bluuebunny/crossref_metadata_2025_split huggingface dataset:
Hyper Th17 responses in IL-17R knockout is regulated by segmented filamentous bacteria (SFB) (MUC4P.857)
Authors: Kumar Pawan , McAleer Jeremy , Elsegeiny Waleed , Armentrout Rachel , Pociask Derek , Kolls Jay 
Published: January 2023

Abstract
Blockade or genetic deletion of IL-17RA resulted in marked increases in IL-17 response by T-cells. We hypothesize that the hyper Th17 response in IL-17 deficient mice is due to alterations of the gut microbiome particularly overgrowth of segmented filamentous bacteria (SFB). Our data shows that Il17ra-/- and Il17rc-/- mice have overgrowth of SFB (10 fold higher than WT.Taconic), suggesting a critical role of IL-17 signaling in SFB colonization. Higher SFB colonization in Il17ra-/- and Il17rc-/- mice results in expansion of IL-17A and IL-22 producing Th17 cells. As further evidence that this expansion was not T-cell intrinsic, we observed similar frequencies of IL-17 producing cells in WT and Il17ra-/- when naïve T-cells were polarized in vitro to Th17 cells. Furthermore, vancomycin depletion of SFB in Il17ra-/- mice resulted into fewer Th17 cells in the lamina propria and spleen, suggesting gut microflora responsible for hyper Th17 response. SFB-colony free WT mice with intact IL-17 signaling can control SFB overgrowth but IL-17 deficient mice could not following SFB inoculation. To further define the role of IL-17 in SFB colonization, we have generated SFB-free Il17ra conditional knockout (Il17rafl/flxe2a cre) mice. SFB-free Il17rafl/flxe2a cre mice are devoid of hyper Th17 responses, and SFB inoculation resulted into acquisition of hyper Th17 phenotype as observed in Il17ra-/-. Our data suggest that IL-17 signaling regulates SFB colonization and associated Th17 responses.
Parenteral immunization with double-mutant heat labile toxin engages CD103+ dermal dendritic cells and redirects T cell immunity to the gut mucosa. (MUC4P.833)
Authors: Frederick Daniel , McLachlan James , Freytag Lucy 
Published: January 2023

Abstract
The vast majority of infectious agents enter hosts through mucosal compartments. Despite this, most current vaccines are administered via parenteral injection, which induce vigorous systemic but only limited mucosal immune responses. Studies have suggested that certain adjuvants can induce mucosal antibody responses regardless of route; however, there is minimal evidence that these injections direct antigen-specific effector T cells to the mucosa, where they are likely needed for protection against mucosal pathogens. We sought to explore the potential of the mucosal adjuvant dmLT (double-mutant heat-labile toxin) to drive antigen-specific T cells to the mucosa after parenteral immunization. By using a combination of peptide:MHCII-specific antibodies and tetramers, we can directly observe the impact of intradermal delivery of dmLT plus antigen, on both antigen-presenting cells (APCs) and antigen-specific T cells. Here we show that this immunization results in T cell upregulation of the gut mucosal homing marker, α4β7, in local draining lymph nodes and mucosal sites when compared to immunization with CpG. Additionally, we observed dmLT preferentially recruits skin CD103+ dendritic cells to the draining lymph nodes after parenteral immunization. These results show that administration of a mucosal adjuvant in the skin results in recruitment of mucosal-like APCs and induce T cell responses in remote, mucosal compartments.
The role of type I interferon signaling in the maintenance of resident memory T cell populations in the lung (MUC5P.867)
Authors: Turner Damian , Farber Donna 
Published: January 2023

Abstract
We have recently described populations of memory T cells that establish residence within specific niches of the lung following respiratory influenza virus (IAV) infection. Lung resident CD4 and CD8 memory T cells (TRM) are highly enriched with IAV-specific cells and exhibited increased CD11a and CD69 expression, which is characteristic of tissue resident populations. Confocal microscopy further revealed that lung TRM are clustered around airways and blood vessels within bronchovascular bundles. We sought to investigate the environmental factors that are involved in the retention of lung TRM. To investigate the role of tonic type I interferon (IFN-α/β) signaling in the maintenance of lung TRM we treated mice (previously infected with IAV) with interferon receptor (IFNAR1) blocking antibody and found that short-term blockade led to the down regulation of CD69 without a reduction in the number of lung TRM. Blocking IFNAR1 for 8 days resulted in a dramatic reduction in the numbers of CD4 and CD8 TRM in the lung as well as lung γδ T cells. We also found that, over time, fewer IFNAR1 deficient T cells were retained within the lung resident niche than wildtype T cells following IAV infection. These results suggest that constitutive IFN-α/β signaling may play a role in the maintenance of lung TRM populations, and may have therapeutic implications for respiratory diseases.
A method for detecting protein expression in single cells using the C1™ Single-Cell Auto Prep System (TECH2P.874)
Authors: Egidio Camila , Ooi Aik , Holcomb Ilona , Ruff Dave , Boutet Stephane , Wang Jing , Ramakrishnan Ramesh 
Published: January 2023

Abstract
Recent improvements in microfluidics and biochemistry have enabled single-cell molecular analysis, providing new insight into the heterogeneity of cell populations. The C1 Single-Cell Auto Prep System is an automated platform that streamlines the isolation and processing of 96 individual, live cells for RNA and DNA analysis. Single-cell protein profiling is a direct complement to genomic analysis as it provides additional insights into key molecular mechanisms and system biology. To enable this, we adapted a highly multiplexed protein detection system (Proseek Multiplex Oncology, Olink Bioscience) for use on the C1 System. We analyzed 192 individual cells from two human promyelocytic leukemia cell lines (HL60 and K562) and compared these result against 16 FACS-sorted cells in tube format. These results indicated that four key proteins (i.e. CASP3, CSTB, CD69 and MPO) are differentially expressed between the two cell lines and vary from cell-to-cell. For example, the human protein MPO, produced by neutrophils for their microbicidal activity, was differentially expression between individual HL60 and K562 cells, suggesting varying levels of neutrophil activation within the cell populations. This work demonstrates highly-multiplexed protein detection at the single-cell level and shows promise to have sufficient sensitivity, reproducibility and scale to effectively interrogate a statistically significant numbers of individual cells.
MicroRNA-dependent modulation of histone acetylation in multiple sclerosis (THER3P.875)
Authors: Guan Hongbing , Singh Narendra , Singh Udai , Rao Roshni , Mrelashvili Davit , Nagarkatti Prakash , Nagarkatti Mitzi 
Published: January 2023

Abstract
Inhibitors of histone deacetylases (HDACs) are of interest for treatment of MS. However, global modifications of DNA and chromatin structure are found to be associated with toxicities and adverse effects. For example, HDAC inhibitor, Trichostatin A, was capable of ameliorating the development of a mouse model of MS (EAE). However, the clinical trials in MS patients showed exacerbation of symptoms. This discrepancy may have been due to the non-specific nature produced by the drug or additional genetic changes. Thus, novel epigenetic approaches capable of targeting a specific set of genes in encephalitogenic T cells are desirable for controlling MS while minimizing adverse effects. We recently found that encephalitogenic CD4+ T cells present with a specific miRNA and HDAC signature characterized by increased expression of let-7e, HDAC-3, -5, -9 and reduced miR-140-5p and the rest of HDAC. Predicted let-7e targeted genes include HDAC-2, -10, and -11. We confirmed that let-7e regulated expression of HDAC-10. Using lentivirus or synthetic oligonucleotides to modulate the expression level of let-7e, we found that down-regulated HDAC-10 accompanied with up-regulation of acetyl-histone-3 and -4, which led to increased encephalitogenic Th1- and Th17-cell development and EAE. Our results indicated that HDAC-10 contributed to the pathogenesis of MS. Thus, use of let-7e oligonucleotides may constitute an alternative to chemical HDAC inhibitors in the treatment of MS.
T cell repertoire renewal following autologous stem cell transplantation for multiple sclerosis (THER3P.879)
Authors: Robins Harlan , Muraro Paolo , Malhotra Sachin , Howell Michael , Phippard Deborah , Desmarais Cindy , de Paula Alves Sousa Alessandra , Griffith Linda , Lim Noha , Nash Richard , Turka Laurence 
Published: January 2023

Abstract
Autologous hematopoietic stem cell transplantation (HSCT) is commonly employed for hematologic and non-hematologic malignancies and also has been used in clinical trials for severe autoimmune diseases to “reset” the immune system and produce a new, hopefully non-autoimmune repertoire. While it has long been feasible to eliminate the vast majority of mature T cells, it has been very difficult to accurately and quantitatively determine the relationship of the regenerated repertoire to the baseline repertoire. Here, in a phase II study of HSCT for poor prognosis multiple sclerosis (MS) patients, we have used high throughput deep TCRβ chain sequencing to directly assess millions of individual TCRs per patient sample. We find that HSCT has distinctive, and separate, effects on the CD4 and CD8 repertoires. In the case of CD4 T cells, the dominant TCR clones present pre-treatment were not detectable following reconstitution, and patients largely developed a new repertoire. In contrast, dominant CD8 clones were not effectively removed, and the reconstituted CD8 repertoire was created by clonal expansion of cells which were present pre-treatment. Importantly, patients who failed to respond to treatment had less diversity in their T cell repertoire early during the reconstitution process. This approach may enable monitoring of pathogenic or protective T cell clones following HSCT and or cellular therapies.
CD11b regulates immunity to tumor and mediates tumor promotion of macrophages recruited by mesenchymal stem cells (TUM4P.922)
Authors: Zhang Jimin , Yuan Zeng-rong , Ren Guangwen , Zhang Liying , L'Huillier Andrew , Shi Yufang 
Published: January 2023

Abstract
Here we report that systemic gadolinium chloride, or GdCl3, treatment, a potent macrophage inhibitor, significantly suppressed melanoma development in mice, which was related to the reduction of CD11b expression. We then directly examined the role of CD11b in tumorigenesis in mice by using CD11b-/- mice, or by blocking CD11b function with anti-CD11b antibody. We found that knockout or blockage of CD11b significantly suppressed the development of melanoma. Histological analysis of melanoma tissue demonstrated that abundant inflammatory cells infiltrated the surrounding region of tumor in CD11b-/- mice but not in wild type mice. This result is consistent with the increased expression of inflammatory cytokines, including TNFα, IL-6, IL-12 and IL-23, in GdCl3-treated macrophages in vitro. More IL-17 was produced in CD4+ T cells co-cultured with GdCl3-treated macrophages compared with no GdCl3 treatment, and administration of a low amount of IL-17 suppressed melanoma growth in mice in vivo, showing that CD11b-mediated immune tolerance in tumor may be through inhibition of IL-17 production in T cells. Furthermore, we reported that tumor-educated or TNFα-pretreated mesenchymal stem cells promoted tumor development by recruiting macrophages, thus we examined the effect of GdCl3 treatment or CD11b knockout and found that the tumor promoting effect was abolished. Therefore, our findings demonstrate that CD11b provides a tolerogenic signal during the immune response to tumor.
The central role of tumor-evoked Bregs in breast cancer metastasis as inducers of metastasis-promoting Tregs and myeloid-derived suppressive cells. (TUM4P.903)
Authors: Biragyn Arya , Bodogai Monica , Lee-Chang Catalina 
Published: January 2023

Abstract
We recently found that, by utilizing metabolites of 5-lipooxygenase, highly metastatic 4T1 breast cancer induces the generation of unique subset of regulatory B cells, tBregs1,2. The role of tBregs in metastasis is that they induce conversion of FoxP3+ regulatory T cells (Tregs) that protect metastasizing cells by inactivating antitumor effector NK cells and CD8+ T cells 1,3. The absence of tBregs or a B-cell deficiency in μMT mice is sufficient to disable the ability of 4T1 cancer cells to metastasize into the lungs. Although in this model the myeloid-derived suppressive cells (MDSC) have been shown to also promote metastasis, surprisingly we failed to associate their importance in metastasis. MDSC are normally expanded (an assumed pro-metastatic requirement) without apparent phenotypic changes in B-cell deficient mice or if tBregs are inactivated. However, adoptive transfer of MDSC from tumor-bearing WT mice, but not μMT mice, is able to restore lung metastasis of 4T1 cancer in μMT mice, suggesting that tBregs may be needed for the education of MDSC. Here we provide the mechanistic proof that tBregs indeed render MDSC pro-metastatic. tBregs target and use relatively small subset of MDSC. Thus, our data not only uncover previously unknown regulatory pathway, but also suggest a cautious new interpretation on the importance of MDSC in metastasis. 1 Olkhanud, P. B. et al. Cancer Res. 2011. 2 Wejksza, K. et al. J.Immunol. 2013. 3 Olkhanud, P. B. et al. Cancer Res. 2009.
Novel small molecule nucleotide homologs (SMNH) activate human macrophages to kill intracellular Mycobacterium tuberculosis. (VAC6P.949)
Authors: Khan Arshad , Singh Christopher , Pandey Rajendra , Sheri Anjaneyulu , Padmanabhan Seetharamaiyer , Iyer Radhakrishan , Jagannath Chinnaswamy 
Published: January 2023

Abstract
Tuberculosis is a major cause of death in mankind and drug resistant tuberculosis has posed a problem for therapy. Immunotherapy is an option for better control of tuberculosis. We discovered two types of novel SMNH compounds which were active as adjuvants since they activated respectively NOD2 and TLR-7 receptors in macrophages. Since these cytosolic pattern recognition pathways lead to bactericidal mechanisms, we tested the efficacy of the SMNHs to activate macrophages to kill mycobacteria. Methods: Human THP1 macrophages were treated with either NOD-2 (SB44, SB44-1) or TLR-7 (SB9922) SMNH followed by infection with M. tuberculosis. Three days later macrophages were lysed and plated for bacterial counts. Cytokines from BMs were estimated using sandwich ELISA and autophagic markers were evaluated using monodansyl cadaverine and fluorescence microscopy. Results: Both NOD2 and TLR-7 activating SMNH compounds reduced the numbers of M. tuberculosis in human macrophages by 1-log10 over 3 days of culture. These SMNHs also reduced the numbers of M. tuberculosis within mouse BMs between 1-2log10 over 3 days. SMNHs activated autophagic processes in macrophages underscoring a new mechanism to kill intracellular M. tuberculosis. Since the main scaffold of SB44 has been used as an adjuvant in human trials, we anticipate that SB44 and SB9922 like compounds can be used as adjuncts to drugs for treating ongoing infections of M. tuberculosis in humans.
Heterologous prime-boost with Ad35/AERAS-402 and MVA85A elicits potent CD8+ T cell immune responses in a phase I clinical trial (VAC7P.969)
Authors: Hokey David , O'Dee Dawn , Graves Andrew , Douoguih Macaya , Pau Maria , Versteege Isabella , Sheehan Sharon , Harris Stephanie , Landry Bernard , Sadoff Jerald , McShane Helen 
Published: January 2023

Abstract
Developing an effective vaccine to prevent pulmonary tuberculosis represents a public health priority. AERAS-402 (adenovirus expressing a fusion of M.tuberculosis antigens 85A, 85B, and TB10.4) and MVA85A (Modified Vaccinia Anakara expressing Ag85A) are TB vaccine candidates that have been independently evaluated in adult and infant studies and shown to be safe and immunogenic. Despite robust immune responses in adults, infant trials have revealed lower than expected immunogenicity for these vaccine platforms, possibly contributing to the failure of the MVA85A vaccine in a recent phase II efficacy trial. Complimentary studies of malaria and HIV vaccines report enhanced immune responses using heterologous vaccine approaches with combinations of adenoviral and pox virus vectors. Here we present the preliminary immunogenicity data from a phase I clinical trial conducted in healthy BCG-vaccinated adults that were vaccinated with one or two doses of AERAS-402 and subsequently boosted with MVA85A. Preliminary data indicate that MVA85A boosts both CD4+ and CD8+ T cell responses. Interestingly, MVA85A, which primarily stimulates CD4+ T cell responses when given alone,robustly boosted AERAS-402 CD8+ T cell responses to Ag85A, with average responses reaching approximately 0.5% to 1% of CD8+ T cells. These data suggest that this vaccine combination is highly immunogenic in adults. Further studies are needed to assess the clinical efficacy of this vaccine combination.
Development of a recombinant BCG based vaccine that prevents the infection with the human metapneumovirus stimulating Th1 immunity (VAC7P.975)
Authors: Palavecino Christian , Céspedes Pablo , Gomez Roberto , Kalergis Alexis , Bueno Susan 
Published: January 2023

Abstract
The human metapneumovirus (hMPV) is one of the leading causes of childhood hospitalization and a major health burden worldwide. Unfortunately, due to an inefficient immunological memory, hMPV infection provides limited immune protection. The inadequate immune response causes severe lung inflammation leading to airway obstruction. It is likely that an effective clearance of hMPV require a balanced Th1 type immune response, involving the participation of activated, IFN-γ secreting, T cells. A recognized inducer of Th1 immunity is Mycobacterium bovis bacillus Calmette-Guerin (BCG), which has been used for decades to protect newborns from tuberculosis. Here, we show that a re-infection worsens the viral illness in Balb/c mice, questioning the safety of vaccines based on attenuated virus. We also observed a Th1 pattern of cytokines animals infected as normal infection curse. The immunization with our recombinant BCG strains expressing the Phosphoprotein (P) from hMPV induces a protective Th1 immunity. The mice were protected from clinical disease such us weight loss, inflammatory cells recruitment in airway and viral replication in the lungs. Our recombinant vaccine also induced the activation of hMPV-specific T cells producing IFN- γ and IL-2, which protect from hMPV infection when transferred to naïve mice. These data strongly support the notion that recombinant BCG induces protective Th1 immunity and could be considered as an efficient vaccine against hMPV.
Optimizing oncolytic vaccinia virus and anti-CTLA4 combination therapy to treat cancer (VAC8P.997)
Authors: Rojas Juan , Sampath Padmavathi , Hou Weizhou , Stephen Thorne 
Published: January 2023

Abstract
Monoclonal antibody therapies, such as anti-CD25 or anti-CTLA4, have proven effective in reducing immunosuppressive cells and allowing T-cell responses in tumors. In addition, clinical trials with Oncolytic Vaccinia vectors are showing promising results thanks to the their ability to directly destroy tumors and elicit a potent immune response against cancer cells. Here we examined the possibility for combining both therapies in the treatment of tumor models. Firstly, we compared anti-CD25 and anti-CTLA4 therapies for synergistic effects in combination with Oncolytic Vaccinia. While both combinations were able to reduce tumor growth, anti-CTLA4 combination therapy produced the greatest antitumor response. Interestingly, when anti-CTLA4 antibody and Oncolytic Vaccinia where administered at the same time, immune stimulation driven by the monoclonal antibody facilitated the recognition of the virus, depleting viral replication and reducing antitumor activity. Instead, a two step approach resulted in enhanced anti-tumor effects: firstly, Oncolytic Vaccinia virus is injected and selective-replication occurs in the tumor site; once the virus reached maximum distribution throughout the tumor, anti-CTLA4 monoclonal antibody was added, resulting in enhanced recognition of tumor cells by the immune system. This combination resulted in potent antitumor activity in several orthotopic mouse models, demonstrating the requirement for both immune stimulation and viral replication.
Glycerol channel aquaporin-9 plays a critical role in the formation of memory CD8 T cells after viral infection (VIR4P.1006)
Authors: Cui Guoliang , Kaech Susan 
Published: January 2023

Abstract
Glycerol channel aquaporin 9 (AQP9) has been reported to be highly expressed in memory CD8 T cells post infection, but its function in memory CD8 T cells formation remains unknown. In this study, we report that AQP9 is vitally required for survival and differentiation of memory T cells after viral infection. AQP9 imports glycerol into cells for fatty acid esterification and triglyceride storage. Loss of AQP9 substantially impaired lipid storage and caused antiviral CD8 T cells sensitive to apoptosis. Interestingly, this defect could be rescued by ectopic expression of glycerol channels or triglyceride synthases. Furthermore, IL-7 induces AQP9 expression dependent on Foxo1 pathway. This study revealed an unexpected role of glycerol in CD8 T cell fate decision by regulating energy storage.
Broad alphaherpesvirus cross-reactivity of human CD4 and CD8 T-cells (VIR4P.1014)
Authors: Koelle David , Jing Lichen , Laing Kerry , Russell Ronnie , Barlow Russell , Haas Juergen , Johnston Christine , Wald Anna 
Published: January 2023

Abstract
Peptide-HLA diversity exceeds the TCR repertoire. The resultant T-cell cross-reactivity implies that prior antigen exposure could modulate the outcome of heterologous infections. The alphaherpesviruses HSV-1, HSV-2, and VZV have unexplained ranges of infection severity, and the reasons for partial efficacy of therapeutic live VZV vaccination are unknown. The viruses share 60 homologous ORFs. Using whole viruses, protein sets covering the viral ORFeomes, and peptides, we studied the cross-reactivity of polyclonal and mono-specific CD4 and CD8 T-cells. Effectors were enriched from PBMC using HSV or VZV by direct or cross-presentation (J. Clin. Invest. 122:654) or with tetramers. Cross-reactivity between HSV and VZV was readily detectable. For example, VZV-driven polyclonal CD4 T-cells lines showed a mean of 31% IFN-gamma/IL-2 reactivity to whole VZV and a mean of 11% reactivity to HSV-1, implying that 35% of T-cells were cross-reactive. Cross-reactivity was confirmed at the protein and peptide levels for both CD4 and CD8 T-cells. Cross-reactive CD8 T-cells directly recognized infected target cells, including non-IFN-treated VZV-infected dermal fibroblasts, despite VZV encoding a TAP-blocking protein. We hypothesize that cross-reactive alphaherpesvirus ORFs are rational subunit vaccine candidates to cover HSV and VZV, and that heterologous infections and resulting T-cell cross-reactivity may contribute to variability in infection severity and vaccine responses.