bluuebunny/crossref_metadata_2025_split huggingface dataset:
Functional subsets of intestinal NKT cells (MUC3P.942)
Authors: Aguiar Cristhiane , Takahashi Daisuke , Câmara Niels , Wingender Gerhard , Engel Isaac , Kronenberg Mitchell 
Published: January 2023

Abstract
NKT cells constitute a distinct population of lymphocytes characterized by their reactivity to glycolipids presented by CD1d. Although they are found at a relatively low frequency, because of their clonal specificity and rapid cytokine production, they have been reported to influence many types of immune responses. Recently NKT cells have been divided into functional subsets analogous to T helper (Th) Th1, Th2 and Th17 cell subsets, called NKT1, NKT2 and NKT17. We have examined if similar functional subsets are found in the mouse intestine. NKT cells are found at a low frequency in intraepithelial lymphocytes (IEL) of the small and large intestine and a higher prevalence in lamina propria lymphocytes (LPL). We analyzed intestinal NKT cells using flow cytometry to explore the transcription factors and surface markers expressed by these cells. We found out that all the functional subsets are represented in intestinal NKT cells, however, the percentages of each subset in the intestine are different from those found in spleen. We also evaluated the response of intestinal iNKT cells to the agonist α-Galactosylceramide. We observed that, in response to in vivo exposure to the potent antigen α-Galactosylceramide, intestinal NKT cells produced IFN-γ, TNF-α and GM-CSF and increased expression of activation markers, as CD69 and CD40L, suggesting that cells with an NKT1 function are most prevalent. We are now exploring how the microbiota affects the functions of intestinal NKT cells.
Identification of highly potent and selective Tyk2 inhibitors for the treatment of autoimmune diseases through structure-based drug design (THER2P.961)
Authors: Masse Craig , Miao Wenyan , Greenwood Jeremy , Shelley Mee , Kennedy-Smith Joshua , Kapeller Rosana 
Published: January 2023

Abstract
The JAK family kinase Tyk2 is essential for IL-12 and IL-23 signaling, which are associated with Th1 and Th17 cell differentiation and activation. The Th1 and Th17 pathways have been implicated in the pathogenesis of psoriasis and inflammatory bowel diseases (IBD) thereby making Tyk2 a highly attractive target for the treatment of these disorders. However, given the high degree of sequence homology between the JAK family kinases, designing potent and selective Tyk2 inhibitors remains a challenge. Using an innovative structure-based approach, we have designed, synthesized and characterized small molecule inhibitors optimized for JAK family selectivity using computational free energy perturbation (FEP) methods and medicinal chemistry SAR. We have identified selective Tyk2 inhibitors with pM activity against Tyk2 (Ki=140-520 pM) and &gt;100 fold selectivity over JAK2 and JAK1 with more moderate selectivity over JAK3. These analogs are orally bioavailable (85%F) with suitable drug-like properties. NDI-031232 was determined to be highly selective across 359 kinases, and is a potent inhibitor of IL-12 induced pSTAT4 in hPBMCs (IC50=17 nM) and IL-12 induced IFNg in human whole blood (IC50=520 nM). NDI-031232 also demonstrated robust efficacy in blocking IL-12-mediated IFNg production in an ex vivo mouse model. Therefore, selective inhibitors of Tyk2 retain the anti-inflammatory activity while reducing potential for dose-limiting side effects observed with non-selective JAK inhibitors.
RGC-32 enhances suboptimal CD8 cytotoxic T cell effector function in acute graft versus host disease and promotes lupus-like disease in chronic graft versus host disease (THER2P.965)
Authors: Rus Violeta , Nguyen Vinh , Badea Tudor , Cudrici Cornelia , Tegla Cosmin , Mekala Armugam , Rus Horea 
Published: January 2023

Abstract
Response Gene to Complement (RGC)-32 is an intracytoplasmatic protein expressed in a variety of cells in response to sublytic complement activation, TGFb and other cytokines. It plays a role in cell growth, fibroblast activation and renal fibrosis, Th17 and plasma cell differentiation. To assess whether RGC-32 expression promotes lupus-like disease, we used WT and RGC-32 KO mice to investigate whether lack of RGC-32 in host cells affects parameters of chronic (c)GVHD in the Bm12→B6 model and whether lack of RGC-32 in donor T cells affects parameters of acute (a)GVHD in the P→F1 model. In F1 mice injected with RGC-32 KO splenocytes, an attenuated phenotype of aGVHD was observed at suboptimal (30 x106) but not optimal (50 x106) donor cell doses as indicated by a significant decrease in donor CD8 engraftment, proliferation and host B cell elimination. Bm12→B6 cGVHD induced in RGC-32 KO hosts displayed an attenuated phenotype as indicated by decreased number of germinal center B cells and plasmablasts, mRNA expression of Prdm1, IRF-4 and Aicda and anti-dsDNA Ab production. These results suggest that RGC-32 expression in host B cells promotes autoimmune parameters in cGVHD while its expression in T cells enhances suboptimal CD8 cytotoxic T cell effector function thus preventing humoral autoimmunity in aGVHD. These data support the idea that RGC-32 blockade has the potential to reverse abnormalities in the CD8 T cell and B cell pathways that contribute to lupus pathogenesis.
Impact of human mutant TGF-β1/Fc protein on memory and regulatory CD4+T cells following lymphodepletion in rhesus monkeys (THER2P.953)
Authors: Guo Hao , Lu Lien , Wang Rijian , Abdulkerim Hayat , Reimann Keith , Thomson Angus , Ezzelarab Mohamed 
Published: January 2023

Abstract
Transforming growth factor beta (TGFβ) is critical for regulatory T cells (Treg) homeostasis and regulation of peripheral tolerance. In rodents, combined human TGFβ1 fusion protein (TGFβ1/Fc) and rapamycin treatment prevents allograft rejection. We evaluated the influence of human TGFβ1/Fc on CD4+T cells in vitro, and in vivo following lymphodepletion in rhesus monkeys. In vitro, TGFβ1/Fc and rapamycin synergistically reduced ki67 expression and proliferation of rhesus memory T cells (Tmem), following CD3/CD28 activation or allo-stimulation. However, increased percentage of Treg and Treg/Th17 ratio were only observed in the presence of interleukin-2 (IL-2). Four monkeys received immunosuppression in the form of Thymoglobulin and rapamycin, without (n=2) or with (n=2) TGFβ1/Fc infusion. In monkeys receiving TGFβ1/Fc, trough levels were maintained at 2-7 ug/mL, and were undetectable by day-45. With no TGFβ1/Fc infusion, rapid recovery of Tmem to pre-depletion levels was observed by day-21, and continued to increase. In contrast, with TGFβ1/Fc infusion, the recovery of Tmem was delayed until day-56. In addition, increased Treg and Treg/Tmem ratio were observed in peripheral blood and lymph nodes. However, Treg/Th17 ratios were reduced in all monkeys. In lymphodepleted monkeys, infusion of human TGFβ1/Fc delays CD4+Tmem recovery, and is associated with increased Treg/Tmem ratio. Combined IL-2 and TGFβ1/Fc infusion may be essential to improve Treg/Th17 ratio after lymphodepletion.
TWEAK receptor-Fc suppresses germinal center formation and pathogenic B cells in lupus mice model via inhibition of TWEAK/Fn14 pathway (THER3P.1002)
Authors: Kim Sung-Min , Min Hong-Ki , Park Jin-Sil , Byun Jae-Kyeong , Lee Jennifer , Kwok Seung-Ki , Park Young-Woo , Cho Mi-La , Park Sung-Hwan 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is autoimmune mediated chronic inflammatory diseases. Half of SLE patients suffer from lupus nephritis, and lupus nephritis is major cause of death in SLE. TWEAK/Fn14 interactions have been identified as pathologic factor in SLE pathogenesis. We conducted TWEAK/Fn14 pathway blocking by TWEAK receptor-Fc in SLE mice model and searched for the probable therapeutic mechanisms. CD19+ B cells were harvested from sanroque mice and TWEAK was administered. Sanroque mice were treated with TWEAK receptor-Fc or control-Fc for 3 weeks. Immunoglobulin (Ig) G, IgG1, IgG2a were measured in the sera of each group. Spleens from each group were stained with antibodies against CD4, B220, GL-7, CD138, and PD-1. Expression levels of mRNAs were determined by real-time PCR. Kidneys were stained with H&E and PAS. Administration of TWEAK promoted B cell differentiation. Treatment with TWEAK receptor-Fc suppressed the levels of IgG, IgG1, and IgG2a in sera and reduced B cell, plasma cell, follicular helper T cell (Tfh) in spleen of sanroque mice. Additionally, renal protective effects of TWEAK receptor-Fc were demonstrated. In conclusion, TWEAK receptor-Fc had beneficial effects on SLE mice model by repressing auto-reactive B cell, plasma cell, Tfh, and renal damage. In futher, TWEAK receptor-Fc would be a potential therapeutic agent in SLE patient.
Soluble RAGE overexpressed mesenchymal stem cell attenuates autoimmune arthritis via induction of Th17/Treg balance and immunomodulatory effects (THER3P.971)
Authors: Lee Seung Hoon , Park Min-Jung , Lee Jung-Ah , Lee Eun-Jung , Kim Eun-Kyung , Cho Mi-La 
Published: January 2023

Abstract
Receptor for advanced glycation end-products (RAGE) plays the essential role in the pathogenesis of autoimmune rheumatoid arthritis (RA). Mesenchymal stem cells (MSCs) have recently shown promise as a therapeutic tool in various types of autoimmune disease models. In the persent study, we constructed stable MSC with sRAGE overexpression by DNA transfection. We hypothesized that sRAGE overexpression MSC may enhance the self-renewing and modulatory effects on RA therapy. We found that MSCs in inflammatory condition increase expression of High Mobility Group Box1 Protein (HMGB1) and IL-1beta, IL-6, VEGF. However, MSCs that overexpressed sRAGE significantly decrease these inflammatory factors. In addition, sRAGE overexpressing MSCs produced higher amounts of immune modulatory factors. MSCs that overexpressed sRAGE displayed enhanced migration activity and autophagy vesicle. Using a co-culture model of MSC and T cell, we showed that sRAGE overexpressing MSCs potently suppressed Th1, Th17 cell differentiation. Systemic infusion of sRAGE overexpressing MSCs inhibited development of arthritis and reduced bone erosion and cartilage destruction. These therapeutic effects were associated with an increase in CD4+FoxP3+ Treg cells and inhibition of Th17 cell formation in the spleen. These data suggest that enhancing the immunomodulatory activity of MSCs and modulating T cell-mediated immunity using gene-modified MSCs may be a gateway for new therapeutic approaches to clinical RA.
The upregulation of HtrA2 ameliorates autoimmune arthritis via inhibition of Th17 formation and STAT3 expression (THER3P.974)
Authors: Lee Seung Hoon , Young-Mee Moon , Seo Hyeon-Beom , Kim Eun-Kyung 
Published: January 2023

Abstract
HtrA serine peptidase 2 (HtrA2) performs pivotal role in maintaining of the mitochondria homeostasis and involves in cellular stress response such as inflammation. Rheumatoid arthritis (RA) appears to have a basis of mitochondrial dysfunction. To identify the role of HtrA2 in RA, we used a mnd2 mutant mice. We hypothesized that HtrA2 overexpression may reduce immune inflammation and reveal therapeutic effects on RA. We observed that inhibitor and siRNA of HtrA2 in T helper cell (Th) 17 condition increase the mRNA expression of interleukin (IL)-17. On the other hand, the overexpression HtrA2 significantly reduces mRNA level of IL-17 and induces cleavage of STAT3. Moreover, p-STAT3 and lymphocyte activation is promoted in mnd2 mutant mice compared to wild type. HtrA2 overexpression attenuates collagen-induced arthritis (CIA) in mice model. Th17 and osteoclastogenesis are suppressed in CIA mice treated with HtrA2 overexpression. Additionally, HtrA2 overexpression inhibited development of plaque formation and activation of Th17 cell and STAT3 in APOE knock-out mice with proteoglycan immunization for animal model of hyperlipidemia based RA. The therapeutic function of HtrA2 in inflammatory diseases is related with the targeting of Th17 development and STAT3 pathway in splenocytes. These results suggest that HtrA2 takes part in immunomodulatory activity and upregulation of HtrA2 may shed light on therapeutic approaches to inflammatory disorder such as RA and hyperlipidemia.
PD-1 blockade does not enhance T cell reactivity to autologous tumor cells ex vivo in breast cancer. (TUM2P.1013)
Authors: Egelston Colt 
Published: January 2023

Abstract
Recent efforts to identify the antigens tumor specific T cells target have suggested PD-1+ T cells target epitopes made up from mutated proteins. To investigate this possibility in patients with breast carcinoma, a cancer with few mutated antigens, we sought out T cells that specifically react and kill autologous tumor cells. We found that PD-1 was highly expressed on both CD4 and CD8 T cells in the tumor and tumor draining lymph nodes (TDLNs) of breast cancer patients and to a lower extent on circulating T cells. To investigate tumor reactivity, we co-cultured CD8 T cells from tumor, TDLN, or PBMCs with autologous cancer cells overnight. Very low numbers of T cells were found to react to autologous tumor cells by measurement of either CD137 expression or IFNy secretion. Performing the same experiment in the presence of PD-1 blockade did not increase reactive T cell numbers. We have also found that there is a correlation of reduced T cell functionality and PD-1 expression in T cell clones even in the absence of PD-1 ligands. This data suggests that PD-1+ T cells in cancers with a high mutation rate may be able to be restored via PD-1 blockade, while PD-1+ T cells in cancers with few mutations may be self-antigen specific T cells that may have undergone irreversible tolerance. Such data has strong implications for checkpoint blockade therapy.
Activating self-tolerant T cells with altered structure of MHC class I (TUM2P.1024)
Authors: Parks Christopher , Hansen Michael , Ju Josephine , Reed Brendan , Kranz David , Bordner Andrew , Pease Larry 
Published: January 2023

Abstract
Self-tolerant T cells can cross-react weakly on self, but are usually incapable of activation. However, when activated these T cells can cross-react effectively on cells expressing self antigens. We have observed that mutants of the MHC class I molecule H2-Kb have structural differences resulting in altered antigen presentation. We hypothesize that structural manipulation of MHC class I can increase stabile amino acid contacts with the TCR. Increasing stabile contacts may decrease the energy barrier required to overcome the threshold for activation, breaking tolerance for self antigens. Using computer modeling techniques, amino acid substitutions at specific sites of MHC class I were predicted to result in stabilization of the TCR:pMHC complex. These altered MHC molecules were made and expressed in cell lines. We have shown that altered MHC enhanced TCR binding. Altered MHC enhanced activation and proliferation of naïve T cells in response to a non-stimulatory, weak antigen. Altered MHC also induced CTL killing against self-antigens. In an in vivo transplant model, protective immunity was induced against a native tumor after an altered MHC expressing tumor challenge. By altering the structure of MHC class I we have activated normally tolerant T cells that are capable of cross-reacting on self.
The quantity of CD26 on helper T cells correlates with the efficacy of adoptive cancer immunotherapy (TUM2P.1008)
Authors: Bailey Stefanie , Nelson Michelle , Bowers Jacob , Majchrzak Kinga , Stefanik Leah , Paulos Chrystal 
Published: January 2023

Abstract
Although Th17 cells cause striking tumor eradication upon transfer into mice bearing established melanoma, their translation into the clinic is barred by a lack of FDA-approved cytokines. Consequently, we sought to determine if IL-17-producing CD4+ cells could be enriched from peripheral blood. We found that functional Th17 cells could be isolated from blood, without the need for polarization, by using the surface marker CD26. Excitingly, very few CD26-rich cells, termed CD26high T cells, displayed striking antitumor activity and survival upon transfer into melanoma-bearing mice compared to CD26negative or bulk CD4+ T cells. Of translational significance, we found that human CD26high cells had heightened cytotoxicity and caused greater cancer cell death in vitro. Interestingly, human CD26high T cells have enhanced expression of cytokines (IL-17, IFN-γ, IL-2, TNFα), transcription factors (RORγ, Tbet), integrins (LFA-1, VLA-1) and chemokines (RANTES). Given the unique phenotype of CD26high T cells compared to cells with decreased CD26 expression, the question of this marker's importance is raised. Our preliminary studies reveal that the enzymatic activity of CD26 may be important for the enhanced function, cytotoxicity and migration of these cells and could possibly be targeted to enhance future therapies. Not only do CD26high T cells give hope for Th17 clinical translation, but also provide a potential mechanism by which to improve the T cells used for cancer immunotherapies.
Quantifying the lower limit of antibody-dependent cell cytotoxicity using a TCRm against a WT1 peptide/MHC epitope (VAC3P.1058)
Authors: Casey Emily , Scheinberg David 
Published: January 2023

Abstract
The WT1 protein is an ideal cancer antigen, but therapeutically targeting this transcription factor has proven difficult. We have discovered a therapeutic TCR mimic antibody that specifically binds to a WT1-derived peptide (RMFPNAPYL) in the context of the MHC class I haplotype HLA-A2. This high affinity mAb, called ESK, has demonstrated significant anti-cancer activity in animal models of human cancers and leukemias. Studies in vitro and in vivo have shown the dominant role of ADCC in ESK activity. While commercially available mAbs target tens to hundreds of thousands of sites on a cell, ESK binds as few as ~500 epitopes and still promotes cancer cell clearance in vivo. Therefore, we believe ADCC can be a more sensitive mode of killing than currently estimated. To probe this question, we developed a biotinylated RMF peptide that can be pulsed onto HLA-A2 on T2 cells at various concentrations and detected using labeled streptavidin probes. By using 3D confocal microscopy photon scanning, we can detect single molecules of the streptavidin probe on a cell surface. Therefore, using a CFP-DEVD-YFP FRET reporter expressed in the pulsed T2 cells, we can measure the number of peptides bound by ESK in the immunological synapse between a target T2 cell and a human NK cell required to induce T2 cell death. Identifying this number will impact patient selection for ESK clinical trials and will influence the development of therapeutic mAbs against other low-density epitopes.
Dietary fibre-mediated effects on the primary antibody response to keyhole limpet hemocyanin differs between sexes (VAC3P.1059)
Authors: Shastri Padmaja , Brooks Stephen , Kalmokoff Martin , Green Judy , Mathias Fernando , Green-Johnson Julia 
Published: January 2023

Abstract
Dietary fibre can mediate effects on mucosal immunity and the gut microbial community. The aim of this study was to examine the impact of dietary fibre intake on immunization efficacy, and to assess potential sex-based differences. Following a 6 week feeding trial, male and female rats fed purified diets containing single sources of fermentable material (cellulose; C, wheat bran; WB, oat bran; OB, resistant starch; RS or oligofructan; OF) were parenterally immunized with 0.3 mg/kg Keyhole Limpet Hemocyanin (KLH). Anti-KLH and total antibody concentrations were evaluated at intervals from tail vein blood, and splenic lymphocyte percentages and cytokine levels were measured at the end of the trial. Circulating anti-KLH IgG, IgG2a and IgG2b levels decreased significantly in females fed OB, while anti-KLH IgG2a levels decreased in males fed either RS or OF-supplemented diets compared to control-fed rats. In contrast to females, males fed OB had significantly lower splenic TGF-b1/IFN-g ratios and higher total IgG antibody levels. Additionally, females fed RS or OF had significantly reduced splenic CD3+ and CD3+4+ percentages while only CD3+ percentages were significantly reduced in males fed OF, relative to their control-fed counterparts. Collectively these results suggest dietary fibre can impact systemic immune responses to immunization, and that these effects can differ between sexes.
Flagellin has differential and route-dependent adjuvant effects when encoded in heterologous systemic or mucosal gene-based prime-boost vaccines (VAC4P.1107)
Authors: Rady Hamada , Dai Guixiang , Huang Weitao , Shellito Judd , Ramsay Alistair 
Published: January 2023

Abstract
Flagellin has been tested as a protein-based adjuvant, with most studies focused on antibody responses. In this study, we have evaluated the adjuvant activity of flagellin for cellular and humoral immune responses in gene-based immunizations. DNA and adenovirus (Ad) vectors were constructed to encode mycobacterial Ag85B and/or flagellin (FliC) of S. typhimurium. DNA-encoded flagellin given to mice via the IM route enhanced antigen-specific splenic CD4+ and CD8+ T cell responses. Boosting either IM or intranasally (IN) with Ad vectors expressing Ag85B without flagellin enhanced antigen-specific antibody and CD4+ and CD8+ T cell responses in both spleen and pulmonary tissues, correlating with improved protection against pulmonary challenge with Mtb. However, inclusion of flagellin in both DNA prime and Ad boosting vaccines induced localized pulmonary inflammation and transient weight loss, along with route-dependent effects on vaccine-induced T cell immunity. The latter included marked reductions in mucosal CD4+ and CD8+ T cell responses following IM DNA / IN Ad prime-boosting, although antibody responses were not diminished. These findings suggest that flagellin has differential and route-dependent adjuvant activity when included in systemic or mucosal gene-based prime-boost immunization. Adjuvant activity for both T and B cell responses and safety were optimal when flagellin was included only in the DNA vaccine priming phase of systemic or mucosal DNA/Ad prime-boosting.
Lipopolysaccharide induced type I interferon is essential for effector CD4 T cell expansion and migration to liver (VAC4P.1108)
Authors: Shinde Paurvi 
Published: January 2023

Abstract
Vaccines that provide long-term protection against a pathogenic infection often use adjuvants to boost the immune response to less immunogenic antigens. Lipopolysaccharide (LPS) is such an adjuvant that enhances CD4 T cell clonal expansion, survival and IFN-γ production. Similarly, engagement of OX40 receptor (CD134) on antigen activated CD4 T cells increases CD4 T cell activation and clonal expansion. Administration of LPS and CD134 costimulation together profoundly increases the Ag-specific CD4 T clonal expansion, survival and memory. It is known that LPS induced TRIF signaling is needed CD4 and CD8 T migration to liver, lung and spleen as well as for IFNγ production. TRIF downstream activates Type I interferon which can upregulate co-stimulatory molecules on APC and mediate CD4 T cell priming and expansion. Hence our goal is to study how LPS induced Type I Interferon together with CD134 costimulation impacts antigen specific CD4 T cell expansion, migration and effector potential in lymphoid and non-lymphoid organs. Our results show that, IFNAR blockade reduced antigen-specific CD4 T cell expansion and migration to liver on days 6 and 12 on immunization with LPS and CD134 agonist. In pLN however, IFNAR blockade reduced T cell expansion and migration on day 6, but by day 12 T cells began to accumulate. Additionally even though Type I interferon signaling was needed for IFNγ secretion by Th1 cells in pLN, it actually reduced IFNγ and TNFα secretion of Th1 cells in liver.