bluuebunny/crossref_metadata_2025_split huggingface dataset:
Negative regulation of pathogenic TH17 cells by IK cytokine ameliorates inflammatory arthritis (THER3P.969)
Authors: Park Hye-Lim , Min Jun-Ki , Lee Sang-Myeong , Moon Su-Jin , Lee Dong-Hee , Nam Jae-Hwan 
Published: January 2023

Abstract
The pathogenesis of inflammatory autoimmune rheumatoid arthritis (RA) is driven by the activation of macrophage, and the development of pathogenic interferon-γ (IFN-γ)- producing and interleukin-17 (IL-17)- producing CD4+ T cells (TH1 and TH17). Here we showed that truncated IK (tIK) cytokine, inhibitor of IFN-γ induced MHC class II expression, ameliorated RA in a mouse model, which was associated with decreases in joint swelling and bone destruction, as well as the incidence. However, the detailed mechanism of tIK-reduced inflammatory arthritis is still unclear. Of note, we found that the splenocytes of tIK-Tg after LPS treatment showed the induction of A20 (also termed as TNFAIP3) compared with those of WT in microarray analysis. We also found that differentiation of CD4+ T cells from tIK-Tg to TH17 cells was decreased. Consistent with this, A20 were highly expressed in CD4+ T cells from tIK-Tg under TH17 cells polarizing condition, which was associated with reduced NF-κB and p38 signaling pathways. Thus, these findings suggest therapeutic and regulatory function for IK cytokine, supporting the strategy for drugs to target pathogenic TH17 cells in inflammatory arthritis.
IL-12p40 homodimer (p40)2 ameliorates experimental autoimmune arthritis (THER3P.975)
Authors: Lee Seon-Young , Jung Young Ok , Kim Ho-Youn , Kim Doo-Jin , Sung YoungChul , Cho Mi-La 
Published: January 2023

Abstract
Interleukin (IL)-23 is the key cytokine that induces the expansion of helper T (Th) 17 cells. It is composed of p19 and p40 subunits of IL-12. The p40 subunit binds competitively to the receptor of IL-23 and blocks its activity. Our aim was to assess the preventive and therapeutic effect of the (p40)2 subunit in autoimmune arthritis animal models. In the present study, by using IL-1receptor antagonist knockout mice (IL-1RaKO) and a collagen-induced arthritis (CIA) model, we inspected the suppressive effect of the IL-12p40 homodimer ((p40)2) on inflammatory arthritis. We demonstrated that the recombinant adenovirus expressing mouse (p40)2 (rAd/(p40)2) model prevented the development of arthritis when given before the onset of arthritis. It also decreased the arthritis index and joint erosions after it was transferred once arthritis was established in the mouse model. (p40)2 inhibited the production of inflammatory cytokines and antigen-specific T cell proliferation. It also induced the CD4+CD25+Foxp3 regulatory T (Treg) cells in vitro and in vivo, whereas the generation of RORgt and Th17 cells was suppressed. The induction process of Treg cells and the suppression of Th17 cells were mediated via the activated STAT5 and suppressed STAT3. Conclusion: Our data suggested that (p40)2 suppressed inflammatory arthritis successfully. This could be a useful therapeutic approach in autoimmune arthritis to regulate the Th17/Treg balance and IL-23 signaling.
PD-L1 blockade and upregulation in an ovarian cancer treatment model (TUM2P.1017)
Authors: Grabosch Shannon , Zhang Lixin , Zeng Feitianzhi , Mony Jyothi , Ma Tianzhou , Tseng George , Edwards Robert , Vlad Anda 
Published: January 2023

Abstract
Objective: We explored mechanisms of PD-L1 upregulation in ovarian cancer cells and tested in vivo the efficacy of intraperitoneal anti-PD-L1 therapy in two different ovarian cancer mouse models. Methods: We have derived several novel murine ovarian cancer cell lines from orthotopic ovarian tumors of triple transgenic mice. 129x SvJ and C57Bl/6 tumor-bearing mice were treated with 200 μg anti-PD-L1 antibody (or rat IgG as control) every two weeks for three doses. Immune gene profiling of splenocytes collected at necropsy was performed using 561 Nanostring probes. Results: Cell cycle analysis of in vitro cultured tumor cells demonstrates that PD-L1 expression increases preferentially on cells in G2/M cells. Exposure to cisplatin modifies PD-L1 levels, although the effect varies according to tumors’ baseline susceptibility. In vivo, we observed murine PD-L1 upregulation on ascites resident tumor cells, an effect reproduced ex vivo upon exposure of tumor cells to ascites fluid. PD-L1 blockade triggers increase in survival in both preclinical models with immune gene signatures pointing to cytotoxic anti-tumor immune response. Conclusion: PD-L1 expression increases in response to inflammatory stimuli from cell-free ascites and shows preference for dividing cells. PD-L1 upregulation varies according to tumor cells’ susceptibility to cisplatin. PD-L1 blockade increases systemic T cell responses and intratumoral T cell accumulation suggesting therapeutic potential in ovarian cancer.
Immunization of CD200R1KO mice post surgical resection of EMT6 breast cancer leads to long-term (&gt;1year) cure of metastatic tumor growth (TUM2P.1022)
Authors: Gorczynski R. , Khatri Ismat , Podnos Anna 
Published: January 2023

Abstract
Attenuated expression of CD200 decreases local tumor growth and metastasis in immunocompetent animals. We explored the ability to cure BALB/c CD200KO or CD200R1KO mice of tumors ≤1cm3 in size by surgical resection of localized tumor, followed by immunization with irradiated EMT6 cells using Monophosphoryl Lipid A (MPLA:25υg/mouse) as adjuvant. Controls developed pulmonary and liver metastases within 30 days of surgery. Protection was seen in both CD200KO or CD200R1KO mice, with no macroscopic lung/liver metastases observed in CD200R1KO mice at day 300. After resection and immunization, draining lymph nodes from controls contained tumor cells cloned at limiting dilution in vitro before pulmonary and hepatic metastasis was seen. Within the limits of detection of the assay used (sensitivity ~1 in 107 cells) no tumor cells were detected in CD200R1KO mice, and reductions were seen in CD200KO mice. Infusion of anti-CD8 mAb into surgically treated and immunized CD200R1KO mice attenuated protection from both macroscopic (liver/lung) and microscopic (assayed by limiting dilution of DLN) metastasis. Adoptive transfer of lymphocytes from treated CD200R1KO mice to surgically-treated control mice, particularly after Treg depletion attenuated metastatic growth of tumor, an effect abolished by pretreatment with anti-CD8 mAb. We suggest CD200:CD200R attenuates a potentially tumor-protective CD8 host response to breast cancer following immunotherapy using MPLA as adjuvant.
HHLA2 and TMIGD2: A new pathway of the B7/CD28 families and novel targets for human cancer immunotherapy (TUM2P.1028)
Authors: Zang Xingxing 
Published: January 2023

Abstract
We have recently identified HHLA2 as a new member of the B7 family that shares 23-33% similarity to other human B7 proteins and phylogenetically forms a subfamily with B7x and B7-H3. HHLA2 is expressed in humans but not in mice. HHLA2 protein is constitutively expressed on monocytes and induced on B cells, and inhibits proliferation and cytokine production of human CD4 and CD8 T cells in the presence of TCR signaling. HHLA2 protein is not detected in most organs, but is widely expressed in human cancers from the breast, lung, thyroid, melanoma, pancreas, ovary, liver, bladder, colon, prostate, kidney, and esophagus. 56% of patients with triple negative breast cancer have aberrant expression of HHLA2 on their tumors, and high HHLA2 expression is significantly associated with regional lymph node metastasis and stage. HHLA2 copy-number gains are present in 29% of basal breast cancers. HHLA2 binds putative receptors expressed not only on resting and activated CD4 and CD8 T cells but also on APCs. Furthermore, we have identified TMIGD2 as one of the receptors for HHLA2. TMIGD2 shares 23-29% similarity to other human CD28 proteins, but is expressed in humans not in mice. Our results identify HHLA2/TMIGD2 as a new pathway of the B7/CD28 families and suggest that the HHLA2 pathway represents a novel immunosuppressive mechanism within the tumor microenvironment and an attractive target for human cancer therapy.
Ibrutinib, a dual Btk/Itk inhibitor, promotes dendritic cell maturation by inducing IFN-α/β. (TUM2P.1037)
Authors: Natarajan Gayathri , Terrazas Cesar , Oghumu Steve , Varikuti Sanjay , Satoskar Abhay 
Published: January 2023

Abstract
Ibrutinib, a Btk and Itk inhibitor, treats hematological malignancies by acting on lymphoid cells. Additionally, its efficacy against myeloid cell-mediated inflammatory diseases suggests that Ibrutinib could target myeloid cells. Previous studies show that Btk is expressed by murine dendritic cells (DCs). Further, Btk deficient DCs are more mature, express higher levels of MHC-II, co-stimulatory molecules and cytokines and thereby enhance CD4+ T cell activation. Hence, we studied the effect of Ibrutinib on DC function by treating murine bone marrow derived DCs with Ibrutinib during their development. We observed that Ibrutinib-treated DCs upregulate the expressions of MHC-II and co-stimulatory molecules such as CD80 and CD40. Further, Ibrutinib-treated DCs differentially regulate cytokine synthesis by producing more IL-10 while reducing IL-12 production. Ibrutinib-treated DCs promote CD4+ T cell proliferation and enhance T cell derived IFN-γ production. We studied the mechanism by which Ibrutinib promotes DC maturation by studying the expression of cytokines that direct DC maturation. We identified that Ibrutinib-treated DC cultures increase the induction of IFN-α and IFN-β which are known to promote DC maturation and IRF-7, a transcription factor which is expressed upon IFN-α/β activation. Hence, Ibrutinib treatment enhances DC maturation in an IFN-α/β dependent manner and should be further explored in the development of DC-based therapeutic vaccines against cancer.
NK cells and CD8+ T cells cooperate to improve therapeutic responses in melanoma treated with CTLA-4 blockade and IL-2 (TUM2P.1039)
Authors: Kohlhapp Frederick , Broucek Joseph , Hughes Tasha , Huelsmann Erica , Zloza Andrew , Kaufman Howard 
Published: January 2023

Abstract
Immunotherapy is a promising approach for the treatment of metastatic melanoma and combination strategies may have potential for improving therapeutic responses. CTLA-4 blockade increases the CD8:Treg ratio in the tumor microenvironment and IL-2 promotes NK and CD8+ T cell function. Thus, we hypothesized combination IL-2 and CTLA-4 blockade would result in improved anti-tumor responses. This hypothesis was confirmed utilizing the poorly immunogenic B16 melanoma model, with CTLA-4 blocking antibody on days 3, 6, and 9 and/or IL-2 twice daily on days 4-8 after challenge, mirroring clinical use of these agents. Combination immunotherapy resulted in delayed tumor growth and prolonged survival compared to either monotherapy. Depletion studies suggested that both CD8+ T and NK cells were required for the therapeutic effects. In this model, combination treatment resulted in fewer tumor-infiltrating CD8+ T and NK cells but more infiltrating Tregs in the tumor microenvironment when compared to monotherapy CTLA-4 blockade. Further, IL-2 therapy was associated with a reduction in the proportion of highly differentiated/exhausted NK cells in the tumor microenvironment. No significant toxicity was observed in the mice. These results support the combination of IL-2 and CTLA-4 blockade in melanoma and a clinical trial based on these data has been initiated.
Generation and characterization of an anti-PD1-resistant lune cancer line (TUM2P.1040)
Authors: Wang Xiaohong , Ye Huiping , Valdecanas David , Welsh James 
Published: January 2023

Abstract
Immunotherapies targeting PD1/PDL1 have shown unprecedented rates of durable clinical responses in patients with melanoma and lung cancer. However, there is a substantial portion of patients who do not respond to these therapies. The goal of this study is to generate an anti-PD1-resistant tumor line and identify mechanisms of resistance, which should provide novel insights to extend the benefit of immunotherapies. A preclinical lung cancer model was generated by inoculation of a murine lung cancer cell line 344SQ (containing p53 and kras mutations) into syngeneic 129SV/ev mice. Anti-mouse PD1 antibodies were administrated at day 4 post tumor inoculation for a total of 5 doses. The anti-PD1 resistant line was successfully generated after sequential in vivo passaging of a nonresponsive tumor followed by anti-PD1 treatment. Upon the anti-PD1 treatment, the parental line showed 70% tumor reduction, whereas the resistant line showed only 12% tumor reduction. PDL1 expression did not alter in the resistant line. Instead, anti-PD1 treatment remarkably increased MHC I expression in the parental line, whereas the resistant line showed significantly lower MHC I and II expression and failed to upregulate MHC I upon anti-PD1 treatment. The parental and resistant lines showed distinct profiles of tumor-infiltrating immune cells and costimulatory molecule expression in vivo. Next we will study if inducing MHC expression through radiation or HDAC inhibitors will overcome anti-PD1 resistance.
Extracorporeal photopheresis promotes IL-1β production (VAC3P.1054)
Authors: Hackstein Holger , Jakobs Christopher , Peric Adriana , Michel Gabriele , Baal Nelli , Bein Gregor , Bruene Bernhard , Hornung Veit , Yakut Erhan 
Published: January 2023

Abstract
Extracorporeal photopheresis (ECP) is a widely used clinical cell-based therapy exhibiting efficacy in heterogenous immune-mediated diseases such as cutaneous T cell lymphoma, graft versus host disease (GVHD) and organ allograft rejection. Despite its documented efficacy in cancer immunotherapy, little is known regarding the induction of immunostimulatory mediators by ECP. Here we show that ECP promotes marked release of the prototypic immunostimulatory cytokine IL-1β. ECP primes IL-1β production and activates IL-1β maturation and release in the context of caspase-1 activation in monocytes and myeloid dendritic cells. Interestingly, IL-1β maturation by ECP was fully intact in murine cells deficient for caspase-1, suggesting the predominance of an inflammasome-independent pathway for ECP-dependent IL-1β maturation. Clinically, patient analysis revealed significantly increased IL-1β production in stimulated leukapheresis concentrates and peripheral blood samples after ECP. Collectively, these results provide evidence for promotion of IL-1β production by ECP and provide new insight into the immunostimulatory capacity of ECP.
Targeting DNA vaccines to dendritic cells using a small peptide to enhance immune response (VAC4P.1110)
Authors: Ye Chunting , Choi Jang Gi , Abraham Sojan , Shankar Premlata , Swamy Manjunath 
Published: January 2023

Abstract
Dendritic cell targeting of antigens greatly enhances immunogenicity. Although there are several reagents (DEC205, Dectin-1, Langerin, Clec9, nanoparticle, etc) to target protein antigens to DCs, there is still no method to target DNA vaccines directly to DCs. Here we show that a small peptide derived from the rabies virus glycoprotein, fused to protamine residues (RVG-P) can target DNA to DCs resulting in enhanced T-cell and humoral responses. Targeted delivery of DNA vaccine encoding the immunodominant Vaccinia virus B8R gene to DCs with RVG-P was able to re-stimulate Vaccinia-specific memory T cells in vitro. Moreover, a single iv injection of B8R gene bound to RVG-P without any adjuvants was able prime a Vaccinia-specific T-cell response that was able to rapidly clear a subsequent Vaccinia virus challenge in mice. Finally, immunization of mice with a DNA vaccine encoding with West Nile virus (WNV) prM and E proteins via RVG-P elicited high titers of WN neutralizing antibodies that protected mice from lethal WNV challenge. Thus, RVG-P provides a tool to target DNA vaccines to DCs to elicit robust T-cell and humoral immune responses.
Mucosal vaccination with an attenuated strain of Mycobacterium tuberculosis induces a strong central memory response and protects non-human primates against lethal challenge (VAC4P.1104)
Authors: Foreman Taylor , Mehra Smriti , Gautam Uma , Rangel-Moreno Javier , Khader Shabaana , Kaushal Deepak 
Published: January 2023

Abstract
There is an urgent need for new vaccine(s) that protect efficiently against Mycobacterium tuberculosis (Mtb), as well as, to better understand the correlates of a protective host response to Mtb infection. Aerosol infection of macaques with the redox-stress deficient mutant, Mtb:Δ-sigH, resulted in the recruitment of significantly more inducible bronchus-associated lymphoid tissue, which is associated with protection from Mtb infection in mice, macaques, and humans. We therefore hypothesized that vaccination with Mtb:Δ-sigH could protect against a lethal Mtb challenge significantly better than a comparable Bacille Calmette-Guerin (BCG) vaccination. A single vaccination of the aerosol delivered, live-attenuated strain completely protected macaques from lethal Mtb challenge. Protection was associated with three-to-four log lower bacterial burden, significantly reduced granulomatous pathology and clinical features of Mtb infection, as well as, a strong T cell central memory response when compared to BCG-vaccinated animals. A higher magnitude, as well as, breadth of polyfunctional T cell responses was detected in the Mtb:Δ-sigH vaccinated animals. This includes an increased recruitment and turnover of central and effector T cells in the lung post vaccination compared to BCG-vaccinated animals. These results point to the protective qualities induced by a live-attenuated mucosal vaccine for protection against lethal Mtb challenge, and its future in TB vaccine initiatives.
Antibody immunotherapy targeting the self-tumor antigen nucleolin (VAC5P.1129)
Authors: Sutkowski Natalie , Sun Wei , Fernandes Daniel 
Published: January 2023

Abstract
Nucleolin is a multifunctional protein found in the nucleus of all cells, and it’s a self-tumor antigen. It is overexpressed on the plasma membrane and cytoplasm of the two most common forms of leukemia acute myeloid leukemia (AML) and chronic lymphocytic leukemia (CLL). The cell surface expression of nucleolin on CLL and AML cells, compared with its nuclear profile in normal cells, makes nucleolin an attractive self-antigenic target for antibody immunotherapy. Accordingly, we have utilized our patented platform technology (US Patent 8,715,743) to generate a panel of fully human monoclonal antibodies (HuMAbs) that target nucleolin (NCL) and potently induce AML cell death, while having no effect on normal blood cells (PCT/US2010/057046). Preliminary data indicated that AML cells were killed independently of complement or antibody dependent cellular cytotoxic mechanisms, possibly by interfering with nucleolin’s cellular function. This might have significant therapeutic implications, since the HuMAbs may retain activity in immunocompromised leukemia patients. We hypothesize that the anti-NCL HuMAbs have both immune mediated and immune independent antitumor activity, eliciting nucleolin specific killing through both complement and cell mediated cytotoxic mechanisms, and through interfering with the integral cellular function of nucleolin. Thus, autoreactive antibodies targeting this self-tumor antigen might be important in the treatment of leukemia.
Characterization of T cell responses induced by Flu SAM(NP)® and SAM(M1)® vaccines (VAC5P.1124)
Authors: Magini Diletta , Buccato Scilla , Mangiavacchi Simona , Giovani Cinzia , Montomoli Emanuele , Brito Luis , Maione Domenico , Geall Andrew , Brazzoli Michela , Bertholet Sylvie 
Published: January 2023

Abstract
Vaccination remains the most cost-effective way to control Influenza outbreaks. However, current Influenza vaccines do not provide efficacious heterosubtypic immunity. Therefore, the development of a successful universal vaccination strategy is urgently needed. To achieve this goal, one approach is focused on the induction of a T cell based response versus more conserved flu antigens, such as the Nucleoprotein (NP) and the Matrix protein 1 (M1). The aim of this study was to evaluate the immunogenicity and protective efficacy conferred by conserved NP and M1 antigens expressed by a self-amplifying RNA-based vaccine (SAM(NP)® and SAM(M1)®). We demonstrate that SAM(NP)® and SAM(M1)® are immunogenic in Balb/c mice. SAM(NP)® formulation induced IFNγ+ and LAMP1+ NP-specific cytotoxic CD8+ T cells as well as multifunctional NP-specific CD4+ T cells. SAM(M1) ® elicited a robust polyfunctional CD4 T helper 1 response. Moreover, we show that SAM(NP)® and SAM(M1)® formulations, when administered separately or in combination, provided differential levels of protection against a lethal challenge with mouse-adapted A/PR/8/1934 (H1N1) Influenza virus. SAM(NP)® and SAM(M1)® formulation induced T cell immune responses, potent mediator of heterosubtypic immunity, therefore, they are promising antigens in the design of a cross-protective flu vaccine. Since the SAM® vaccine technology offers several advantages , it could potentially speed up the development of universal flu vaccine candidates.
Anatomic redistribution of natural killer cells within secondary lymphoid organs during virus infection (VIR1P.1138)
Authors: Moran Michael , Rydyznski Carolyn , Waggoner Stephen 
Published: January 2023

Abstract
Natural killer (NK) cells are innate lymphocytes that classically function to restrict microbial infection and tumor development. NK cells are also important regulators of adaptive immunity. Specifically, we found that NK cells inhibited antiviral humoral immunity by restricting the germinal center (GC) response via a perforin-dependent cytotoxic mechanism during the initial few days of lymphocytic choriomeningitis virus (LCMV) infection. We hypothesize that this NK cell suppression of antiviral B cells requires redistribution of activated NK cells to regions of secondary lymphoid tissues where T-B cell interactions shape the GC reaction. NK cells were predominately localized within the red pulp of the spleens of uninfected mice, but infection with LCMV triggered a redistribution of NK cells to the white pulp. At day 3 p.i, many NK cells were present in T cell rich areas, most notably at the T-B cell interface where follicular helper T cells (TFH) typically reside. In addition, a small population of NK cells penetrated deep into the B cell follicles. The redistribution of NK cells was transient, with most NK cells returning to the red pulp by day 6 p.i. We speculate that white pulp localization is stimulated by altered expression of chemokine receptors by NK cells, and that this proximity to the GC facilitates direct interactions between NK cells and TFH or GC B cells, thereby contributing to diminished GC responses in the presence of NK cells.