bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of tumor-derived exosomes in the formation of a pre-metastatic niche in cancer
Authors: Morrissey Samantha M. , Kong Xiangyu , Noe Jordan , Yan Jun 
Published: January 2023

Abstract

Purpose
Determine if tumor derived exosomes (TDE) can drive cancer progression through polarization of immunosuppressive cell phenotypes capable of restricting αβ T cell functioning. Preliminary data shows that exosome stimulation differentially upregulates PD-L1 (programmed-death-ligand-1) expression on peritoneal macrophages and lung resident γδ T cells while also upregulating PD-1 expression on αβ T cells. The ultimate goal is to assess whether elimination of TDE can synergize with current PD-1/PD-L1 treatments by removing antagonizing factors and improving clinical outcomes.

Methods
TDE were isolated using serial ultracentrifugation from Lewis Lung Carcinoma cells. Exosome stimulated peritoneal macrophages and γδ T cells were co-cultured with ova-transgenic –I (CD8+) T-cells to assay for suppressive function. Anti-PD-1 antibody was added to the culture to confirm that the PD-1/PDL-1 axis mediates the immune suppression. The effect of TDE on γδ T cell metabolism was also investigated. Lastly, human peripheral blood samples from lung cancer and anti-PD-1 treated lung cancer patients were compared via flow cytometry for γδ/PD-L1 expression profiling.

Results
The results suggest that TDE play a role in tumor metastasis by polarizing γδ T cells and macrophages to an immunosuppressive PD-L1+ phenotype capable of restricting αβ T cell effector functioning.

Discussion/Conclusion
TDE could be directly antagonizing the effects of immune checkpoint therapies. Therefore, future experiments will focus on eliminating exosome secretion by using siRNARab27 inhibition and the small drug compound, cambinol, concurrently with anti-PD-1 or anti-PD-L1 monoclonal antibody in efforts to increase progression free survival.
Immunogenicity of diffuse intrinsic pontine glioma (DIPG)
Authors: Marquez-Manriquez Juan P. , Fernandez Lorena Montijo , BRISENO JOSE ANTONIO MATUTE , MONTANO GEOBANNI TORRES , VERBURG MARK L. , CAMACHO-HERNANDEZ ALEJANDRO , LUCERO-DIAZ PEDRO-ALEJANDRO 
Published: January 2023

Abstract

Background
Little information exists about the potential immunogenicity and immunotargets for DIPG. Most of the pediatric neurotumors are mainly unexplored immunologically. We studied twenty repository samples of DIPG tumors. We selected two relevant immune markers to demonstrate immune infiltration such as CD8 and Tbet and eight biologically and clinically relevant proteins found in aggressive neoplasias including DIPG. According with this preliminary data DIPG is immunogenic and six of eight relevant proteins were overexpressed. This data will open the possibility to treat this lethal tumor with active antigen-specific immunotherapy either alone and in combination.

Methods
N=20 repository DIPG pediatric tumors were analyzed by immunohistochemistry (IHC). Moreover we analyzed by granzyme B and IFN-gamma ELISPOT the naturally occurring immune response against predicted Th1 and CD8 epitopes from the eight proteins and we performed univariate and multivariate analysis using SAS/SAT and prism.

Results
six of eight bad prognosis proteins were overexpressed as following RCAS1 (80%), Fascin-1 (90%), EGFR (55%), VCP (85%), Ape-1 (95%) and Sox2 (70 %). We found 35% of expression of CD8 cells and 40% of Tbet, which is an interesting finding in terms of immune infiltration in DIPG. CD8 antigen specific by Granzyme ELISPOT against Ape-1, RCAS1 and Fascin-1 were significant as well (P=0.0001), (p=0.005) and (9=0.0001), respectively.

Discussion
DIPG is immunogenic according with our preliminary data and we will expand the number of samples in order to validate a large repository and move to the clinic. We predicted immunogenic peptides for each protein using repository PMBC’s and most of them were recognized by autologous PBMCS’s.
MR image-guided focused ultrasound immune modulation for glioma therapy
Authors: Sheybani Natasha Diba , Witter Alexandra R. , Bullock Timothy N. , Price Richard J. 
Published: January 2023

Abstract
Glioblastoma (GB) is the most common and malignant brain tumor. Despite standard treatment with surgery, radiation and chemotherapy, its diffuse nature and proclivity for recurrence renders it largely intractable. Immunotherapy (ITx) approaches (e.g. anti-PD1) may hold promise for treating GB; however, the blood-brain (BBB) and blood-tumor (BTB) barriers hinder delivery of systemically administered ITx drugs. A potential approach to enhancing ITx delivery is MRI-guided focused ultrasound (FUS), a non-invasive technique that, when combined with concomitant systemic injection of microbubbles (MB), can transiently disrupt the BBB/BTB and mechanically perturb the tumor microenvironment. Here, we investigate whether localized BBB/BTB disruption with FUS+MB enhances anti-tumor immune responses and inhibits tumor growth, as a prelude to eventual combination with immune checkpoint blockade. One week after FUS+MB (peak negative acoustic pressure=0.6 MPa) treatment of a murine glioma model stably transfected with luciferase (GL261-luc2), CD86 mean fluorescence intensity on dendritic cells (DC) increased ~3-fold in deep cervical lymph nodes, intratumoral CD4+ T cells doubled, and intratumoral CD8+ T cells increased by ~17% (by flow cytometry). Serial bioluminescence imaging of tumors revealed significant reduction in total photon flux as early as 6 days following FUS+MB (p=0.0106), indicating tumor growth inhibition. We conclude that FUS+MB can promote DC maturity and potentially mediate adaptive immunity against glioma, independent of drug delivery. Ongoing studies entail combining FUS+MB with anti-PD-1 delivery to evaluate whether an allied treatment approach can promote an even more robust anti-glioma response.
A modest change in regulation of NOD1 expression has a major impact on inflammation and gastric cancer
Authors: Subramanian Naeha , Rommereim Leah M. , Dutta Bhaskar , Akhade Ajay Suresh , Hutcheon Carolyn , Rostomily Clifford , Lounsbury Nicolas , Savan Ram , Fraser Iain D.C. , Germain Ronald N. 
Published: January 2023

Abstract
Altered protein dosage by defects in single genes leads to haploinsufficiencies and monogenic disorders, but the impact of small changes in gene expression on multifactorial disease is unknown. Here we show that a persistent small (~1.5 fold) increase in expression of NOD1 (Nucleotide-binding oligomerization domain-containing protein 1), a key innate sensor of bacterial infection, precipitates a large physiological effect with dramatically altered cellular function associated with carcinogenesis. Inhibition of miR-15b and miR-16 microRNA function leads to a ~1.2–1.4 fold increase in NOD1 protein concentration, with even slightly greater increases leading to ligand-independent, switch-like NOD1 activation. miRNA regulation of NOD1 plays a crucial role in limiting the development of an inflammatory state is impaired in gastric cancer with a small increase in NOD1 being associated with greater early patient mortality. Overall, our data show that tight control of NOD1 expression by miRNA prevents this sensor from exceeding a physiological switching checkpoint that promotes persistent inflammation and lethal cancer progression, and reveal the impact of a single and modest cellular alteration on cancer. These findings also have broader implications for understanding how small expression changes caused by genetic variation impact development of complex diseases like autoimmunity and cancer.
SphK1/S1P Axis Regulate PPARγ Levels to Program Metabolically Fit Anti-Tumor T Cells
Authors: Chakraborty Paramita , Thyagarajan Krishnamurthy , Chatterjee Shilpak , Selvam Shanmugam Panneer , Ogretmen Besim , Mehrotra Shikhar 
Published: January 2023

Abstract
Metabolic competition with highly glycolytic tumor and overcoming tumor-induced suppression are prerequisites for achieving prolonged tumor control by adoptive T cell therapy (ACT). Given the importance of sphingosine-1-phosphate (S1P) signaling that are independent of immune cell trafficking, we evaluated if altering sphingosine kinase (SphK) mediated S1P level have benefits for anti-tumor T cell response. A comparison of melanoma epitope gp100 reactive T cell receptor transgenic pMel vs. pMel-SphK1−/− mice derived splenic T cells showed better control of subcutaneously established murine B16-F10 melanoma, increased infiltration at the tumor site with activated pMel-SphK1−/− T cells. Additionally, pMel-Sphk1−/−T cells also exhibited enhanced recall response tracked in different organs after secondary antigen exposure. Mechanistically, we found that attenuation of SphK1/S1P signaling results in higher Sirt1 (an NAD+ dependent protein deacetylase) activity, lower PPARγ activity and higher retention of FOXO1 in the nucleus. While higher nuclear localization of FOXO1 regulates higher migration of SphK1−/− CD8+ T cells, lower PPARγ activity endows them with increased ability to use their stored lipid content even in starved condition and decreased iTreg generation. Owing to their efficacious utilization of lipid as fuel, Sphk1−/− T cells persist longer in nutrient deprived tumor microenvironment. In reciprocal studies, endogenous production of S1P in Sphk1−/− T cells (transfected with WT Sphk1 vector) enhanced PPARγ expression and thereby reversed their phenotype in terms of iTreg generation and lipid utilization. Overall, these data highlight that strategies targeting SphK/S1P axis improves efficacy of ACT.
Homologous and heterologous protection against influenza B virus following live attenuated influenza virus (LAIV) immunization
Authors: Santos Jefferson , Finch Courtney , Sutton Troy , Obadan Adebimpe , Aguirre Isabel , Wan Zhimin , Lopez Diego , Geiger Ginger , Gonzalez-Reiche Ana Silvia , Ferreri Lucas , Perez Daniel R. 
Published: January 2023

Abstract
Influenza B virus (IBV) is considered a major human pathogen, responsible for seasonal epidemics of acute respiratory illness. Two antigenically distinct IBV hemagglutinin (HA) lineages co-circulate worldwide with limited antigenic cross-reactivity between each other. Licensed seasonal influenza vaccines are available as inactivated, recombinant, or live attenuated influenza virus (LAIV) vaccines. Although LAIV vaccines mimic a natural infection that is thought to provide better cross-protection than inactivated vaccines, the former are not currently recommended by the CDC for use in the US due to the sub-optimal efficacy. In order to improve LAIV vaccines and following the footsteps of similar work performed by our laboratory using influenza A viruses, we sought to develop an IBV LAIV vaccine. Thus, we created an IBV LAIV using a two-strategy combination that incorporates temperature sensitive (ts) mutations and an epitope tag in the PB1 gene segment. Such modifications resulted in a LAIV candidate with ts characteristics in vitro. We assessed vaccine safety following intranasal immunization and found that IBV LAIV displayed an attenuated (att) phenotype in vivo with virus replication restricted to the upper respiratory tract. Immunization experiments further showed that a single dose of IBV LAIV candidate conferred sterilizing immunity against lethal homologous challenge. Single immunization with IBV LAIV also provided complete protection against heterologous challenge as evidenced by reduced weight loss and lung viral titers. The ability of LAIV to induce cross-protective immune responses may have important implications for the development of universal influenza vaccine approaches.
Synergistic anti-tumor efficacy of combined TLR3 and CD40 neoantigen vaccine requires Batf3-dependent dendritic cells
Authors: Hoki Toshifumi , Yamauchi Takayoshi , Odunsi Kunle , Ito Fumito 
Published: January 2023

Abstract
Therapeutic cancer vaccine therapy targeting neoantigens has yielded encouraging results, yet overall clinical responses to the treatment have thus far limited. A better understanding of the mechanisms that regulate priming of neoantigen-specific T cells is required to improve vaccine strategy. To this end, we used murine MC38 colon adenocarcinoma cells that harbor a single-epitope mutation within the Adpgk protein with the neo-epitope presented in MHC-I H-2Db molecules. MC38 tumor-bearing mice were treated with Adpgk mutant epitope in combination with toll-like receptor (TLR) 3 agonist, anti-CD40 agonist, or both. Therapeutic vaccination with the Adpgk mutant peptide combined with TLR3 and anti-CD40 agonists (TLR3/CD40) significantly improved survival compared with either TLR3 or CD40 stimulation alone. This was associated with expansion and differentiation of CD8+ T cells in the periphery as well as in the tumor microenvironment. Significantly increased terminally differentiated neoantigen-specific CD8+ T cells in blood and spleen were identified using the tetramer staining assay. Interestingly, frequency of neoantigen-specific CD8+ T cells in blood was inversely correlated with the tumor volume, suggesting epitope spreading and broadening T-cell clonality induced by TLR3/CD40 stimulation. Importantly, therapeutic efficacy of neoantigen/TLR3/CD40 vaccination was abrogated in Batf3 knockout mouse, suggesting Batf3-dependent dendritic cells play a critical role in priming adaptive immunity. These studies provide the rational basis for the use of combined TLR3 and CD40 agonists together as essential adjuvants to optimize vaccines designed to elicit therapeutic immunity against cancer neoantigens.
Ectromelia-encoded protein B22 restricts CD4+ T cell activation.
Authors: Forsyth Katherine S. , Fisher Devin , Hersperger Adam , Eisenlohr Laurence Crane 
Published: January 2023

Abstract
Orthopoxviruses encode many immunomodulatory proteins that profoundly interfere with various aspects of the immune system. Ectromelia (ECTV) is the orthopoxvirus that naturally infects mice, affording the opportunity to study viral-host interactions that have co-evolved. Importantly, disease progression closely mimics that of monkeypox and smallpox in humans. CD4+ T cells are one immune cell type crucial for the control of ECTV and have important roles in both direct cytolytic killing of infected cells and helping B and CD8+ T cell activation. Here we show that ECTV infection of antigen presenting cells inhibits their ability to activate CD4+ T cells directly. However, the inhibitory mechanism is not straightforward as neither down-regulation of surface MHC-II nor soluble factors are responsible. A promising candidate protein for this inhibition is B22, which is located on the plasma membrane and has homologs in most virulent strains including smallpox and monkeypox, but not vaccine strains. Indeed, we have determined that B22 contributes substantially to ECTV virulence. Here we show that B22 is both necessary and sufficient to restrict CD4+ T cell activation, with experiments ongoing to elucidate the specific mechanism of inhibition.
Primary human B cells and plasma cells exhibit different sensitivities to vaccinia virus binding and infection
Authors: Shepherd Nicole , Lan Jie , Li Wei , Xing Yanyan , Rane Sushmita , Yu Qigui 
Published: January 2023

Abstract
Previous reports from our lab and others have shown differential vaccinia virus (VACV) binding to and infection of primary human leukocyte subsets, and that VACV preferentially binds and infects antigen-presenting cells (APCs). Although various groups have examined VACV infection in monocytes, macrophages, and dendritic cells, few studies have evaluated VACV binding and infection in primary B cells, a main type of APCs. In this study, we evaluated VACV binding to and infection of primary human B cells at various stages of differentiation. We observed plasmablasts were resistant to VACV binding, while B cells in other differentiation stages exhibited no defects in VACV binding. However, VACV preferentially infected memory and activated B cells whereas plasma and mature-naïve B cells were resistant to VACV infection. Additionally, VACV infection in total B cells was abortive, halted at the late viral gene expression stage. Furthermore, B cell function, measured by cytokine production, was not significantly affected by VACV infection within 24 h post-infection, but revealed the trend of decreased and increased cytokine production in 8 h and 24 h post-infection, respectively, when comparing uninfected and infected cells. Understanding VACV infection in primary human B cells at various stages of differentiation is important for the development of a safer smallpox vaccine and better vectors for vaccines against cancers and other infectious diseases.
Mechanisms of Zika virus antibody neutralization
Authors: Zhao Haiyan , Crowe James E. , Diamond Michael S. , Fremont Daved H. 
Published: January 2023

Abstract
Zika virus (ZIKV) infection during pregnancy has emerged as a global public health problem because of its ability to cause severe congenital disease. To gain insights into the humoral immune response against ZIKV, we set out to characterize a panel of neutralizing monoclonal antibodies (mAbs) with the goal of correlating B-cell epitopes with neutralizing potency. We developed six mouse mAbs against ZIKV including four that are ZIKV specific and neutralized infection of African, Asian, and American strains to varying degrees. The crystal structures of four of these mAbs in complex with DIII domain of the viral E protein revealed three spatially distinct epitopes, corresponding to the lateral ridge, C-C′ loop, and ABDE sheet regions. Of these, antibodies that target the lateral ridge epitope can confer protection in a lethal mouse model of ZIKV infection. Currently, we are focusing on a subset of human mAbs from healthy subjects with prior ZIKV infection. One of these human neutralizing mAbs binds an epitope in domain II, exhibits broad inhibitory activity in vitro, and protects pregnant and non-pregnant mice from ZIKV infection. Other neutralizing mAbs target DIII and show significant neutralizing activity in vitro. Strikingly, mechanistic studies indicate that the most potent murine and human mAbs in our panels are all capable of blocking pH triggered membrane fusion, and confocal microscopy studies indicate that they can traffic into target cells in complex with virions. These studies provide a path for developing prophylactic or therapeutic antibodies for use in pregnancy and designing epitope-specific vaccines against ZIKV.
Mature anti-insulin B cells survive and present antigen in the absence of Bruton’s tyrosine kinase.
Authors: Nyhoff Lindsay E. , Clark Emily S. , Khan Wasif N. , Kendall Peggy L. 
Published: January 2023

Abstract
Bruton’s tyrosine kinase (Btk) is a tec-family kinase present in B lymphocytes and innate immune cells. Btk is an important regulator of autoreactive B cells. In the non-obese diabetic (NOD) mouse model of type 1 diabetes (T1D), Btk-deficiency is protective against disease development and results in significant loss of anti-insulin IgG, even as total IgG is preserved. Anti-insulin B cells drive T1D by presenting antigen to autoreactive T cells. In a transgenic model, conventional Btk-deficiency reduces anti-insulin B cells by 95%, effectively blocking their development. However, the ability of mature anti-insulin B cells to survive or present antigen without Btk was unknown. We induced deletion of Btk using a loxP-flanked Btk mouse model paired with tamoxifen-inducible Cre. Surprisingly, these anergic anti-insulin B cells survive without Btk, as normal numbers of mature B cells were maintained in transgenic Btkflox/Cre-ERT2 animals after tamoxifen treatment. Btk-negative anti-insulin B cells also remained able to internalize and present antigen to cognate T cells and to phosphorylate phospholipase C γ2 in response to anti-IgM. These findings show that though anti-insulin B cells require Btk for their development, it is not required for mature anti-insulin B cell survival. In addition, our finding that Btk-negative anti-insulin B cells can present antigen may have implications for the use of Btk-inhibition in autoimmunity driven by this mechanism.
Ikaros regulates B-cell tolerance
Authors: Schjerven Hilde , Rodriguez Princess , Hagen Danielle , Rasmussen Marit , Ayongaba Etapong F. , Frietze Seth 
Published: January 2023

Abstract
B-cell development and activation are tightly regulated at multiple steps to ensure a protective immune response, and at the same time avoiding harmful self-reactivity and autoimmunity. Ikaros is a transcription factor that is critical for B-cell development, as demonstrated by the complete lack of B-cells in Ikaros-null mice. Furthermore, Ikaros is shown to play important roles also at later stages of B-cell development, but the precise roles of Ikaros at different stages of B-cell development is still not fully understood. In recent years, Ikaros (encoded by the IKZF1 gene) has been linked to autoimmune disease in humans through both genome-wide association studies (GWAS) as well as recent reports of germline IKZF1 mutations in patients with autoimmune disease. However, the mechanisms underlying altered Ikaros function and the development of autoimmunity are not known. We previously developed Ikaros-mutant mouse models with targeted deletions of the exons encoding the DNA-binding zinc finger 1 (ZnF1) or ZnF4, and found that both mutants have B cells, but display selective partial defects at different stages of B-cell development. We recently found that the Ikzf1-ZnF4-mutant strain displays very high levels of serum Anti-Nuclear Antibodies (ANA) at young age, a hallmark of autoimmune disease. This indicates that Ikaros (and specifically exon 6 encoding ZnF4) is required to regulate B-cell tolerance, and we hypothesize that Ikaros regulates this, at least in part, by establishing a chromatin structure that sets restrictions on B-cell responses to limit autoreactive B cells. We are using our Ikaros-mutant mice to further study the role of Ikaros in B-cell development and tolerance, and will present our results to-date at the meeting.
TLR7 and IFNγ signaling converge at STAT1 in B cells to control germinal center mediated autoimmunity
Authors: Chodisetti Sathi Babu , Domeier Phillip P. , Choi Nicholas , Rahman Ziaur S.M. 
Published: January 2023

Abstract
Toll like receptor 7 (TLR7) is required for the development of spontaneous germinal center (Spt-GC) mediated autoimmunity. Accordingly, autoimmune-prone B6.Sle1b mice carrying an extra copy of TLR7 in the Yaa locus (named B6.Sle1bYaa mice) exhibit significantly elevated Spt-GC responses that strongly correlate with exacerbation of lupus disease. However, the underlying molecular mechanisms in TLR7 driven autoimmunity are not clearly defined. Here we report that overexpression of TLR7 or stimulation with its agonist, imiquimod increases IFNγR and STAT1 expression, and the strength of IFNγR-STAT1 signaling in B cells. B6.Sle1bYaa mice deficient in IFNγR (Sle1bYaa.IFNγR−/−) fail to develop Spt-GCs, resulting in markedly reduced serum autoantibody (autoAb) titers and renal pathology. Further, B cell-intrinsic IFNgR or STAT1 deficient mice fail to generate Spt-GC responses upon imiquimod treatment. Interestingly, TLR7 stimulation induces STAT1 phosphorylation at serine-727 in B cells. Consistently, lupus-prone B6.Sle1b mice with alanine substitution for serine-727 in STAT1 (Sle1b.STAT1-S727A) show significantly reduced Spt-GC responses, resulting in decreased number of ANA-producing antibody forming cells and serum autoAb titers. Finally, using mixed bone marrow chimeras, we find that B cell-intrinsic STAT1-S727 phosphorylation is required for increased autoAb titers in B6.Sle1b mice. Mice with S727A mutation in STAT1, however, respond normally to T-dependent protein immunization. Together these data highlight the importance of a crosstalk between IFNγ and TLR7 signaling in B cells at STAT1 in Spt-GC responses and lupus-like autoimmunity.