bluuebunny/crossref_metadata_2025_split huggingface dataset:
PD-L1+PD-L2+ monocytes from patients with hepatocellular carcinoma may be associated with a poor prognosis
Authors: Yasuoka Hidetaka , Asai Akira , Matsui Masahiro , Okamoto Norio , Nishikawa Tomohiro , Ohama Hideko , Tsuchimoto Yusuke , Fukunishi Shinya , Higuchi Kazuhide 
Published: January 2023

Abstract
The prognosis of patients with hepatocellular carcinoma (HCC) is poor. Anti–programmed death-1 (PD-1) antibodies (Abs) has been approved to treat HCC. Some PD-1 ligands (PD-L1 and PD-L2) negative tumors respond to anti–PD-1 Abs. This response may be caused by the expression of PD-1 ligands on non-tumor cells. PD-1 ligands were recently found to be expressed on monocytes (CD14+ cells) from diverse cancer patients. In this study, we investigate the expression of PD-1 ligands on CD14+ cells from HCC patients and the role of CD14+ cells in an antitumor response.
From October 2017 to June 2019, 67 patients with HCC were enrolled. The expression of PD-L1 and PD-L2 on CD14+ cells was analyzed by flow cytometry. Their prognosis was evaluated by overall survival (OS). Tumoricidal activities of both CD14+ and CD8+ cells were calculated by LDH release assay.
In the results, CD14+ cells from the patients expressed PD-L1 (4.5–95.5%) and PD-L2 (1.6–90.2%). We set cut-off values as the mean values of PD-L1 (50.2%) and PD-L2 (50.5%) expression. In addition, according to the cut-off value, we classified patients as those either with PD-L1+PD-L2+CD14+ cells or other types of CD14+ cells. The OS of patients with PD-L1+PD-L2+CD14+ cells was significantly shorter than that of patients with other types of CD14+ cells (p = 0.0145). The tumoricidal activity of CD8+ cells from patients with PD-L1+PD-L2+CD14+ cells was suppressed by co-cultivation with syngeneic CD14+ cells. Furthermore, anti–PD-1 Ab restored the tumoricidal activity of CD8+ cells.
In conclusions, some patients with HCC have PD-L1+PD-L2+CD14+ cells that suppress their antitumor response of CD8+ cells. These inhibitory functions of CD14+ cells may be associated with a poor prognosis in such patients.
Adipocyte PD-L1 suppresses anti-tumor immune response and promotes breast cancer progression
Authors: Wu Bogang , Sun Xiujie , Gupta Harshita B. , Yuan Bin , Chiang Huai-Chin , Hu Yanfen , Curiel Tyler J , Li Rong 
Published: January 2023

Abstract
PD-L1 has become a major target in anti-cancer immunotherapy, however the overall response rate still remains relatively low among most types of cancers, notably breast cancer. There is an unmet need to boost efficacy of immune checkpoint blockade therapies. While both tumor and host immune cell-derived PD-L1 are implicated in immune suppressive functions of PD-L1, the exact immunosuppressive contribution of PD-L1 from different host tissues is little studied. Here we show that PD-L1 expression is significantly higher in human breast adipose tissue versus stromal vascular fractions. In vitro adipogenesis of mouse pre-adipocytes significantly up-regulates PD-L1 versus pre-adipocytes. In vitro co-culture shows that adipocyte PD-L1 suppresses T cell activation and response to anti-PD-L1. In an adipocyte-specific knockout (KO) mouse model, we show that syngeneic mammary tumors grow slower in KO than wildtype hosts. Immunophenotyping shows that tumors grown in KO mice have higher CD8+and CD4+ T cell infiltration as well as more CD8+ Prf+ cytotoxic T cells. Tumor tissue RNA-seq analysis reveals that genetic ablation of adipocyte PD-L1 confers a distinct transcriptomic signature of T cell activation and tumor killing. Our current findings uncover a previously unappreciated source of immune suppressive PD-L1 in the breast cancer microenvironment and could inform novel therapeutic strategies through targeting tumor-surrounding adipose tissue for treating breast cancer.
Intracranial tumors lead to sequestration of T cells in the bone marrow
Authors: Lorrey Selena J , Chongsathidkiet Pakawat , Wilkinson Daniel S , Champion Cosette , Fecci Peter 
Published: January 2023

Abstract
Glioblastoma (GBM) is the most common primary, malignant brain tumor and is universally fatal. Immunotherapeutic intervention has shown some success in immunogenic solid tumors, but little efficacy in GBM. Tumor-imposed T cell deficits remain a major obstacle. Recently, we demonstrated that GBM patients exhibit local and systemic T cell dysfunction, severe lymphopenia and contracted lymphoid organs. These observations are recapitulated in multiple murine models of GBM. Although T cell numbers are decreased in blood, lymph nodes, and spleen, we find large numbers of T cells sequestered in the bone marrow of both patients and mice with GBM. T cell sequestration occurs not only in the setting of GBM, but with nearly any cancer placed intracranially. Intriguingly, none of the tumors tested, including GBM, induce sequestration of T cells in the bone marrow when implanted subcutaneously. These data suggest that the capacity to sequester T cells resides within the brain and is usurped by tumors of the intracranial compartment. Such capacities may well represent a novel mechanism of immune privilege. Sequestration of T cells follows loss of sphingosine-1 phosphate-receptor 1 (S1P1) from the T cell surface. Understanding the drivers of S1P1 loss will provide crucial insight for how brain tumors facilitate systemic immune alterations and permit the development of more effective immune-based strategies for brain tumor patients.
A peptide presentation by an MHC tetramer reduces the expansion of the peptide-specific T cells and the reduced expansion is reversible by PD-1 blockades
Authors: Ouerkaxi Wushouer , George Marybeth , Daftarian Pirouz 
Published: January 2023

Abstract

Background
Peptide-MHC (pMHC) tetramers have been widely used in infectious disease and immune-oncology area to detect antigen-specific T cells. However, little is reported about their application in the context of pMHC presentation to stimulate T cells. Here we compared the expansion of antigen-specific T cells stimulated by an unlabeled pMHC tetramer and the peptide. We also analyzed the relationship among the programmed death-1 (PD-1), lymphocyte activation gene-3 (LAG-3), PD-L1, CD80 and CD38 expression on antigen-specific and non-specific T cells.

Results
Compared to the peptide stimulation, the pMHC tetramer induced lower expansion of the peptide-specific T cells. PD-1, PD-L1, CD80, and LAG3 expression was significantly higher on peptide-specific T cells than on non-specific T cells. Interestingly, peptide-specific T cells expanded by pMHC tetramer induced lower numbers of dysfunctional PD-1+CD38high CD8+ T cells. Furthermore, blockade of the PD-1 pathway by using an FDA approved antibody reversed such a restriction of expansion.

Conclusion
The pMHC tetramer stimulation resulted in the reduced expansion of peptide-specific CD8 T cells when compared to the peptide stimulation. Since the impaired expansion was reversed by the anti-PD1 therapeutic antibody, this artificial stimulation method could be used as a functional in vitro immune-oncology drug screening tool.
Aluminum hydroxide and calcium phosphate adjuvants in adsorbed tetanus and diphtheria vaccines: Immunologic vs histopathologic effects
Authors: Bagriacik Emin Umit , Oruklu Nihan , Yaman Melek , Karakus Resul 
Published: January 2023

Abstract
We aimed to study effects of aluminum hydroxide (AH) and calcium phosphate (CAP) adjuvants in potentiation of new tetanus and diphtheria vaccines that have been developed in our laboratory. AH gel or CAP nanoparticles were prepared in vaccine formulations in adsorbed tetanus or tetanus-diphtheria (Td) vaccines. Vaccine formulations were injected to 8-week old female Balb/c mice. Only phosphate buffered saline (PBS) was injected to control animals. Tissue samples, peripheral blood samples and spleen cells were collected from each mouse. Serum was separated by centrifugation of peripheral blood. Spleen cells were stained with anti-CD4-FITC, anti-CD8-PE, and anti-CD19-APC monoclonal antibodies. ELISA was performed to test presence of anti-tetanus or anti-diphtheria antibodies. We found that either a single dose or an additional booster doses did not change significantly the composition of lymphocyte distribution in AH and CAP adjuvant injected mice in comparison to the control animals. No significant difference was detected for CD4/CD8 ratio and for CD19-expressing cell number. However, presence of IgM and IgG antibodies that were specific for both tetanus and diphtheria toxins increased significantly following injection of vaccines. Granulomas and cyst formation after subcutaneous injections of CAP-containing vaccines at the site of injection were evaluated. Supported by Gazi University BAP, No: 64/2018-04.
Single-Cycle Adenoviruses (SC-Ads) as Platforms for Immunostimulation
Authors: Barry Michael A , Barry Mary , Parrett Brian , Lu Shao-Chia , Zell Brady , Matchett William 
Published: January 2023

Abstract
Most adenovirus vaccines are replication-defective RD-Ads. In contrast, replication-competent adenovirus (RC-Ad) replicate antigen genes 10,000-fold in each cell to amplify immune responses. While RC-Ads vaccines are more potent than RD-Ad, they risk causing adenovirus infections in humans. We engineered “single-cycle” Ad (SC-Ad) vectors that still amplify transgenes, but that do not produce infectious progeny. RD-, SC-, and RC-Ads were tested by single immunization against influenza, Ebola virus, HIV-1, Zika virus, HCV, CMV, Clostridium difficile, and cancers in mice, hamsters, and macaques and antibody and T cell responses were measured. SC-Ad expressing GFP, influenza HA, or HIV gp140 generated antigens with 33 to 100-times less virus than RD-Ad. SC-Ad generated vaginal antibodies that persisted beyond 6 months after single intranasal immunization. Intramuscular injection of 1010 vp of SC-Ad6 expressing HIV-1 gp160 generated env antibodies in sera within 4 weeks of a single immunization in rhesus macaques. SC-Ad expressing a C. difficile toxin A and B immunogen generated toxin-binding antibody reciprocal titers above 105 that persisted in excess of a half of a year after a single immunization. Single immunization with SC-Ad protected vaccinated animals against toxin and against challenge with C. difficile spores. When SC-Ads were used to amplify the expression of immune stimulatory proteins in the context of cancer, single intratumoral injection of SC-Ad expressing 4-1BBL or GMCSF mediated significant reductions in tumor growth and extension of survival without the need for exogenous cancer antigen expression. These data suggest that SC-Ads may have utility against infectious diseases and cancer.
IRF5 Drives Potentiated IFN-beta and IL-12 production by dendritic cells in response to combinatorial CpG- and MPLA- delivery on microparticles
Authors: Jhita Navdeep , Toy Randall , Pradhan Pallab , Blanchard Emmeline , Narang Neha , Cortes Gabriela , Keenum Mark Cole , Santangelo Philip J , Shayakhmetov Dmitry M , Roy Krishnendu 
Published: January 2023

Abstract
Vaccines have enabled control of many deadly infectious diseases. However, efficacy in host protective immune responses vary greatly. Adjuvants targeting Toll like and cytosolic receptors can greatly enhance antigen-specific immunity. In this regard, pathogen-like micro-particles (PLP) co-loaded with TLR4 (MPLA) and/or TLR9 (CpG) ligands was used as a novel platform to deliver combinatorial adjuvants. We analyzed molecular signaling pathways activated in bone-marrow-derived dendritic cells (BMDCs) upon stimulation with PLPs, specifically leading to IFNb and IL-12p70 production. Stimulation of BMDCs with PLPs containing only MPLA or CpG resulted in low-level or no IFN-b and IL-12p70 release. However, stimulation of BMDCs with PLPs containing both MPLA and CpG triggered production of very high amounts of IFN-b and IL-12p70. Analysis of signaling pathways responsible for these results demonstrated IFN-b and IL12-p70 production in BMDCs harvested from WT, IRF3-KO, and IRF7-KO mice, but not from IRF5-KO animals. Exploration of IRF5 phosphorylation kinetics showed MPLA-loaded PLP transiently stimulated IRF5 (0.5 to 2 hours post stimulation) with dual adjuvant PLPs displaying a greatly prolonged p-IRF5, detectable even at 4 hours post stimulation. We are currently analyzing molecular regulators of p-IRF5 stability and kinetics of its translocation to the nucleus. Our study reveals an unexpected role for IRF5 in driving a highly potentiated production of immune-stimulatory cytokines from dendritic cells in response to dual MPLA and CpG adjuvants. These findings may help develop novel vaccines with improved efficacy when combined with PLPs loaded with MPLA and CpG adjuvants that activate IRF5-dependent cytokine production.
DNA priming vaccination to induce T cell responses to subdominant conserved HIV epitopes
Authors: Felber Barbara K. , Lu Zhongyan , Hu Xintao , Valentin Antonio , Rosati Margherita , Mullins James I. , Pavlakis George N 
Published: January 2023

Abstract

Background
HIV sequence diversity and epitope immunodominance exerted by the variable regions are hurdles in the development of an effective AIDS vaccine. We developed novel DNA vaccine regimens to redirect T cell responses to highly conserved regions (CE) in the HIV proteome.

Methods
Rhesus macaques were primed with DNA expressing CE immunogen and boosted with a combination of DNAs expressing CE and/or full-length (FL) immunogens. Different prime-boost protocols were compared. The CE prime/CE+FL boost vaccination co-delivered in the same anatomical site was compared to delivery of CE and FL DNA in contralateral sites.

Results
CE prime/CE+FL boost regimens induced high and broad CTL responses which cannot be achieved using FL DNAs only. Thus, priming with CE DNA profoundly alters the immunodominance exerted by the variable non-CE regions, promoting responses to subdominant CE. This vaccine regimen is currently tested in clinical trials (HVTN 119, ACTG A5369). Comparison of same site vs contralateral vaccine delivery showed that separating CE and FL DNA resulted in increased breadth of T cell responses to non-CE while maintaining CTL responses to CE. Separating the immunogens at the prime allows for induction of independent primary adaptive responses by targeting the two vaccine components to distinct lymph nodes and preventing interference.

Conclusions
HIV DNA vaccine regimens comprising CE as prime and native immunogens as boost induce responses to subdominant epitopes, maximizing breadth and focusing the immune response against vulnerable regions of the virus. Separating the immunogens results in broader epitope recognition. This has implications for many vaccine protocols promoting induction of protective responses.
Understanding the early steps of the adaptive immune response to differentially-effective vaccines during chronic schistosomiasis
Authors: Bement Zachary James , Norwood Stephanie K , Shollenberger Lisa M 
Published: January 2023

Abstract
Schistosomiasis is the second most burdensome parasitic disease in the world after malaria and is classified as a neglected tropical disease. Schistosomiasis is caused by an immunomodulatory helminth that can induce a Th2 biased response, which can cause Tc1 vaccine failure. Interestingly, the use of pathogenic Listeria monocytogenes vaccine vectors can overcome this failure through an unknown mechanism. Despite the promise of Listeria-vectored vaccines, there is reluctance among the vaccine community and the public to incorporate this vector into immunization programs, due to safety concerns. Here we ask what the question: Why do Listeria vaccines function when other methods, such as DNA vaccination, fail? To answer this question, we use bone-marrow derived dendritic cells (BMDCs) in vitro to model the development of vaccine immunity during the early steps of the adaptive immune response. To compare the immune environment in naïve and schistosome-infected vaccine recipients, we compare responses from both reactogenic DC1 and tolerogenic DC2 dendritic cells, respectively. The DC1 and DC2 subtypes are exposed to a DNA vaccine for HIV, a Listeria vaccine expressing the HIV-1 Gag protein, or left untouched. Through immunological and transcriptomic approaches, we examine these cells for their activation state and functionality; these results will help to elucidate the mechanism/s by which Listeria vectors can overcome schistosome-induced vaccine failure and identify unique molecular vaccine adjuvants functional in schistosome-endemic areas.
The prevention and augmentation of tetravalent recombinant measles-vectored dengue vaccine in dengue-2 infected AG129 mice
Authors: Pan Chien-Hsiung , Lin Tsung-Han , Yan Jia-Ying 
Published: January 2023

Abstract
Dengue is an emerging mosquito-borne disease and the use of prophylactic vaccine is still limited. We have previously developed the tetravalent dengue vaccine (rMV-TDV) by a recombinant measles virus (MV) vector expressing envelope protein domain III (ED3). In this study, we used AG129 mice to evaluate the protective and/or pathogenic immune responses induced by rMV-TDV. Immunization of rMV-TDV has induced a significant neutralizing antibody response against all serotypes of DENV, as well as a significant CD4-dependent IFN-γ response biased to DENV-3, compared to the vector controls. After DENV-2 challenge, rMV-TDV-immunized mice showed a significant lower viremia and no increase of inflammatory cytokines comparing to the vector controls, which had a ~100 times higher viremia and a significant increase of IFN-γ and TNF-α 3 days after challenge. A robust DENV-2 specific IFN-γ response detected in rMV-TDV-immunized mice suggested that pre-existing DENV-3 biased T-cell responses didn’t cross-react to DENV-2 but an anamnestic DENV-2 specific T-cell response was recalled. Prior to the challenge, only DENV-3 specific IgG demonstrated a significant higher avidity than other serotypes, whose specific IgG avidity increased after DENV-2 challenge, particular in DENV-1 and -2. However, the passive transfer of rMV-TDV immunized sera plus splenocytes resulted in the higher viremia and the significant increase of inflammatory cytokines in AG129 recipient mice after DENV-2 challenge than the vector control donors did. It suggested that while the immunity by rMV-TDV waned, the potential risks for the severe disease increased. Our data shed the lights on the role of a balanced CD4 T-cell response in the prevention of severe dengue.
Neutralizing monoclonal antibodies against Venezuelan equine encephalitis virus envelope glycoprotein E2 protect from lethal encephalitis
Authors: Kafai Natasha M. , Sukupolvi Soila , Gardner Christina L. , Klimstra William B. , Diamond Michael S. 
Published: January 2023

Abstract
Venezuelan equine encephalitis virus (VEEV) is a mosquito-borne New World alphavirus responsible for severe morbidity and mortality in humans and equines. The development of therapeutics against VEEV is critical due to its potential for outbreak and aerosolization as a bioterrorist agent. However, effective and widely accessible antiviral agents for VEEV do not exist. Humoral immunity is an important defense against many alphavirus infections, and we thus generated a panel of 17 highly neutralizing murine VEEV-specific monoclonal antibodies (mAbs) using the VEEV vaccine TC-83 and attenuated chimeric SINV-VEEV Trinidad donkey (TRD) subtype IAB. Nine of these mAbs demonstrate 50% effective inhibitory concentrations (EC50) values below 100 ng/ml when tested against enzootic subtype ID and epizootic VEEV subtypes IAB and IC virion envelope glycoproteins. To define specific viral epitopes important for mAb binding, mapping studies including competition assays, HDX, alanine-scanning mutagenesis, and viral escape mutants are ongoing. Preliminary data suggests the epitopes for these mAbs are found in both the A and B domains of the surface virion envelope 2 glycoprotein. Additionally, several of these mAbs confer prophylactic and therapeutic protection in mice aerosol challenged with virulent TRD VEEV. Mechanism of action studies are underway to define the steps in the virus life cycle (viral attachment, entry, fusion, and egress) at which our most potent protective antibodies act. Identification of specific epitopes targeted by these protective mAbs will better inform the development of targeted immunotherapeutics and vaccines against VEEV and related encephalitic alphaviruses.
A recombinant subunit Lassa virus vaccine elicits strong antibody responses
Authors: To Albert , Lai Chih-Yun , Wong Teri Ann , Namekar Madhuri , Lehrer Axel T 
Published: January 2023

Abstract
Due to the epidemic potential and high case fatality rate, the WHO has declared the development of a Lassa virus (LASV) vaccine as a high priority for preventative use in endemic regions. We used a recombinant subunit vaccine platform to generate recombinant LASV glycoprotein (GP) in an insect cell expression system and tested its immunogenicity in an outbred mouse model. Briefly, LASV GP was secreted from a stably transformed Drosophila S2 cell line and the clarified culture supernatant was purified using immunoaffinity chromatography (IAC). SDS-PAGE gel analysis showed that the purity of the antigen was sufficient for immunogenicity testing. The eluted LASV GP was composed of GP1 and GP2 subunits, the GP1–2 heterodimer, as well as trace amounts of oligomer. Swiss Webster mice immunized intramuscularly with 3 doses of purified recombinant LASV GP, formulated with various adjuvants, showed maximal GP-specific antibody response after two immunizations. We concluded that we have successfully developed a production method for highly-purified, insect-cell expressed recombinant LASV GP capable of eliciting high antibody titers when formulated with a suitable adjuvant after only two doses. These results indicate the potential for recombinant GP to be an efficacious vaccine candidate. Further work on structural enhancements of the antigen and in-depth characterization of antibody and cell-mediated responses is ongoing.
Development of a DNA vaccine for SFTSV that confers complete protection against lethal infection in ferrets
Authors: Kwak Jeong-Eun , Kim Young-Il , Park Su-Jin , Jeong Moonsup , Maslow Joel N. , Song Min-Suk , Jung Jae U. , Shin Eui-Cheol , Choi Young Ki , Park Su-Hyung 
Published: January 2023

Abstract
Severe fever with thrombocytopenia syndrome (SFTS) is a newly emerging tick-borne infectious disease caused by the SFTS virus (SFTSV) belonging to the Phenuiviridae family. The WHO lists SFTSV as one of the most dangerous viral pathogens, and considers it likely to cause wide epidemics in the near future. The incidence of SFTSV infection has increased from its discovery in 2012 through 2018 with a mortality rate of 10–20% and the major clinical symptoms of SFTSV infection are fever, vomiting, diarrhea, thrombocytopenia, leukopenia and multiple organ failure. However, no effective vaccines are currently available for SFTSV. Here, we describe the development of a SFTSV-specific DNA vaccine, its immunogenicity, and its protective efficacy against SFTSV lethal challenge. Vaccine candidates induced both a neutralizing antibody response and multifunctional SFTSV-specific T cell response in mice and ferrets. To investigate the vaccine efficacy in vivo, we applied a recently developed ferret model of lethal infection that recapitulates fatal clinical symptoms in SFTSV infection in humans. Vaccinated ferrets were completely protected from lethal SFTSV challenge without developing any clinical signs. Moreover, we found that anti-envelope antibodies play an important role in protective immunity and non-envelope-specific T cell responses also can contribute to protection against SFTSV infection. This study provides important insights into the development of an effective vaccine, as well as corresponding immune parameters, to control SFTSV infection.