bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of a RANKL ELISPOT assay as a tool for periodontal disease research (TECH2P.913)
Authors: Zhang Jinrong , Cole Leah , Brown Anna , Ma Bing , Anosova Natalie , McCluskey Jacqueline 
Published: January 2023

Abstract
Periodontal disease (PD) results from the immune response to the bacterial components of the sub-gingival plaque. PD progresses from gingival inflammation, to bone resorption and ultimately tooth loss. Receptor activator of nuclear factor-kappa B ligand (RANKL) is critical for regulating inflammatory bone resorption. Studies have shown that activated T and B cells are one of the main sources of RANKL in PD. The ELISPOT assay is widely used tool that can measure antigen specific cellular immune responses. We developed a RANKL specific ELISPOT assay for quantifying the number of RANKL secreting cells in the murine Porphyromonas gingivalis induced bone loss model. Various potential positive controls were tested for their ability to induce RANKL secretion including IL-7, ionomycin, LPS, and α-CD3 combined with α-CD28. We found that the combination of α-CD3 and α-CD28 was the most effective positive control for cells collected from either murine lymph nodes or spleens. Gingipains are trypsin-like cysteine proteases and have been implicated as important virulence factors for Porphyromonas gingivalis. When we performed the RANKL ELISPOT using the purified native gingipain complex as the recall antigen, we were able to detect more RANKL secreting cells from the mandibular lymph nodes of Porphyromonas gingivalis challenged animals than in the buffer challenged control animals.
Isolation of broadly neutralizing antibodies from single human memory B cell against human influenza A viruses (TECH2P.922)
Authors: Fu Ying , Zhang Zhen , Sun Jiusong , Zhu Quan 
Published: January 2023

Abstract
During the past five years, broadly neutralizing antibodies (BnAbs) represent promising prophylactic and therapeutic treatments against influenza. Here, we have developed a novel method to isolate broadly neutralizing antibodies against influenza A virus from human memory B cells. Antigen-specific memory B cells were isolated from human PBMCs using tetramerized H3 (A/Brisbane/10/2007) hemagglutinin (HA) trimers. The H3-reactive single memory B cells were sorted into plates and stimulated in vitro. More than 40% sorted B cells produced on average 200 ng/ml IgG in the supernatant after 14 days. Supernatants from the expanded B cells were measured for their heterosubtypic binding specificity and neutralizing activity by MSD or a highly sensitive neutralization assay. The selective clones were recovered by single cell RT-PCR. Through screening 2688 memory B clones from 7 individuals, 11% clonable memory B cells were reactive with H3 hemagglutinin. Among them, H3/H7, H3/H7/H1 and H3/H7/H1/influenza B heterosubtypic binding population were 16%, 6.9% and 0.35%, respectively. A new broadly neutralizing Ab, 3I14 encoded by the IGHV3-30 germline gene, was further characterized and shown to possess cross-reactive binding and neutralization activity against both group 1 and group 2 influenza A viruses. Epitope mapping and competition assay revealed a highly conserved epitope located in the HA stalk. Passive transfer of this antibody also conferred in vivo protection.
An innovative systems perspective of protein-protein interactions in the physiologic T cell antigen receptor signaling network (TECH2P.929)
Authors: Neier Steven , Smith Stephen , Davis Tessa , Sinnwell Jason , Nevala Wendy , Enninga Elizabeth , Markovic Svetomir , Eckel-Passow Jeanette , Sciallis Gabriel , Wieland Carilyn , Torgerson Rochelle , Neuhauser Claudia , Schrum Adam 
Published: January 2023

Abstract
Signaling networks and the protein interactions that define them are of great interest, but techniques to assess these networks in unmanipulated primary human cells are limited. We have developed and applied a novel technological and analytical platform to measure proteins in shared complexes detected by exposed surface epitopes (PiSCES). Immunoprecipitation reactions are performed in multiplex using microsphere beads to measure 20+ proteins and 210+ distinct pair-wise protein associations that are important for T cell antigen receptor signaling. Jurkat T cells stimulated with superantigen generated an extensive PiSCES signature using this method and provided a dataset from which we optimized an analytical package. The system is currently being utilized to address the hypothesis that T cells isolated from skin lesions of autoimmune alopecia areta (AA) patients will have a unique PiSCES signature compared to those isolated from non-AA donors. To investigate this, we have isolated non-engineered primary cells from AA patient skin lesions and healthy controls, and have begun to identify differences in their PiSCES signatures, either directly after isolation or after stimulation with anti-CD3 and anti-CD28. We conclude that this novel PiSCES approach is capable of generating protein-protein interaction signatures that provide a unique view of TCR signalosome activity solicited from physiologic non-engineered primary human patient samples.
MIF inhibition as novel treatment for autoimmune myocarditis and dilated cardiomyopathy (THER7P.956)
Authors: Nalawade Saisha , Jamison Braxton , Casabar Julian , Maldonado Daniel , Forsthuber Thomas 
Published: January 2023

Abstract
Myocarditis is an inflammatory disease of the myocardium and a major cause of sudden death in young adults. It is characterized by the presence of immune infiltrates in the myocardium and often progresses to dilated cardiomyopathy (DCM). Despite the inflammatory nature of this autoimmune disease, immunosuppressive treatments with glucocorticoids (GCs) such as dexamethasone (Dex) have not been very effective in preventing myocarditis and progression to DCM. In addition, some patients develop resistance to GCs. We hypothesized that macrophage migration inhibitory factor (MIF) may play a role in resistance to GCs, as it is the only known pro-inflammatory cytokine to be induced by GCs. Importantly, MIF counter-regulates GC-mediated immunosuppression. Using the experimental autoimmune myocarditis (EAM) model, we observed that MIF-/- mice treated with Dex were highly resistant to disease and progression to DCM. In addition, we observe lower expression of CCL3 mRNA in MIF-/- mice treated with Dex compared with wild-type mice during the onset of EAM, which indicates that MIF promotes recruitment of inflammatory cells to the myocardium. Our results suggest that therapeutic inhibition of MIF may increase the efficacy of GC treatment. This study will allow us to better understand the mechanism by which MIF affects treatment of myocarditis by inhibiting the therapeutic effects of GCs.
The spleen is the primary site for Breg induction and is required for anti-TIM-1 mediated tolerance (TRAN2P.963)
Authors: Mohib Kanishka , Ding Qing , Cherukuri Aravind , Rothstein David 
Published: January 2023

Abstract
IL-10+ B cells (Bregs) have potent regulatory activity and can transfer tolerance in auto/allo immune models. We showed that: TIM-1 (T1) is an inclusive Breg marker; T1+ B cells transfer allograft tolerance; and anti-T1 induces IL-10+ T1+ Bregs. However, little is known about where Bregs are induced or act in vivo. We found that most Bregs reside in the spleen (85%) vs LN (1%) or marrow (15%), and few are found in islet allografts. Anti-T1 increased Bregs in the spleen from 2% to 7% of B cells, but had no effect in LN (0.5%) or marrow (1%). In splenectomized (spl-X) mice, anti-T1 did not increase Bregs in LN or marrow. Apoptotic cells (ACs) can induce Bregs in vivo. ACs increased Bregs from 2% to 5% in spleen and from 0.5% to 2% in LN. However, after spl-X, ACs did not increase Bregs in LN. Hence, Bregs are induced in the spleen, but then may migrate. Next, we examined the importance of the spleen for Breg-mediated allograft survival (GS). Anti-T1 was unable to induce long-term GS in spl-X islet recipients vs. controls (MST 24d vs. 100d; p=0.002). To confirm that anti-T1 acts through Bregs, we used inducible hCD20ERT2-CreXIL-10fl/fl mice. In such “B cell-IL-10KO” mice, anti-T1 was unable to prolong GS compared to controls (MST 20d vs. &gt;50d). Moreover, LTβR-KO mice (lack LNs) that received alloreactive T cells, exhibited longer islet GS (MST 18d) vs spl-X LTβR-KO recipients (MST 7d p=0.02). Thus, our data elucidate a vital role for the spleen in Breg induction and function.
Effect of selective CD28 blockade with FR104 alone or combined with CD154 blockade in monkey cardiac allograft transplantation. (TRAN2P.967)
Authors: Azimzadeh Agnes , Zhang Tianshu , Braileanu Gheorghe , Sun Wenji , Kronfli Anthony , Sievert Evelyn , Harris Donald , Parsell Dawn , Dahi Siamak , Kubicki Natalia , Woodall Jhade , Uluer Mehmet , Cheng Xiangfei , Schwartz Evan , Quinn Kevin , Vanhove Bernard , Pierson Richard 
Published: January 2023

Abstract
Selective CD28 blockade, which preserves CTLA-4-dependent immune regulation, is a promising approach to induce tolerance. FR104, a novel CD28 antagonist, was evaluated in monkey heart transplant recipients that received αCD28 alone (n=3, FR104) or in combination with αCD154 (n=5, hu5C8), dosed to achieve receptor coverage for 84 days; results were compared to αCD154 alone (n=4). Acute (AR) and chronic (CAV) rejection scores were quantified histologically. Median graft survival time (MST) with FR104 (167d), hu5C8 (133d), and FR104+hu5C8 (173d) was not statistically different between groups. Rejection was seen in 1 of 3 animals with FR104 monotherapy despite CD28 receptor saturation, but not with additional CD154 blockade. CAV was decreased with FR104 (0.7±0.3) and FR104+hu5C8 (0.5±0.3) compared to hu5C8 (1.6±0.7 p&lt;0.05). AR tended to be lower with FR104 (0.5±0.4, p=0.047) or combined therapy (0.8±0.4, p=0.07) on biopsy/explanted functional grafts relative to hu5C8 (1.4±0.6). Blood Tregs frequency on d7-21 was increased (fold change vs. baseline) with FR104 (1.5±0.2), hu5C8 (1.4±0.3), and FR104+hu5C8 (2.3±0.1). CD28 saturation became undetectable by d150 in blood and LN; subsequent graft failure was associated with alloantibodies. We conclude that selective non-activating CD28 blockade protects primate cardiac allografts from pathogenic alloimmunity, an effect enhanced by simultaneous CD154 blockade, supporting the hypothesis that CD28 blockade is potentially tolerogenic.
Histamine receptor 2 blockade reduces breast tumor development and metastasis (TUM9P.1010)
Authors: Vila-Leahey Ava , Oldford Sharon , Marignani Paola , Marshall Jean 
Published: January 2023

Abstract
Histamine has multiple immunomodulatory roles that impact innate and acquired immunity. Histamine receptor antagonists are widely used clinically to treat allergic disease and gastrointestinal disorders, but their ability to alter effective immune function to breast tumors is unknown. We hypothesized that histamine receptor antagonist treatment could alter immune function in mouse models of breast cancer, therefore altering the growth and spread of breast cancer. Two injectable breast tumor models (the 4T1-BALB/c and E0771-C57BL/6 model) and a spontaneous breast cancer model (STK-/-/NIC model) were utilized. Histamine receptor antagonists were administered in the drinking water prior to tumor cell injection or at time of weaning, respectively. The immune cell populations in the spleen and bone marrow of tumor-bearing mice were analyzed. Ranitidine and famotidine (Hrh2 antagonists) but not selective Hrh1 or Hrh4 antagonists significantly inhibited metastasis in the 4T1 model. Ranitidine treatment also caused significant primary tumor regression in the E0771 breast cancer model. In the STK-/-/NIC model, ranitidine increased latency and decreased the number of tumors per mouse at week 26. Bone marrow and spleen analysis revealed that oral ranitidine treatment significantly and selectively decreased monocyte populations. These studies suggest that HRH2 antagonists may have potential as a beneficial immunomodulatory agent in the prevention or treatment of breast cancer.
NK cell effector functions are differentially regulated by hypoxic conditions (TUM9P.1014)
Authors: Lim Seon Ah , Moon Yunwon , Park Hyunsung , Lee Kyung-Mi 
Published: January 2023

Abstract
NK cells provide the ﬁrst line of defense against tumor and viruses, without the requirement for prolonged pre-activation. However, anti-tumor function of NK cells is progressively dampened in the terminal stages of cancer patients, largely associated with the tumor-induced immune evasion mechanism. Hypoxic tumor environment provides once such mechanism providing NK cell tolerance, and become a potential therapeutic problem to overcome in the patients of solid cancer for NK immunotherapy. To better understand the effect of hypoxia on NK cells, we assessed the anti-tumor function of NK cells in response to decreasing concentration of O2 concentration. As expected, NK cells cultured in severe hypoxia, 0.5% O2, demonstrated diminished cytotoxicity and IFNg against A375 melanoma targets and failed to expand to a larger number ex vivo. However, to our surprise, NK cells cultured in mild hypoxia, 1.5% O2, were expanded to a much higher level and mounted increased anti-tumor effector function, both in normoxic and hypoxic conditions. NK cells grown in mild hypoxia demonstrate upregulation of major activating NK cell receptors, NCRs, while inhibiting apoptosis during ex-vivo expansion. Thus, NK cells respond differentially to O2 concentration, and mild hypoxic conditions can indeed facilitate the expansion and function of NK cells that can overcome NK cell intolerance in the patients undergoing NK cell immunotherapy.
Regulatory B cells accumulate in tumor-draining lymph nodes and promote tumor growth (TUM9P.1015)
Authors: Ganti Sheila , Albershardt Tina , Iritani Brian , Ruddell Alanna 
Published: January 2023

Abstract
Tumor metastasis to regional lymph nodes is a reliable marker for determining cancer progression and is associated with poor prognosis. However, mechanisms utilized by the tumor to modify the tumor-draining lymph node (TDLN) to facilitate metastasis remain to be identified. TDLNs from B16-F10 melanoma-bearing mice exhibit considerable B cell accumulation and lymphatic sinus growth (lymphangiogenesis) which precede and predict metastasis of solid tumors. Phenotypic characterization of B cells in mice bearing B16-F10 melanoma in the rear footpad identified preferential accumulation of T2-MZP B cells in the TDLN. Comparison of TDLNs and non-draining LNs of tumor-bearing mice with LNs and spleens from naïve mice determined that this pattern of B cell accumulation was restricted to the TDLN. B cell-deficient and immunocompetent mice reconstituted with T2-MZP B cells but not with other B cell subsets displayed accelerated tumor growth, demonstrating that T2-MZP B cells possess regulatory activity. Unlike splenic regulatory B cells, however, TDLN T2-MZP B cells did not exhibit increased IL-10 secretion, nor did they promote the conversion of naïve CD4+ T cells to CD4+Foxp3+ Tregs in the TDLN. These findings demonstrate that tumors initially signal via the lymphatic drainage to stimulate the preferential accumulation of T2-MZP regulatory B cells. This local response may be an early and critical step in generating an immunosuppressive environment to permit tumor growth and metastasis.
Enterotoxigenic Bacteroides fragilis induces oncogenic regulatory T cells (TUM9P.1000)
Authors: Geis Abby , Dejea Christine , Fan Hongni , Wu Xinqun , Wu Shaoguang , Huso David , Sears Cynthia , Housseau Franck , Pardoll Drew 
Published: January 2023

Abstract
Enterotoxigenic Bacteroides fragilis (ETBF) is an occasional inducer of inflammatory diarrhea and acute colitis, and preliminary studies have recently demonstrated an association with inflammatory bowel disease and colorectal cancer in humans. The pathogenesis of ETBF can largely be attributed to the secretion of a toxin, BFT (B. fragilis toxin), which has a profound effect on the colonic microenvironment. BFT promotes cleavage of E-cadherin, activates NF-kB and Wnt/b-catenin signaling, and stimulates the secretion of pro-inflammatory cytokines, such as IL-6, IL-8, and TNF-a. The inflammation associated with ETBF is characterized by robust IL-17 cytokine secretion, and in MinAPC/+ mice, ETBF induces IL-17-dependent distal colon tumorigenesis. In an attempt to elucidate the immunologic differences between the distal and proximal colon, we found ETBF induced a greater density of Foxp3+ Tregs in the distal colon. Furthermore, those Tregs initiate the oncogenic IL-17 response to ETBF. While investigating the Treg and IL-17 responses to ETBF, we found that depletion of Tregs in ETBF-colonized C57BL/6 Foxp3DTR-GFP MinAPC/+ mice mitigated intestinal IL-17 production as well as colonic neoplasia. Antibody blockade of IL-2, but not genetic ablation of IFN-γ, restored ETBF-induced microadenoma formation. These findings unveil a new mechanism of cancer-induced inflammation whereby an overt Treg response to intestinal bacteria promotes colon tumorigenesis.
Oral rapamycin (eRapa) requires IFN-γ and promotes γδ T cell cytotoxicity to prevent carcinogen and inflammation-induced dermal cancer (TUM9P.1003)
Authors: Dao Vinh , Pandeswara Srilakshmi , Liu Yang , Hurez Vincent , Liu Aijie , Curiel Tyler 
Published: January 2023

Abstract
Rapamycin, a mammalian target of rapamycin (mTOR) inhibitor, prevents distinct cancers in mice and prolongs life, making it a candidate cancer prevention (and life extension) agent. We hypothesized that eRapa cancer prevention includes immune effects, as mTOR is pivotal in immunity. We tested immune effects in carcinogen (DMBA) and inflammation (TPA)-induced dermal carcinogenesis with eRapa daily at 14 ppm, which protected 100% from skin cancer in wild type (WT) mice. Tumor mTORC1 was not suppressed, supporting eRapa immune effects. γδ T cells and IFN-γ are protective in this model. In δ TCR KO (no γδ T cells) and IFN-γ KO mice, eRapa cancer protection was lost, confirming immune mechanisms. IFN-γ KO mice had significantly reduced CD69+Lamp1+GzB+CXCR3+ epidermal γδ T cells and CXCL10, suggesting IFN-γ is required for epidermal γδ T cell migration (via CXCR3/CXCL10), activation and cytolysis. Bone marrow (BM) chimeras showed eRapa skin cancer protection and maximum γδ T cell epidermal infiltration requires IFN-γ signals in BM (e.g., γδ T cells) and non-BM (e.g., epidermal) cells. Intratumor injections of WT but not Prf1 (perforin) KO γδ T cells regressed DMBA/TPA tumors in δ TCR KO mice on eRapa but not control, showing eRapa boosts anti-tumor γδ T cell cytotoxicity. Our data confirm eRapa cancer prevention through immune mechanisms, challenging the paradigm of mTOR effects solely on cancer cells for prevention, and suggesting novel immune-mediated life extension mechanisms.
Epigenetic modulation of CD1d-mediated antigen presentation by B cell lymphomas (TUM10P.1054)
Authors: Tiper Irina , Webb Tonya 
Published: January 2023

Abstract
Histone deacetylases (HDACs) are a family of enzymes that regulate diverse cellular events such as gene expression, cell proliferation, and immune pathways through deacetylation of their protein targets. We hypothesize that tumors use epigenetic mechanisms to dysregulate CD1d-mediated antigen processing and presentation, which leads to a functional impairment in the ability of natural killer T (NKT) cells to recognize and destroy cancerous cells. We examined CD1d-mediated antigen presentation to NKT cells following treatment with HDAC inhibitors (HDACi). Consistent with previous studies, we found that preventing deacetylation by treatment with Trichostatin A, a pan-HDACi, enhanced both CD1d and MHC class II-mediated antigen presentation. Similarly, treatment of B cell lymphomas with Panobinostat resulted in increased CD1d-dependent NKT cell responses, due at least in part to a dose-dependent increase in CD1d-cell surface expression. In addition, treatment with HDCAi results in a decrease in STAT3 expression as well as IL-10 secretion by B cell lymphomas. Overall, our studies demonstrate the efficacy of HDACi in restoring anti-tumor responses to B cell lymphomas through both cell-intrinsic and extrinsic factors. Collectively, these results suggest that HDACi may work to enhance the immune response and increases the immunogenicity of the tumor itself.
Humoral dysregulation among injection drug users: implications for HIV vaccine strategies (VAC11P.1103)
Authors: Kobie James , Samuel Memorie , Zheng Bo , Piepenbrink Michael , Bunce Catherine , Kiebala Michelle , Maggirwar Sanjay , Morse Diane , Keefer Michael 
Published: January 2023

Abstract
The primary objective of a preventative HIV vaccine is the induction of a persistent humoral response that mediates sterilizing immunity. Although modest, yet encouraging, a reduction in infections in low-risk individuals was obtained in the RV144 HIV vaccine efficacy trial; for high-risk individuals, including injection drug users (IDU) no substantial reduction in infection has been observed in efficacy trials. The impact of injection drug use on humoral responses is poorly defined, particularly in the context of HIV prevention. Through a cross-sectional assessment of peripheral B cells and plasma antibodies, we have identified substantial alterations in the HIV-/HepC- active heroin IDU population. IDU exhibited significantly increased total B cells compared to healthy subjects, that were associated with significantly increased memory B cells and plasmablasts. Additionally, although plasma total IgG levels were similar between IDU and healthy subjects, IDU had significantly higher titers of IgG3 and IgG4 subclass plasma antibodies. Total IgM plasma antibodies were also increased in IDU. This humoral dysregulation may be the consequence of a pro-inflammatory environment as evident by significantly increased plasma sCD40L in IDU. These results demonstrate extensive differences in the steady-state humoral profile of IDU, which could have the potential to severely influence the quality of the B cell and antibody response, and ultimate efficacy of future HIV vaccine strategies.
Polyguanine-conjugated antigens for scavenger receptor targeting and self-adjuvanting vaccines (VAC13P.1125)
Authors: Ciccotelli Jo Erika , Toussaint Helene , Erdos Geza , Donahue Cara , Watkins Simon , Falo Louis 
Published: January 2023

Abstract
We have recently developed a novel vaccine strategy that involves conjugating antigens to polyguanine sequences to fabricate aggregated particulates with exposed scavenger receptor ligands for in vivo targeting to antigen presenting cells. We have previously demonstrated that polyguanine-conjugated ovalbumin (Poly(dG)-OVA) leads to enhanced antigen-specific T cell and antibody responses. In this study, we show that exposing dendritic cells (DCs) to Poly(dG)-OVA results in increased antigen internalization that is inhibited by scavenger receptor ligands. Further, DCs internalizing Poly(dG)-OVA complexes exhibit increased cytokine secretion and inflammatory marker expression. In vivo, intradermal injection of Poly(dG)-OVA results in complex persistence in the skin for up to 7 days. Intradermally immunized mice demonstrated increased immune infiltrates in the skin and a persistent inflammatory cytokine profile. In these immunized animals, Poly(dG)-OVA particles reach the draining lymph nodes directly and in association with migratory DCs. We further demonstrate that Poly(dG)-OVA immunization of mice with established OVA-expressing melanoma tumors exhibited delayed tumor growth and improved survival. Taken together, these results suggest that immunization with polyguanine-conjugated antigens can induce more effective antitumor immunity by targeting DCs through scavenger receptors and inducing proinflammatory DC activation.