bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antigen-specific nanoparticle tolerance treatment actively induces both FoxP3- and IL-10-dependent regulatory mechanisms
Authors: Podojil Joseph R , Cogswell Andrew , Chiang Ming-Yi , Neef Tobias , Murthy Tushar , Boyne Michael , Elhofy Adam , Miller Stephen D 
Published: January 2023

Abstract
There are many autoimmune diseases where pathologic self T cells are the underlying cause of disease symptoms and progression. T cells are found at the site of tissue destruction, resulting not only in the exacerbation of disease, but also the release of self-epitopes in the context of inflammation resulting in the activation of additional T cell populations perpetuating disease. Published data have shown treatment with antigen-containing biodegradablepoly(lactide-co-glycolide) (PLGA) nanoparticles, i.e. tolerogenic immune-modifying particles (TIMP), is both safe and induces antigen-specific tolerance in mouse models of autoimmunity and allergy, as well as in a celiac disease Phase I/IIa clinical trial. This study further investigated the role and mechanism of TIMP-induced tolerance. The present data show that TIMP treatment modulated T cells specific for spread epitopes associated with disease progression, but not encapsulated within the TIMP, i.e. tissue-specific bystander suppression. The PLP139–151 TCR transgenic model systems was utilized to determine the cellular and molecular mechanisms driving antigen specific tolerance mediated by tolerogenic nanoparticle treatment. These data show antigen-specific TIMP treatment induced both FoxP3+ iTregs and IL-10+ Tr1 regulatory phenotypes within the antigen-specific T cell populations in both naïve mice and mice pre-primed with antigen/CFA. Additionally, both functional Tregs and IL-10 are required for TIMP-induced tolerance. Treatment thus activates various antigen-specific Treg subsets capable of regulating responses to disease-relevant autoepitopes not encapsulated within the TIMP via release of immunoregulatory cytokines within the inflammatory site.
Supported by funding provided by Cour Pharmaceuticals Development, Inc.
Liver macrophage subsets differentially regulate metastasis in pancreatic cancer
Authors: Thomas Stacy K , Beatty Gregory L 
Published: January 2023

Abstract
Metastasis is the main cause of morbidity and mortality in patients with cancer. In pancreatic ductal adenocarcinoma (PDA), the liver is the most common site of metastasis. During PDA progression, the liver microenvironment becomes altered to support metastatic seeding and colonization by disseminated tumor cells (DTC). Macrophages (MΦ), including liver-resident Clec4f+ Kupffer cells (KCs) and bone marrow derived MΦ (BMDMs), are the dominant immunological components of this niche, yet their precise roles in regulating metastasis remain poorly defined. Here, we sought to understand the interaction of DTCs with liver MΦ as they seed and colonize the liver. To model liver metastasis, mice received intraportal (iPo) injection of murine PDA cells with analysis 2 or 14 days later to examine metastatic seeding and outgrowth, respectively. Liver metastatic burden and immune cell populations were quantified by flow cytometry and immunohistochemistry. Liver MΦ were depleted using clodronate encapsulated liposomes (CEL) and KCs were depleted using Clec4fDTR mice administered diphtheria toxin. DTCs were found to interact with BMDMs more often than KCs during the seeding phase of metastasis. During colonization, KCs became excluded from metastatic colonies whereas BMDMs actively infiltrated. Although depletion of liver MΦ and KCs specifically did not affect metastatic seeding, MΦ depletion during the colonization phase inhibited metastatic colony outgrowth. Interestingly, BMDM infiltration into lesions was largely unaffected by CEL treatment, whereas KCs were uniformly depleted. This finding suggests that MΦ residing outside of metastatic lesions may be critical to supporting metastatic outgrowth.
Supported by grants from NIH (T32 HL0439, F30 CA257287, R01 CA245323)
The role of complement-mediated signaling during antigen presentation
Authors: Bird Caryn Lishele , Pacholczyk Gabriela , Johnson Theodore 
Published: January 2023

Abstract
Local production of complement C3a and C5a at the immune synapse by activated dendritic cells (DCs) enhances T cell responses in a variety of experimental systems. We have shown that complement C3 plays a mechanistic role in driving synergistic anti-tumor effects of blocking indoleamine 2,3-dioxygenase (IDO) during radiation and chemotherapy in a murine brain tumor model. We hypothesize that, when anti-tumor responses are allowed to happen, locally produced complement C3 acts as a costimulation factor for activation of T cells. To study the role of complement C3 in promoting T cell responses against a nominal tumor antigen (gp10025–33 peptide expressed by B16F10 melanoma), we used T cells from syngeneic pmel-1 mice (TCR transgenic with CD8 T cells that recognize the cross-presented gp10025–33 peptide). We found florid deposition of complement C3 in B16F10 tumors when IDO-blockade was added to a pmel-1/gp10025–33 based vaccine. To model homeostatic conditions of the tumor microenvironment, we used an in vitro co-culture system with gp10025–33 peptide in stringent limited supply. Under these conditions, pmel-1 CD8 T cells stimulated by DCs from complement C3 knock-out (C3-KO) mice had decreased activation marker expression, proliferation, and cytokine effector function, relative to pmel-1 CD8 T cells stimulated by DCs from wild-type (WT) mice. These findings suggest that suppression of local complement production by DCs in the tumor microenvironment may inhibit anti-tumor immune responses.
Supported by grants from Alex’s Lemonade Stand Foundation, Press On Foundation, and Hyundai Hope On Wheels.
Impact of constitutive FLT3 signaling on dendritic cell development and function in a genetically-engineered mouse model of Acute Myeloid Leukemia
Authors: Flynn Patrick A , Coy Jesse L , Romine Kyle Alexander , Kosaka Yoko , Lind Evan 
Published: January 2023

Abstract
Acute myeloid leukemia (AML) is a hematological cancer with a very poor prognosis. FLT3-ITD mutations that cause constitutive FLT3 signaling are commonly seen in patients. Our lab and others have shown that AML patient samples exhibit hallmarks of immune exhaustion such as T cell dysfunction, increased MDSCs, and increased Tregs that associate with worse survival. In solid tumor models DCs have been shown to be important for anti-tumor T cell activity and we aim to further describe the role of DCs in the immune response to leukemia. FLT3 signaling is critical for DC development but the effects of FLT3-ITD on DCs and how it contributes to leukemogenesis remains unclear. In a novel FLT3-ITD driven AML mouse model we found that DCs from these mice have elevated pFLT3, indicating constitutive FLT3 signaling. Moreover, significantly expanded DC progenitors and DCs in the bone marrow and spleen are observed. Analysis of serum cytokines identified increased levels of IL-27, IL-10, and IL-17a in leukemia mice compared to healthy controls, suggesting that the expansion of DCs in these leukemia mice may be driving pathogenic expansion of T helper subsets. Furthermore, we measured higher levels of circulating Th2, Th17, and Treg phenotypes in the blood compared to healthy controls. Thus, we hypothesize that FLT3-ITD DCs are a significant contributor to T cell skewing in AML resulting in poor anti-tumor responses. We are using scRNA-seq methods to identify transcriptional changes in FLT3-ITD DCs that lead to the phenotypes we identified. Understanding how FLT3-ITD DCs contribute to AML immune suppression is critical to interpreting leukemia etiology and the development of targeted therapies such as immune checkpoint blockade or small molecule inhibitors.
Long-term sterile immunity induced by an adjuvant-containing live-attenuated AIDS virus
Authors: Yasutomi Yasuhiro 
Published: January 2023

Abstract
Antigen 85B (Ag85B) is one of the most dominant proteins secreted from most mycobacterial species, and it induces Th1-type immune responses as an adjuvant. In the present study, a new strategy to improve live-attenuated SHIV was examined by genetically engineered virus that co-expresses an immunostimulatory molecule Ag85B such as an adjuvant. We genetically constructed a live nef-deleted simian-human immunodeficiency virus to express the adjuvant molecule Ag85B (SHIV-Ag85B) and assessed vaccine effects in cynomolgus macaques. The SHIV-Ag85B could not be detected 4 weeks after injection in cynomolgus macaques, and strong SHIV-specific T cell responses were induced in these macaques. Moreover, most of macaques inoculated with SHIV-Ag85B showed protective effects against the intravenous challenge of pathogenic SHIV89.6P, and viral RNAs remained undetectable for more than 200 weeks. Remarkably, eradication of SHIV89.6P was confirmed by an adoptive transfer experiment and CD8+ cell depletion study. In these macaques, SHIV antigen-specific CD8+ T cell responses with polyfunctionality were rapidly induced in the acute phase of SHIV89.6P challenge. The results suggest that SHIV-Ag85B elicited strong sterile immune responses against pathogenic SHIV and that it may lead to the development of a live vaccine and cure for AIDS virus infection.
Supported by Health Science Research Grants from the Ministry of Health, Labor and Welfare of Japan, Ministry of Education, Culture, Sports, Science and Technology, Japan, Japan Agency for Medical Research and Development, AMED (19ak0101047h0004, 19fk0410025h0001, 19ak0101068h0003 and 19fk0410011h0702).
Exposure to SARS-CoV-2 spike protein induces antigen specific IgE and basophil activation
Authors: Zhang Tianyi , Magazine Nicholas , McGee Michael Conrad , Huang Weishan 
Published: January 2023

Abstract
The spike glycoprotein of SARS-CoV-2 (S) has been used as a main target antigen for vaccine design. There are concerns of vaccine-induced allergies, but mostly about allergic responses to the adjuvant. We, however, found that repetitive exposure to SARS-CoV-2 spike, not Nucleocapsid (N) protein, can induce robust type 2 inflammation in the airway of mice, which is exacerbated by the presence of house dust mite allergens. This type 2 inflammation is accompanied with elevated IgE and neutralizing antibody production. To further test whether spike glycosylation influences these responses, we compared the immunogenicity of insect cell expressed spike (in-S), mammalian cell expressed spike (m-S) and deglycosylated spike (dg-S). With intranasal treatment, in-S induced high levels of anti-spike/S1 specific IgE, while m-S and dg-S induced detectable NAb titers as determined by pseudo-virus (PV) neutralization assays. Moreover, exposure to in-S induced a significant amount of IgE+ basophils in the spleens and lungs of the mice. These spike-induced phenotypes are dependent on the TLR4/MyD88 and STAT6 signaling pathways. When delivered via intramuscular immunization following the priming/boosting protocol, spike protein induced high levels of antigen-specific IgE and IgE+ basophils. Of note, in-S induced the highest level of anti-spike/S2 IgE. However, interestingly, deglycosylation of spike antigen resulted in enhanced induction of NAbs and spike-specific CD8+ T cells following intramuscular immunization. Together, our data suggests that vaccination with SARS-CoV-2 spike may induce antigen-specific allergic response, while immunization with the deglycosylated spike protein may help enhance protective immune responses.
Supported by National Institutes of Health (R01 AI151139, R56 AI146226 and P20 GM130555). W. Huang also received research support from MegaRobo Technologies Corporation.
Hybrid immunity and vaccine breakthrough lead to robust humoral response and antibodies that effectively neutralize SARS-CoV-2 variants
Authors: Bates Timothy A , McBride Savannah K , Leier Hans C , Lyski Zoe L , Messer William B , Curlin Marcel E , Tafesse Fikadu G 
Published: January 2023

Abstract
Current COVID-19 vaccines significantly reduce overall morbidity and mortality and are vitally important to halting the pandemic; individuals who previously recovered from COVID-19 display enhanced immune responses after vaccination (hybrid immunity) compared to their naïve-vaccinated peers. However, the effects of vaccine breakthrough infections on humoral immune response remain to be determined. Here, we measure neutralizing antibody responses from 104 vaccinated individuals including those with breakthrough infections, hybrid immunity, and no infection history. We find that human immune sera collected after either a breakthrough infection or a vaccination post-natural infection will both broadly neutralize SARS-CoV-2 variants to a similar degree. While age negatively correlates with antibody response after vaccination alone, no such association was found in breakthrough or hybrid immune groups. Together, our data suggest that the additional antigen exposure from natural infection substantially boosts the quantity, quality, and breadth of humoral immune response regardless of whether it occurs before or after vaccination.
Supported by grants from the M.J. Murdock Charitable Trust, OHSU Innovative IDEA grant (1018784), NIH (R01AI145835, T32HL083808), and an unrestricted grant from the OHSU Foundation.
Characterizing oncolytic potential and safety of a G-protein pseudotyped vesiculovirus in sarcoma
Authors: Dumbauld Chelsae R , Seetharam Mahesh , Nagalo Bolni Marius , Borad Mitesh 
Published: January 2023

Abstract
Several vesicular stomatitis virus (VSV)-based oncolytic therapies have advanced to phase II clinical trials for treatment of solid tumors. As most antibodies produced after VSV infection are directed towards the surface glycoprotein (G), switching this G-protein of VSV to that of a novel virus could enable repeat dosing in cancer patients by evading adaptive immunity. We aim to study whether a hybrid virus expressing the G-protein of a closely related and non-pathogenic vesiculovirus exhibited oncolytic efficacy across sarcoma models.
We compared cytotoxic effect of VSV, VX1 (wild-type novel virus), and VxG (hybrid) post infection in sarcoma lines. We quantified viral progeny production over time and compared real-time rate of viral-induced apoptosis. Sarcoma lines were tested for production of type-I interferon (IFN) and protection from VxG-induced cell death by exogenous IFNβ. We completed in vivo efficacy studies of intratumoral administration of VxG in xenograft and syngeneic models. From syngeneic tumors, we performed immunophenotyping on residual tumors treated with VxG compared to controls.
VxG was observed to be well tolerated and efficacious. In a xenograft model, a single low-dose administration of VxG completely inhibited tumor growth compared to controls. Lower tumor burden was observed in syngeneic tumors treated with VxG compared to controls. Comparisons of tumor infiltrating lymphocytes between residual VxG treated and control tumors will be completed.
We conclude that the novel hybrid virus VxG has potent oncolytic effects in multiple models of sarcoma and low toxicity in immunocompetent mice. Future endeavors will focus on improved viral engineering and further pre-clinical studies.
Supported by the Mayo Clinic Graduate School of Biomedical Sciences
Novel RNA-based Cancer Vaccine for AML immunotherapy
Authors: Liu Haiyan , Tham Guo Hong , Ren Xi , Yang Henry 
Published: January 2023

Abstract
Acute Myeloid Leukemia is a rapidly progressing malignancy relying primarily on chemotherapy as the frontline treatment option. Success of personalized neoantigen vaccines in melanoma patients has garnered great attention in the field of immunotherapy. We hypothesize that personalized cancer vaccine, in the form of mRNA, can be a viable treatment strategy for AML patients. Next generation sequencing was performed on both C57BL6Jinv mouse’s and C1498 cell line’s nucleic acid materials. Three different neoepitope prediction pipelines were utilized to identify potential immunogenic neoepitopes. Candidate neoepitopes within intron polyadenlyation sites (iPAS), retained intron (RI) sites and somatic mutation (SM) sites were found. A total of 285 SM-derived neoepitopes were identified. Top 38 highly expressed genes were selected for further analysis. Using Sanger sequencing, we confirmed 37 out of the 38 mutations. A total of 5 iPAS-derived neoepitopes were identified. A total of 26 RI-derived neoepitopes were identified in our first RNAseq analysis and a separate 476 RI-derived neoepitopes were identified in our second RNAseq analysis. With these three categories of candidates, we narrowed down our selection and did a preliminary screen for immunogenic epitopes using a total of 71 synthetic peptides in an ELISpot assay. Conventional ELISpot assay’s results were however non-reproducible, thereby propelling us to adopt Culture ELISpot assay. Subsequently, 10 neoepitope candidates were short-listed for further in-vivo evaluation. mRNA vaccine was synthesized and evaluated in a murine AML model. T cell responses against the 10 neoepitopes were analyzed to provide pre-clinical evidence for patient studies.
Supported by NMRC CS-IRG grant
Immunogenicity and efficacy of a multiple antigen-presenting system (MAPS™) COVID-19 vaccine in Non-human Primates
Authors: Besin Gilles , Cieslewicz Brian , Makrinos Daniel , Haridas Renuka , Burke Heidi , Stevenson Taylor , Malley Richard 
Published: January 2023

Abstract
In response to the COVID-19 pandemic, the international scientific and biopharmaceutical community has rallied with unprecedented speed to develop effective vaccines to combat this novel pathogen. Despite this remarkable success, the rise of SARS-CoV-2 variants, some of which appear more resistant to the effects of vaccination, highlights the potential need for additional COVID vaccines.
We used the MAPS technology, in which proteins are presented on a polysaccharide polymer to induce antibody, Th1, Th17 and CD8+ T cell responses, to engineer a COVID-19 vaccine. The COVID-19 MAPS vaccine contained a fragment of the S-RBD sequence of the original strain. Nonhuman primates (NHP) were immunized twice with the COVID-19 MAPS for assessment of immunological responses. NHPs were then challenged with SARS-CoV-2 and protection was assessed by measuring the viral load in the lung and in the nasopharynx.
The COVID-19 MAPS vaccine generated robust neutralizing antibodies in all three animal species as well as Th1, Th17 and cytotoxic CD8 T-cell responses in NHPs. Following viral challenge, MAPS-immunized NHPs had significantly lower viral load both in the nasopharynx and the lung than control animals. This difference was appreciable in the earliest stages following challenge.
A COVID-19 MAPS vaccine containing the RBD of the D614G SARS-CoV-2 strain was very immunogenic in mice, rabbits and NHPs, generated de novo T cell responses and was highly protective against viral challenge in NHPs. These results, as well as the plug-and-play nature of the MAPS technology, in which different RBD variants can be included in a single vaccine preparation, suggest that the MAPS platform could be used to make a vaccine to target multiple variants of SARS-CoV-2.
The Reaction of Normal and Rabbit Pox Immune Adult Rabbits to Vaccination with Vaccine Virus
Authors: Rosahn Paul D. , Hu C. K. , Pearce Louise 
Published: January 2023

Summary and Conclusions
Vaccination with vaccine virus of a rabbit colony of approximately 1500 animals was carried out as a prophylactic measure against rabbit pox. The observations made on the adult population are discussed in the present paper.
Of 233 presumably susceptible animals, 195 or 83.7 per cent gave a positive local reaction to vaccination. Generalized lesions, comprising in particular cutaneous and mucocutaneous papules, adenopathy, and in males an orchitis, developed in 78 animals, an incidence of 33.5 per cent. Nine of these cases had had a negative vaccination. Clinical evidence of susceptibility to vaccinal infection was thus demonstrated in 204 presumably susceptible rabbits, an incidence of 87.6 per cent.
Of 310 rabbits which had recovered from rabbit pox ten months previously and were, therefore, presumably refractory to vaccination with vaccine virus, 294 or 92.3 per cent gave completely negative results. A positive reaction to vaccination developed in 21 animals, however, and there were 3 cases of generalized lesions in animals with a negative local vaccination site, a result which shows that the refractory state induced by rabbit pox may eventually disappear.
An outstanding feature of the results concerned the different character of the reaction of female as compared with male rabbits. A slight, even a negative, local reaction to vaccination was often observed in females and there were comparatively few cases of generalized lesions. This type of reaction was especially striking in the case of pregnant animals. The differences between the lower incidence of both local and generalized lesions in pregnant rabbits as compared with males were found to be statistically significant.
Studies of the Early Stage of Resistance to Pneumococcic Infection
Authors: Schlesinger Helen , McBroom Josephine 
Published: January 2023

Summary
The early stage of resistance to infection by a type I pneumococcus may be elicited within 48 to 72 hours in response to intracutaneous treatment with the homologous type I vaccine, but not in response to the heterologous type III vaccine.
Small quantities of mouse-protective substances are demonstrable in the blood of slightly more than half of the resistant animals by the sixth and tenth days following a single intracutaneous immunizing injection; they are not present in demonstrable quantities as early as the second and third days following this treatment. No increase in agglutinins could be demonstrated during the early stage of resistance.
Sensitization to Ragweed-Extract and the Production of Antibodies by Means of Adjuvants
Authors: Kulka Anna M. , Hirsch Dorothy 
Published: January 2023

Summary and Conclusions
Ragweed-pollen extract combined with a lanolin-like substance and paraffin oil containing killed tubercle bacilli induces sensitization and antibody-formation against pollen-extract far more vigorously than pollen-extract alone. Sensitzation in both groups of animals was noted earlier than the presence of circulating antibodies. Animals with the highest cutaneous sensitiveness had also the highest precipitative titer. The presence of passively transferable antibodies against ragweed-pollen could be demonstrated in animals injected with pollen-extract plus adjuvants.