bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tumor released autophagosomes regulate M2-like macrophage polarization (TUM6P.974)
Authors: Wen Zhifa , Liu Hongxiang , Zhou Meng , Wang Li-xin 
Published: January 2023

Abstract
Tumor cells secrete mediators that modulate macrophage activation and polarization into M2 type tumor associated macrophages (TAMs), which contribute to tumor progression. However, the mechanisms that regulate the polarization are poorly defined. We have previously reported that the enrichment of autophagosomes in tumor cell conditioned medium and malignant ascites from patients. The present study investigated the hypothesis that tumor released autophagosomes promoted M2 polarization. Here we isolated autophagosomes released from murine liver cancer cells using affinity purification. In vivo mouse experiments demonstrated that intraperitoneal injected autophagosomes were efficiently internalized by peritoneal macrophages, which correlated with upregulation of M2 markers. In vitro, coculture of autophagosomes with bone marrow-derived macrophages (BMDMs) also induced macrophage polarization into a M2-like phenotype. Furthmore, this effect of autophagosomes was largely abolished in BMDMs from MyD88 KO mice. Taken together, our findings suggest that, under stress conditions, tumor released autophagosomes may promote tumor progression via regulation of TAMs in tumor microenvironment, providing new insight into TAMs polarization. Keywords: Tumor released autophagosomes, Macrophage, Polarization, MyD88.
Plasminogen activator inhibitor-1 increases migration of monocytes to the tumor and skews their differentiation towards M2 macrophage phenotype (TUM6P.1002)
Authors: Kubala Marta , Placencio Veronica , DeClerck Yves 
Published: January 2023

Abstract
Macrophages exhibiting M2 (pro-tumorigenic) phenotype have been linked to cancer progression. Plasminogen activator inhibitor-1 (PAI-1) is overexpressed by a variety of tumors and indicates poor clinical outcome. This pro-tumorigenic role has been linked to pro-angiogenic and anti-apoptotic functions. Here we demonstrate that PAI-1 is an orchestrator of monocyte migration towards tumor cells and of their M2 polarization. We observed that PAI-1 KO mice xenotransplanted with HT1080 PAI-1 KD cells exhibited tumors less infiltrated with macrophages compared to WT mice (p-value: 1.27E-09). In vitro, recombinant PAI-1 (40 nM) increased monocyte migration by an average 4-fold and increased the expression of CD163 (a M2 marker) by 10-fold (Median Fluorescence Intensity 3 to 30). PAI-1 induced monocyte production of IL-6 and IL-10 to 4000 pg/mL and 600 pg/mL, respectively, and activated STAT3 in the cells. Supporting the critical role for IL-6 and STAT3 in PAI-1 effect on M2 polarization in monocytes, we show that both blocking the IL-6 receptor (tocilizumab) and STAT3 activation (ruxolitinib) in monocytes treated with PAI-1 blocked CD163 expression. Our data point to a potential new pro-tumorigenic activity of PAI-1 via an effect on monocyte migration and polarization.
Scavenger receptor A is involved in tumor associated immune suppression (TUM6P.958)
Authors: Guo Chunqing , Wang Xiang-Yang 
Published: January 2023

Abstract
Tumors activate a plethora of immunosuppressive mechanisms to counteract effective immune responses, e.g. establishing T cell tolerance against tumor antigens or inducing the expansion of T-cell suppressive cell populations, in particular myeloid derived suppressor cells (MDSCs). Our previous studies have demonstrated that scavenger receptor A (SRA) dampened the intrinsic immunogenicity of antigen presenting cells. To determine the potential involvement of SRA in tumor-mediated immune suppression, we examine the expression of SRA on MDSCs during tumor progression and its impact on the suppressive activity of MDSCs. We show that expansion of MDSCs in tumors and peripheral lymphoid organs (e.g., spleen) is dependent on tumor size. SRA is predominantly expressed on monocytic MDSCs (not granulocytic population), which shows a positive correlation with tumor development. Regardless of tumor growth stages, SRA expression in tumor infiltrating MDSCs is substantially higher compared with their counterparts in lymphoid tissues. Interestingly, tumor growth-dependent elevation of SRA expression correlates with the T-cell suppressive activity of MDSCs. Additionally, the absence of SRA strongly promotes the therapeutic efficacy of adoptively transferred tumor-reactive T cells. Our findings suggest that SRA may participate in tumor-mediated immunosuppressive mechanisms and contribute to the immune evasion during cancer progression.
Effector TCD8 use different homing receptor/ligand interactions to enter subcutaneous and intraperitoneal B16 melanoma tumors (TUM7P.1022)
Authors: Woods Amber , Srivinasan Nithya , Engelhard Victor 
Published: January 2023

Abstract
Immunotherapies have the potential to control cancer, but are currently only effective in patients with a preexisting TCD8 infiltrate. TCD8 infiltration into tissues relies on interactions between homing receptors on TCD8 and ligands on the vasculature. However, those required for entry into tumors are incompletely characterized. Using B16 melanoma expressing the strong antigen ovalbumin, the homing receptor ligands VCAM-1 and E-selectin were more highly expressed on SC tumor vasculature, while MadCAM-1 was expressed more highly on IP tumor vasculature. B16 parental tumors and tumors grown in Rag1KO animals expressed negligible levels of VCAM-1 and MadCAM-1, suggesting that activated adaptive immune cells that previously infiltrated the tumor drove their expression. Our previous work showed that anatomical site of TCD8 priming results in differential expression of α4β1, α4β7, and E-Selectin Ligand (ESL) on effectors. We showed that α4β1+ effectors enter SC and IP tumors via interaction with VCAM-1, while ESL+ effectors preferentially enter SC tumors via interaction with E-Selectin. Unexpectedly, α4β7 and MadCAM-1 did not mediate entry into IP tumors, apparently because MadCAM-1 was not expressed on the luminal surface. Our results suggest that the commonly used SC vaccination route generates effectors that may not home efficiently to visceral metastases. Vaccination targeting non-skin draining LN may generate effectors that infiltrate visceral metastases more efficiently.
Adoptive transfer of autologous tumor-infiltrating lymphocytes in metastatic human papilloma virus-induced carcinoma (VAC7P.1040)
Authors: Stevanovic Sanja , Campbell Tracy , Draper Lindsey , Langhan Michelle , Somerville Robert , Wunderlich John , Kwong Mei Li , Kerkar Sid , Yang James , Sherry Richard , Kammula Udai , Rosenberg Steven , Hinrichs Christian 
Published: January 2023

Abstract
Given the constitutive tumor-specific expression of high risk human papilloma virus (HPV) E6 and E7 oncoproteins in HPV+ carcinomas, we evaluated whether adoptive transfer of autologous tumor-infiltrating lymphocytes (TIL) selected for HPV E6/E7 reactivity could mediate regression of metastatic HPV+ carcinomas. TIL were selected preferentially for HPV E6/E7 peptide pool reactivity, when possible. Patients received a single TIL infusion and interleukin-2 to tolerance after non-myeloablative chemotherapy. The treatment regimen led to objective clinical responses in 6/23 patients, including 4/15 cervical cancer, 1/5 anal cancer and 1/3 head and neck cancer patients. Two complete responses occured in cervical carcinoma that are ongoing 20 and 26 months after treatment. The other four responses were partial, three lasted from 3 to 5 months, and one is ongoing at 3 months. The frequency of HPV reactive T cells in the TIL infusion of 17 patients analyzed thus far (including five with a clinical response), as measured by IFNg ELISPOT and CD137 upregulation, correlated with clinical response (P=0.0238 and P=0.0485). In addition, the magnitude of HPV reactivity as measured by IFNg production correlated with clinical response (P=0.0003). Regression of metastatic HPV+ carcinomas can occur after TIL therapy in some patients. Exploratory studies suggest a correlation between HPV reactivity of the infusion product and clinical response.
Adoptive therapy of ROR1+ murine solid tumors with chimeric antigen receptor-modified T cells (VAC7P.1045)
Authors: Srivastava Shivani , Riddell Stanley 
Published: January 2023

Abstract
Adoptive therapy with T cells modified to express chimeric antigen receptors (CARs) holds promise for treating epithelial cancers if target molecules can be identified and strategies developed to overcome the immunosuppressive mechanisms of solid tumors. Receptor tyrosine kinase-like orphan receptor 1 (ROR1) is expressed in many blood and solid tumors but is absent from vital adult tissues, making it a potential tumor-specific target for therapy. We developed ROR1-specific CARs comprised of an extracellular single chain antibody linked to various intracellular signaling modules. T cells modified to express ROR1-CARs exerted potent anti-tumor effects against ROR1+ leukemia and lymphoma in vivo, but their effectiveness against solid tumors is not known. To test this, we used the orthotopic 4T1 mouse mammary tumor model that recapitulates the immunosuppressive microenvironment and metastatic pattern of human breast cancer. We transduced 4T1 cells with firefly luciferase (ffluc) to allow in vivo imaging of tumor growth, along with mouse or human ROR1. Murine CD8+ T cells modified to express ROR1-CARs lysed ROR1+ but not ROR1- 4T1 cells in vitro, while unmodified T cells were unable to lyse either ROR1+ or ROR1- 4T1 cells. We are currently evaluating the anti-tumor activity of ROR1-CAR T cells against 4T1-ffluc/ROR1 cells in vivo to define optimal CAR constructs and cell compositions and identify what immunosuppressive pathways to target to improve anti-tumor activity.
Retroviral transfer for a chimeric antigen receptor to generate regulatory T-like cells for the suppression of undesired immune responses. (VAC7P.1036)
Authors: Stavrou Maria , Pule Martin , Holler Angelika , Shlomchik Warren , Stauss Hans 
Published: January 2023

Abstract
Adoptive transfer of antigen specific regulatory T (Treg) cells was shown to be beneficial for the suppression of autoimmunity and Graft versus Host Disease (GvHD). To create a cellular regimen for the targeted suppression of unwanted immune responses, we engineered mouse MHC-class II specific Treg-like cells. The restricted expression of MHCII to professional antigen presenting cells and presence at high levels at all inflammatory sites, allow the employment of MHCII specific Treg-like cells for suppression in various inflammatory settings. A mouse MHCII-specific Chimeric Antigen Receptor (CAR) was used to redirect the specificity of T cells via retroviral transduction, while Foxp3 gene transfer into purified CD4+ cells leads to the acquisition of the Treg-like phenotype. Incorporation of a suicide mechanism within the engineered Treg-like cells, for their in vivo selective deletion, when desired, will ensure the short-term nature of suppression. The functionality of the generated CAR and the specificity of responses elicited by CAR bearing T cells were validated in vitro. In subsequent preliminary in vivo experiments, intravenous injection of C57BL/6 mice with syngeneic mouse MHCII-specific CD4+ T cells, led to GvHD-like toxicity, while no signs of toxicity were observed when Treg-like cells of the same specificity were transferred alone or in 1:1 mix with mouse MHCII specific CD4+ T cells. These data suggest the suppressive potential of the engineered Treg-like cells.
A fusion protein containing the adenovirus type 2 fiber trimerizing and knob domains linked to HIV-1 gp120 exhibits increased nasal immunogenicity in rabbits (VAC8P.1046)
Authors: Staats Herman , Jones Dorothy , Maddaloni Massimo , Pickup David , Permar Sallie , Abraham Soman , Pascual David 
Published: January 2023

Abstract
We previously reported that the inclusion of the trimerizing and knob domains from adenovirus type 2 fiber (Ad2F) in protein immunogens enhanced their mucosal binding and immunogenicity when delivered nasally. This study was performed to determine if Ad2F would enhance the nasal immunogenicity of HIV-1 gp120 from the Clade C Env C.1086. A fusion protein with Ad2F at the C-terminus of HIV-1 gp120 was used as gp120-Ad2F. NZW rabbits were nasally immunized with 100, 200 or 300 µg of gp120 or equimolar doses of gp120-Ad2F fusion proteins adjuvanted with the cationic mast cell activating peptide mastoparan 7 on days 0 and boosted on days 28 - 34. Intramuscular immunization with 100 µg gp120 adjuvanted with an MF59-like adjuvant served as the positive control. Serum anti-HIV-1 gp120 IgG titers were measured 2 weeks after the final boost. While only the 200 and 300 µg doses of gp120 delivered intranasally induced serum anti-gp120 IgG similar to those induced by IM immunization, all doses of gp120-Ad2F fusion proteins induced serum anti-gp120 IgG titers similar to those induced by IM immunization. While the 100 µg dose of gp120 delivered nasally induced serum anti-gp120 IgG titers significantly lower than that induced by 300 µg of gp120 delivered nasally, all doses of gp120-Ad2F induced similar anti-gp120 IgG titers. Our results demonstrate that the addition of Ad2F to HIV-1 gp120 enhances its nasal immunogenicity, lessening the amount of gp120 required for vaccination.
Mycobacterium tuberculosis Ag85C, MPT51, HspX, and the fusion recombinant CMX proteins or rBCG CMX vaccine modulate the innate immune response in a pro and anti inflammatory manner (VAC9P.1103)
Authors: da Costa Adeliane , de Rezende Danilo , Santos Bruno , Machado Renato , Kipnis André , Junqueira-Kipnis Ana Paula 
Published: January 2023

Abstract
Introduction: Mycobacterium tuberculosis produces several proteins that have been associated with its virulence and therefore intensively studied as target for subunit vaccine against tuberculosis. Ag85C, MPT51 and HspX were shown to be immunogenic and antigenic proteins. Also, a fusion protein (rCMX) developed using part of these individual molecules induced specific Th1 and Th17 and poly functional Th cells, that were related to tuberculosis protection. Objectives: This study aimed to understand how this immune response was induced. Methodology: Pulmonary, peritoneal, alveolar, and bone marrow derived macrophages from BALB/c mice were stimulated with the recombinant antigens: Ag85C, MPT51, HspX, CMX or with BCG or rBCG CMX vaccines. Results: All macrophage types responded to Ag85C and CMX with elevated NO production when compared with unstimulated macrophages, while the response to MPT51 and CMX produced urea at similar levels as M2 derived macrophages. HspX did not stimulate macrophages. Pulmonary macrophages infected with BCG produced NO similarly to M1 macrophages, whereas rBCG CMX infection induced urea comparable to M2 macrophages. Conclusion: Ag85C, CMX and MPT51 proteins induced M1, mixed and M2 macrophage phenotypes, respectively. However when CMX was produced within rBCG CMX, a predominant M2 phenotype was observed. The results suggest that the used antigens can modulate the immune response similarly to that shown by rBCG CMX in vivo.
Engineering enhanced vaccine cell lines to eradicate vaccine preventable diseases: the polio endgame (VAC9P.1107)
Authors: Tripp Ralph , van der Sanden S. M.G. , Wu W , dybdahl-Sissoko N , Weldon William , Brooks P , O'donnell J , Jones L , Brown C , Tompkins Stephen , Oberste M , Karpilow J 
Published: January 2023

Abstract
Advances in genomic screening and gene editing technologies provide a means to create the next generation of vaccine manufacturing cell lines. A genome-wide RNAi screen was used to identify gene knock-down events that enhance poliovirus replication with the aim of increasing vaccine yield and reducing the costs and time associated with vaccine production. Primary screen hits were validated with several attenuated and wild poliovirus strains in a Vero cell line employed in vaccine production. Multiple genes were identified that upon silencing increased titers &gt;20-fold, and in some instances, knockdown of a gene or gene combination increased virus titers by &gt;50-fold. Importantly, stable knockout of top candidates from the screen and validation studies using CRISPR technology provided equivalent or improved viral production of multiple serotypes. Several of these gene knockdowns also enhanced titer of enterovirus 71, a clinically relevant virus for which vaccines are being targeted. Host gene silencing events that facilitate poliovirus replication mapped to several cell pathways, and in all cases, gene silencing did not affect poliovirus antigenicity. This study suggests that modulating host genes to increase vaccine production can significantly enhance vaccine production at reduced costs, a finding that should greatly facilitate global implementation of inactivated poliovirus vaccine for polio eradication, and change the paradigm in mammalian vaccine cell line production.
Antiviral inducing properties of staphylococcal enterotoxin mimetic peptides (VAC9P.1065)
Authors: Mujtaba Mustafa 
Published: January 2023

Abstract
Superantigens, like the staphylococcal enterotoxins, activate vast numbers of T-cells to produce large amounts of cytokines, including interferon-gamma (IFN-γ), and eventually cause these cells to undergo apoptosis. The objective of this research was to design mimetic peptides of staphylococcal enterotoxins that are not toxic to cells, but can produce antiviral activity in cells via production of IFN-γ. Based on the amino acid sequences of staphylococcal enterotoxins (SE) A, B, C, and toxic shock syndrome toxin, peptides were designed to mimic the antigenic sites of these superantigens. Whole protein superantigens (SEA) at concentrations ranging from 1.0 to 10.0ng/mL stimulated T-cell proliferation and induced IFN-γ production. Superantigen concentrations at or above 100ng/mL showed cellular toxicity. Of the eight mimetic peptides tested, SEA 3 was the only peptide that induced IFN-γ production, as determined by the IFN-γ ELISA kit, in HPBMC but did not induce cellular proliferation. SEA1, SEA2, and TSST peptides induced cellular proliferation but no IFN-γ stimulation. The peptides showed no toxicity directly on HeLa cells or HPBMC at 100 ug/mL or lower. Cell supernatant from the SEA3 peptide treated HPBMC also had antiviral activity against vesicular stomatitis virus. Thus, this study showed that mimetic peptides of superantigens could be developed that can induce T-cells to produce IFN-γ without the cellular toxicity associated with superantigens.
Zebrafish diverse immunoglobulin domain-containing proteins share structural features and ligands with mammalian CD300 and TREM family members and are linked to a MHC class I gene cluster (VET1P.1126)
Authors: Rodriguez Nunez Ivan , Haire Robert , Cannon John , Litman Ronda , Litman Gary , Yoder Jeffrey 
Published: January 2023

Abstract
Bony fish encode multiple multi-gene families of innate-type immune receptors that are comprised of immunoglobulin (Ig) domains and are of unknown function. One of these families, the diverse immunoglobulin (Ig) domain-containing protein (DICP) genes, recently have been identified and characterized from three chromosomal loci in zebrafish. DICPs possess one or two Ig ectodomains and include membrane-bound and secreted forms. The membrane-bound DICPs include putative inhibitory and activating receptors. Recombinant DICP Ig domains bind lipids with varying specificity, a characteristic shared with mammalian CD300 and TREM family members. Here, we employ PCR based strategies to investigate the expression of DICPs and their variability between individuals from three different zebrafish lines. We demonstrate that certain DICPs display restricted expression in adult tissues while others are ubiquitously expressed. In addition, transcripts of a subset of DICPs are detected during embryonic development suggesting roles in innate immunity or other embryonic processes. Finally, the observation that the expression of certain DICPs is linked to their genetic background led to the identification of a new DICP haplotype that is linked to an alternative MHC class I Z lineage haplotype on chromosome 3. These findings highlight the complexity and dynamic nature of the DICP family within zebrafish and it is likely that differences in DICP haplotype may result in altered immune phenotype.
Alterations in MHCII expression on B cells isolated from cattle naturally infected with bovine leukemia virus. (VET1P.1115)
Authors: Frie Meredith , Coussens Paul 
Published: January 2023

Abstract
Bovine leukemia virus (BLV) is a deltaretrovirus that infects B cells and is widespread in US dairy herds. 83% of US dairy herds are infected with BLV and the within-herd infection burden is estimated to be as high as 46%. BLV infection causes reduced milk production and decreased longevity, which leads to economic losses of $520 million every year. However, BLV is predicted to cause immune suppression in infected cattle, which would increase the economic losses incurred as a result of BLV infection in dairy cattle. To characterize cellular mechanisms of immune suppression in BLV-infected cattle, the expression of MHCII in B cells from naturally infected dairy cattle was investigated. PBMCs were isolated from healthy dairy cattle or naturally infected dairy cattle and the MHCII expression on B cells was assessed in unstimulated or pokeweed mitogen-stimulated culture conditions. qRT-PCR and flow cytometry were used to determine the change in MHCII expression on B cells isolated from BLV-infected cattle. BLV-infected cattle have a significantly higher mean relative percentage of B cells expressing MHCII, but significantly reduced mean fluorescence intensity of MHCII. This could suggest that B cells from BLV-infected cattle are less efficient at presenting peptide antigens to CD4+ T cells.
Interaction between feed management systems and omega-3 fatty acid supplementation on bovine immunization parameters (VET1P.1116)
Authors: Clarke Sandra , Fredeen Alan , Glover Kathleen , Fillmore Sherry , MacLaren Leslie , Enright Catherine , Green-Johnson Julia 
Published: January 2023

Abstract
A trial was designed to investigate impacts of omega-3 fatty acid feeding regimes on bovine immunization responses. Dietary supplements contained either fish oil or microalgae as omega-3 fatty acid sources, and cattle were divided into 2 feed management systems; they either grazed pasture or were kept in barn tie stalls and fed total mixed rations (TMR). Cattle were immunized with keyhole limpet hemocyanin (KLH), a standard T cell dependent antigen. Dietary supplements and feeding systems influenced the response of cattle to immunization with KLH. Anti-KLH IgM, IgG and IgA antibody levels in serum were significantly higher in cattle grazing pasture than those fed TMR. Cattle consuming microalgae had consistently higher anti-KLH IgG2 and IgM antibody levels compared to cattle fed fish oil. Effects of omega-3 fatty acid supplements and TMR versus pasture-based diets were also apparent in cytokine profiles from spleen and cecal tissues. Levels of transforming growth factor-beta (TGF-β) were highest in cecal tissue of pasture-grazing cattle (P &lt; 0.0001), and spleen TGF-β levels were highest in pasture-grazing cattle consuming microalgae (P = 0.0073).These findings suggest that effects of omega-3 supplements on bovine immunity are influenced by the feed management system, with heightened effects on antibody production observed with a pasture system relative to a confinement and TMR-based diet.