bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pembrolizumab treatment increases K+ channel function and calcium fluxes in cytotoxic T cells of head and neck cancer patients
Authors: Newton Hannah , Gawali Vaibhavkumar S , Chimote Ameet A , Lehn Maria , Palackdharry Sarah , Hildeman David , Janssen Edith , Wise-Draper Trisha , Conforti Laura 
Published: January 2023

Abstract
Head and neck cancer (HNC) is the sixth most common cancer worldwide with a five-year survival of 50%. Immunotherapy is emerging as a promising treatment modality in HNC. Pembrolizumab, an anti-PD-1 antibody, is an immunotherapy currently approved in metastatic HNC and in clinical trials for curative intent. Although there are promising responses to Pembrolizumab, its effect on cytotoxic CD8+ T cell function is not understood. T cell cytotoxicity is critically important in the elimination of tumors and in the tumor microenvironment, cytotoxicity depends on ion channel activity which is defective in HNC patients. Therefore, to elucidate mechanisms of Pembrolizumab, we studied potassium channel (KCa3.1 and Kv1.3) functionality and Ca2+ fluxes in CD8+ peripheral blood T cells (PBTs) of naïve HNC patients before/after Pembrolizumab treatment (n=22) as well as tumor infiltrating lymphocytes (TILs) functionality. Untreated HNC patients were controls (n= 13). Pembrolizumab-treated patients were categorized as responders (R) or non-responders (NR) based on pathological response. We then performed electrophysiological and Ca2+ flux experiments. We observed that Pembrolizumab increased KCa3.1 currents in PBTs (29.8%, p=0.026). Moreover, after treatment, R PBTs also had increases in Kv1.3 currents and Ca2+ fluxes (22.1%, p=0.047; 16.5%, p&lt;0.001; respectively). In TILs, Kv1.3 currents and Ca2+ fluxes were increased in Pembrolizumab-treated patients (64%, p=0.002; 34.8%, p=0.004; respectively). These data show that Pembrolizumab increases the fitness of CD8+PBTs and TILs in relation to ion channel functionality, especially in R, and thus could lead to better understanding of the Pembrolizumab mechanism of action/resistance.
Novel non-segmented negative-sense RNA virus-based vaccine platforms for Zika virus
Authors: Li Anzhong , Lu Mijia , Yu Jingyou , Harder Olivia , Attia Zayed , Shan Chao , Liang Xueya , Xue Miaoge , Shi Pei-Yong , Peeples Mark E. , Liu Shan-Lu , Boyaka Prosper N , Niewiesk Stefan , Li Jianrong 
Published: January 2023

Abstract
Zika virus (ZIKV) is a mosquito-borne flavivirus that can cause Congenital Zika Syndrome (including microcephaly), Guillain-Barré syndrome, and other severe neurological disorders. Despite major efforts, there is no FDA-approved vaccine for ZIKV. Current efforts to develop ZIKV subunit vaccines have been exclusively focused on pre-membrane (prM) and envelope (E) proteins. However, the role of other viral proteins such as nonstructural protein 1 (NS1) in protection is poorly understood. Here, we developed two novel ZIKV vaccine candidates using non-segmented negative-sense RNA viruses (vesicular stomatitis virus, VSV; measles virus, MeV) as a vector. ZIKV prM-E-NS1 or prM-E gene was inserted into the attenuated VSV or MeV vector, and recombinant viruses expressing prM-E-NS1 or prM-E were recovered. The VSV- and MeV-based vaccine candidates were highly attenuated, induced strong ZIKV-specific antibody and T cell immune responses in animal models, and provided complete protection against ZIKV challenge in both immunocompetent and immunodeficient mice. Compared with vaccine candidates expressing only prM-E, ones with NS1 induced significantly higher levels of antibody and/or T cell responses and provided better protection against ZIKV infection, suggesting that NS1 plays a regulatory role in immune response and protection. In addition, we found that NS1, alone, is capable of conferring protection against ZIKV infection even though it did not induce neutralizing antibody. Our results demonstrated that prM-E-NS1 was superior to prM-E as a vaccine antigen. In summary, we have developed safe and highly efficacious ZIKV vaccine candidates, which are ready for pre-clinical trials in nonhuman primates and humans in the future.
Mucosal Vaccination with a Novel Live Attenuated Brucella melitensis Mutant Protects via IFN-γ-producing Memory CD8+ T Cells in Concert with B Cells
Authors: Goodwin Zakia I , Yang Xinghong , Hoffman Carol , Pascual David W 
Published: January 2023

Abstract
A B. melitensis mutant vaccine expressing mCherry (znBM-mC), when applied mucosally, conferred nearly complete protection against pulmonary challenge with wild-type (wt) BM 16M than did BM Rev-1 vaccine. To identify cellular subsets and mechanisms involved in inducing and maintaining protection post-BMDM vaccination, we hypothesize that both arms of the adaptive immune system are required for optimal protection. Mucosal vaccination with znBM-mC induced both effector memory and resident memory CD8+ T cells in the lungs, which were the primary source of IFN-γ. In the absence of IFN-γ, IL-17 significantly increased, but was insufficient for protection. In the absence of B cells, the total number of lung CD8+ T cells and proinflammatory cytokine production were significantly reduced resulting in diminished systemic protection. Reconstitution with naïve or immune B cells restored some protection in znBM-mC-vaccinated B cell-deficient mice. B cell involvement begins as early as 5 days post-nasal vaccination. Total CD19+ B cells significantly increased in BALB/c head and neck lymph nodes (LNs), lower respiratory LNs, and bronchio-alveolar lavage fluid. mCherry+ BALB/c B cells were elevated in the lungs and spleen correlating to a significant increase in CD8+ T cell expansion in the spleen compared to B cell-deficient mice. In-vitro uptake of znBM-mC by B cells resulted in their activation evidenced by increased CD80, CD86, CD11b, and CD24 expression. Moreover, purified immune CD8+ T cells co-cultured with znBM-mC-infected B cells showed enhanced T cell proliferation. Although B cells become infected with Brucella, these are important for optimal T cell immunity. Work is supported by NIH AI123244 & AI125516.
Vaccine adjuvant choice regulates antigen-specific B cell activation and phenotype
Authors: Prior John Timothy , Taylor Justin J. , McLachlan James B 
Published: January 2023

Abstract
There has been a global rise of antibiotic resistant bacteria that are causing a significant increase in morbidity and mortality. Vaccines offer one of the best ways to combat and prevent these infections. Antibody responses are often the most effective outcomes of vaccine-induced immunity; however, the role of different vaccine adjuvants has not been fully explored in terms of antigen-specific B cell responses. Using a B cell antigen tetramer, we sought to determine how a novel adjuvant, double mutant heat labile toxin from enterotoxigenic E. Coli (dmLT), compared with those currently used in vaccines. We show that either intradermal or intramuscular immunization of mice with a model vaccine formulated using dmLT induces greater antigen specific B cell numbers in the injection site draining lymph nodes and spleen compared to antigen alone. These B cells were also antibody isotype switched and were phenotypically germinal center B cells. We further demonstrate that vaccine-specific B cells migrate from the injection site draining lymph nodes to more distal lymphoid organs. When we compared dmLT to the most commonly used human vaccine adjuvant, alum, we found dmLT was better at inducing a B cell response. Lastly, we show that dmLT is a more potent B cell adjuvant compared to the TLR4 agonist monophosphoryl lipid A (MPL-A) and that the combination of these two adjuvants is superior to either adjuvant alone. Interestingly, MPL-A induced greater antigen specific IgG compared to dmLT despite dmLT inducing a more robust B cell expansion. These results demonstrate that dmLT is a potent B cell activating adjuvant and that combining dmLT with existing adjuvants has the potential to greatly improve vaccine responses.
Vaccine and abscopal effects result from in situ vaccination with nanosecond electric pulse treatment for breast cancer in mice
Authors: Guo Siqi , Nanajian Anthony , Scott Megan , Beebe Stephen J. , Ruedlinger Brittaney 
Published: January 2023

Abstract
An electrical engineering technology, nanosecond electric pulse (nsEP), has been studied to ablate local tumor in a non-thermal and minimally invasive manner. Recently, several groups including our lab discovered that animals with complete tumor ablation following nsEP treatment were protected from a second tumor challenge. Therefore, nsEP tumor ablation is also a novel in situ vaccination approach. Breast cancer is considered a low immunogenic tumor with only a small subset of patients responsive to immune checkpoint inhibitors. We found nsEP treatment resulted in not only a potent in situ vaccine effect in a poorly immunogenic 4T1 mouse breast cancer model but also an abscopal effect, a rejection of distant untreated tumor lesion. We further examined the impact of nsEP on cancer cell death, tumor microenvironment (TME) modification, the dynamic changes of local and systemic immune response. 4T1 cells treated with nsEP did not show apoptotic death markers but released damage-associated molecular patterns, including calreticulin, high mobility group protein B1, and ATP. In contrast to abundant immune suppressor cells in untreated breast TME, nsEP treatment led to a significant reduction of both myeloid-derived suppressor cells and regulatory T cells. Conversely, the ratio of tumor associated macrophages M1-like verse M2-like was reversed. An increase of tissue-resident memory T cells was observed in draining lymph nodes. In summary, our results suggest that nsEP is a novel non-drug immunogenic cell death inducer and in situ vaccination approach. Further elucidation of its mechanisms and the development of an in vivo nsEP delivery system suitable for patients is critical to warrant this novel technology toward a clinical trial.
Secreted heat shock protein gp96-Ig vaccine induces malaria specific intrahepatic CD8 T cell responses
Authors: Strbo Natasa , Padula Laura , Fisher Eva , Wijayalath Wathsala , Patterson Noelle B. , Huang Jun , Ganeshan Harini , O’Neill Katelyn , Garcia Denisse , Etobayeva Irina V. , Edgel Kimberly A. , Podack Eckhard R , Sedegah Martha , Villasante Eileen F. 
Published: January 2023

Abstract
A highly-efficacious and durable malaria vaccine is an essential tool for global malaria eradication. One promising strategy to develop such a vaccine is to induce robust CD8+ T cell-mediated protective immunity against malaria liver stage parasites.
We developed and tested a novel malaria vaccine platform based on the secreted form of the heat shock protein gp96 immunoglobulin, (gp96-Ig) to induce malaria antigen-specific liver resident memory CD8+ T cells. Gp96-Ig has ideal properties as an antigen carrier and vaccine adjuvant to activate antigen presenting cells (APCs) and effectively chaperone antigens for cross presentation via MHC I to CD8+ T lymphocytes. Our study shows that vaccination of mice and non-human primate (NHP) animal models with HEK-293 cells transfected with gp96-Ig and two well-known Plasmodium falciparum (Pf) vaccine candidate antigens, circumsporozoite protein (CSP) and apical membrane antigen 1 (AMA1), induce liver infiltrating, antigen-specific, cytotoxic memory CD8+ T cell responses. The majority of the CSP and AMA specific intrahepatic CD8+ T cells expressed CD69 and CXCR3, hallmarks of tissue resident memory T cells (TRM). Also, we found an increased frequency of intrahepatic CD8+ T cells secreting IL-2, which is relevant for maintenance of effective memory responses.
Our findings are strongly supportive of a novel gp96-Ig-based malaria vaccine as a unique systemic and liver-homing, liver-stage antigen-specific CD8+ cytotoxic T lymphocyte (CTL) vaccine strategy.
Superior CD8 T cell-mediated protection against malaria parasite liver stage infection after vaccination with Replication Competent Genetically Attenuated Parasites (RC-GAP) and impact of blood stage infection on immunity
Authors: Patel Hardik , Minkah Nana K. , Sheikh Amina A. , Reynolds Laura , Vaughan Ashley M. , Kappe Stefan H.I. 
Published: January 2023

Abstract
Plasmodium sporozoites (SPZ) are transmitted by mosquito bite, infect the liver, grow and differentiate as intrahepatocytic liver stages (LS) and ultimately form the first generation of red blood cell-infectious merozoites, which initiate the symptomatic blood stage infection. A vaccine against the clinically silent pre-erythrocytic SPZ and LS stages is attractive as it confers protection against infection before onset of symptomatic blood stage infection. Immunization with replication-deficient attenuated SPZ constitutes a promising vaccination strategy to engender protective humoral and CD8 T cell responses, with tissue resident memory CD8 T cell (TRMs) mediating sterilizing immunity in animal models. Attenuated parasite vaccines have undergone clinical testing and it was observed that sterilizing immunity is high in malaria naïve subjects but lower in malaria pre-exposed subjects residing in malaria-endemic regions. Previous studies in rodent models indicate that the ongoing blood stage infection suppresses protection after immunization with replication-deficient irradiated sporozoites. We have shown that replication competent late liver stage arresting genetically attenuated parasites (RC-GAP) confer superior protection when compared to irradiated sporozoites. We show that this protection is dependent on CD8 TRMs in the liver. However, whether the blood stage infection also suppresses the RC-GAP parasite vaccine mediated protective immunity remains unknown. Here, we examined whether blood stage infection either before or during RC-GAP immunization affects vaccine induced protection in the Plasmodium yoelii/Balb/c mouse model and how this correlated with perturbation of memory T and B cell responses.
A small molecule based immunotherapy targeting IL-10/STAT3 praxis to augment the potential of rBCG30 vaccine against experimental tuberculosis
Authors: Ahmad Faraz , Umar Mohd Saad , Khan Nazoora , Gupta Pushpa , Gupta Umesh Datta , Owais Mohammad 
Published: January 2023

Abstract
Several recent studies have contributed to our understanding of the need for host directed immunotherapeutic strategies against Mycobacterium tuberculosis (Mtb), either concomitantly with classical chemotherapy or as standalone intervention. Nevertheless, most of these studies were focused on enhancing chemotherapeutic potential of standard anti-tuberculosis chemotherapy and only few were aimed towards improving prophylactic immunity offered either by BCG or its potential replacements. In the present study we tried to address the potential gap with the aim to improve anti-TB immunity offered by 2nd generation rBCG30 vaccine. We attempted to bolster rBCG30 invoked anti-TB immunity in mice through modulation of the IL-10/STAT3 interaction mediated anti inflammatory program. An immunomodulator of IL-10/STAT3 signaling axis was administered in immunized mice to reinforce the anti-pathogen capabilities of mononuclear phagocyte system with expectation to improve T cell mediated secondary immune responses. We envisage that modulation of this crucial praxis may result in reduced expansion of pathogen permissive AAMs/AADCs (Alternatively Activated Monocytes/Macrophages/DCs) and augmented abundance of pro-inflammatory CAMs/CADCs (Classically Activated Monocytes/Macrophages/DCs), the cells that resist establishment of successful Mtb infection. The study was also aimed towards generating optimum activated APCs early either after vaccination or infection, which can process and present mycobacterial antigens more efficiently to T cells.
Th17 responses to Pseudomonas aeruginosa vaccines are defective in cystic fibrosis mice
Authors: Merakou Christina , Bonney Sarah , Liao Tiffany , Priebe Gregory P. 
Published: January 2023

Abstract
Pseudomonas aeruginosa causes chronic lung infections in people with cystic fibrosis (CF) associated with lung function decline. Since levels of IL-17 are high in the CF airway, it is unclear whether vaccines that induce Th17 responses in CF hosts would be effective, or potentially harmful. Our prior work showed that vaccine-induced Th17 cells confer antibody-independent protection against acute P. aeruginosa pneumonia in mice and that the P. aeruginosa protein PopB stimulates protective Th17 responses. In this study, we tested whether vaccine-induced Th17 responses are abnormal in CF mice.
CF mice (delF508 in FVB/N background) and littermate controls were immunized intranasally either with live-attenuated P. aeruginosa vaccines or with PopB mixed with curdlan as adjuvant. We tested Th17 responses by measuring IL-17 secretion from splenocytes stimulated with antigens in vitro. Protective efficacy was assessed using a murine acute pneumonia model.
Our results demonstrate that CF mice show 2-fold lower IL-17 recall responses in the spleen after IN immunization with a live-attenuated vaccine in comparison to wild-type mice (P&lt;0.05 by ANOVA). CF mice were not protected when challenged with a LPS-heterologous strain, suggesting the absence of Th17-mediated immunity when LPS-directed opsonic antibodies are not available. CF mice immunized with PopB also showed defective Th17 responses in the spleen compared to wild-type or heterozygous mice (2-fold lower, P&lt;0.05 by ANOVA).
CF mice have low Th17 responses to P. aeruginosa vaccines. Correction of the underlying CF defect may be needed to achieve vaccine-induced protection against P. aeruginosa in the setting of CF.
The regulation of transcription factor T cell factor 1 (TCF1) in vaccine-elicited CD8+ T cells
Authors: Ivanova Daria Leonidovna , Kilgore Augustus , Thompson Scott , Kedl Ross 
Published: January 2023

Abstract
Our lab has developed a subunit vaccination regimen that induces a highly robust T cell response. These vaccine-elicited CD8+ T cells (Tvac) are phenotypically, functionally and metabolically distinct from infection-elicited CD8+ T cells (Tinf). In contrast to Tinf, almost all Tvac rapidly acquire a memory phenotype (CD127hi, KLRG1lo). This suggests that Tvac could be differentially transcriptionally programmed to commit to a memory, rather than effector, cell fate. The transcription factor TCF1 (encoded by tcf7) is essential for CD8+ T cell memory in response to infection. Since high TCF1 expression is also important for the generation of stem-like T cells with long-term survival capacity and anti-tumor properties, it is now recognized as a promising target for immunotherapies. However, little is known about how TCF1 is regulated in response to subunit vaccination. We find that, in striking contrast to Tinf, TCF1 expression is actually increased in Tvac after vaccination. As others have shown, we also find that increasing inflammation during vaccination by the addition of TLR9 agonist CpG reduces TCF1 expression in Tvac, skewing their memory phenotype towards the effector phenotype (CD127lo, KLRG1hi) and compromising their long-term memory. While the loss of TCF1 in this context has conventionally been ascribed to inflammatory receptors, utilizing reporter Nur77GFP mice, we also observe an altered duration and frequency of TCR triggering in the context of inflammation. Our data indicates a complex, previously unappreciated interplay between inflammatory and TCR-mediated signals in the regulation of TCF1.
Immunomodulatory effects of topotecan augment the effectiveness of chemotherapy-immunotherapy combination
Authors: Lee Jeongmi , Shin Kwang-Soo , Koh Choong-Hyun , Jeon Insu , Kang Tae-Seung , Song Boyeong , Kang Chang-Yuil 
Published: January 2023

Abstract
Combination therapy is a cornerstone of anticancer therapy, suggesting potential therapeutic benefit to nonresponders to single-agent treatment. BVAC, a B-cell-and-monocyte-based vaccine loaded with alpha-galactosylceramide (α-GC) and tumor antigen as reported in our previous studies, can be successfully combined with other treatments.
Here we found out a novel synergistic regimen in TC-1 tumor model which combines BVAC with topotecan, a chemotherapeutic agent as a topoisomerase I inhibitor. We demonstrated some significant alteration in the immunoprofiles of topotecan-treated mice. We observed increased tumor recruitment of monocytes and CD8+ T cells, and decreased frequencies in regulatory T cells in tumor. Accordingly, we are currently investigating how topotecan-induced changes in immune cell composition could lead to improvement of antitumor effect. These data indicate that topotecan not only has cytotoxicity to tumor but can contribute to modulating tumor immune microenvironment.
We anticipate that this study could provide a rationale for combination of cancer vaccine and chemotherapy and define future optimal treatment regimens in cancer patients.
Novel minigene vaccine stimulates CD8+ memory inflation to control growth of murine colorectal tumor
Authors: Kwok Jonathan , Kroon Anna Maria Theresa , Chinnakannan Senthil , de Lara Cathy , Gileadi Uzi , Cerundolo Vincenzo , Klenerman Paul , Lee Lian Ni 
Published: January 2023

Abstract

Fundamental science
Memory “inflation” describes the sustained accumulation of expanded pools of antigen-specific CD8+ T-cells, which occurs in certain persistent viral infections e.g. cytomegalovirus, and after vaccination with replication deficient adenoviral vectors. Inflating T cells are sustained at high frequencies, possess an effector memory phenotype and home to non-lymphoid tissues – characteristics which are desirable in anti-tumor responses. Use of minigene (epitope only) vaccines allows targeting of responses and reliable generation of strong memory inflation. To explore this, adenoviral vectors were designed to stimulate an anti-tumor response against the dominant antigen in CT26 tumors, AH1, either as a full length or peptide immunogen. Groups of mice were immunized with the minigene AH1 vector. Tumor growth and immune responses were monitored using prophylactic or therapeutic regimens.

Results
Our lab has previously demonstrated that minigene vaccines generate strong inflationary responses compared to a full-length vector. Compared to the controls, tumors from minigene-immunized mice grew at a significantly slower rate, and the intra-tumoral environment was enriched for functional effector cells. The presence of AH1-specific CD8+ T cell populations in peripheral blood correlated overall with the anti-tumor effect.

Conclusions
Inflationary CD8+ T cells can be targeted against tumor antigens using our adenovirus minigene approach. Such responses show anti-tumor activity in prophylactic and therapeutic settings. Further studies to assess the impact of increasing the breadth of targeting and the impact of checkpoint inhibition are ongoing.