 14. Herein, in silico simulation approaches were used to identify the human ISRAA. The data showed that ISRAA gene constitutes a sequence within human chromosome 10 in Zmiz1 gene intron with no similarity between coding sequences. Even though, this mouse protein showed potential biological activities on human cells. Titration of mouse ISRAA on human cells depicted dose-dependent effects since 5μg exhibited positive apoptosis while 50pg revealed significant proliferation (p&lt;0.0005) and increased IFN-γ production. Cells from immunosupressed patients did not proliferate after PHA stimulation, but demonstrated significant proliferation to 50 pg ISRAA (p&lt;0.005). A concentration of 500ng ISRAA showed cytotoxic effects on two sets of tumor cell lines, but not on healthy cells. Furthermore, a predicted active site of ISRAA gene was constructed and a mutant ISRAA lacking this motif was produced and tested. This motif was found to have 72% similarity with the interspecies conserved signal peptide motif of TNFR1. Mutant ISRAA showed no biological activity compared to its wild type. Thus, functional interspecies non-coding conserved domains exist, and ISRAA effects on immunocompromised cells and tumor cell lines can be explored in future therapies to treat immunosuppressive and cancer disorders.
Development of a vaccine to attenuate viral exacerbation of asthma (P4276)
Authors: White Maria , Debley Jason , Lindell Dennis 
Published: January 2023

Abstract
Viral exacerbation of asthma exacerbation is a major cause of emergency department visits and hospitalization. Thus, vaccination against causative viruses would be an effective method for the attenuation of viral exacerbation for asthmatics. Because live/attenuated vaccines are not safe for asthmatics, inactivated viral vaccines need to be developed. We hypothesize that the innate immune response produces pro-inflammatory signals that determine the outcome of the adaptive immune response. We are evaluating the potential of an intranasal inactivated respiratory syncytial virus vaccine consisting of inactivated virus in conjunction with liposome (delivery agent) and TLR agonists (immune stimulatory agent) to attenuate viral asthma exacerbation. In vitro, bone marrow dendritic cells stimulated with inactivated RSV and TLR7/9 agonists induced more CXCL10, IFNβ, and DLL4 compared to inactivated RSV alone. In vivo, mice vaccinated with liposome-RSV and TLR 7/9 agonists enhanced viral clearance, without inducing an IL-33 or IL-17a response. Using a cockroach allergen RSV-exacerbated asthma mouse model, vaccinated mice had reduced viral load and increased activated lymphocytes and RSV specific antibodies. However, mucus secretion was also increased, indicating that further refining of the vaccine will be necessary. However, our data indicate successful vaccination against a virus clinically associated with asthma exacerbation in a well-defined animal model of asthma.
Long-term immune responses to AS03-adjuvanted, low antigen dose influenza vaccines in BALB/c mice (P4299)
Authors: Yam Karen , Brewer Angela , Gupta Jyotsana , Allen Elizabeth , Ward Brian 
Published: January 2023

Abstract
During the 2009 pandemic influenza outbreak, a vaccine formulated with the oil-in-water adjuvant AS03 and 25% of the usual antigen dose was selected for administration to Canadians. Since the long-term (LT) immune response to this vaccine remains unclear, our objective is to study AS03-adjuvanted low antigen dose influenza vaccines in mice with a focus on LT immunity. We hypothesize that LT memory following unadjuvanted full-dose vaccine will be superior to adjuvanted low-dose vaccination. Mice received 2 IM injections of influenza A/Uruguay H3N2 split vaccine formulated with 3µg antigen only or low antigen dose with AS03 adjuvant. At 3 weeks post-boost, serum hemagglutination inhibition (HAI) titers were: 276 (3µg unadjuvanted); 1365 (0.03µg+AS03); and 1681 (0.003µg+AS03). Lower antigen doses (0.00003-0.0003µg) with AS03 failed to consistently produce detectable antibodies. LT studies with these same formulations (3µg, 0.03µg+AS03, 0.003µg+AS03) are ongoing for up to 18 weeks post-boost; we are examining spleen and bone marrow cells (eg: ELISpots, cytokine profiles, B cell subsets by flow cytometry). At 1, 3 and 6 weeks post-boost, splenocytes of mice immunized with antigen alone produced higher levels of IL-4 and IL-10 than mice immunized with adjuvanted low-dose vaccines. Conversely, splenocytes from the adjuvanted low-dose groups produced higher levels of IL-2 than those from mice immunized with antigen alone. These differences in LT immunity may affect vaccine efficacy.
Systems biology assessment of human immune responses after seasonal trivalent inactivated influenza vaccine (P4307)
Authors: Hoek Kristen , Howard Leigh , Allos Tara , Samir Parimal , Niu Xinnan , Creech Buddy , Edwards Kathryn , Link Andrew 
Published: January 2023

Abstract
Systems biology represents a novel approach to comprehensively study the human immune response to vaccines at the global transcriptional and proteomic level. However, most systems vaccinology approaches utilize total PBMCs in their analyses. In this context, responses of underrepresented immune cell types in the blood are potentially obscured by the predominant cells in the PBMC fraction, and the contribution of PMNs is completely ignored. To investigate the contribution of individual cell types in the immune response following vaccination, we developed a rapid and efficient method for purifying large numbers of T cells, B cells, monocytes, NK cells, myeloid DCs and neutrophils from fresh venous human blood for systems vaccinology studies. This optimized protocol was applied to adult volunteers vaccinated with 2011-12 seasonal TIV. 100mL blood was obtained prior to and on days 1, 3, and 7 post-vaccination. Whole blood, PBMC and PMN fractions were subjected to phenotypic analysis by flow cytometry. Immune cells were fractionated and processed for RNA and protein extraction in a single day. RNA-Seq and quantitative proteomics were performed on purified cells in order to determine individual expression profiles. Our results show significant variation in the phenotypes and expression profiles of immune cells at each time point. This innovative systems approach is currently being utilized to evaluate vaccine safety and efficacy in an adjuvanted influenza clinical trial.
Involvement of the Aim2 inflammasome pathway in generating antibody responses elicited by DNA vaccination. (P4313)
Authors: Suschak John , Wang Shixia , Remington Karen , Fitzgerald Katherine , Lu Shan 
Published: January 2023

Abstract
Recent human study data suggests that DNA priming immunization can substantially enhance the protective antibody responses elicited by conventional influenza vaccines, but their mechanisms remain completely unknown. The current study explored the roles of innate immunity in regulating acquired immunity elicited by influenza vaccines. Innate pattern recognition receptors (PRRs), and their downstream signaling molecules, were investigated for their effect on DNA vaccination. Of particular interest is the absent in melanoma 2 (AIM2) protein pathway and its effects on the antigen-specific antibody response. AIM2 is a cytosolic DNA sensor important in both the maturation of the pro-inflammatory cytokines, IL-1b, IL-18, and an inflammatory form of cell death known as pyroptosis. Aim2 knockout mice and wild type controls were immunized with a DNA vaccine expressing the hemagglutinin antigen (HA) of H1 subtype influenza virus. Mouse serum anti-H1HA antibodies and B cell responses revealed that deletion of the Aim2 pathway led to a significant reduction of the anti-HA antibody response. Interestingly, non-antigen specific cytokine profile analysis did not demonstrate a dramatic change in cytokine signaling between wild type and knockout mice. Furthermore, deletion of IL-1 signaling did not significantly affect the anti-HA antibody responses. These results suggested that the Aim2 pathway plays an important role in shaping the antigen specific antibody response to DNA vaccines.
Therapeutic immunization with EBNA-1 elicits responses to novel T cell epitopes in latently Lymphocryptovirus infected rhesus macaques (P4323)
Authors: Silveira Eduardo , Fogg Mark , Leskowitz Rachel , Ertl Hildegund , Wiseman Roger , O'Connor David , Lieberman Paul , Wang Fred , Villinger Francois 
Published: January 2023

Abstract
Epstein-Barr virus (EBV) is a gamma-Herpesvirus associated with lymphoma or nasopharyngeal carcinoma in immunocompromised patients. Lymphocryptovirus (LCV) infection of rhesus macaques replicates several characteristics of the human EBV infection. EBNA-1 is a nuclear virus protein expressed in all phases of EBV/LCV infections, essential for survival, malignant transformation, tumor progression and escape from host immune response. In spite of its importance for viral replication and survival, EBNA-1 is poorly immunogenic during lytic or latent infection. However, since protection against select experimental infections was achieved after enhancement of antigen immunogenicity through immunization, we hypothesized that eliciting EBNA-1-specific responses during chronic latent infection could increase control of EBV/LCV in vivo. We therefore attempted to enhance and broaden LCV EBNA-1 responses via immunization with an AdC68-LCV-EBNA-1 vector in groups of rhesus macaques that were latently infected with LCV but exhibiting low baseline responses. Following immunization, several CD4+ and CD8+ T cell epitopes were identified through IFN-g ELISPOT assays either as novel or markedly expanded responses. One such novel expanded CD8+ T cell epitope was mapped to a 9-mer peptide restricted by Mamu B*029, and a tetramer was developed for staining of specific CD8+ T cells. The study provides proof of concept that EBNA-1 may be a suitable vaccine target.
Vaccination with recombinant PIV5 expressing influenza neuraminidase provides protection against homologous and heterologous influenza virus infection (P4291)
Authors: Tompkins Stephen , Mooney Alaina , Li Zhou , Gabbard Jon , Dlugolenski Daniel , Johnson Scott , Tripp Ralph , He Biao 
Published: January 2023

Abstract
Vaccination is considered the most effective measure for controlling influenza virus infection, however current vaccination methods are inadequate. Virus-vectored vaccines offer an appealing alternative. Parainfluenza virus 5 (PIV5), a non-segmented, negative-stranded RNA virus (NNSV) in the family Paramyxoviridae is an appealing vector candidate as it has a stable genome without a DNA phase in its life cycle, is readily grown to high titers in approved vaccine cell lines, and infects many mammals without causing disease. Neuraminidase (NA), a glycoprotein found on the surface of influenza A virus as well as virus-infected cells is more conserved than the hemagglutinin surface glycoprotein, the primary antigen in current influenza vaccines. This increases the likelihood of achieving broad protection if used as a vaccine antigen. We show that vaccination with PIV5 expressing the NA from H5N1 highly pathogenic avian influenza (HPAI; rPIV5-N1 (H5N1)) or pandemic H1N1 (rPIV5-N1 (H1N1)) primes robust NA-specific antibody and T cell responses and confers complete protection against homologous (NA-matched) influenza virus challenge. Moreover, immunization also confers significant cross-protection against challenge with heterologous influenza virus within the same NA type. These results suggest that NA can be an effective vaccine antigen and protect against even a stringent challenge, such as with H5N1 HPAI virus, making it an appealing candidate pandemic influenza vaccine.
Dual functions of PD-1 in CD4+ T cell responses by sublingual mucosa-mediated antigen application (P3210)
Authors: Ohno Tatsukuni , Zhang Chenyang , Yagita Hideo , Azuma Miyuki 
Published: January 2023

Abstract
[Objective] PD-1 is a co-inhibitory T cell receptor and binds two ligands, B7-H1 and B7-DC. Although the B7-H1:PD-1 pathway is involved in the induction of peripheral tolerance, immunostimulatory effects of B7-H1 or B7-DC have been demonstrated. The sublingual mucosa (slm) has been used as a route for sublingual immunotherapy to induce tolerance against allergens. On the other hand, the slm is also used as vaccination sites to induce immune responses against pathogens. The mechanisms by which slm-mediated immune responses are controlled are unclear. In this study, we investigated the involvement of PD-1 in OVA-specific CD4+ T cell responses by slm-mediated antigen application with and without mucosal adjuvant. [Results and Discussion] CFSE- labeled DO11.10 T cells were transferred into immunodeficient mice and OVA was painted onto slm with or without cholera toxin (CT). In the presence of CT, blockade of PD-1 by mAb or deficiency of PD-1 inhibited Ag-specific CD4+ T cell proliferative responses, whereas without CT, similar PD-1 blockade enhanced the proliferation. PD-1 is comparably induced on Ag-specific T cells with and without CT. Blockade of B7-H1 did not clearly affect T cell responses even in the absence of CT. CT markedly inhibited the induction of B7-H1 in slm DCs. Our results suggest the existence of PD-1-mediated immunostimulatory pathway in the presence of CT. CT may affect the availability of B7-H1 and B7-DC on slm DCs and regulate the dual function of PD-1.