bluuebunny/crossref_metadata_2025_split huggingface dataset:
TLR8/7 combined withTLR3 or TLR4 enhances DC-NK driven effector T cells
Authors: Nouri-Shirazi Mahyar A , Tamjidi Saba , Nourishirazi Erika 
Published: January 2023

Abstract

Background
Most current prophylactic vaccines confer protection primarily through humoral immunity. Indeed, aluminum salts that have been used extensively as adjuvants primarily enhance Th2-driven antibody responses. Therefore, new vaccines formulation is moving toward a careful selection of adjuvants that also elicit significant Th1 responses or CTLs. Various TLR ligands have been tested as potential adjuvants in clinical and preclinical studies with some efficacy. These studies suggest that combining more than one of TLR ligands enhance the magnitude of immune responses to cancer and infectious disease. In order to evaluate the synergic effect of TLR agonists for effective induction of cellular immunity, we investigated the effects of individual and/or combined TLR agonists on monocyte-derived DC maturation, DC-NK cross talk and ultimately T cells differentiation.

Results
Among the TLR agonists tested, we found TLR8/7 in combination with TLR3 or TLR4 synergistically optimize DC maturation and IFN-γ secretion from NK cells co-cultured with DCs. Moreover, co-culture of DC-NK-T treated with TLR8/7 and TLR3 or TLR4 increased the frequency of IFN-γ producing effector T cells when compared to Aluminum salt-treated cultures.

Conclusions
Our data suggests that enhancement of a desired cellular immunity requires a selection of TLR agonists with defined synergistic effects. NIH-NIAID 1R03AI103750-01A1
Anti-cotinine CAR-modified T cells provides a novel switchable CAR platform using cotinine-conjugated adaptor molecules
Authors: Park Hyung-Bae , Kim Hyori , Oh Yu Mi , Kim Ki-Hyun , Lee Ji Eun , Chung Junho , Choi Kyungho 
Published: January 2023

Abstract
T cells engineered with chimeric antigen receptor (CAR T cells) have been reported to provide potent anti-tumor effects in various preclinical experimental settings. Subsequently, CAR T cells against some hematologic malignancies showed favorable results in early clinical trials. However, there are various obstacles to overcome in generalizing this CAR T platform. Usually CAR T cells can target only a single tumor antigen although generation of CAR T cells takes considerable labor and expenses. Also, excessive activation of CAR T cells often leads to adverse side effects that need to be controlled. Switchable CAR T platforms using anti-hapten CAR T cells and hapten-conjugated adaptor antibodies was proposed as a novel way to target multiple antigens using single CAR T cells. Also, removal of hapten-conjugated adaptor antibodies can limit adverse side effect caused by CAR T cell recognition of the antigen. Here, we report a novel switchable CAR T platform using a chemical hapten, cotinine. Anti-cotinine CAR T cells generated using anti-cotinine single chain Fv recognized cotinine-conjugated adaptor molecules such as aptamers and antibodies. Especially, anti-cotinine CAR T cells complexed with cotinine-conjugated antibodies could recognize tumor antigens on the surface of tumor cells, which led to activation of CAR T cells and resulting cytotoxicity on tumor cells. Thus, we propose that anti-cotinine CAR T platform may provide useful tools for a novel multi-targeting CAR T therapy.
Role of necroptotic cancer cells in anti-cancer immunity
Authors: Krysko Dmitri , Aaes Tania Løve , Vandenabeele Peter 
Published: January 2023

Abstract
Successful immunogenic apoptosis in experimental cancer therapy depends on the induction of strong host anti-tumor responses. Given that tumors are often resistant to apoptosis, it is important to identify alternative molecular mechanisms that elicit immunogenic cell death.
Necroptosis is one form of regulated necrosis and is mediated by RIPK1, RIPK3, and its substrate mixed lineage kinase domain-like and has been reported to contribute to inflammation under pathological conditions. Necroptosis is commonly induced by ligand-dependent activation of certain members of the TNFR and TLR families. To circumvent the use of cell death-inducing ligands or other immune active molecules that would not allow us to evaluate the immune response resulting uniquely from necroptosis induction, we opted for the use of inducible Tet-On systems for the expression of the downstream necroptotic effector proteins, allowing us to induce necroptosis independently of any receptor activation. In our genetic model a direct dimerization of FADD combined with inducible expression of RIPK3 promotes necroptosis. We report that necroptotic cancer cells release DAMPs and promote maturation of dendritic cells, the cross-priming of cytotoxic T cells, and the production of IFN-γ in response to tumor antigen stimulation. Using both FADD-dependent and FADD-independent RIPK3 induction systems, we demonstrate the efficient vaccination potential of immunogenic necroptotic cells. Our study broadens the current concept of immunogenic cell death and opens doors for the development of new strategies in cancer therapy.
Dissecting the role of Phosphotidylinositol 3-Kinase (PI3K) δ in the germinal center reaction through a mouse model of hyperactivated Pik3cd
Authors: Preite Silvia , Cannons Jennifer L. , Gomez-Rodriguez Julio , Kapnick Senta M. , Handon Robin , Reilley Julie , Notarangelo Luigi , Uzel Gulbu , Schwartzberg Pamela L. 
Published: January 2023

Abstract
The catalytic p110δ subunit of PI3K drives AKT pathways orchestrating activation and differentiation of T and B cells. Patients with gain of function mutations in Pik3cd (encoding for p110δ) exhibit a primary immunodeficiency characterized by lymphopenia, lymphadenopathy, recurrent and chronic infections, and occasionally lymphoma. They have reduced circulating naïve and increased effector T cell numbers, fewer class switched memory B cells and inefficient responses to vaccination. Nonetheless, in patient peripheral blood we find increased frequencies of T follicular helper (Tfh) cells. We have generated a mouse model expressing constitutively active p110δ that recapitulates many features of the human disease. Pik3cdMut mice, both in steady state condition and after immunization, display increased Tfh and germinal center (GC) B cell numbers in secondary lymphoid organs. Moreover, naïve mutant transgenic T cells (OT-II) preferentially acquire Tfh cell phenotype when adoptively transferred to WT mice, indicative of a T cell intrinsic defect. Nonetheless, despite increased Tfh and GC B cells, antigen specific B cell responses are reduced, particularly antigen-specific IgG. In contrast, we find the appearance of autoantibodies in Pik3cdMut mice. We further find that mutant OT-2 cells differentiate to Tfh cells independently from ICOS-ICOSL interactions. In line with this, mutant T cells show higher phosphorylation and inactivation of FoxO1, potent inhibitor of Tfh cell differentiation, that is phosphorylated and inhibited by AKT activation downstream of ICOS. Our results demonstrate that hyperactivated PI3Kδ leads to increased generation of Tfh cells and germinal centers, yet with defective in vivo function.
MicroRNA-21 in Natural Killer T cell α-galactosylceramide activation
Authors: Frias Adolfo B , Buechel Heather Marie , Piccirillo Ann Renee , D’Cruz Louise M 
Published: January 2023

Abstract
Natural Killer T (NKT) cells are innate-like lymphocytes that rapidly produce cytokines upon activation. However, following primary stimulation with the potent ligand α-galactosylceramide (αGC), NKT cells become hyporesponsive, defined as a severe reduction in effector cytokines upon re-stimulation. The purpose of this project is to investigate the role of microRNAs (miRNAs) during NKT cell transition from effector response, expansion and hyporesponsiveness. MiRNAs are ~22bp non-coding RNAs that associate with the RNA-induced silencing complex (RISC) to inhibit the translation of target transcripts. We performed a Nanostring microRNA analysis to determine global expression of microRNAs in NKT cells after α-GC stimulation. MicroRNA-21 (miR-21) was found to be highly upregulated in splenic NKT cells at 3-hours, 3-day, and 30-days post-activation compared to unstimulated controls. Our preliminary data show that MiR-21KO/KO splenic and liver NKT cells have a lower frequency at day-3 compared to WT-littermate controls. We hypothesize that miR-21 regulates the NKT cell transition from activation to hyporesponsiveness and that miR-21KO/KO NKT cells exhibit an interrupted hyporesponsive state.
Effector CD4+ T cells can convert to a CD8+, MHCII-restricted T cell lineage
Authors: Wong Elizabeth , Nyanhete Tinashe , Frisbee Alyse , Ni Qingshan , Zhang Baojun , Payne Tamika , Liu Siqi , Li Chaoran , Chen Liang , Zhuang Yuan , Wan Ying , He You-Wen , Moody Tony , Tomaras Georgia D , Li Qi-Jing 
Published: January 2023

Abstract
CD4+ and CD8+ T cells are conventionally considered as separate lineages characterized by distinct surface markers, function and MHC restriction. However, CD4+ T cells have a recently-appreciated capacity to acquire CD8+ T cell-like characteristics upon TCR-mediated activation. We have identified a novel, CD4+ T cell-derived effector population in mice that is CD4−CD8αβ+ and MHCII-restricted. These cells express a unique lineage-intermediate transcriptional program (ThPOKlo-int Runx3hi-int), upregulate granzyme B/IFN-γ expression and exhibit antigen-specific cytotoxicity. Following acute infection with LCMV Armstrong in wild-type mice or with Listeria monocytogenes in MHCII-restricted, TCR-transgenic mice, we have used CD4+ T cell lineage tracing, MHCII tetramer staining and TCR repertoire sequencing techniques to demonstrate that ex-CD4+ T cell responses can be induced in vivo and originate from bona fide CD4+ T cells. We also report ex-CD4+-like T cells in human HIV patients that are CD8+ and exhibit MHCII-blockable antiviral activity. This population is diminished in elite controller patients upon loss of virus control, indicating that it may contribute to protection against HIV. Finally, we have generated preliminary evidence that ex-CD4+ T cell generation is controlled by an autophagy-related mechanism. CD4+ T cell-specific deficiency in either of the autophagy regulators Vps34 or Atg7 results in dramatic loss of CD4 expression upon antigen stimulation, concomitant with skewing towards the CD8+ transcriptional program in Vps34-deficient cells. We propose that the ex-CD4+ T cell lineage dramatically extends the paradigm of αβ T cell lineage plasticity and may indicate new antiviral vaccine strategies.
Lipopolysaccharide-induced CD300b receptor binding to Toll-like Receptor 4 alters signaling to drive cytokine responses that enhance septic shock.
Authors: Voss Oliver H. , Krzewski Konrad , Coligan John E. 
Published: January 2023

Abstract
LPS, present in gram-negative bacteria membranes, causes strong immune responses following detection by TLR4 on immune cells. Excess immune cell activation resulting from severe infection initiates a pro-inflammatory cytokine storm that leads to a more severe immunopathology, like septic shock, and subsequently, death. Intriguingly, Cd300b−/− mice were observed to be more resistant to septic shock than WT mice, however the mechanism responsible for this phenomenon remained elusive. Here, we identify LPS as a ligand for CD300b, and demonstrate that CD300b-expressing macrophages (Mφ) are key cells augmenting septic shock.
We show that CD300b and DAP12, associate with TLR4/CD14 upon LPS binding, resulting in the dissociation of MyD88/TIRAP from the complex and the recruitment and activation of Syk and PI3K. This leads to a PI3K/AKT-dependent inhibition of the MEK1/2-ERK1/2-NFκB pathway, subsequently resulting in a reduced IL-10 production by Mφ. In addition, CD300b also enhances TLR4/CD14-TRIF-IRF3 signaling, resulting in elevated IFN-β levels. Compared to WT mice, Cd300b−/− mice demonstrate decreased serum levels of pro-inflammatory cytokines and elevated levels of the IL-10 anti-inflammatory cytokine. Neutralization of IL-10 in Cd300b−/− mice diminishes their survival advantage over WT animals. Importantly, we demonstrate that an anti-CD300b Ab treatment suppresses pro-inflammatory cytokine and enhances anti-IL-10 production in Mφ, thus reducing the mortality of septic mice.
These findings provide evidence for a previously unappreciated regulation of TLR4 signaling by CD300b in myeloid cells, and highlight CD300b as a potential therapeutic target for clinical intervention to manage septic shock in humans.
Influence of aging on the phenotype and function of alveolar macrophages
Authors: Lafuse William P , Rajaram Murugesan VS , Curry Heather M , Nealer Connor , Canan Cynthia H , Moliva Juan I , Torrelles Jordi B , Turner Joanne , Schlesinger Larry S 
Published: January 2023

Abstract
Aging is associated with a decline in immune function (immunosenescence), resulting in increased susceptibility to infectious diseases, malignancies, and autoimmunity. However, the immunosenescence paradigm does not explain increased circulating pro-inflammatory cytokines and increased tissue expression of pro-inflammatory genes with aging. These observations have led to a second paradigm, termed inflammaging, in which chronic, low grade inflammation develops with increasing age. Published studies have shown that inflammaging also occurs in the lung. However, the influence of inflammaging on alveolar macrophage (AM) phenotype and function is largely unexplored. In this study, we examined the phenotype and function of AMs isolated from young (3 months) and old (18 months) C57BL/6 mice. Phenotypic analysis of AMs by flow cytometry showed that in young mice there is a major AM population that is CD11c+CD11b− and a minor population that is CD11c+CD11b+. In young mice, this minor population represents 1.3% of AMs. However, in old mice the percentage of this minor population is significantly higher (4.9%, p &lt;0.0001). The CD11c+CD11b+ AMs have higher expression of several cell surface markers, e.g. MHC II and CD64, indicating that these AMs are in a more prime or activated state. Functional studies of these AMs stimulated with LPS, PamCSK4, or infected with Mycobacterium tuberculosis show significant spontaneous production of IFN-β, CCL2, and IL-10 that is prolonged in old mice and increased further by stimulation with ligands or infection. Our studies indicate that inflammaging in the lung primes AMs, resulting in robust production of key cytokines upon stimulation that potentially alter host susceptibility.
Feeding immunity: The regulation of circadian immune responses by mealtimes
Authors: Geiger Sarah Sofia , Traba Javier , Sack Michael N. , O’Neill Luke A. J. , Siegel Richard M. 
Published: January 2023

Abstract
Both, mice and humans have a striking circadian variability in the responsiveness to innate immune stimuli and in mice, susceptibility to sepsis induced by acute stimulation of the immune system with LPS; and disruption of the circadian rhythm is associated with inflammatory diseases, such as cancer and obesity. The intrinsic cellular clock exists in immune cells and can control some aspects of innate immunity, but what extrinsic signals, termed ‘zeitgebers’ or time-givers, entrain the circadian behavior of immune response are not known. We have investigated the relative contributions of light and the feeding cycle to innate immune responses by dissociating the light cycle and access to food through time restricted feeding (TRF), allowing access to food either during the dark - and natural feeding time of mice - or the light phase. Using RNA profiling we found that the circadian clock in the liver is entirely programmed by food, whereas in the spleen, mainly light influences the expression of clock genes. Strikingly, in this setting susceptibility to LPS-induced morbidity was entrained by the food, not the light cycle, suggesting that food is the ‘zeitgeber’ for circadian susceptibility to LPS-induced sepsis. While the exact mechanism is under thorough investigation, daily mortality patterns following LPS stimulation could not be attributed to corticosteroids or ketone bodies, both of which possess anti-inflammatory properties and exhibit strong food dependency. Understanding the interface between nutrition and circadian rhythmicity may provide new insights with which to design strategies to prevent and improve metabolic and inflammatory diseases in individuals at risk.
Palmitic acid promotes t-bet and gata-3 mRNA degradation in NKT cells via regulated IRE1a-dependent decay
Authors: ko jaesung , Koh Jae Moon , So Jae-Seon , Chung Doo Hyun 
Published: January 2023

Abstract
Long chain fatty acids (LCFAs) exert pro-inflammatory effects in vivo. However, little is known regarding the effect of LCFAs on invariant (i) NKT cell functions. Here, we report an inhibitory effect of saturated LCFAs on transcription factors in iNKT cell. Among the saturated LCFAs, palmitic acid (PA) specifically inhibited IL-4 and IFN-g production and reduced gata-3 and t-bet transcript levels in iNKT cells during TCR-mediated activation. In iNKT cells, PA was localized and induced dilation in the endoplasmic reticulum and increased the mRNA levels of downstream molecules of IRE1 RNase. Moreover, PA increased the degradation rates of gata-3 and t-bet mRNA, which were restored by IRE1 inhibition, indicating that gata-3 and t-bet are cleaved via regulated IRE1a-dependent decay (RIDD). A PA-rich diet suppressed IL-4 and IFN-g production by iNKT cells in wildtype mice, thereby attenuating arthritis. This study demonstrates that a saturated LCFA induced RIDD-mediated t-bet and gata-3 mRNA degradation in iNKT cells, thereby suppressing cytokine production.
Investigating modulation of inflammation in atherosclerosis by complement protein C1q
Authors: Pardo Paulina Valeria , Flores Abraham , Wong Ashley J. , Ho Minh-Minh , Fraser Deborah A. 
Published: January 2023

Abstract
Atherosclerosis is a chronic inflammatory disease characterized by the accumulation of fatty streaks within arterial walls. In hyperlipidemic conditions, cholesterol accumulates in macrophages disrupting cholesterol trafficking and leading to an inflammatory environment. C1q plays a dual role in atherosclerosis. Activation of complement by C1q exacerbates disease by release of pro-inflammatory activation fragments C3a and C5a. However, we have shown in vitro that C1q has numerous protective roles in atherosclerosis such as improving macrophage cholesterol ingestion and efflux, and polarizing macrophages towards a resolving, anti-inflammatory phenotype. Here we investigate if C1q programs similar macrophage inflammatory responses in vivo. We hypothesize that C1q polarizes macrophages to a protective phenotype in the early stages of atherosclerosis by increasing anti-inflammatory cytokines and decreasing pro-inflammatory cytokines, leading to a reduction in disease markers such as cholesterol and triglyceride levels and atherosclerotic lesion size. To investigate, atherosclerosis model, LDLr−/−, mice that are C1q sufficient or deficient were fed a high fat Western diet for 12 weeks. Changes in plasma cyto/chemokine levels are measured by luminex multiplex assay. Cholesterol and triglyceride levels are measured by colorimetric assay. Atherosclerotic lesions are measured in H&E stained aortic sinus. Preliminary data suggests that C1q modulates disease markers as predicted. By understanding the role of C1q in atherosclerosis it may be possible to design improved treatment strategies.
A novel role for C5a in regulating B1 cell homeostasis and function
Authors: Karsten Christian M , Bröker Katharina , Figge Julia , Manz Rudolf A , Köhl Jörg 
Published: January 2023

Abstract
B1 cells are innate-like B cells producing natural IgM (nIgM) antibodies without previous antigen exposure. They ensure first line humoral defense and long-lasting immunity against T-cell independent antigens found in encapsulated bacteria. B-1 cells are predominantly located inside body cavities and in lower numbers in spleen and bone marrow. Previous findings suggest that their egress is contolled through several integrins and the chemokines CXCL12 and CXCL13. B1 cell need to migrate from the peritoneal cavity to the spleen to become nIgM producing plasma cells. The role of complement in B1 cell homeostasis and migration has not been explored. Here, we determined the impact of C5a and the C5a receptors (C5aR) 1 and 2 on B1 cell biology. We found significantly reduced B1 cell numbers in the peritoneal cavity of C5aR1−/− or C5aR2−/− as compared with wildtype mice that were associated with increased B1 cell numbers in the spleen. Consequently, C5aR1−/− mice had elevated serum levels of nIgM against phosphorylcholine and different polysaccharides of S. pneumoniae under steady state conditions. Using a GFP-C5aR1 reporter mouse, we further demonstrate that C5aR1 expression on B1 cells is upregulated whereas the CXCL13 receptor CXCR5, CXCR4, IgM and aM (CD11b) integrin expression is downregulated before B1 cells leave the peritoneal cavity upon TLR2 stimulation in vivo. In line with this observation, we found that in C5aR1−/− mice, TLR2-driven B1 cell egress was diminished, as B1 cell numbers in C5aR1−/− mice were unchanged whereas they markedly dropped in wildtype mice. Thus, our data suggest a novel role for C5a in the regulation of B1 homeostasis in the peritoneal cavity, nIgM production and TLR-driven egress from the peritoneum into the spleen.
Obesity-induced changes to the immune landscape in human renal cell carcinoma subjects
Authors: Norian Lyse A , Norris Katlyn E , Wald Gal , Thomas Lewis J , Bertrand Laura A , Buchta Claire M , Boi Shannon K , Nepple Kenneth G , Brown James A 
Published: January 2023

Abstract
Immunotherapy for advanced renal cell carcinoma (RCC) has shown tremendous promise. However, clinical responses remain &lt;30% and the reasons are unclear. Obesity affects ~35% of U.S. adults and is a major risk factor for RCC. We reported previously that obesity impairs immunotherapeutic efficacy in a murine renal cancer model, due to increased myeloid suppressor cells and weakened effector CD8 T cell responses. Based on these results, we hypothesized that obesity would also induce detrimental shifts in the immune landscape in treatment-naive subjects with confirmed RCC. In an IRB-approved study, we consented 86 RCC subjects with obesity (mean BMI = 38.4) and 77 normal to over-weight RCC subjects (mean BMI = 25.0), plus tumor-free controls, into a prospective study to examine intra-tumoral and systemic leukocytes by multi-parameter flow cytometry, plasma proteins via multiplex, and tumor gene expression by Nanostring. Numerous systemic cell populations (ex: CD45RO+ PD1+ CD4 or CD8 T cells, and HLA-DR- CD14− CD11b+MDSC, all p&gt;0.05) were unaltered by obesity in RCC subjects. Peripheral CD14+ inflammatory monocytes and Foxp3+ Tregs were decreased in RCC subjects with obesity (both p&lt; 0.05). However, obesity altered plasma proteins to favor RCC progression, with increased angiopoietin, VEGF-A and –C, and IL-8 (all p&lt; 0.05). Within renal tumors, obesity did not alter CD4 or CD8 T cell percentages or phenotypes (CD45RO, PD1, CD56). Nanostring analysis of renal tumors revealed multiple obesity-related changes in pro-tumorigenic genes. Thus, obesity in RCC subjects had surprisingly few effects on cellular immunity, but multiple effects on protein mediators that may shift the overall immune landscape toward one that favors tumor progression.