bluuebunny/crossref_metadata_2025_split huggingface dataset:
Neutralizing CXCL13 attenuates neuropsychiatric manifestations in lupus-prone mice
Authors: Huang Michelle , Stock Ariel D , Putterman Chaim 
Published: January 2023

Abstract
The pathogenesis of neuropsychiatric systemic lupus erythematosus (NPSLE) remains unclear, despite the large number of lupus patients affected by neuropsychiatric manifestations. We recently showed that MRL/lpr mice, a classic SLE mouse model with neuropsychiatric symptoms, develop tertiary lymphoid structures (TLS) (a.k.a. ectopic germinal centers). TLS structurally and functionally resemble lymph nodes and form at the site of chronic inflammation to create more localized and specialized immune responses, which may promote a local immune response against self-antigen and thus contribute to the pathogenesis. Since CXCL13 is a key chemokine that drives lymphoid formation, we injected female MRL/lpr mice intraperitoneally with an anti-CXCL13 antibody, an IgG isotype control antibody, or PBS three times weekly for 12 weeks starting at 6–8 weeks of age. Cognitive dysfunction and affective deficits were assessed at the end of treatment. Anti-CXCL13 mAb treated MRL/lpr mice showed significant improvement in spatial memory, recognition memory, and depression-like behavior as compared to the two control groups (p=0.034, p=0.013, and p=0.0087, respectively). Furthermore, mice receiving the antibody by intracerebroventricular delivery showed similar results (p=0.055, p=0.20, and p=0.018, respectively). There were, however, no apparent differences in indicators of systemic disease, including autoantibody titers. Our results suggest that TLS play an important role in the pathophysiology of affective deficits and cognitive dysfunction, and that CXCL13 neutralization may be a potential therapeutic strategy to target NPSLE.
Transferable Immune Reactive Microbiota Determine Clinical and Immunologic Outcome of FMT in UC
Authors: Lima Svetlana Ferreira , Gogokhia Lasha , Viladomiu Monica , Woo Viola , Marderstein Andrew , Putzel Gregory , Scherl Ellen J , Brown Su-Ellen , Hambor John , Rosenthal Melissa , Jacob Vinita , Crawford Carl , Chou Lance , Longman Randy 
Published: January 2023

Abstract

Background
Over two million people worldwide suffer from ulcerative colitis (UC). Biologic therapy has significantly improved treatment, but nearly two-thirds of patients attenuate response. Fecal microbiota transplant (FMT) is an emerging therapy for the treatment of UC, but the microbial mechanism responsible for clinical response is poorly understood. Using samples from our pilot FMT study (Jacob V, et al 2017), we aim to define the core transferable microbiota (CTM) in UC patients responsive to FMT therapy and its therapeutic mechanism.

Methods
IBD disease activity scores were used to define clinical response. Metagenomic sequencing of donor, recipient, and 4 week post-FMT fecal samples was performed to define the CTM and strain level transferability. To define the transferable immune-reactive microbiota (TIM), IgA-seq was also performed on donor and recipient samples. Patient TIM strains were isolated and tested in gnotobiotic mouse models to evaluate their impact on mucosal immunity and colitis.

Results
We defined a CTM associated with clinical response to FMT. CTM strain tracking confirmed that clinical response correlated with strain transferability. We defined a core TIM by IgA-seq that correlated with clinical response. In humanized mouse models, TIM induced IgA in a T cell independent manner. Colonization of germ-free mice with a core TIM strain, Odoribacter splanchnicus, robustly induced mucosal Th17 and RORgt+/Foxp3+ iTreg cells and reduced the severity of transfer T cell colitis.
Our data highlights a core TIM in UC responders to FMT and the mechanistic impact of it in shaping mucosal immunity and guiding the response to UC. This provides a framework for rational selection of TIM for microbial-therapy in IBD.
Myelin-specific CD8 T cells require IFNγ responsiveness to optimally suppress CNS autoimmunity
Authors: Boyden Alexander W. , Brate Ashley A. , Karandikar Nitin J. 
Published: January 2023

Abstract
Investigating the complex cellular interplay controlling immunopathogenic and immunoregulatory responses is critical for understanding multiple sclerosis (MS) and for developing successful immunotherapies. Our group has demonstrated that central nervous system (CNS) myelin-specific CD8 T cells unexpectedly harbor striking regulatory capacity in both mouse and human. In particular, PLP178–191-specific CD8 T cells (PLP-CD8) robustly suppress the MS mouse model experimental autoimmune encephalomyelitis (EAE). We have recently shown this depends on PLP-CD8 elaborating IFNγ and perforin in a coordinated suppression program over time. However, the cellular target and downstream effects of CD8 T cell-elaborated IFNγ remains poorly understood. Here, we show that while WT PLP-CD8 were robustly suppressive in IFNγR-deficient mice, IFNγR-deficient PLP-CD8 exhibited defective suppression in WT mice. Further, compared to WT counterparts, IFNγR-deficient PLP-CD8 were defective in suppressing disease in IFNγ-deficient recipients, a scenario in which the only IFNγ WT PLP-CD8 “see” is their own. These data indicate that myelin-specific CD8 T cells must sense their own IFNγ in an autocrine/paracrine manner to optimally perform autoregulatory function in vivo. Further mechanistic dissection showed that IFNγR-deficient PLP-CD8 exhibited altered degranulation/granzyme profiles, altered migration in recipients, and potential deficits in killing capacity. Collectively, this work suggests that myelin-specific CD8 T cells require IFNγ-responsiveness to optimally regulate CNS autoimmunity and may help elucidate future adoptive immunotherapeutic approaches in MS.
Metformin reduces PD-L1 expression in the tumor and enhances the efficacy of vaccine generated CD8 T cells in a murine model of triple negative breast cancer
Authors: Munoz Luis E , Huang Haley Lei , Guin Rohini Neha , Bommireddy Ramireddy , Selvaraj Periasamy 
Published: January 2023

Abstract
Immunotherapy has emerged as a promising treatment of breast cancer, however, response rates to therapy remain far from ideal. We have developed a new platform for personalized vaccine immunotherapy that utilizes tumor tissue to generate tumor membrane vesicles (TMVs). These TMVs can be incorporated with immunostimulatory molecules such as IL-12 and B7-1 and used for immunization. In this study, we investigated the combinatorial effect of TMV-based vaccine immunotherapy, to induce an anti-tumor immune response, with the type 2 diabetes drug metformin, which has been reported to exert anti-tumor effects by promoting CD8 T cell activity in the tumor microenvironment. Our results show that TMV vaccination inhibits primary tumor growth in preclinical tumor models, while the combination with metformin resulted in greater inhibition of primary growth. TMV vaccination reduced the metastatic burden in the lungs while the combination with metformin further abrogated metastatic spread. Interestingly, TMV vaccination alone induced a significant increase in T cell infiltration to the tumor, and combination with metformin caused a decrease in the accumulation of T cells, despite having better control of tumor growth. Further, we show that metformin can reduce PD-L1 expression within the tumor microenvironment and result in more IFN-γ producing CD8 T cells within the tumor, providing a mechanistic insight for the improvement of TMV-based vaccine immunotherapy.
Combining a novel dual RAF/MEK inhibitor with immunomodulation to promote an anti-tumor response
Authors: Dong Lauren , Choi Hyejin , Budhu Sadna , Mehanna Nezar , Falik Nouraiz , Rosen Neal , Merghoub Taha , Wolchok Jedd D 
Published: January 2023

Abstract
The therapeutic scope of MEK inhibitors (MEKis) is currently limited to use in BRAF mutant melanoma. Therefore, we aim to develop new strategies to extend their usage to MEKi resistant RAS mutant cancers, which represent an unmet clinical need. A strategy we investigated is to balance the positive and negative immunomodulatory effects of MEKis for optimal combination with immunotherapy. In Ras mutant murine lung cancers, CH5126766 (CKI27) is novel due to its ability to inhibit both RAF and MEK, preventing the rebound of ERK that normally results from the relief of negative feedback in the MAPK pathway. We observed that CKI27 increased MHC expression on tumor cells and T cell mediated killing. Yet, CKI27 also decreased T cell proliferation, activation, and cytolytic activity. Implementing a break for T cells to recover with intermittent dosing of CKI27 partially relieved these inhibitory effects. Further combination with co-stimulatory agonist antibodies targeting OX40 and GITR completely alleviated these T cell toxicities and increased combination efficacy with checkpoint blockade antibody anti-CTLA-4. Understanding the immunomodulatory effects of combining CKI27 with immunotherapy will elucidate the mechanism behind their increased efficacy. This will allow us to make more informed decisions in dosing regimens, overcoming resistance, and generating long-term immune responses in current and future clinical trials treating patients with RAS mutant cancers.
Androgen receptor targeted therapy sensitizes tumor bearing mice to effective immunotherapy
Authors: Polesso Fanny , Hipfinger Christina , Sehrawat Archana , Moran Amy 
Published: January 2023

Abstract
Sex hormones impact both the innate and adaptive immune response, contributing to differences in autoimmune diseases, response to infection, and cancer incidence between males and females. Androgens are known to exert a general immunosuppressive effect. Historically, immune checkpoint blockade (ICB) has not proven to be efficacious in patients with prostate cancer despite standard of care consisting of androgen deprivation therapy (ADT). Upon disease progression, men will be treated with second line ADT therapy; some of which can prevent androgen receptor transcriptional activity. One such drug is enzalutamide. Given T cells express AR, we hypothesized that enzalutamide treatment would relieve the immunosuppressive effects of androgens on T cells, rendering them more responsive to immunotherapy. In the studies presented, we sensitize castration resistant prostate cancer tumor bearing mice to effective ICB with a combination of castration and enzalutamide. This combination therapy leads to ~40% tumor rejection and long-term survival; an effect that is dependent on CD8 T cells. These tumor infiltrating CD8 T cells made 6-fold more IFNγ within the tumor in the triple therapy group compared to control group. Surprisingly, this combination therapy showed efficacy in the androgen independent sarcoma tumor model MCA-205. Using in vitro and in vivo assays, we show that enzalutamide has a direct effect on T cell function. Taken together, our data suggests that ADT, and specifically enzalutamide treatment, can sensitize tumor bearing animals to effective immune checkpoint blockade and allow for tumor control by T cells.
DNMTi plus anti-4-1BB therapy reverses suppressive tumor microenvironment and reduces tumor burden in ovarian cancer
Authors: Gomez Stephanie , Palmer Erica , Kobyra Julie , Hadley Melissa , Chiappinelli Katherine B. 
Published: January 2023

Abstract
Novel therapies are urgently needed for ovarian cancer (OC), the fifth deadliest cancer in women. Better OC prognosis is associated with high tumor-infiltrating CD8 T cells, but OC is generally characterized by an immunosuppressive tumor microenvironment (TME), with less than 10% of patients responding to immune checkpoint blockade therapy in clinical trials. DNA methyltransferase inhibitors (DNMTi) activate transcription of repetitive elements to induce a type I interferon response in OC, which recruits and activates host immune cells to fight the tumor. Additionally, changes in DNA methylation in T cells regulate T cell memory. Thus, DNMTi increase anti-tumor immunity through 1) tumor IFN response and 2) reversal of T cell exhaustion.
To test the hypothesis that combination epigenetic and immune therapy could increase the anti-tumor response in OC, we treated an immunocompetent mouse model of OC with DNMTi +/− anti-CD137. The ID8 MOSE P53−/− model grows intraperitoneally and produces ascites, mimicking human OC which is 90% P53 mutant. The TME of this model is immunosuppressive and it is not responsive to immune checkpoint blockade therapy. At 9 weeks post tumor injection, we observed the following survival percentages: 50% for the combination group, 30% for the DNMTi group, and 0% survival for the both the anti-CD137 and mock groups. We also observed statistically significant increases in %CD3+IFNg+ T cells in the spleen for the DNMTi + anti-CD137 group compared to the mock group. We hypothesize that anti-CD137 treatment is activating immune cells recruited by DNMTi. Future experiments will manipulate cytokine expression to delineate mechanism. We thus describe a novel immunotherapeutic combination for ovarian cancer.
Characterizing the mechanism of action of a HER2xCD3 bispecific T-cell engager (BiTE) antibody in 2D and 3D immune cell killing assays using a combined live-cell imaging and flow cytometry workflow
Authors: Bevan Nicola , McBain Kirsty , Kelsey Lauren , Szybut Clare , Trezise Del , Dale Tim 
Published: January 2023

Abstract
Solid cancers have proved to be a challenge for immunotherapies and targeted approaches are needed. Evaluation of these therapies using relevant models plays a large part in their development. A workflow that combines live-cell imaging and flow cytometry was used to characterize immune cell killing induced by a BiTE antibody.
Immune cell co-culture assays were set up in 2D or 3D formats with green nuclear labeled AU565 HER2 positive breast cancer cells and various effector cell types. Immune cell activation was induced using HER2xCD3 BiTE or CD3/CD28 Dynabeads. Images were captured every 4 h with IncuCyte and quantified for numerous cellular parameters. Cytokine analysis of supernatant samples was performed using a multiplex QBead panel alongside assessment of immune cell activation markers using the iQue3.
Quantification of PBMC mediated cytotoxicity showed a comparable 70 ± 2% and 62 ± 1% reduction in spheroid growth with BiTE (10 ng mL−1) and Dynabead (1:1 bead-to-cell ratio) activation. In contrast to Dynabeads, BiTE activation did not induce immune cell shape change or upregulation of T cell activation markers; 1 ± 0.2% CD69 expression was observed on CD3+CD8+ cells, compared to 53 ± 0.6%. Further evaluation found BiTE activated isolated CD56+ cells induced less target cell death compared to PBMCs and granzyme B secretion was not detected, suggesting a lack of involvement of NK cells. Release of IL-6 and CCL2 cytokines indicate immune cell mediated cytotoxicity via an inflammatory pathway.
These data suggest HER2xCD3 BiTE activity may not be mediated through T cell or NK cell subtypes. At present the mechanism of target cell killing is unclear, however further studies combining imaging and flow cytometry may provide the required insight.
Heterosubtypic human antibody response to influenza H7N9 in survivors of natural infection
Authors: Gilchuk Iuliia M , Dong Jinhui , Li Sheng , Turner Hannah L , Irving Ryan P , Ward Andrew B , Crowe James E 
Published: January 2023

Abstract
H7N9 avian influenza virus causes severe infections in humans after cross-species transmission from bird reservoirs. Previous studies revealed that infection with H7N9 elicits a heterosubtypic polyclonal antibody (Ab) response between H7 and H3 or H1 hemagglutinins (HAs) in humans. However, relatively little is known about the functional activity, targeted epitopes, and breadth of reactivity of human monoclonal Abs (mAbs) against H7N9. In this study, we isolated 18 H7-reactive mAbs from survivors of natural H7N9 infection. The majority (14 of 18) of H7-reactive mAbs within the panel possessed neutralizing activity, mediated by inhibition of attachment or egress of the virus. Binding analysis of the isolated mAbs against fourteen influenza virus HAs representing group 1 and 2 revealed cross-reactivity for diverse strains within the H7 subtype and an extended response to H15 HA. Seven identified cross-reactive mAbs targeted the HA head domain, including neutralizing and non-neutralizing mAbs. Two neutralizing mAbs that are specific to the HA stem region revealed heterosubtypic reactivity against H7, H15 and H3 HAs. Further analysis identified the epitopes for heterosubtypic mAbs. This study provides insights into the basis for heterosubtypic antibody responses against H7N9 in human infection survivors and may inform efforts to incorporate novel epitope targets in broadly protective influenza vaccines.
Codelivery of antigens with TLR7/8 agonists leads to an improved immune response to influenza vaccination
Authors: Lathrop Stephanie K , Massena Casey J , Davison Clara J , Schoener Roman , Burkhart David J , Evans Jay T 
Published: January 2023

Abstract
When designing an effective and safe vaccine, inducing a protective immune response to a pathogen without also inducing unacceptable levels of immunopathology is a major challenge. One promising strategy is the use of purified or recombinant antigens along with adjuvants that specifically engage pathogen-associated molecular pattern (PAMP) receptors. This targeted approach aims to minimize immune-mediated reactogenicity. Our group has developed libraries of novel toll-like receptor (TLR)7 and/or TLR8 ligands which show a range in receptor activity. Previous studies have shown that the high potency of TLR7/8 adjuvants leads to unacceptable toxicity in human clinical trials. One strategy to overcome this toxicity is direct adjuvant-antigen conjugation, which serves to limit systemic distribution of the adjuvant through the codelivery of the antigen and adjuvant to APCs. We have demonstrated that covalent conjugation of a TLR7/8 agonist to an antigen results in improved humoral and cell-mediated responses, with decreased systemic and local inflammation. This approach allows for the use of a reduced adjuvant dose by preventing the engagement of PAMPs in the absence of antigen. We hypothesize that antigen/adjuvant codelivery boosts the engagement of CD4+ T cells and elicits the necessary cytokines to promote a Th1-related switch to antigen-specific IgG2a antibodies. In addition, engagement of TLR7/8 appears to improve cross-presentation, thereby boosting the CD8+ T cell response, which is desirable for improved immunity to influenza and other viruses. We find this approach has great potential to enhance vaccine efficacy, while reducing the immunopathology often associated with highly active TLR7/8 adjuvants.
Efficacy of agonist αOX40 and antagonist αCTLA-4 cancer immunotherapy is increased by ibrutinib treatment through enhanced Eomes expression and T-cell response
Authors: Emerson Dana A , Redmond William L 
Published: January 2023

Abstract
Antagonistic monoclonal antibodies targeting individual checkpoint receptors on T cells, such as CTLA-4 and PD-1, are used to treat patients with metastatic disease; however, only a subset of patients respond. Combination therapies, such as CTLA-4 blockade with agonist αOX40 mAb, are synergistic in preclinical models. αOX40/αCTLA-4 therapy synergistically enhances expression of Eomesodermin (Eomes), a transcription factor that drives CD8 T cell differentiation and memory function. We hypothesize that Eomes expression plays a critical role in the synergistic efficacy of αOX40/αCTLA-4 therapy. Compared to Eomeslo CD8 T cells, Eomeshi CD8 T cells induced by αOX40/αCTLA-4 therapy express lower levels of checkpoint receptors (PD-1, TIM-3, LAG-3) and higher levels of tumor trafficking and activation markers (CXCR3, CD44), and intratumoral cytokines (IFN-γ, TNF-α). CD8 T cell-specific Eomes−/− tumor-bearing mice did not respond to αOX40/αCTLA-4 therapy, demonstrating the necessity of Eomes expressing CD8 T cells. Interleukin-2 inducible T cell kinase (ITK) negatively regulates T cell Eomes expression following T cell receptor ligation. Thus, we asked whether enhancing T cell Eomes expression by combining ibrutinib, an FDA-approved ITK inhibitor, with αOX40/αCTLA4 therapy would further improve anti-tumor responses by increasing Eomeshi CD8 T cells. This triple combination increased CD8 T cell-specific Eomes expression, tumor-specific CD8 T cells, Ki-67 (proliferation), and Th1 cytokines (IFN-γ, TNF-α) leading to reduced tumor growth and improved survival over αOX40/αCTLA-4 alone, making ibrutinib/αOX40/αCTLA-4 a potential novel triple combination therapy.
CD4+ T cells are dispensable for induction of heterologous tier 2 HIV neutralizing antibodies in rhesus macaques
Authors: Kobie James , Barnette Philip , Pandey Shilpi , Spencer David , Basu Madhubanti , Piepenbrink Michael , Keefer Michael , Hessell Ann J , Haigwood Nancy L , Sarkar Sanghita 
Published: January 2023

Abstract
Induction of a sustained and broad neutralizing antibody (Ab) response is a major goal in developing a protective HIV vaccine. Many HIV Envelope (Env)-specific broadly neutralizing antibodies isolated to date exhibit substantial somatic hypermutation, and their development in HIV infected individuals has been correlated with T follicular helper responses. This has bolstered the rational that a robust CD4+ T helper cell response will likely be required for the induction of such HIV broadly neutralizing antibody responses by vaccination. Using the VC10014 DNA-protein co-immunization vaccine platform consisting of gp160 envelope plasmids and gp140 trimeric envelope proteins derived from a clade B HIV-1 infected subject who developed broadly neutralizing serum Abs, and which has been previously shown to induce Tier 2 heterologous neutralizing Abs in rabbits and rhesus macaques, we determined the influence of CD4+ T cell depletion during the vaccination regimen on the characteristics of the Env-specific humoral response in rhesus macaques. Both CD4+ depleted and non-depleted animals developed Tier 2 heterologous neutralizing plasma antibodies, with no inferiority in titers among the CD4+ T cell depleted animals. Similarly, there was no diminishment of titers of HIV Env-specific cross-clade binding antibodies, antibody dependent cellular phagocytosis or antibody dependent complement deposition in the CD4+ depleted animals. These results suggest that HIV neutralizing antibodies can be induced in the absence of robust CD4+ T cell help, which may have implications for developing effective HIV vaccine strategies.