bluuebunny/crossref_metadata_2025_split huggingface dataset:
Killed Klebsiella pneumoniae treatment reduces tumor burden in murine melanoma
Authors: Costa Amanda M , Perekslis Thomas , Alexander Matthew P , Sanders Kiah , Gunn Hal , Mullins David W 
Published: January 2023

Abstract
Acute infection in a cancer-bearing organ has been associated with improved prognosis and enhanced survival, although the anti-cancer immune mechanisms remained undefined. In collaboration with Qu Biologics, we hypothesized that stimulating pathogen-specific immunity, using bacteria that have previously infected specific organs, may induce protective anti-cancer immune responses in those target organs. We evaluated Klebsiella-mediated anti-cancer efficacy in metastatic-like B16 melanoma using heat-killed Klebsiella pneumoniae (Qu Biologics’ drug product QBKPN Site Specific Immunomodulator [SSI]). Subcutaneous injection of QBKPN SSI significantly reduced tumor burden. Furthermore, subcutaneous treatment with heat-killed E. coli, (QBECO SSI) reduced lung tumor burden but not as substantially as with QBKPN. This suggests that subcutaneous immune induction using a lung specific-pathogen activates a lung-specific antitumor response.
We next asked whether pre-exposure to K. pneumoniae would improve anti-tumor efficacy. We exposed mice to live K. pneumoniae via intratracheal infection prior to subcutaneous injections with QBKPN SSI. Pre-exposure to K. pneumoniae significantly enhanced QBKPN SSI-induced anti-tumor immunity and control of metastatic-like B16 melanoma in the lungs. The anti-tumor efficacy in exposed mice correlated with an influx of monocytes and neutrophils, but did not correlate with an influx of T cells into the lungs. Collectively, these data suggest pre-exposure to K. pneumoniae may induce tissue resident memory cells that mediate tumor cytolysis, and thus explain the lack of T cell infiltration into the lungs.
Critical roles of Noxa and Mcl-1 in the signaling mechanisms of bortezomib in TRAF3-deficient mouse B lymphoma and human multiple myeloma cells
Authors: Luo Chang , Han Yeming , Liu Yingying , Edwards Shanique K E , Xie Ping 
Published: January 2023

Abstract
Bortezomib, a clinical drug for multiple myeloma (MM) and mantle cell lymphoma, exhibits complex signaling mechanisms, which vary depending on the cancer cell type and the critical genetic alterations of each cancer. In the present study, we investigated the signaling mechanisms of bortezomib in mouse B lymphoma and human MM cells deficient in a new tumor suppressor gene, TRAF3. We found that bortezomib robustly induced up-regulation of the pro-apoptotic protein Noxa, cleavage of the anti-apoptotic protein Mcl-1, and accumulation of the oncoprotein c-Myc. Interestingly, bortezomib exhibited contrasting roles on canonical versus non-canonical NF-kB pathways. Overexpression of Noxa was sufficient to induce apoptosis and inhibit proliferation in human MM cells by inducing cleavage of Mcl-1 and caspase 3. Furthermore, we demonstrated that oridonin (an inhibitor of NF-kB1 and NF-kB2) or AD 198 (a drug targeting c-Myc) drastically potentiated the anti-cancer effects of bortezomib in TRAF3-deficient malignant B cells. Taken together, our findings provide new insights into the signaling mechanisms of bortezomib, which would aid the design of novel bortezomib-based combination therapies. Our results also provide a rationale for clinical evaluation of the combinations of bortezomib and oridonin (or other inhibitors of NF-kB1/2) or AD 198 (or other drugs targeting c-Myc) in the treatment of lymphoma and MM, especially in patients containing TRAF3 deletions or relevant mutations.
Altered host immune responses following infection with miR-150 expressing influenza viruses
Authors: Xia Yingju , Izzard Leonard , Ye Siying , Stambas John 
Published: January 2023

Abstract
Current influenza vaccines offer considerable protection for the majority of the population. However, vaccine efficacy amongst high risk populations, such as the elderly, is variable. Novel approaches are therefore required to improve vaccine efficiency in these susceptible populations. MicroRNAs (miRNAs) play important roles in the regulation of immune responses by modulating gene expression. Our work describes the generation and delivery of engineered influenza viruses with the ability to express defined immunomodulatory miRNAs. MicroRNA-150 (miR-150) has been shown to be involved in the development of T cell and B cell. Furthermore, evidence suggests that circulating miR-150 can be actively transported to target cells where it can influence gene expression. In this study, we first investigated the ability of recombinant influenza viruses to deliver immunomodulatory miR-150, and alter cytokine production in vitro. miR-150 was inserted into an artificial intron between the NS1 and NEP genes of the PR8 H1N1(PR8-150) and X31 H3N2 (X31-150) influenza A viruses and was rescued using reverse genetics. Mature miR-150 expression was confirmed using qPCR and by knockdown of its target gene, c-Myb, using a dual-luciferase assay. Moreover, cytokine mRNA expression levels (IL-6, IFN-β, CXCL10 and CCL5) were enhanced following infection with PR8-150 in A549 cells. Mice infected intranasally with recombinant X31-150 virus showed significantly higher numbers of neutrophils, macrophages and influenza specific CD8+ T cells post infection compared to X31 wild type controls. These results demonstrate that recombinant influenza viruses can be used to deliver functional miRNAs with the ability to modulate immunity both in vitro and in vivo.
Seasonal vaccination with the inactivated influenza vaccine generates antibody profiles that are distinct from those generated by the live-attenuated influenza vaccine
Authors: Shagal Ayelet , Monto Arnold , Hertz Tomer 
Published: January 2023

Abstract
Vaccination is an effective strategy for preventing influenza infection and is now recommended annually by the CDC for all individuals age 6 months and above. There are currently two licensed seasonal influenza vaccine formulations in the US: the inactivated influenza vaccine (TIV) and the live-attenuated influenza vaccine (LAIV). A previous clinical trial (FluVacs) conducted in 2007–2008 comparing the efficacy of these two vaccines in adults aged 18–65 reported that both vaccines were efficacious, but the TIV had an overall higher efficacy than the LAIV. We hypothesized that the two vaccines will also generate distinct antibody signatures partially due to the difference in the reported efficacy. We therefore compared the antibody profiles generated by the two vaccines. Profiles were generated using antigen microarrays (AMs) which are a high-throughput binding assay. We generated peptide AMs spotted with overlapping 20mer peptides from a seasonal vaccine strain (2012) and used serum samples of 150 participants from the FluVacs study taken at day 21 post vaccination and at the end of the season. We also used baseline samples to account for previous antibody memory responses. We found that both vaccines generated relatively weak responses when compared to baseline. However, in-line with our hypothesis, the antibody titers generated by the TIV vaccine were higher than those of the LAIV. We then used unsupervised clustering to cluster the post-vaccine responses of all trial participants. We found distinct clusters, each corresponding to a different response pattern. In conclusion we used a novel assay to profile influenza vaccine induced antibody responses and found that the TIV and LAIV vaccines generate different antibody profiles
Immunization with a VCG-based vaccine protects against Chlamydia-induced infertility in the mouse genital infection model
Authors: Eko Francis , Pais Roshan , Omosun Yusuf , He Qing , Blas-Machado Uriel , Igietseme Joseph U. , Fujihashi Kohtaro 
Published: January 2023

Abstract
Chlamydia trachomatis genital infections constitute a major public health challenge due to the significant morbidity that includes pelvic inflammatory disease, ectopic pregnancy and tubal factor infertility. An effective vaccine is required to protect against infection and prevent development of upper genital tract pathology. Previous studies showed intramuscular immunization with a Vibrio cholerae ghost (VCG)-based chlamydial vaccine induced cross protective immunity in mice but its ability to protect against upper genital tract pathology was not evaluated. We sought to assess the ability of rectal immunization with this vaccine to protect against upper genital tract pathology following infection with a heterologous chlamydial serovar in the mouse model. Female mice were rectally immunized with VCG expressing select C. trachomatis serovar D outer membrane proteins and challenged intravaginally with live serovar E chlamydiae 2 weeks postimmunization (PI) to evaluate cross protection. To evaluate upper genital tract pathology, mice received a repeat challenge infection 8 weeks after initial challenge. Rectal immunization elicited high levels of antigen-specific Th1 cell-mediated and humoral immune responses in mucosal and systemic tissues. Furthermore, immunization induced cross-protective immunity that reduced the length and intensity of genital chlamydial shedding and protected against Chlamydia-induced infertility and tubal pathologies. The ability of the VCG vaccine candidate to elicit heterotypic genital tract immunity and protect against Chlamydia-induced upper genital tract pathology is remarkable and highlights its potential for development as a universal chlamydial vaccine candidate.
Secreted heat shock protein gp96-Ig vaccine for malaria prophylaxis
Authors: Strbo Natasa , Romero Laura , Podack Eckhard R , Villasante Eileen F , Edgel Kimberly A , Strbo Natasa 
Published: January 2023

Abstract
Proposed vaccine principle relies on secreted gp96-Ig chaperoning Plasmodium falciparum (Pf) AMA1 and PfCSP sporozoite proteins that are efficiently taken up and cross presented by activated DC via MHC I to CD8 CTL, thereby stimulating an avid, antigen specific, cytotoxic T cell response. The generation of a powerful, cytotoxic anti-sporozoite CD8 CTL response by the vaccine is expected to provide prophylactic immunity against malaria by killing infected liver cells, thereby preventing blood stage infection.
We performed mouse immunogenicity experiments using 293-gp96-IgPfAMA1-PfCSP vaccine cells that produce gp96-Ig and express PfAMA1 and PfCSP. One million 293-gp96-IgPfAMA1-PfCSP cells were injected in B6 mice by intraperitoneal (IP), subcutaneous (SC), intramuscular (IM) and intradermal (ID) routes. Control mice received 293-gp96-Ig. Five days later, mice were sacrificed. Total spleen cells were cultured overnight in medium only or with pool of overlapping PfAMA1 and PfCSP peptides. Frequency of PfAMA1- and PfCSP-specific CD8 T cell responses in the spleen was measured by intracellular cytokine staining assay and analyzed by flow cytometry.
We found that vaccine cells delivered by IP and SC routes induced the strongest PfAMA1- and PfCSP-specific immune response systemically (1.8±0.4% and 1.3±0.3% PfAMA1- and PfCSP-specific spleen CD8 T cells, respectively). In addition, after 293-gp96-IgPfAMA1-PfCSP vaccination frequency of liver-infiltrating CD8+ T cell was significantly increased (70% of all liver CD3+ cells were CD8+CD44+CD62L− T cells).
Our findings are strongly supportive of the novel gp96-Ig malaria vaccine as unique systemic and liver-homing, sporozoite specific CD8 CTL vaccine strategy.
Fc-dependent effector functions of neutrophils induced by hemagglutinin stalk-specific antibodies
Authors: Mullarkey Caitlin E , Bailey Mark J , Golubeva Diana , Azarm Kristopher , Nachbagauer Raffael , Miller Matthew S , Palese Peter 
Published: January 2023

Abstract
Broadly neutralizing antibodies that recognize the conserved hemagglutinin (HA) stalk have emerged as exciting new biotherapeutic tools to combat both seasonal and pandemic influenza viruses. Our general understanding of the mechanisms by which stalk-specific antibodies achieve protection is rapidly evolving. It has recently been demonstrated that broadly neutralizing HA stalk-specific IgG antibodies require Fc-FcR interactions for optimal protection in vivo. Here we explore the relationship between such stalk-specific antibodies and neutrophils. Neutrophils represent a critical innate effector cell population and play an important role in orchestrating downstream adaptive responses to influenza virus infection. Yet, at present the interplay of HA stalk-specific IgG, Fc-FcR engagement, and neutrophils has remained largely uncharacterized. Using an in vitro assay to detect the production of reactive oxygen-species (ROS), we show that both human and mouse monoclonal HA stalk-specific IgG antibodies are able to induce the production of ROS in neutrophils, while antibodies specific to the HA head domain do not. Furthermore, our results indicate that the production of ROS is dependent on Fc receptor engagement and phagocytosis. We went on to assess the ability of monoclonal HA stalk-specific IgA antibodies to induce ROS. We found that monoclonal IgA lead to a greater induction of ROS as compared to monoclonal IgG. Our data demonstrate this activity is dependent on the engagement of FcαR1. Taken together our findings describe a novel FcR-dependent effector function induced by HA stalk-specific IgG and IgA antibodies.
Responses to fibroblast-adapted human cytomegalovirus (HCMV) vaccines conform to CD8 T cell recognition paradigms
Authors: Murray Susan E , Nesterenko Pavlo , Munks Michael , Smart Savannah , Vanarsdall Adam , Lee Ronzo , Cox Josephine , McVoy Michael , Adler Stuart P , Hill Ann B 
Published: January 2023

Abstract
Recently, a rhesus CMV-based SIV vaccine that clears SIV from half of vaccinated monkeys was shown to elicit unconventional CD8 T cell responses to both SIV and RhCMV (IE1) antigens in all immunized monkeys. These responses were characterized by i) recognition of an unusually large number of peptide epitopes within each antigen, and ii) MHCII restriction by ~2/3 of responding CD8 T cells (Hansen et al, Science, 2013). Unconventional responses were only elicited by fibroblast-adapted rhCMV strain 68-1 that lacks ability to infect cells other than fibroblasts due to loss of pentameric viral glycoprotein expression.
To determine whether fibroblast-adapted human CMV (HCMV) vaccines also elicit unconventional CD8 T cell responses, we exploited an ongoing phase I clinical trial of live HCMV vaccines. These vaccines have similar fibroblast-restricted tropism to rhCMV68-1, although the precise genetic lesions differ. We analyzed CD8 T cell responses in 12 seroconverters by stimulating PBMC with overlapping peptide pools covering 20 HCMV antigens. Seven subjects in this low dose trial (100 or 1000 pfu) made consistent, measurable CD8 T cell responses. All 7 responded to IE1; 3 also responded to 1–2 additional antigens. We have mapped responses to individual 15mers in 3 subjects, and all revealed highly focused responses to 1–3 epitopes within IE1. In one HLA-B08 positive subject, the two responses mapped to known immunodominant B08-restricted IE1 epitopes. We have mapped 4 other epitopes to date; of these, 3 are 9mers and are likely conventional, but one may be a 13mer. Thus far, in contrast to RhCMV in monkeys, the CD8 T cell response to fibroblast-adapted HCMV vaccines in humans appears to be predominantly conventional.
Supported by NIH R21 AI116107
Optimizing antiviral humoral responses the natural (killer) way
Authors: Rydyznski Carolyn E. , Waggoner Stephen N 
Published: January 2023

Abstract
Many vaccine efforts to prevent deadly viral diseases aim to generate high-affinity, class-switched virus-specific antibodies, which arise mainly from germinal center (GC) reactions. However, vaccine-induced GCs are sometimes inadequate to prevent infections with major human pathogens, including HIV. An improved understanding of the mechanisms regulating the GC reaction will improve future vaccines. We discovered that natural killer (NK) cells suppress the generation of GC B cells and follicular helper T (Tfh) cells in a perforin-dependent manner following acute viral infection. Remarkably, a fraction of activated NK cells up-regulate the B-cell follicle homing receptor, CXCR5, which potentially facilitates interactions between cytotoxic NK cells and GC-inducing cells in or near B-cell follicles. We performed RNAseq analysis of this unique subset of NK cells to uncover potential mechanisms of humoral immune suppression. We also interrogated qualitative changes in antigen-specific B cells induced by NK cells following immunization with the hapten-carrier immunogen, NP-KLH. NK cells restricted the NP-KLH-induced GC response and number of NP-specific B cells. In the absence of NK cells, there was an increase in NP-specific plasma cells in the bone marrow at early time points and a higher ratio of high- to low-affinity NP-specific IgG in sera at late time points, suggesting that NK cells impair the quality of the humoral response that develops following immunization. To date, our data provide evidence that NK-cell suppression of GCs represents a previously underappreciated regulatory axis, potentially mediated by a unique subset of NK cells, which could be manipulated to enhance future vaccine efforts.
A unique subset of CD11c+ B cells upregulate FasL expression in response to IL-12p40 during lethal dose influenza virus (IAV) infections.
Authors: Zacharias Zeb Raplh , Legge Kevin L 
Published: January 2023

Abstract
Our recent work has detailed a novel regulation of the IAV-specific CD8 T cell response during lethal dose IAV infections. During such infections, FasL+ plasmacytoid dendritic cells (pDC) within the regional lymph nodes (LN) target and eliminate the developing Fas+ effector CD8 T cell response limiting immunity and driving severe disease. While our studies have shown that pDC are sufficient to drive this mechanism (i.e. transfer of wild type pDC into gld mice results in elimination of activated CD8 T cells), antibody targeted elimination of pDC in wild type mice does not completely reverse CD8 T cell apoptosis. This suggested that an additional cell type might contribute to the overall loss of CD8 T cells during lethal/high pathogenic IAV infections. Here we describe a CD11c+B cell population that shares some common markers with both pDC and several defined B cell subsets, but can be distinguished from both based on unique marker expression. Our data suggests that these CD11c+ B cells are recruited to the LN in greater numbers compared to pDCs and appear to express FasL upon exposure to IL-12p40 homodimer in lethal dose IAV infections. Further our results suggest that FasL+ expression with BST2+ expression within CD11c+ B cells. Our current studies are focused on further determining the factors regulating this CD11c+B cell population and its relative contribution to the overall FasL-dependent control of the magnitude of the CD8 T cell response during lethal dose IAV infections.
TCF1 regulates antiviral T follicular helper (Tfh) cell responses through a negative feedback loop with IL-2 and Blimp1
Authors: Wu Tuoqi , Shin HyunMu , Moseman E. Ashley , Ji Yun , Huang Bonnie , Harly Christelle , Sen Jyoti M , Berg Leslie J , Gattinoni Luca , McGavern Dorian B , Schwartzberg Pamela L 
Published: January 2023

Abstract
Th1 cells and T follicular helper cells (Tfh cells), are generated early after viral infection. However, the molecular pathways that drive the differentiation of these two distinct subsets remain unclear. Here, we demonstrate that a molecular circuit involving TCF1, Blimp1, and IL-2 regulates the differentiation of viral-specific Tfh and Th1 cells. Tcf7 gene (encodes TCF1) is actively transcribed in naïve and Tfh cells but strongly repressed in Th1 cells. TCF1 expression was negatively correlated with CD25 and Blimp1 expression, and was suppressed by IL2 signaling and Blimp1. Notably, loss of Tcf7 in T cells led to defective Tfh-cell and GC responses after viral infection. Tcf7-deficient Tfh cells had reduced Tfh transcription signatures and increased Th1 transcription signatures. Moreover, Tcf7-deficient Tfh cells had lower mitochondrial mass and membrane potential, and showed reduced expression of genes related to oxidative metabolism. Il2ra and Prdm1 (encodes Blimp1) were ectopically expressed in Tcf7-deficient Tfh cells and were bound by TCF1, suggesting the existence of a negative feedback loop linking TCF1 to IL-2 and Blimp1. Over-expression of Bcl6 rescued Tfh defects caused by Tcf7 deficiency, demonstrating that TCF1 is upstream of the Bcl6-Blimp1 axis. Finally, we found that Tcf7-deficiency did not cause evident defect in Tfh responses in Th2 immunization models. Therefore, TCF1 specifically regulates antiviral Tfh responses.
Regulatory T-cells are essential to promote proper CD4 T-cell priming upon mucosal infection
Authors: Lund Jennifer M , Soerens Andrew , Costa Andreia 
Published: January 2023

Abstract
Regulatory T-cells (Tregs) limit autoimmunity and immunopathology using a variety of suppressive mechanisms, but their roles during pathogen-directed immune responses remain unclear. Following Herpes Simplex virus-2 (HSV-2) infection, mice lacking Tregs fail to control viral replication, pointing to a role for Tregs in facilitating productive immune responses. Using adoptive transfer of TCR transgenic CD4 T-cells into Treg-sufficient or Treg-depleted mice prior to HSV-2 infection, we found that Tregs are required for timely accumulation of HSV-2-specific CD4 T-cells within the infected tissues. Further, Tregs are critical for appropriate trafficking of dendritic cells (DCs) from the vaginal mucosa to the dLN, which results in fully effective CD4 T-cell priming, activation, and ultimately migration to the infected tissues. Using CTLA-4 conditional knockout mice, we demonstrate that Tregs impact DC migration through a CTLA-4-mediated mechanism. Together, our data highlight the critical role of Tregs in proper potentiation of adaptive immune responses to microbial infection.
Immunosequencing reveals diagnostic signatures of pathogen infection and HLA type in the T-cell Receptor Repertoire
Authors: DeWitt Will , Emerson Ryan O. , Vignali Marissa , House Steven , Desmarais Cindy , Rieder Mark J. , Carlson Christopher S. , Gravley Jenna , Hansen John A , Robins Harlan S. 
Published: January 2023

Abstract
We analyzed the T-cell response to CMV by sequencing rearranged TCRs in 528 subjects (240 CMV+ and 288 CMV−). We assessed concordance between ~80 million unique TCRs and CMV serostatus, looking for significant associations. We identified 104 CMV-associated TCRs at FDR ≈ 0.20. Training a binary classifier on these features, we predict CMV serostatus in a cross-validation procedure with a diagnostic odds ratio of 17 on the same cohort. A second independent cohort of 120 subjects with known CMV serostatus yielded a diagnostic odds ratio of 31.
Next, we studied the HLA-restriction of each CMV-associated TCR by assessing enrichment of particular HLA types among subjects that carry each CMV-associated TCR. Of 104 CMV-associated TCRs, 41 were HLA-restricted at p ≤ 10−3, and none of these were significantly associated with multiple alleles in any locus.
We find concordance between our data and previously-reported CMV-specific TCRs: most previously-reported CMV-specific TCRs are present in our data, however only 5 CMV-associated TCRs have been previously reported. Of these, four were significantly HLA-restricted in these data, and agreed with previous findings.
We investigated the association of TCRs with each HLA allele present in the cohort, with significant results for several higher frequency alleles. Cross validation with binary classifier training resulted in high accuracy prediction for these alleles, indicating that HLA type can be inferred from immunosequencing data.
We demonstrate the validity of association studies using immunosequencing for detection and HLA-association of public T-cell responses to infection, and report that assessing the presence of associated T-cell responses can serve as a powerful diagnostic classifier.