bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tumor membrane vesicle (TMV)-based cancer vaccine induces dendritic cell maturation and downstream T cell activation
Authors: Munoz Luis E. , Shafizadeh Yalda , Margaroli Camilla , Bommireddy Ramireddy , Selvaraj Periasamy 
Published: January 2023

Abstract
Breast cancer remains the most commonly diagnosed cancers among American women, with over 250,000 new cases expected per year. Breast cancer can be subdivided into different categories depending on the expression of receptors, most commonly into ER+, PR+, HER2+, and TNBC. Clinically, small molecule inhibitors of the hormone receptors and antibodies for HER2 have been developed to target these receptors and inhibit tumor signaling or eliminate these cells via phagocytosis and ADCC. Despite this, tumor cells that develop resistance to therapy remain a significant unmet need as they tend to be more aggressive and metastasize. We developed a novel immunotherapy approach that addresses the high degree of inter-patient and intra-tumor heterogeneity by making tumor membrane vesicle (TMV) vaccines out of individual tumor samples and modifying them with GPI-linked molecules like IL-12 and B7-1 to induce an antitumor immune response. In the present study, we investigated whether GPI-GM-CSF and GPI-IL-12 on TMVs from HER2+ murine breast cancer can target and activate antigen presenting cells. Here, we show that TMVs containing with GPI-GM-CSF and GPI-IL-12 can be phagocytosed by both macrophages and dendritic cells, induce upregulation of MHC-I, MHC-II and CD86, and inflammatory cytokine production in vitro. Further, when the DCs activated with our TMV vaccine were co-cultured with syngeneic T cells, they induced the upregulation of T cell activation markers. In summary, our results indicate that TMVs containing GPI-IL-12 and GPI-GM-CSF can enhance uptake and maturation of APCs and induce a strong T cell response downstream making it a potential therapeutic approach for HER2+ breast cancer.
Vaccine targeting antigens associated with cancer stem cells/epithelial to mesenchymal transition inhibits triple negative breast cancer growth
Authors: Cecil Denise L. , Curtis Benjamin , Gad Ekram , Corulli Lauren , Disis Mary L. 
Published: January 2023

Abstract
Triple negative breast cancer (TNBC) is characterized by higher relapse and poor survival rates compared to other breast cancer types and is marked by a cancer stem cell (CSC)/epithelial to mesenchymal transition (EMT) signature. We questioned whether a vaccine targeting CSC/EMT associated proteins could prevent development of TNBC. We identified five overexpressed proteins found in TNBC and associated with different CSC/EMT pathways. We used a scoring system derived from three common algorithms for predicting class II MHC binding to select Th epitopes for CD105, CDH3, MDM2, SOX-2 and YB-1. Those epitopes that induced a selective Th1 (IFN-gamma) immune response in human population based screening were included in a multi-antigen vaccine. Epitopes were highly homologous between mouse and man. Immunization with the multi-antigen vaccine induced Th1 specific for all five antigens compared to adjuvant control (p&lt;0.05 for all). Immunity persisted 3 months after the final vaccine (p&lt;0.05 for all). Vaccination generated an antigen-specific cytotoxic T-cell (CTL) immune response to three antigens (p&lt;0.05). The vaccine increased effector memory T-cells by 2-fold compared to control (p=0.03). Tumor growth in vaccinated FVB-Tg(C3-1-TAg) mice, a basal breast cancer model, was inhibited by over 80% compared to control mice (p&lt;0.0001). We observed significantly more CTL (p=0.04) and Th1 (p&lt;0.001) infiltrating the tumor with a concomitant reduction in cells expressing the stem cell marker Sca-1 (p=0.003) after immunization with vaccine compared to control. This multi-antigen vaccine targeting CSC/EMT antigens is currently being investigated in a Phase I clinical trial (NCT02157051).
Development of vaccines for treatment of opioid abuse using carrier proteins suitable for pharmaceutical manufacturing
Authors: Huseby Kelcher April M. , Gradinati Valeria , Limkar Ajinkya , Baruffaldi Federico , Lees Andrew , Pravetoni Marco 
Published: January 2023

Abstract
In 2016, over 50,000 people died in the US from heroin and prescription opioid related overdoses. Current standard of care is not sufficient. Therapeutic vaccines against opioids are safe, long-lasting, and cost-effective alternatives to opioid-based pharmacotherapy. Opioid vaccines elicit opioid-specific antibodies that prevent opioid distribution to the brain and opioid-induced behavior and toxicity. To ensure translation, we have developed candidate vaccines that include components suitable for good manufacturing practices and have the ability to be characterized according to FDA guidelines. In this study, we synthesized three oxycodone-based (OXY) haptens to assess linker and bioconjugation chemistry in vaccine efficacy against oxycodone in mice. Then, we tested efficacy of lead OXY haptens conjugated to novel carrier proteins consisting of E. coli-expressed diphtheria cross-reactive material (EcoCRM™) and nontoxic tetanus toxin fragment (rTTHc). Haptens using tetraglycine linkers coupled with carbodiimide chemistry elicited the greatest oxycodone specific serum IgG antibody titers and significantly reduced oxycodone distribution to the brain. Oxycodone vaccines containing EcoCRM™ and rTTHc, adsorbed on alum adjuvant, were effective in inducing expansion of oxycodone-specific plasma and germinal center B cells, blocking oxycodone distribution to the brain, and were easily characterized by mass spectrometry. The OXY-EcoCRM™ and OXY-rTTHc were equivalent, or superior, to vaccines containing keyhole limpet hemocyanin protein, tetanus toxoid and CRM197, making them candidate immunogens for further development.
Combination of a DepoVax™ peptide vaccine with a lentivector vaccine induces strong antigen-specific immune responses and provides effective tumor control in murine models
Authors: Hrytsenko Olga , Weir Genevieve M. , Stanford Marianne M. 
Published: January 2023

Abstract
Many strategies are being investigated to induce and sustain anti-tumor immune responses through vaccination. Viral vectors have been used to induce robust cellular immune responses, however due to immune responses against the vector they loose potency with boosting vaccinations. In this study, we investigated a heterologous prime boost strategy that combines lentivirus and peptide vaccination to induce robust anti-tumour immune responses in a preclinical model.
DepoVax™ is a water-free vaccine formulation that facilities strong and sustained immune responses to peptide antigens. We showed that while homologous immunizations with DepoVax peptide vaccine containing HPV16 E749–57 T cell peptide antigen (DPX-FP) elicit potent antigen-specific responses, heterologous prime-boost immunizations with DPX-FP and lentivector vaccine expressing HPV16 E7 protein could further enhance systemic immune responses detected by IFN-γ ELISPOT. The synergy between two vaccines is influenced by the order of vaccine administration, dose, and delivery route. Using an HPV-expressing murine tumor model (C3), we found that heterologous priming with the DPX-FP vaccine and boosting with E7 lentivector vaccine enhanced tumor control and increased survival rates as compared to the treatments with either vaccine alone. RT-qPCR profiling demonstrated that the vaccine-draining lymph node cells of mice primed with DPX-FP and boosted with E7 lentivector vaccine had an altered immune gene signature with an increased expression of cytotoxic T lymphocyte markers (Gzmb and Tbx21). Taken together our results support a rationale for combining DepoVax based therapy with lentivector vaccines in future clinical trials.
Development of an Effective Cancer Vaccine Platform Using Attenuated Salmonella typhi
Authors: Xu Xin , Metelitsa Leonid S. 
Published: January 2023

Abstract
Poor antigen delivery is one of the major limitations of modern cancer vaccine vectors. To overcome this challenge, we exploited Salmonella Pathogenicity Island 2 and type III secretion system to deliver tumor-associated antigen (TAAs) into the cytosol of antigen-presenting cells in situ. In a recent report, we demonstrated that an attenuated strain of S. typhimurium, which was engineered to express SPI2-regulated oncoprotein survivin (SVN), induced potent CD8 T-cell-mediated antitumor response that was curative in a murine model of highly aggressive lymphoma. In further development of this technology for clinical use, the vaccine has been transferred from an experimental strain to a clinically validated strain of S. typhi, CVD908. To adapt CVD908 to stably express recombinant antigens without antibiotic-dependent selection, we used a recently reported plasmid stabilization system that encodes the single-stranded binding protein (SSB), an essential protein in DNA metabolism, which was deleted from the bacterial chromosome. The SPI2-regulated expression cassette was then cloned into the SSB plasmid, so that the resultant construct maintained bacterial vector stability while expressing and translocating TAAs. We found that CVD908Δssb vector effectively infects human dendritic cells in vitro and translocates recombinant SVN and MYCN oncoproteins into their cytosol. Furthermore, CVD908Δssb remains stable in mice and induces generation of antigen-specific CD8 T cells. Therapeutic vaccination of tumor-bearing mice with SVN or MYCN vaccine produced potent antitumor activity in murine models of lymphoma or neuroblastoma. The results justify clinical testing of CVD908Δssb-based SVN and MYCN vaccines in cancer patients.
Molluscum contagiosum virus subverts MHC-I antigen presentation by targeting tapasin for degradation
Authors: Harvey Ian B. , Wang Xiaoli , Fremont Daved H. 
Published: January 2023

Abstract
Molluscum contagiosum virus (MCV) is an extremely contagious poxvirus and one of the most common human skin pathogens. It is capable of producing persistent skin lesions that lack an inflammatory response, suggesting potent immune evasion strategies. However, the lack of an animal model or cell line to propagate the virus hampers investigations of the underlying mechanisms of pathogenesis. Here we demonstrate that MCV-encoded MC80 subverts MHC-I antigen presentation to evade CTL recognition. The MC80 ORF is conserved in the genome of all known MCV strains and encodes a type I TM protein with sequence similarity to classical MHC-I proteins. We find that expression of MC80 in human and mouse cells results in retention of host MHC-I in the ER and consequent surface MHC-I downregulation. Mechanistically, we find that MC80 can directly interact with tapasin via its ectodomain and target Tpn for ER-associated degradation in a TM and cytoplasmic tail dependent manner. The degradation of Tpn leads to a loss of TAP and impedes assembly of MHC-I with peptide. Our findings reveal a novel strategy for viral sabotage of the peptide loading complex and provide evidence that MCV actively subverts adaptive immune surveillance.
CRABP1 and 2 differentially regulate HCV infection via modulation of lipid droplet abundance.
Authors: Bang Bo R. , Li Meng , Tsai Kuen-Nan , Aoyagi Haruyo , Machida Keigo , Aizaki Hideki , Ou Jing-Hsiung James , Saito Takeshi 
Published: January 2023

Abstract
Vitamin A is an essential nutritional constituent that governs a broad spectrum of cell biology through the activation of its nuclear receptor complexes. The active metabolite of vitamin A, all-trans retinoic acid (ATRA), binds to the chaperone molecule, cytoplasmic retinoic acid binding protein (CRABP) 1 or 2 to regulate its signaling pathways. Given the extremely high homology, CRABP1 and 2 have been predicted to play a redundant role. However, a few studies by others suggested that CRABP1 and 2 might differentially regulate the cellular response to ATRA.
Although the antiviral effect of ATRA is well recognized, the roles of CRABPs in the regulation of viral pathogens have never been studied. In this study, we employed hepatitis C virus (HCV) as a study model to assess the importance of CRABPs. Of great interest, CRABP1 expression enhanced HCV infection while CRABP2 expression resulted in the suppression of HCV lifecycle. Our follow up studies collectively indicated that CRABPs regulate the efficiency of HCV infection through the modulation of lipid droplet (LD) abundance wherein HCV establishes the replicase complex.
The transcriptome analysis revealed that CRABP1 expression downregulated the genes regulated by ATRA and is associated with the expression of genes involved in the steatosis of the liver. In contrast, CRABP2 expression augmented ATRA response, resulting in the inhibition of LD accumulation. Moreover, the chemical inhibition of the CRABP1 and 2 pathways resulted in the abrogation of their pro- and antiviral effect, respectively.
In conclusion, our study revealed a novel link between the cellular response to the micronutrients and its influence on the viral infectious disease.
HIV specific B-cell memory persists in seronegative early treated children and is dominated by IgM-memory responses
Authors: Cotugno Nicola , Morrocchi Elena , Pepponi Ilaria , Rocca Salvatore , Bernardi Stefania , Rinaldi Stefano , Di Cesare Silvia , Cameron Mark J. , Pallikkuth Suresh , Rossi Paolo , Ananworanich Jintanat , Pahwa Savita , Palma Paolo 
Published: January 2023

Abstract
Early initiation of ART in HIV vertically infected infants influences specific immunity by limiting Ag exposure and reducing the viral reservoir size. Currently these patients represent ideal candidates for testing immune therapeutic strategies towards HIV-remission. However a proportion of these early ART treated children (ET) show an undetectable HIV Ab response. It is still unknown whether such profile is associated with a lower ability of these children to mount HIV specific responses once the Ag is re-encountered.
In the present study, we investigated whether HIV specific B-cells persist in seronegative (SN) patients, and the associated gene signatures after re-encountering the virus in vitro. We enrolled 20 ET (6 SN and 14 seropositive, SP), who initiated ART at a mean age of 4.4±3.6 mo. and under durable viral control (&gt; 2 years). We found that gp140+ B-cells, analyzed by FACS, persist in SN patients with a similar frequency of SP. Further analysis revealed a higher percentage of HIV-specific IgM cells in SN CD27+ IgD+ memory subset compared to SP (P=0.002). In addition we investigated by multiplexed RT-PCR, gene expression dynamics after in vitro stimulation with HIV peptides mix. Our results on sorted HIV specific IgM+ cells showed up-regulation of Blimp1 expression only in SP (fold change=1.9; p=0.03), suggesting an impairment of plasma cell differentiation in IgM+gp140+ B-cells in SN patients. Our results firstly reveal that HIV specific B-cell response persists in SN ET, and predominantly resides in the IgM memory compartment. We hypothesize that such responses could be targeted by novel strategies aiming at HIV remission in HIV early treated children.
Establishing the HIV reservoir: HIV-susceptible cells and the signals that recruit them
Authors: Packard Thomas Alexander , Luo Xiaoyu , Grimmett Zachary W. , Herzig Eytan , Roan Nadia R. , Greene Warner C. 
Published: January 2023

Abstract
HIV infection rapidly causes localized inflammation, which contributes to viral pathogenesis by recruiting cells to become infected. At the same time, the stable HIV reservoir forms swiftly and persists over the life of the individual, likely due to infection of long-lived central memory CD4 T cells. The elimination/control of the reservoir is critical to an HIV cure. We sought to understand how host inflammatory responses to HIV drive the formation of the viral reservoir.
Our early studies found that CD4 T cells rapidly produce the chemoattractant CCL2 following HIV infection, and this host response is dependent on HIV reverse transcription but not integration of viral DNA. We proposed that CCL2 produced early in response to HIV lures cells bearing the receptor—CCR2—to the zone of infection. Using mass cytometry, we identified a novel population of human lymphoid CCR2+ CD4 T cells that co-express CCR5 and have a central memory phenotype, along with markers associated with the HIV reservoir. We found that CCR2/5+ cells migrate in response to CCL2, fuse to R5 and X4-tropic HIV and support productive and latent infection. To measure the HIV reservoir in vivo, we sorted CCR2/5+ cells from PBMCs of HIV-infected ART-suppressed individuals and measured integrated HIV proviral DNA. Our preliminary findings show that CCR2/5+ cells have significantly higher levels of integrated HIV than CCR-negative memory (12-fold, p = 0.046) or naïve cells (28-fold, p = 0.012).
We propose that HIV hijacks the host response to the virus to recruit CCR2+ cells, which may become latently infected to form the long-lived reservoir, providing a link between inflammation and reservoir seeding. We are now testing if blockade of CCL2 disrupts formation of the HIV reservoir.
T cells Protect the Brain from a Nasal Virus Infection by Engaging Local Myeloid Cells that Cross-Present Antigen
Authors: Moseman E. Ashley , Ciesinski Alexa F. , McGavern Dorian 
Published: January 2023

Abstract
Pathogens that gain entry into the CNS elicit a uniquely tailored response from the immune compartment by modulating cytolytic and inflammatory functions to confer viral clearance with minimal brain damage. Neurotropic viruses such as vesicular stomatitis virus (VSV) infect neurons in the olfactory epithelial barrier and pass via sensory axons directly into the brain. Within the olfactory bulb, VSV-infected neurons contact a variety of CNS neurons; however, the infection rarely invades the brain. While type I interferon responses are crucial for initial viral control, little is known about how the local adaptive immune response prevents fatal viral neuroinvasion. In this study, we sought insights into how cytotoxic lymphocytes (CTL) prevent the distal spread of infection from the olfactory bulb. Without T cells, virus containment is compromised and VSV traverses caudally to hindbrain regions. Utilizing intravital two-photon microscopy and other techniques, we characterized the mechanism by which T cell interactions within the olfactory bulb facilitate barrier protection. Interestingly, chimeric mice deficient in MHC class I on brain-resident cells, as well as microglia-depleted mice, showed reduced antiviral T cell calcium signaling upon engagement of target cells, suggesting that microglia can cross-present viral antigen and indirectly facilitate viral clearance from adjacent virally infected neurons. This study has revealed a novel mechanism of T cell-mediated viral control in neurons that involves engagement of resident myeloid cells that acquire antigen but are not infected by the virus. This in turn provides barrier protection against a virus that attempts to enter the CNS via the nasal route.
Eastern equine encephalitis virus evades induction of the host immune response through miR-142-3p restriction of myeloid cell replication
Authors: Trobaugh Derek , Sun Chengqun , Klimstra William 
Published: January 2023

Abstract
Eastern equine encephalitis virus (EEEV) is an alphavirus that causes the most virulent mosquito-borne disease in North America. We have previously determined that a myeloid cell-specific miRNA, miR-142-3p, binds to the 3′ non-translated region of EEEV suppressing virus replication and the induction of systemic IFN-α/β, promoting severe disease. Deletion of the miR-142-3p binding sites (virus 11337) rescues virus replication in myeloid cells, induces systemic IFN-α/β production, and leads to virus attenuation in vivo. We hypothesized that miR-142-3p restriction of EEEV replication in myeloid cells suppresses the adaptive immune response. Infection of C57BL6 mice with the myeloid tropic 11337 virus lead to increased cellularity in the popliteal lymph node (PLN) within 12 hours, predominantly comprised of CD11b+ F4/80+ Ly-6C+ inflammatory macrophages. Recruitment of inflammatory macrophages was significantly delayed in the non-myeloid tropic WT infected mice. NK cells and neutrophils were also recruited to the PLN after 11337 infection but not WT infection. In addition, 11337 infection of myeloid cells lead to increased cytokine and chemokine mRNA levels in the PLN compared to WT infection. In the spleen, 11337 infection lead to an increased frequency and number of epitope-specific CD8+ T cells compared to WT infection; however, the frequency was similar in the PLN. On day 6, epitope-specific CD8+ T cells expressed both KLRG1+ and CD127+during a WT infection while 11337 infection lead to the generation of short-lived effector cells (KLRG1+ CD127−). These results demonstrate that miR-142-3p restriction of EEEV replication in myeloid cells limits the induction of the host immune response and the generation of effector CD8+ T cells.
Tracing self-reactive B cells in normal mice
Authors: Kuraoka Masayuki , Nojima Takuya , Reynolds Alexander E. , Kitamura Daisuke , Kelsoe Garnett H. 
Published: January 2023

Abstract
Self-tolerance removes or inactivates autoreactive B cells in primary and secondary lymphoid organs. Studies have used transgenic mouse models or characterizing recombinant antibodies from single B cells to study B-cell tolerance. These approaches are limited, however, by the restricted diversity of transgenic B-cell repertoires or the effort necessary to generate significant numbers of recombinant antibodies. To overcome these difficulties, we used a single-cell culture approach, which supports high-throughput analysis of the B-cell repertoire in pre-tolerance, post-tolerance, and tolerizing anergic B-cell compartments. This approach revealed the affinity “setpoints” of the tolerance checkpoints, the stochastic nature of B-cell tolerance, and subsets of autoreactive B cells that are persistent in periphery. We propose that these anergic compartments may be the origin of systemic autoimmunity and a potential target for HIV-1 vaccines that elicit neutralizing antibodies cross-reactive with self-antigens.
Programmed death-1 restrains the germinal center reaction in type 1 diabetes
Authors: Martinov Tijana , Spanier Justin , Swanson Linnea , Fife Brian T. 
Published: January 2023

Abstract
Programmed death-1 (PD-1) is a T cell inhibitory receptor important for tolerance maintenance. PD-1 deficiency or blockade accelerates autoimmune diabetes in non-obese diabetic (NOD) mice, but the mechanism remains unclear. Autoantibody production is a hallmark of autoimmunity, and has also been reported in patients treated with PD-1 blockade, suggesting that PD-1 might regulate this process. Autoantibody production results from B cell:CD4 T cell interactions in the germinal center of the lymph node. The dynamics and regulation of the germinal center response in spontaneous autoimmunity and in the face of PD-1 deprivation are not well understood, primarily due to an inability to track self-specific lymphocytes. To bridge this knowledge gap, we used tetramers to phenotype islet-specific CD4 T cells and B cells in mice. PD-1-deficient mice, and NOD mice treated with anti-PD-1, had increased insulin autoantibodies, as well as increased insulin-specific germinal center T follicular helper CD4 T cells and germinal center B cells compared to controls. This work provides a mechanistic explanation for autoantibody onset following PD-1 blockade in the clinic, and has important implications for cancer immunotherapy and autoimmunity.