bluuebunny/crossref_metadata_2025_split huggingface dataset:
Adenosine generation limits radiation-induced tumor immunogenicity by abrogating recruitment and activation of CD103+ DCs
Authors: Wennerberg Erik , Cronstein Bruce , Formenti Silvia C , Demaria Sandra 
Published: January 2023

Abstract
Radiation therapy (RT) induces release of ATP in the tumor micro environment (TME) triggering recruitment and activation of dendritic cells (DCs), including CD103+ DCs, identified as the key DC subsetresponsible for cross-presentation of tumor antigens to CD8+ T cells. However, CD39 and CD73-expressing cells in the TME hydrolyze ATP into adenosine that potently inhibits DCs and CD8+ T cells. Here, we tested the hypothesis that adenosine generation limits the ability of RT totrigger anti-tumor immunity.
Wild type (WT) or BATF3−/− mice (CD103+DC-deficient) were inoculated s.c. with TSA tumor cells (day 0) and assigned to treatment with: (1) control Ab; (2) anti-CD73 Ab (100 μg) (3) RT (20 Gy); (4) RT + anti-CD73 Ab. Antibodies were administered i.p. on day 11, 14, 17 and 20. RT was given locally as single 20 Gy dose on day 12. On day 18, tumors were analyzed by flow cytometry for DC and T cell infiltration. Mice were monitored for tumor progression. DCs generated in vitro by culture of bone marrow from WT mice with Flt3L (&gt;90% CD103+) were labeled with CFSE and intravenously injected in BATF3−/− recipient mice. Tumors were harvested after 48h and analyzed by flow cytometry for infiltration of CFSE+ DCs.
In tumors of RT- but not sham-treated mice, anti-CD73 mAb increased infiltration of CD103+DCs and enhanced CD8+T cell/Treg ratio. Importantly, CD73 blockade had no anti-tumor effect by itself but significantly improved RT-induced tumor control. The therapeutic effect of RT + CD73 blockade was abrogated in BATF3−/− mice. Moreover, CD73 blockade increased recruitment of adoptively transferred CD103+DCs to irradiated tumors in BATF3−/− mice. Overall, data suggest that adenosine is a critical regulator of RT-induced immunogenicity.
Phagocytic plasticity of macrophage towards healthy self cells: inflammatory activation elicit self-attacking phenotype in macrophages lacking SIRPα-CD47 restraint
Authors: Bian Zhen , Shi Lei , Liu Yuan 
Published: January 2023

Abstract
The interaction between macrophage signal-regulatory protein alpha (SIRPα) and its ligand CD47 serves as a critical self-recognition mechanism, in which their interaction triggers inhibitory signaling via SIRPα cytoplasmic ITIMs and prevent macrophage phagocytosis toward self-cells. Despite this mechanism being suggestively imperative, CD47−/− mice demonstrate no, or only mild, macrophage phagocytosis toward self-cells, suggesting additional activation and inhibitory mechanisms controlling macrophage behavior. Studying our recently established SIRPα −/− mice, as well as CD47−/− mice, we identify that macrophage phagocytosis toward self cells requires activation by inflammatory cytokines/factors. Ex vivo phagocytosis assays demonstrate that macrophages from SIRPα −/− or CD47−/−, and even WT mice, are generally incompetent to attack self-cells, which bear no ‘eat-me’ signal and thus do not activate macrophage phagocytosis. However, treating these macrophages with IL-17, IL-6, IL-1β, TNFα and TLR agonists, but not IFNγ, dramatically initiates potent phagocytosis toward self-cells, for which only the CD47-SIRPα interaction restrains. Mechanistic studies suggest that a PKC-Syk-mediated signaling pathway, to which IL-10 conversely inhibits, is required for activating macrophages toward self cells. Application of activated SIRPα −/− macrophages in targeting CD47-expressing cancer cells also display potent eradication of B16, LLC, MC38, and EL4 cancer cells. These findings significantly expand our knowledge of macrophage phagocytic plasticity under different conditions and also provide insights into strategies for enhancing transplantation tolerance and macrophage-based cancer eradication.
VAT-associated ST2+Tregs from omentum support tolerance to peripheral tumors
Authors: Meza-Perez Selene , Silva-Sanchez Aaron , de la Luz Garcia-Hernandez Maria , Mudunuru Uma , Simpler Thomas S. , Randall Troy D. 
Published: January 2023

Abstract
Tumor metastasis to theomentum is associated with poor clinical outcomes. Although the omentum hasimmune function due to the activities of milky spots, the role of this organ inanti-tumor immunity has not been completely described.
By implanting tumor cells in mice we found that peritoneal tumors are not rejected and cause an increasein visceral adipose tissue (VAT)-associated regulatory T cells in the omentumas well as low frequency of tumor specific CD8 T cells. Additionally, peritoneal tumors impaired the immunity to secondary dermal tumors by limiting the expansion of tumor specific CD8 T cells that mediate tumor rejection.
To better understand this phenomenon, we studied the VAT-associated Tregs, which can be divided in ST2+ and ST2- (ST2 a component of the IL-33R). Particularly, VAT-associated ST2+ Tregs are not found in conventional lymphoid organs, nonetheless we find that following peritoneal tumor inoculation ST2+ Tregs are mobilized and home to secondary dermal tumors. To test whether the ST2+ Tregs were responsible for preventing secondary dermal tumors, we inoculate mice lacking ST2 (ST2−/−) with peritoneal tumors and challenged them with dermal tumors; interestingly, we found that secondary dermal tumors where rejected in ST2−/− mice whereas tumors in wild type mice continue to grow. Finally, to corroborate our findings we used a mouse strain that develops spontaneous ovarian tumors that metastasize to the omentum. By treating these mice with anti-ST2 we observed a reduction in tumor weight, decrease in ST2+Tregs and an increase in tumor specific CD8+ cells in omentum and ovaries. Together these data suggest that omental ST2+ Tregs have an important role promoting and maintaining tolerance in tumor metastasis.
Follicular CD8 T cells in autoantibody-mediated disease.
Authors: Valentine Kristen M , Davini Dan , Al-Kuhlani Mufadhal , Gravano David M , Sindi Suzanne , Hoyer Katrina K 
Published: January 2023

Abstract
The role of CD8 T cells, unlike CD4 T cells, in antibody-mediated autoimmune disease remains largely unexplored. CD8 T cells act both in a cytotoxic CD8 T cell role and helper-like role during autoimmunity. CD4 T follicular cells (Tfh), interact with B cells to promote proliferation, activation and antibody class switching. In autoimmunity, dysregulated self-reactive CD4 Tfh can promote autoantibody production. We have identified a novel population of CD8 T cells within the germinal center that resembles CD4 Tfh cells during systemic autoantibody-mediated disease. IL-2 deficient (IL-2-KO) mice develop systemic autoantibody-mediated disease including autoantibodies against red blood cells (RBC) resulting in autoimmune hemolytic anemia. We show that the absence of either CD4 or CD8 T cells prevents the development of anti-RBC antibodies and reduces lymphoproliferation in IL-2-KO mice. RNA-sequencing of IL-2-KO CD8 T cells revealed an increase in the expression of genes associated with CD4 Tfh lineages (Bcl-6, CXCR5, PD-1, ICOS and IL-21) compared with wild type (WT) CD8 T cells. Flow cytometry of expanded CXCR5+PD-1+ CD4 Tfh cells and CXCR5+PD-1+ CD8 T follicular (Tfc) cells confirms the protein expression of these Tfh markers and the production of IL-21 in IL-2-KO late stage disease (day 18–21). In vitro culture of IL-2-KO CD8 Tfc cells induced antibody class switching in WT B cells at levels similar to that induced by adjuvant-induced WT CD4 Tfh cells. In comparison to CXCR5- PD-1-CD8 T cells, CD8 Tfc cells expressed higher levels of CD40L. Our results indicate that CD8 T cells contribute to autoimmune disease in part through differentiation into follicular T cells that function to induce B cell differentiation and antibody class-switch.
Dual TCRα expression alters agonist selection, redirecting Treg-biased insulin-specific thymocytes into the conventional T cell lineage
Authors: Schuldt Nathan J , Auger Jennifer L. , Spanier Justin , Martinov Tijana , Breed Elise R. , Fife Brian T , Hogquist Kristin Ann , Binstadt Bryce Anthony 
Published: January 2023

Abstract
Despite accounting for ~10% of the T cell population in mice and humans, the role of dual TCR-expressing T cells in immunity remains poorly understood. It has been hypothesized that dual TCR T cells pose an “autoimmune hazard” by allowing self-reactive TCRs to escape negative selection. We revisited this hypothesis using the non-obese diabetic (NOD) mouse model of type 1 diabetes (T1D). We bred NOD mice hemizygous at both TCRα and β (TCRα+/− β+/−) loci, rendering them incapable of producing dual TCR T cells. We found that NOD mice lacking dual TCRα expression were resistant to developing diabetes resulting from an increased insulin-specific Treg:Tconv ratio. Furthermore, early depletion of CD25-expressing cells was sufficient to induce diabetes in single TCR T cell NOD mice demonstrating that these mice are capable of developing diabetes and that Treg cells are responsible for their observed resistance to diabetes. Further investigation uncovered a previously-unrecognized impact of dual TCRα expression on agonist selection. Specifically, we found in single TCR T cell mice that single positive (SP) thymocytes had a higher Treg:CD4 ratio indicating increased Treg commitment, that insulin-specific thymocytes exhibited higher CD5 expression suggesting increased TCR signaling, and that deletion of SP thymocytes was reduced. Altogether these data demonstrate that dual TCRα expression perturbs agonist selection of the insulin-specific thymocyte subset, resulting in skewing of this population toward the diabetogenic conventional CD4+ T cell lineage and away from the regulatory T cell lineage. Thus, dual TCR expression can promote autoimmunity by reducing the ratio of protective Treg cells to pathogenic T cells.
CD27 co-stimulation increases regulatory T cell survival and limits progression of atherosclerosis.
Authors: Winkels Holger , Meiler Svenja , Bürger Christina , Reim Sigrid , Weber Christian , Borst Jannie , Ley Klaus , Lutgens Esther , Gerdes Norbert 
Published: January 2023

Abstract
Co-stimulatory signals are important shaping adaptive immunity which is pivotal to all stages of atherosclerosis. The interaction of the co-stimulatory molecules CD27 and CD70 modulates Treg development. Furthermore, T cell activation, proliferation, and also differentiation are affected by CD27-CD70 interactions. Thus, we hypothesized that the deficiency of CD27 will lead to exacerbated atherosclerosis.
Cd27−/− mice were crossed with Apoe−/− mice. Cd27−/−Apoe−/− and littermate controls (Cd27+/+Apoe−/−) were sacrificed at the age of 18 and 28 weeks. Bone marrow of Cd27−/−Apoe−/− and littermate controls was transplanted into Apoe−/− recipient mice. Lesion size, histology and cellular composition were analyzed in the ascending aorta.
Human and murine T cells co-localize with CD27 in atherosclerotic lesions. At early stages of atherosclerosis, Cd27−/−Apoe−/− mice displayed bigger and more complex atherosclerotic lesions. Flow cytometry revealed a significant decrease in abundance of splenic and aortic Tregs and increased apoptosis of Treg in the thymus of Cd27−/−Apoe−/− mice. In contrast, at later stages of atherosclerosis (28 weeks), splenic and lesional Treg content and atherosclerotic burden were similar. Transplantation of Cd27−/−Apoe−/− bone marrow, in contrast to control, induced increased atherosclerotic plaque size and a profound pro-inflammatory plaque phenotype accompanied by reduced frequency of aortic and splenic Tregs. Furthermore, the number of circulating leukocytes was increased.
Taken together, CD27 deficiency impairs thymic Treg development which exacerbates early atherogenesis. However, the role of CD27–CD70 interactions does not affect later stages of atherosclerosis.
A new approach to examine the ability to control HCMV in patients after haploidentical stem cell transplantation
Authors: Anikeeva Nadia , Grosso Dolores , Flomenberg Neal , Sykulev Yuri 
Published: January 2023

Abstract
We utilized the recently developed approach (1) to examine HCMV-specific responses of CD8 T cells in patients who underwent haploidentical stem cell transplantation (HSCT) by analyzing the pattern of Ca2+ signaling in individual T cells assembled into a continuous monolayer after stimulation with HCMV peptide epitopes. Three groups of responding CD8+ T cells were identified: (i) T cells that rapidly develop sustained Ca2+ signaling typical for T-cell responses to a strong agonist peptide, (ii) T cells developing delayed but sustained Ca2+ signaling that is characteristic for T-cell responses to a weak agonist peptide, and (iii) T cells that revealed oscillating Ca2+ signaling indicative of suboptimal T cell activation. T cells showing sporadic and transient Ca2+ spikes were considered unresponsive. In patients whose immune system was still rebooting after transplant, HCMV was reactivated and this was mirrored by a very low fraction of HCMV-specific T cells, which mount rapid and sustained Ca2+ signaling, while the majority of responding T cells exhibited delayed or oscillating Ca2+ signaling. In contrast, a significant fraction of HCMV-specific T cells from patients who control the virus revealed rapid and sustained Ca2+ signaling. The ability to monitor patients’ immune system response to HCMV enables us to tailor treatment to better keep CMV in check.
Improved single T cell mRNA and TCR α and β chain detection using optimized chemistry and dead cell quality control for RNA-seq on the Fluidigm C1 system
Authors: egidio camila , Ooi Aik , Gonzales Michael , Brockman Joel , Chen Shuwen , Durruthy-Durruthy Robert , Rogers Clare , Ray Manisha 
Published: January 2023

Abstract
The ability to link T cell clonality with the transcriptional profile at the single-cell level provides deep insight into antigen-specific responses and the dynamics of clonal expansion and plasticity. Recent reports show that while it is possible to obtain the transcriptome profile and paired α, β TCR chain information from RNA-seq data for a single T cell, the process is technically challenging. T cells have low RNA content and RNA rapidly degrades during cell preparation for experimental setup.
We investigated two key experimental parameters, chemistry and workflow, on the gene expression profile and the ability to detect paired TCR α, β chains for single human and mouse T cells, using the C1™ system (Fluidigm®). The sensitivity and overall performance of the Clontech® SMART-Seq® v4 kit were compared with those of the SMART-Seq v1 kit. Two workflows were used to process the cells on C1: a four-hour workflow that included live/dead cell staining and imaging to allow cell viability assessment and exclusion of dead cells from downstream steps, and a two-hour workflow that excluded those steps, decreasing processing time and potentially improving RNA preservation. For both mouse and human T cells, SMART-Seq v4 chemistry outperformed v1 for reads aligned to RefSeq, bases aligned to coding and intronic sequences, and reconstruction of paired α, β TCR chains. Additionally, we observed 73% detection of paired α, β TCR chains when dead cells were excluded through imaging analysis versus only 38% detection from the shorter workflow without dead cell exclusion. We conclude that the use of an improved chemistry, coupled with a C1 workflow that allows viability assessment of single cells, improves detection of α, β TCR chains in parallel with mRNA profiling.
MiR-155 and LP-BM5 retrovirus-augmented Monocytic myeloid derived suppressor cells, are players in a profound murine immunodeficiency-causing syndrome
Authors: Green Kathy A , Green William R 
Published: January 2023

Abstract
We have shown augmentation of monocytic MDSCs (M-MDSCs) during a profound immunodeficiency causing LP-BM5 retrovirus infection of B6 mice. Enriched splenic M-MDSCs preparations in suppression assays, inhibited T-, and the much less reported B-, cell responsiveness. This inhibition of T-cell proliferation and IFN-gamma production was almost completely iNOS/NO dependent, whereas suppression of B-cell responses was only ~50% iNOS/NO dependent. To explore additional mechanism(s) in this inhibition of B-cell responsiveness, we reported that VISTA, a negative checkpoint regulator, is involved in LP-BM5 M-MDSC suppression of ex vivo B-cell responses. In light of the reported involvement of miR-155 in B-cell and myeloid cell function, we are now also evaluating M-MDSCs from 5 week LP-BM5 infected miR-155 k.o.s. Our results show miR-155 k.o. M-MDSCs are substantially less (30–50%) suppressive compared to M-MDSCs from infected Wt. mice, and at 10 w.p.i. the k.o.s exhibit a different kind of immunodeficiency: 1) spleen cells are significantly more responsive to B-cell polyclonal activators, 2) significantly less splenomegaly and 3) have fewer resident splenic M-MDSCs. In an initial experiment, involving adoptive transfer of M-MDSCs from donor LP-BM5 infected Wt. B6 to infected miR-155 k.o.s, we were able to significantly augment miR-155 k.o.: 1) splenomegaly, 2) splenic B-and T-cell immunodeficiency, and 3) the number of residing splenic M-MDSCs, to levels almost comparable to those of Wt. infected mice. These results highlight involvement of multiple and unique suppressive pathways in the under-studied area of MDSC suppression of B-cell responses, and were compatible with a role for M-MDSC in LP-BM5-induced immunodeficiency.
RNA Virus hijacks TRIM29 to suppress the host innate immune response
Authors: Xing Junji , Zhang Zhiqiang 
Published: January 2023

Abstract
The host response to RNA virus infection relies on an intrinsic innate immune response to restrict viral replication. Antiviral signaling activation of innate immunity requires strict modulation to avoid damage to the host from exacerbated cytokines and inflammation. However, the precise mechanism of how antiviral signaling activation is strictly controlled still remains obscure. Here, we found the E3 ligase TRIM29 was specifically expressed in poly I:C induced human myeloid dendritic cells (mDCs). TRIM29 was highly induced in human mDCs in response to poly I:C and played negative role to type I interferon (IFN) production. The deficiency in TRIM29 resulted in enhanced type I interferon responses to poly I:C and reovirus infection. Importantly, challenge of wildtype mice with reovirus led to lethal infection. In contrast, deletion of TRIM29 protected the mice from this developing lethality. Additionally, the heart, intestine, spleen, liver and brain of TRIM29−/− mice exhibited attenuated viral loads of reovirus. Meanwhile, the heart of TRIM29−/− mice showed elevated type I IFN production. Mechanistically, TRIM29 was shown to interact with MAVS, the key adaptor in the RNA sensing pathway, and induce its ubiquitination and degradation. Taken together, RNA virus hijacks the E3 ligase TRIM29 to shut down the host innate immune response for immune evasion.
A robust microparticle platform for a STING-targeted adjuvant that enhances both humoral and cellular immunity during vaccination
Authors: Junkins Robert D. , Johnson Brandon M. , Gallovic Matthew D. , Collier Michael A. , Watkins-Schulz Rebekah , Cheng Ning , David Clement N , McGee Charles E. , Sempowski Gregory D , Shterev Ivo , McKinnon Karen , Bachelder Eric M. , Ainslie Kristy M. , Ting Jenny P-Y 
Published: January 2023

Abstract
Most FDA-approved adjuvants for infectious agents boost humoral but not cellular immunity, and have poorly-understood mechanisms. Stimulator of interferon genes (STING) is an exciting adjuvant target due to its role in anti-viral immunity; however, a major hindrance is STING’s cytosolic localization which requires intracellular delivery of its agonists. As a result, STING agonists administered in a soluble form have elicited suboptimal immune responses. Delivery of STING agonists via particle platforms has proven a more successful strategy, but has only been assessed at doses that are likely cost-prohibitive to clinical translation for vaccines against infectious diseases. Hence, alternative platforms for the intracellular delivery of STING-agonists are needed. We evaluated the adjuvant activity of a potent STING agonist, encapsulated in acid-sensitive acetalated dextran (Ace- DEX) polymeric microparticles (MPs) which target antigen-presenting cells for intracellular release. This formulation was superior to all particle delivery systems tested, achieved protective immunity in vivo at doses of STING agonist 50-fold lower than previous reports, and elicited no observable toxicity in animals. Compared to soluble agonist, the STING agonist Ace-DEX MPs enhanced type-I interferon responses up to 1000-fold in vitro and 50-fold in vivo, caused up to 104-fold increases in antibody titers, enhanced Th1-associated responses, and expanded germinal center B cells and memory T cells. It also provided protection against a lethal influenza challenge. Thus, Ace-DEX MP-encapsulated STING agonist represents a novel and feasible vaccine adjuvant of humoral and cellular immunity.
Changes in immune gene transcription associated with phases of lactation in a model marsupial, Monodelphis domestica
Authors: Fehrenkamp Bethaney D , Miller Robert D 
Published: January 2023

Abstract
All mammalian neonates are highly dependent upon milk for nourishment and immune protection. This is especially true for marsupials, a lineage of mammals with limited placental development and an increased reliance on an extended lactation period. Most newborn marsupials do not receive passive maternal immunity in utero and therefore are entirely dependent upon factors within the milk for immune protection until capable of mounting their own responses. Early exposure to potential pathogens, prior to the development of a functional immune system, requires a complex strategy for providing immunological protection. Preliminary transcriptome sequencing results revealed two distinctive peaks in transcription of immune transfer related genes within the mammaries throughout the course of lactation. Additional time points were investigated using quantitative real-time PCR targeting specific immune markers. Lymphocyte markers (CD3E and CD79b) were elevated in a biphasic pattern of expression throughout lactation, suggestive of two waves of lymphocytes entering the mammaries throughout lactation. Immunohistochemistry analyses are underway to investigate the presence of lymphocytes within the mammaries throughout lactation and correlation with gene expression patterns. These investigations have the potential to impact the understanding of the role lactation plays in neonatal immune development from an evolutionary perspective.