bluuebunny/crossref_metadata_2025_split huggingface dataset:
Neuropilin-1 Amplifies Regulatory T cell Function in Human Cancer
Authors: Chuckran Christopher A , Cillo Anthony R , Moskovitz Jessica , Overacre-Delgoffe Abigail E , Somasundaram Ashwin , Kirkwood John M , Luketich James , Pennathur Arjun , Coffman Lan , Modugno Francesmary , Edwards Robert , Schoen Robert E , Ferris Robert L , Bruno Tullia C , Vignali Dario AA 
Published: January 2023

Abstract
Regulatory T cells (Tregs) maintain peripheral tolerance but also contribute to cancer progression by dampening anti-tumor immunity. Knocking out neuropilin-1 (NRP1) on murine Tregs attenuates tumor growth by rendering intratumoral Tregs non-suppressive. Whereas NRP1 is constitutively expressed on all mouse Tregs, NRP1 is infrequently expressed on Tregs in the peripheral blood of healthy humans. We hypothesized that (1) NRP1 expression is upregulated in cancer patients, (2) NRP1 marks highly suppressive human Tregs, and (3) sustained Treg activation initiates NRP1 expression.
NRP1+ Tregs are enriched in tumors across numerous cancer types but largely devoid in non-cancerous inflamed and healthy tissues. Intratumoral NRP1 expression negatively correlates with disease-free survival in head and neck squamous cell carcinoma (HNSCC; p = 0.03; Hazard ratio = 5.4). Intratumoral NRP1+ Tregs upregulate a suppressive module indicated by co-expression of TIGIT, CCR8, and multiple TNF receptor super family members. Correspondingly, NRP1+ Tregs potently suppress CD8+ T cell proliferation in vitro, which can be disrupted with NRP1-specific blockade. Furthermore, Treg activation in vitro drives NRP1 expression, and sustained TCR signals are required to maintain NRP1 expression.
We conclude that NRP1+ Tregs are selectively enriched in human cancer and oppose anti-tumor immunity. Therefore, destabilizing intratumoral Tregs using NRP1 blockade may complement other T cell therapies such as anti-PD1 blockade.
Live attenuated influenza virus vaccine protects obese mice
Authors: Estrada Leonardo D , Vazquez-Pagan Ana G , Schultz-Cherry Stacey 
Published: January 2023

Abstract
Fifty percent of the United States population is predicted to be obese (body mass index &gt; 30) by 2030. Obese individuals are considered a high-risk group for developing severe influenza disease. To compound this, studies have shown that inactivated influenza vaccination has decreased efficacy in this high-risk population. However, the efficacy of live-attenuated vaccines has not been investigated to date. Therefore, we sought to determine if a cold-adapted A/California/04/2009 (caCA/09) H1N1 influenza virus vaccine protected obese mice from lethal influenza virus challenge. In these studies, we demonstrate that intranasal administration of caCA/09 vaccine protected genetically and diet-induced obese (DIO) mice from a lethal challenge with wild-type influenza virus (CA/09) 4 weeks post-vaccination. Protection was not due to route of administration, as intranasal vaccination with inactivated caCA/09 virus was not protective. Unlike non-vaccinated obese mice, caCA/09 vaccinated obese mice cleared the virus and controlled viral spread, as determined by lung TCID50 and immunohistochemistry, respectively. Protection of obese mice lasts at least 17 weeks post-vaccination. Mechanistically, we found that effector memory CD4 and CD8 T cells were increased in lungs of vaccinated mice that may provide heterosubtypic protection. Indeed, the vaccinated obese mice were protected from a lethal challenge with an H5 deselected influenza virus. In conclusion, live-attenuated virus vaccination is a viable option to protect obese hosts against increased disease severity and mortality.
Immune responses to an inactivated influenza vaccine in Indigenous Australians
Authors: Hensen Luca , Rowntree Louise , Damelang Timon , Chung Amy , Wheatley Adam , Auladell Maria , Koutsakos Marios , Nguyen Oanh , Loh Liyen , Aban Malet , Hurt Aeron , Kent Stephen , Flanagan Katie , Plebanski Magdalena , Nelson Jane , Everitt Anngie , Miller Adrian , Davies Jane , Tong Steven , Kedzierska Katherine 
Published: January 2023

Abstract
Indigenous Australians are highly susceptible to severe influenza disease during seasonal and pandemic infections. Current antibody-based vaccines targeting humoral immunity are the most effective way to combat influenza infections but show reduced efficacy in Indigenous Australians. The underlying mechanisms are still unknown. We recruited 166 Indigenous & non-Indigenous donors to elucidate adaptive immune responses induced by the inactivated quadrivalent influenza vaccine. Participants were bled at baseline, day 7 and day 28 post-vaccination. We analysed hemagglutination inhibition titres against the vaccine and previously circulating strains, analysed IgG1 and IgG3 allotypes, evaluated total IgG glycosylation and performed Luminex assays to dissect antibody responses against viral HA, NA and NP proteins. Additionally, we assessed influenza-specific B cell responses with fluorescent HA probes, antibody-secreting cells and circulating Tfh cells. More than 15,000 datapoints were collected to compare adaptive immune responses between Indigenous and non-Indigenous donors. We found that the majority of immune readouts were comparable between the two groups. However, interestingly, we found significantly lower pre-vaccine titres and lower back-boosting to pre-pandemic H1N1 viruses in Indigenous compared to non-Indigenous donors. This might explain, at least in part, high susceptibility of Indigenous Australians to the 2009 pandemic. Our comprehensive dataset will be used to understand and improve optimal vaccine responses in Indigenous and non-Indigenous populations globally.
Combined CpG and MPL as vaccine adjuvants improve duration of protection induced by single dose influenza A virus protein vaccination
Authors: Lampe Anna T , Clark Madeline , Brown Deborah M 
Published: January 2023

Abstract
Despite vaccination, it is estimated more than 37 million people experienced influenza illness in the US during the 2018–19 season. Efficacy of the influenza A virus (IAV) vaccine may be improved with adjuvants that activate innate immunity. We examined the potential of combined pattern recognition receptor agonists CpG and MPL to increase vaccine induced protection. When administered with a single, low dose intramuscular hemagglutinin protein vaccine, mice receiving CpG+MPL experienced less weight loss after homologous challenge eight weeks after vaccination compared to mice receiving single agonists. However, when mice were challenged earlier after vaccination no significant improvements were observed in weight loss or viral titers, suggesting that combined CpG+MPL improves duration of protection. Despite not inducing excessive injection site damage, CpG+MPL elevated systemic inflammatory cytokines and chemokines 24 hours after vaccination compared to agonists alone. CpG and/or MPL increased monocyte derived migratory dendritic cell numbers and CD4 T cell proliferation in the draining lymph nodes (DLN) three days after vaccination. CpG+MPL also enhanced early T cell activation, measured by CD69 expression, in the DLN compared unadjuvanted vaccines. Additionally, at four weeks, mice vaccinated with CpG+MPL displayed a trend of elevated IgG2a/c antibody production compared to mice receiving single agonists. However, CD4 and CD8 T cell activation and cytokine production in the DLN after homologous challenge was not significantly impacted by CpG+MPL. Together these results suggest that CpG+MPL induces enhanced innate immune responses that contribute to extended duration of protection against homologous challenge.
The Complex of Poly-γ-Glutamic Acid and Alum Induces Cross-Protective Immunity of Pandemic H1N1 Influenza Vaccine
Authors: Nguyen Quyen Thi , Kwak Chaewon , Kim Jaemoo , Yang Jihyun , Poo Haryoung 
Published: January 2023

Abstract
Vaccination is the most effective strategy to protect against infectious diseases, and adjuvants are key vaccine components that improve antigen-specific immune responses. Here, we developed a new adjuvant named PGA/Alum by combining poly-γ-glutamic acid (γ-PGA) with alum and investigated its ability to enhance the immunogenicity and the cross-reactive efficacy of pandemic H1N1 (pH1N1) influenza vaccine antigen. PGA/Alum highly induced in vitro activation and antigen processing of dendritic cells, and enhanced antigen delivery to draining lymph nodes and antigen-specific immunogenicity in mice using OVA as a model antigen. Also, OVA-specific antibody production, cytotoxic T lymphocyte (CTL) activity, and antibody-dependent cellular cytotoxicity (ADCC) were greatly increased by PGA/Alum. These abilities of PGA/Alum improved the protective efficacy of pH1N1 vaccine antigen by increasing hemagglutination-inhibition titers, enhancing ADCC and CTL activity, and speeding viral clearance following homologous viral challenge. Importantly, PGA/Alum significantly enhanced the cross-protective efficacy of pH1N1 vaccine against heterologous viruses [A/Puerto Rico/8/34 (H1N1) and A/Hong Kong/1/1968 (H3N2)] by inducing cross-reactive ADCC and CTL activities. Together, our results strongly suggest that PGA/Alum may be a promising vaccine adjuvant for preventing influenza and other infectious diseases.
Photo-regulated control of cytokine signaling via bioinspired, polymer-induced latency
Authors: Dreaden Erik C , Perdue Lacey A. , Do Priscilla , Chyong Andy , Salaita Khalid , Lesinski Gregory , Porter Christopher C 
Published: January 2023

Abstract
Cytokine signaling is challenging to study or therapeutically modulate as the effects of these protein are often pleotropic and context-dependent. One mechanism by which cytokines with diverse effects can, however, transmit cell type-specific information is via expression in an inactive, or latent, form in which the protein is sterically shielded by a binding partner that is later displaced or degraded (e.g. TGF-β1 association with GARP or LAP). Inspired by this precision, we hypothesized that photo-reversible chemical modification could impart orthogonal specificity to cytokines not expressed in a latent state, such as IL-2, IL-15, or IL-12.
Here, we describe a strategy whereby cytokines are modified with photo-labile polymers that mimic the induction of latency while appended, then de-shield to recover protein activity in response to monochromatic light exposure, thus controlling the magnitude, duration, and location of cytokine signals. Using this strategy, we show that latent IL-2 and IL-15 activity can be photo-modulated as much as 103-fold and that polymer-induced IL-2 latency, alone, can bias affinity away from immunosuppressive CD25 binding. We further show that protein de-repression can be achieved with spatial resolution approaching that of a single immune cell and demonstrate the feasibility of transcutaneous IL-2 photoactivation using engineered tissue phantoms. We find that such modifications also prolong the circulation of IL-12 in vivo, thus obviating the need for frequent, high dosing in therapeutic settings.
Future extensions of this approach could enable multicolor, optical reprogramming of cytokine signaling networks and lead to immunotherapies that are more tissue-specific and patient-personalized.
T cells exhibit prolonged immunity against melanoma tumors expressing high, but not low affinity antigens
Authors: Rivera Reyes Amalia M , Wyatt Megan M. , Dwyer Connor J. , Knochelmann Hannah M. , Smith Aubrey S , Rangel Rivera Guillermo O , Paulos Chrystal M 
Published: January 2023

Abstract
We seek to identify the intrinsic properties in T cells from patients responsive or not to immunotherapy. Therapeutic efficacy is seen in patients where the immune system attacks ‘hot’ tumors due to their high mutation burden. ‘Cold’ tumors, in contrast, are not effectively recognized by T cells due to low mutation burden or expression of self-antigen. To simulate this clinical scenario, we used two different melanoma models: 1) B16F10 expressing a low affinity peptide (mgp100 = i.e. self-antigen), which represents a cold tumor or 2) B16F10 expressing a high affinity peptide (hgp100 = i.e. neoantigen), which represents a hot tumor. In these tumor-bearing mice, we infused pmel-1 CD8+ T cells expressing a TCR that recognizes gp100 peptide on B16F10 melanoma. We hypothesized that the function and persistence of adoptively transferred (ACT) cells would be increased in responsive tumors compared to unresponsive. Our findings show that pmel-1 were less exhausted and persisted more in mice bearing responsive tumors. In these mice, efficiency was observed as shrinkage of large subcutaneous tumors post ACT treatment and enhanced long-term survival. Interestingly, mice bearing hot tumors had metabolically improved pmel-1 cells, demonstrated by their ability to take up glucose and mitochondrial health. Mice that were cured from bearing responsive tumors were protected when rechallenged with both cold and hot tumors simultaneously, implicating a memory response. Herein, we underscore how tumor antigen affinity can drastically change the fate and memory of T cells.
Harnessing antiviral memory T cells for tumor immunotherapy
Authors: Rosato Pamela , Wijeyesinghe Sathi P , Ning Jianfang , Gavil Noah Veis , Wu Shaoping , Chen Clark , Vezys Vaiva , Masopust David 
Published: January 2023

Abstract
Overcoming the immunosuppressive tumor microenvironment remains a major impediment to successful cancer immunotherapy. Virus-specific memory T cells are positioned throughout the entire body to sense reinfection or recrudescence. Mouse models have demonstrated upon reencountering cognate antigen, these tissue resident memory T cells (TRM) induce a local immunostimulatory environment that activates and recruits innate and adaptive arms of the immune system, and we extend these functions to include recruitment of circulating antibody. Like healthy tissue, we observe that mouse and human tumors are commonly surveyed by virus-specific memory CD8+ T cells. This was seen in a range of tumor types including traditionally ‘immune-privilege’ tissues such as glioblastoma. Given the described immunostimualtory functions of antiviral TRM in healthy tissue, we tested if we could leverage antiviral CD8+ T cells in tumors as an immunotherapy. Local delivery of adjuvant-free peptide derived from previously encountered viruses successfully reactivated antiviral T cells within melanoma and glioblastoma tumors. This arrested growth of checkpoint blockade-resistant and poorly immunogenic tumors in mice. Antiviral T cell reactivation triggered antigen presentation and cytotoxic pathways within the tumor, activating T cells, dendritic cells and natural killer cells. Viral peptide treatment of ex vivo human tumors demonstrated upregulation of immune activation gene expression profiles similar to those observed in mice. Lastly, viral peptide therapy renders resistant mouse tumors susceptible to PD-L1 blockade. Thus, re-stimulating known antiviral immunity may provide a novel therapeutic approach for a broad range of tumors.
From biTEs to a novel switchable chimeric antigen receptor platform for gated tumor targeting
Authors: Bachmann Michael P , Berndt Nicole , Arndt Claudia , Hoffmann Anja , Koristka Stefanie , Loureiro Liliana , Feldmann Anja 
Published: January 2023

Abstract
Over the past decades we followed the idea of gated targeting of tumor cells starting with modular bispecific antibodies (bsab) followed by T cells genetically modified with modular chimeric antigen receptors (CARs). During these studies we developed a series of bispecific T cell engagers (BiTEs) and in addition the modular UniCAR platform(s). Recently for two of our BiTEs and a first UniCAR complex clinical phase 1 studies could be started. Based on these developments we now established a next generation switchable adaptor CAR platform technology termed RevCARs and characterized it with respect to functionality in vitro and in vivo, possibility of steering of CAR T cell activity (reversible and repeated switching on and off), cytokine response, pharmacokinetic, and imaging with positron emission tomography (PET) in experimental mice: In order to develop polycistronic vectors encoding for more than one CAR (e.g. one signaling CAR and one costimulatory CAR) we were forced to reduce the size of the individual CAR genes. For this purpose minimized CAR genes each using as extracellular domain just a short peptide epitope sequence instead of an antibody domain are fused in a single vector and used for transduction of T cells leading to a defined expression ratio optimally adjusted for “AND” and “OR” gated targeting. The specificity and activation of these RevCARs is mediated via bsab based targeting molecules (revTMs) directed to specific cell surface tumor antigens (TAAs). Due to the small size of revTMs RevCAR T cells can easily and repeatedly be turned on and off simply by starting or stopping of their infusion. In summary, we will present and update the current status of our preclinical and clinical work related to retargeting of T cells to tumor cells.
IL-6 fuels durable memory for Th17 cell-mediated responses to tumors
Authors: Knochelmann Hannah , Dwyer Connor , Smith Aubrey S , Wyatt Megan , Rivera Guillermo Rangel , Lesinski Gregory , Li Zihai , Rubinstein Mark , Paulos Chrystal M 
Published: January 2023

Abstract
Accessibility of T cell transfer therapies is hindered by cost and time required for product development. We found that shortening ex vivo expansion of Th17 cells licenses a proinflammatory cell product which induces cytokine storm with high levels of systemic IL-6 in tumor-bearing hosts. Unique from other helper subsets, briefly expanded Th17 cells eradicate large tumors in low doses and generate long-lived memory against tumor rechallenge, suggesting a benefit to the inflammatory state. As the toxicity of cytokine release is managed in patients through IL-6 blockade, we addressed the physiologic impact of IL-6 on efficacy and durability of Th17 cell therapy in this immunocompetent model. We hypothesized that peak IL-6 induced by Th17 cells was critical to their durable memory properties. To address this, we used the TRP-1 transgenic mouse model where CD4+ T cells express a TCR that recognizes TRP-1 of B16F10 melanoma. Acute IL-6 blockade post Th17 transfer did not impact the primary antitumor response, yet abrogated long-term responses by promoting tumor relapse upon secondary challenge. Mechanistically, IL-6 blockade reduced pSTAT3 and Bcl2 in transferred T cells and promoted a greater frequency of FoxP3+ Treg cells in the peripheral blood and tumor-draining lymph nodes. Finally, serum IL-6 was inversely correlated with engraftment of tumor-specific Tregs from the transferred product. Overall, short-term expanded Th17 cells uniquely induce IL-6, which promotes Th17 survival, suppresses engraftment of tumor-specific Tregs, and is critical to durable memory. This work may suggest that the universal strategy to inhibit IL-6 during cytokine release syndrome may come at the expense of long-term efficacy for cell therapy.
Engineering adoptive T cell therapy to co-opt Fas ligand-mediated death signaling in solid tumors
Authors: Anderson Kristin G , Oda Shannon K , Bates Breanna M , Chiu Edison Y , Burnett Madison G , Suarez Gutierrez Magdalia L , Garcia Nicolas M , Daman Andrew W , Greenberg Philip D 
Published: January 2023

Abstract
Over half of patients diagnosed with high grade serous ovarian cancer will die within five years, highlighting the need for therapy innovation. Engineering T cells to target proteins uniquely overexpressed in tumors has the potential to limit tumor growth without toxicity—mesothelin (Msln) is a rational immunotherapy target as it contributes to the malignant/invasive phenotype and has limited expression in healthy cells. T cells were engineered to express a high-affinity Msln-specific T cell receptor (TCRMsln) and adoptively transferred into a disseminated ID8VEGF murine model, which recapitulates many features of human ovarian cancer. TCRMsln+ T cells preferentially accumulated within established tumors, delayed tumor growth, and significantly prolonged survival. However, T cell persistence and anti-cancer activity were limited by elements in the tumor microenvironment (TME), including Fas Ligand (FasL) that can induce apoptosis in infiltrating lymphocytes expressing Fas receptor (Fas).
To overcome this potential T cell evasion mechanism, we generated a panel of immunomodulatory fusion proteins (IFPs) containing the Fas extracellular binding domain fused to a co-stimulatory domain, replacing the natural death domain. Relative to T cells modified with only TCRMsln, T cells that expressed both TCRMsln and a Fas IFP preferentially infiltrated tumors and expanded/persisted in the TME of tumor-bearing mice. Moreover, adoptive immunotherapy with TCRMsln+IFP+ T cells significantly prolonged survival in tumor-bearing mice, relative to TCRMsln+ only T cells. As many solid tumors overexpress FasL, IFPs may provide an opportunity to enhance engineered adoptive T cell therapy against many malignancies.
Enhanced Fc-mediated complement activity with broadly neutralizing antibody 10E8v4 does not improve viral outcomes following SHIV challenge in rhesus macaques
Authors: Spencer David A , Goldberg Ben , Dufloo Jérémy , Pandey Shilpi , Cheever Tracy , Bruel Timothée , Schwartz Olivier , Ackerman Margie , Hessell Ann J 
Published: January 2023

Abstract
Fc modified broadly neutralizing antibodies (bNAbs) are being developed for both prophylactic and therapeutic treatment of HIV with some success in extending half-life, but in vivo benefits of enhanced FcγR interaction remain unclear. Here we sought to improve bNAb efficacy by increasing Fc-mediated complement activity. A panel of Fc variants was screened for complement enhancement of 10 bNAbs. Modified 10E8v4, which weakly neutralizes SHIVSF162p3 (IC80 30 mg/ml) and does not mediate ADCC, was selected for macaque studies. Compared to wildtype, the EFTAE modification resulted in greater affinity for multiple FcγRs, &gt;2-fold enhanced complement deposition, viral lysis, and antibody-mediated complement dependent cytotoxicity, but reduced serum half-life. Subcutaneous infusion with a sub-protective 5 mg/kg dose of 10E8v4 followed by a single high dose intrarectal challenge with SHIVSF162p3 showed unmodified 10E8v4 treated animals displayed lower plasma viremia compared to the complement/FcgR dual knockout (LALA) and controls. Unexpectedly, the EFTAE group displayed higher plasma viremia than controls, and viral DNA was significantly enhanced in tissues at necropsy (9 wpi, p &lt;0.0001). Viremia in EFTAE treated animals inversely correlated with plasma neutralization titer at time of challenge; however, elevated viremia was not seen in animals treated with 10 and 20 mg/kg Ab, suggesting that complement mediated enhanced infection was mitigated by greater neutralizing activity. These are the first in vivo results substantiating in vitro reports of complement dependent enhanced infection mediated by non-neutralizing HIV Abs and indicate that future studies modulating complement induction by therapeutic bNAbs are warranted.