bluuebunny/crossref_metadata_2025_split huggingface dataset:
Dissecting anti-tumor immunity in glioblastoma
Authors: Chen Dan , Varanasi Siva Karthik 
Published: January 2023

Abstract
Glioblastoma multiforme (GBM) is the deadliest form of brain cancer, for which there is currently no effective treatment. Despite success of immunotherapies such as checkpoint inhibitors (CPI) in treating some cancers, less than 8% of GBM patients responded to anti-PD-1 therapy. Thus, we need to understand what mechanisms underlie tumor resistance to immunotherapies. In our GBM mouse model, we have found that anti-CTLA-4 treatment greatly improved survival and suppressed tumor growth, but surprisingly, its efficacy was lost in mice that were depleted of CD4+ but not CD8+ T cells, suggesting that CD4+ T cells are critical for protection against GBM. This matches clinical data showing decreased populations of CD4+ T cells in patients with adverse clinical outcomes such as high-grade GBM. Our preliminary results from flow cytometry and single-cell RNA sequencing (sc-RNAseq) also suggested enhanced cytokine production (IFNγ, TNFα, IL-2) in CD4+ T cells after anti-CTLA-4 treatment. Similarly, we found that microglia, the local sentinels of the central nervous system, also play a significant role in tumor control, as indicated by faster GBM growth and reduced number of tumor-infiltrating CD4+ T cells when mice are depleted of microglia by a CSF1R inhibitor (PLX3397). Additionally, microglia in GBM may serve as potential antigen-presenting cells, as shown by their high MHC class II (MHC-II) expression. Therefore, we postulate that microglia are necessary to sustain anti-tumor immunity by CD4+ T cells. Importantly, through a more detailed exploration of how CD4+ T cells and microglia interact to impact tumor control, our study will provide new insights for developing therapeutic strategies against “difficult-to-treat” cancers such as GBM.
Target the activin receptor 1c on CD4+ T cells to achieve anti-tumor therapeutic effects.
Authors: Zheng Ying , Lebid Andriana , Fu Juan , Patel Chirag , Wang Xiaoxu , Pardoll Drew 
Published: January 2023

Abstract
Activin signaling molecules and their receptors have not been substantially studied in the context of the immune system and cancer. We show here that tumor-bearing mice had elevated Activins levels, which correlated directly with tumor burden. Likewise, cancer patients had elevated plasma Activins compared to healthy controls. Importantly, our in vitro studies suggested that Activins promoted differentiation of naive wild type CD4+ cells into induced FoxP3-expressing Tregs. Database and qRT-PCR analysis of sorted major immune cell subsets in mice revealed that Activin Receptor 1C (ActR1c) was uniquely expressed on Tregs. ActR2b, as paired to ActR1c, was found to be highly upregulated during iTreg differentiation. In humans, these receptors were expressed on CD4+ Foxp3− cells. Thus, while Activins may have many cellular targets, signaling through these receptors on CD4+ cells likely promotes Treg-mediated immunosuppression. In vivo B16 melanoma and MC38 colon tumor studies have demonstrated that mice lacking ActR1c were more resistant to cancer progression compared to wild type mice. This phenotype correlated with reduced expression of the FoxP3 transcription factor in CD4+ cells. The anti-tumor effect was T-cell dependent, as T cell-deficient mice reconstituted with naïve ActR1c knock out CD4+ cells had delayed tumor progression. In vitro, naïve CD4+ cells lacking ActR1c had a defect in their ability to gain FoxP3 expression when Activin A was present and when TGF-β was limited. Thus, blocking Activins signaling through this receptor is a promising and disease-specific strategy for preventing the accumulation of immunosuppressive iTregs in cancer.
Supported by grants from NIH (R01AI137046)
The role of lymphocyte subsets in preventing tuberculosis following intravenous vaccination with BCG
Authors: Simonson Andrew W , Bucsan Allison N , Zeppa Joseph J , Winchell Caylin G , Myers Amy J , Sutton Matthew , Ameel Cassaundra L , Darrah Patricia A , Roederer Mario , Seder Robert A , Flynn JoAnne L 
Published: January 2023

Abstract

Background
We previously demonstrated in non-human primates (NHP) that intravenous (IV) administration of BCG induces substantial T cell responses and often sterilizing immunity against Mycobacterium tuberculosis (Mtb) infection. Here, we investigated the role of T cell populations in this unmatched response in the context of necessary immune mechanisms for protection against tuberculosis (TB).

Methods
Beginning 20 weeks after vaccination and through the end of the study, rhesus macaques were infused with antibodies to deplete CD4, CD8α, or CD8β expressing T cells, or with control IgG/saline. NHPs inoculated with ~15 CFU Mtb Erdman via bronchoscope were monitored for 8 weeks. Granulomas, areas of disease, lung tissue and thoracic lymph nodes (LNs) were harvested at necropsy using FDG PET CT as a map to identify inflamed lesions. Immune responses were analyzed by spectral flow cytometry and homogenates were plated for colony forming unit (CFU) quantification.

Results
Successful depletion of expected cell types was seen in blood, airways, LNs and lung tissue. Depletion of CD4 T cells ablated IV BCG mediated protection in all NHPs. CD8α and CD8β depleted NHPs retained varying levels of protection, leading to mixed outcomes by bacterial burden. Preliminary results indicate the loss of protection following CD8α (both innate and adaptive cells) and CD8β (mainly adaptive cells) depletion is similar, suggesting a key role for conventional CD8 T cells in vaccination.

Conclusions
Our results provide critical insight regarding roles played by CD4 and CD8 T cell populations in terms of defining desired responses for vaccination against progressive TB. Functionality of these cells in immunized NHPs will clarify the purposes of T cell subsets.
Supported by grants from Bill and Melinda Gates Foundation and NIH NIAID (75N93019C00071)
Adjuvanted multivalent vaccine targeting clinically relevant Pf bacteriophage creates cross-reactive antibodies
Authors: Román-Cruz Valery Cristina , Miller Shannon , Schoener Roman , Lukasiewicz Chase , Secor Patrick , Burkhart David , Evans Jay T 
Published: January 2023

Abstract
Pseudomonas aeruginosa (Pa) is a bacterial pathogen associated with chronic diabetic wounds as well as potentially deadly lung infections in cystic fibrosis patients. Pa is listed by the WHO as “Priority 1” in the Priority Pathogens list and results in hundreds of deaths each year. Pa diversity presents a challenge in developing a vaccine or therapeutic capable of resolving infections caused by different Pa clinical isolates. Filamentous Pf phages (a virus) infect the majority of Pa strains. Additionally, the presence Pf phage is a key virulence factor contributing to Pa infections. Pf is a filamentous-lysogenic phage that is primarily comprised of repeats of coat protein (CoaB). A consensus peptide derived from the coat protein was identified and used as the antigen for our phage vaccine targeted vaccine. This conjugate vaccine targeting Pf phage induced protection against Pa wound challenges in mice. Additionally, our team has implemented the use of a novel TLR4 agonist to further enhance antibody response against Pf coat protein intact Pf phage virions. Based on their amino acid sequence homology, clinical isolates of Pa and associated Pf phage were divided into two major clades. We prepared CRM-197 conjugates using the consensus peptides from both clades. Vaccination with these multivalent conjugates in combination with a novel synthetic TLR4 agonist (adjuvant) boosts antibody response against the peptides and promotes antibody cross-recognition between peptides. Creating a safe and effective vaccine that can mount a robust immune response against both clades of Pf phage is an important advancement towards the development and clinical testing of phage based vaccine targeting deadly Pa infections.
Supported by R01 AI138981-01
Differential sex-specific immune responses following Prime-and-Trap vaccination alters protection against malaria in mice
Authors: Duncombe Caroline J , Watson Felicia N , Kalata Anya C , Shears Melanie J , Murphy Sean C 
Published: January 2023

Abstract
Generating liver resident-memory CD8+ T cells (TRM cells) is critical for effective liver-stage malaria vaccine. The role of biological sex in liver-stage vaccine protection is understudied. Here we report sex-specific immune responses and protection outcomes for the two-step heterologous ‘Prime-and-Trap’ liver-stage malaria vaccine designed to induce liver TRM cells. The Prime-and-Trap strategy combines sporozoite antigen DNA priming with a single intravenous dose of liver-homing radiation-attenuated sporozoites (RAS) to direct and “trap” activated and expanding T cells in the liver. This strategy induces robust liver CD8+ TRM cell responses and confers sterile protection in the P. yoelii (Py) rodent malaria model in female mice. However, when tested in male mice, it induces weaker liver CD8+ TRM cell responses and little to no protection from sporozoite challenge. We examined sex-divergent immune responses in the spleen and liver following each step of the Prime-and-Trap regimen. We found that sexually dimorphic activation of certain immune pathways drives attenuated downstream retention of TRM cells in the liver, leading male mice to generate a sub-optimal immune response that ultimately fails to protect from Py sporozoite challenge. Our findings emphasize the importance of incorporating sex as a variable for evaluation in the design of robust liver-stage malaria vaccines.
Supported by grants from NIH (R01 AI141857).
Role of p16 expressing cells in formation and function of T cell memory with age
Authors: Torrance Blake , Lorenzo Erica C , Panier Hunter A , Cadar Andreia N , Martin Dominique E , Bartley Jenna M , Haynes Laura 
Published: January 2023

Abstract
Aging results in the accumulation of senescent cells that have been shown to cause dysfunction in many contexts but the effect on the function of T cell immunity is still unclear. Here, we aimed to probe the effects of clearing senescent cells on T cell responses to influenza infection. We utilized a powerful p16 trimodality reporter mouse model (p16-3MR) that includes cassettes encoding luciferase, RFP, as well as herpesvirus thymidine kinase (HSV-TK) all under the control of p16 promoter. p16 is commonly upregulated in senescent cells so this model allows us to detect senescent cells via luciferase activity, RFP expression, and selectively delete those cells by treating with ganciclovir (GCV). In aged p16-3MR mice following flu infection, we observed a transient induction of RFP+ p16 expressing cells at two weeks post infection. We hypothesized that p16 expressing cells are important for resolution of inflammation and fostering the effector to memory transition. Aged p16-3MR mice lacking p16 expressing cells following GCV treatment developed fewer flu-specific tissue resident memory CD8 T cells (Trm, CD103+ CD69+) in the lung compared to control treated aged counterparts. Interestingly, deletion of p16 expressing cells also increased the proportion of flu-specific Trm cells in the draining lymph node 30 days post infection. As a result, these mice were less effective in controlling viral burden following a secondary challenge. Conversely, during primary infection, deletion of p16 expressing cells induced a more youthful response in aged mice. This suggests that targeting senescent cells is a double-edged sword that may potentiate primary responses to infection while severely limiting the ability to form durable and protective memory.
Supported by grants from NIH (R21 AG060707, R21 AG071292)
T cell-intrinsic IL-17RA signaling in the spleen supports the establishment of chronic gammaherpesvirus infection.
Authors: Jondle Christopher Ned , Tarakanova Vera of 
Published: January 2023

Abstract
Human gammaherpesviruses, such as Epstein-Barr virus (EBV), are human specific gammaherpesviruses that establish lifelong infection in &gt;95% of adults and are associated with multiple cancers, including B cell lymphomas. These viruses have a natural tropism for B cells and usurp B cell differentiation to drive a robust polyclonal germinal center response, needed to establish a latent reservoir in memory B cells. The viral and host factors that influence the gammaherpesvirus-driven germinal center response are not clearly defined.
Murine gammaherpesvirus 68 (MHV68) is a natural rodent pathogen that is genetically and biologically related to human gammaherpesviruses and provides a tractable animal model to study viral and host factors that impact chronic infection. Utilizing MHV68, we discovered that IL-17RA signaling is proviral and supports the establishment of chronic gammaherpesvirus infection. Specifically, loss of IL-17RA signaling during MHV68 infection significantly attenuated viral latency, reactivation, and the germinal center response.
Importantly, the mechanism by which IL-17A signaling promotes MHV68-driven germinal center response and latency remained unclear. To gain mechanistic insight, we generated a mouse model of T cell specific IL-17RA deficiency. Excitingly, loss of IL-17RA specifically in T cells during MHV68 infection attenuated viral latency, reactivation, and the gammaherpesvirus-driven germinal center response in the spleen, similar to what was observed in the global IL-17RA−/− model. As T cells are not infected by MHV68, our finding unveils a novel crosstalk between T cell intrinsic IL-17A signaling and gammaherpesvirus infection of B cells, including virus-driven B cell differentiation.
Supported by American Cancer Society Postdoctoral Award (134165-PF-19-176-01-MPC, C.N.J.)
Influenza A virus-specific memory T cell persistence and functional recall are retained during pregnancy
Authors: Malavet Valeria Flores , Satchmei Ali , Prokop Emily , Strutt Tara M 
Published: January 2023

Abstract
Influenza A virus (IAV)-specific memory T cells induced by infection or through vaccination with live attenuated cold adapted influenza virus (LAIV) vaccine strains are well known to be able to confer protection against secondary homotypic as well as heterotypic IAV challenge. Whether IAV-specific memory T cell responses induced by LAIV vaccination can persist and retain their capacity to protect the maternal-fetal unit when generated prior to the altered physiological state of pregnancy, during which systemic levels of several hormones with putative immunomodulatory functions increase, is largely unknown. We show here that LAIV A/Alaska/6/77 CR-29 (H3N2)-vaccination prior to pregnancy induces IAV-specific memory CD4 and CD8 T cells that do indeed persist and functionally respond during recall in a manner similar to memory cells present in non-pregnant mice. This recall response can fully protect pregnant dams as well as their offspring from the adverse outcomes of lethal heterotypic infection with A/Puerto Rico/8/1934 (PR8-H1N1) virus. Using an adoptive transfer model and congenic HNT TcR transgenic CD4 T cells specific for PR8, we verify the observations observed with endogenous memory T cells that the recovery, surface marker phenotype, transcription factor profile, and TNF, IFN-g, and IL-2 multi-cytokine producing capacity of memory CD4 T cells responding to infection are largely unaltered by the pregnancy environment. These observations thus support that LAIV-vaccination prior to pregnancy results in the generation of IAV-specific memory T cells that have the potential to universally protect the maternal-fetal unit from the adverse outcomes of seasonal as well as pandemic strains of IAV.
Supported by grants from NIH (R21HD093948), the University of Central Florida Foundation and the Florida Education Fund McKnight Doctoral Dissertation Fellowship.
Phenotypical changes in peripheral NK cells and their impact on the clinical outcomes of patients with severe pandemic influenza A(H1N1)
Authors: Ramírez-Noyola Jazmín Ariadna , Fernández-López Luis Alejandro , Pacheco-Hernández Lynette Miroslava , Ignacio-Cortés Sergio , Gómez-García Itzel Alejandra , Garcia-Padrón Beatriz , Ramírez-Martínez Gustavo , Zúñiga Joaquín , Choreño-Parra José Alberto 
Published: January 2023

Abstract
NK cells are crucial antiviral effectors. However, their role against emerging respiratory pathogens has not been extensively addressed in humans. Here, we characterized the phenotype of peripheral NK cells in patients with severe pandemic influenza A(H1N1) and age- and sex-matched healthy controls (HC) using an extended panel of flow cytometry antibodies against markers of activation, cytotoxicity, cytokine production, immune memory, and surface receptors. Enrolled patients were grouped according to their clinical outcome (survival vs. death). The study included 42 influenza patients (73% men) with a median age of 48 years, from which eight (26%) died during the follow-up period. All patients were admitted to the intensive care unit and required mechanical ventilation. Individuals with influenza differentiated from HC by reduced numbers of total, CD56bright, CD56dimCD16+, CD27+ memory-like, NKp46+, and perforin+ NK cell subpopulations, but increased CD69+ NK cells. Notably, lower percentages of CD27+ memory-like NK cells correlated with increased alkaline phosphatase levels, a readout of lung damage in patients with acute respiratory distress syndrome. Strikingly, a higher expression of the senescence marker CD57 in CD27+ memory-like NK cells and expansion of double-positive CD27+CD57+ NK cells was characteristic of deceased influenza patients, but not survivors. Finally, despite being reduced in the overall influenza group with respect to HC, increased percentages of IFN-γ+ NK cells distinguished death vs. survival outcomes. Collectively, our results demonstrate that alterations in the phenotype of NK cells influence with the progression and outcomes of patients with severe pandemic influenza A(H1N1) infection.
Supported by grants from CONACyT (FORDECyT/10SE/2020/05/14-06, FORDECyT/10SE/2020/05/14-07) and SECTEI CDMX (SECTEI/050/2020) to JZ. JAR-N received a scholarship from CONACyT (CVU-1097402) for achieving her M.S.
Development of uveitis in a mouse model of spontaneous autoimmunity correlates with frequency of autoantigen-specific regulatory T cells
Authors: Yin Mianmian , Hiam Kamir J , Proekt Irina , Chan Jackie , Hu Yongmei , Lowell Clifford A , Gould Douglas B , Spitzer Matthew , Anderson Mark S , DeFranco Anthony L 
Published: January 2023

Abstract
B6 AireGW/+ Lyn−/− mice are a model of spontaneously autoimmunity in which 50% of mice develop uveitis by 9 weeks of age, and the remaining mice do not develop autoimmunity. Uveitis results from CD4+ T cells responding to the retina protein interphotoreceptor retinoid-binding protein (IRBP). To define cellular mechanisms that determine whether mice developed autoimmunity or not, we did single-cell RNA sequencing (scRNA-seq) of eye-draining lymph node (LN) CD4+ T cells that recognize the P2 epitope of IRBP (amino acid 271–290). Uniform manifold approximation and projection (UMAP) analysis showed that these CD4+ T cells included distinct subsets that were Ly6c1hi, Pdcd1hi and Treg, with the Treg population being overrepresented in the mice without disease. Pseudotime analysis indicated that the Ly6c1hi T cell subset represented an earlier stage in activation, whereas the Pdcd1hi subset represented a later stage. The two types of effector CD4+ T cells of mice with and without disease largely clustered together relative to their gene expression. Similar analysis of P2-specific CD4+ T cells from retinas with inflammation indicated a progression from proliferative (Mki67hi) cells to a Lag3hi subset to a Lag3hi/Pdcd1hi subset. In mice without uveitis, depletion of Treg by treatment of AireGW/+ Lyn−/− Foxp3DTR+/Y mice with diphtheria toxin resulted in rapid expansion of P2–specific CD4+ T cells in the draining LN, development of inflammation in the retina, and immune activation in other locations leading to lethality. Ongoing experiments are further testing the hypothesis that the fraction of Treg within the IRBP-specific CD4+ T cells determines whether or not these genetically susceptible mice develop eye-specific autoimmunity.
Supported by AI138479
Interstitial lung disease patients exhibit augmented germinal center responses in lung lymph nodes and increased serum reactivities to novel autoantigens
Authors: Yoon Young me , Velez Tania , Upadhyay Vaibhav , Vazquez Sara , Lee Cathryn T , Blaine Kelly , Decker Donna , Guzy Robert , Adegunsoye Ayodeji , Strek Mary , Noth Imre , Anderson Mark S , DeRisi Joseph , Shum Anthony , Sperling Anne I 
Published: January 2023

Abstract
Interstitial lung disease (ILD), a group of disorders characterized by scarring of the lungs, often leads to fatal impairment of respiratory function. While some patients exhibit features of autoimmunity, the role of the immune system within this heterogeneous group of disorders remains a matter of debate. We previously reported the enlargement of lung-draining lymph nodes (LLN) correlates with decreased survival in ILD patients, suggesting that activation of the adaptive immune system may contribute to pathophysiology. To investigate the composition and functional state of cells in the enlarged LLN, we performed flow cytometry analysis of LLN cells from endstage ILD patients undergoing transplantation (n=53) and control organ donors (n=36). We found that T cells from the ILD LLN exhibit effector memory phenotypes and significantly upregulate activation molecules including HLA-DR, CD38, CD154 and CD137, which are indicative of an ongoing immune response. Analyses of CD4 T cells revealed that Foxp3+CD25− regulatory T cells are specifically increased in ILD LLN. Moreover, follicular helper T cells and germinal center (GC) B cells are highly enriched, suggesting that antibody responses are activated in ILD LLN. To test the hypothesis that autoantigens may drive the augmented GC response, we screened circulating antibodies from ILD patients at two medical centers against the entire human peptidome using phage immunoprecipitation sequencing. We defined novel and shared autoantigens and experimentally validated antibody bindings to 5 protein antigens. Collectively, our findings suggest that antigen-specific immune responses may be involved in the pathophysiology and identify novel autoantigens for subsets of ILD.