bluuebunny/crossref_metadata_2025 huggingface dataset:
Targeting both IL-17 and CXCR2 in the gut leads to the suppression of arthritis and joint inflammation in the autoimmune arthritis mice driven by a mycoplasma superantigen
Authors: Mu Hong-Hua
Published: January 2023

Individuals with autoimmune susceptible genes infected with environmental microbial agents might amplify an autoimmune condition by either exacerbating an ongoing disorder, including a relapse, or by leading to chronic progressive disease. Previously we have established an autoimmune arthritis model by infecting mice with the Mycoplasma arthriditis. M. arthritidis produces a potent superantigen (SAg) MAM, which was found to be responsible for the arthritis resembling human RA. In the present study we found that MAM can also activate intestinal immune cells in mice. Because the mechanisms leading from systemic autoimmunity to joint-specific autoimmune disease are unknown, we questioned if the autoimmunity initiates at mucosal sites. To address this question, by injecting mice with collagen II (CII), or with CII+MAM. We found that IL-17 and chemokine receptor CXCR2 were highly expressed within the intestinal lamina propria (LP) in CII+MAM mice but not in traditional CIA mice. In CII+MAM mice, overexpression of IL-17 and CXCR2 in the gut was shown to be strongly related to the arthritis severity. Massive infiltration of inflammatory leukocytes and the upregulation of chemokines, CXCL1, CXCL2, CXCL12, CCL2 and CCL5 in the joint tissues were detected and associated with the increased IL-17 and CXCR2 in the intestinal LP following CII+MAM immunization. Suppression of both IL-17 and CXCR2 at same time by blocking antibodies greatly alleviated arthritis. We also found that neither IL-17 nor CXCR2 alone is sufficient to trigger arthritis. Thus the results suggest that the excessive IL-17 and CXCR2 in the gut are related to the exacerbation of arthritis in a manner by which the inflammatory signals can be connected between the gut and the joints.
CD6 modulation ameliorates immune complex-mediated glomerulonephritis
Authors: Chalmers Samantha A. , Garcia Sayra J , Herlitz Leal , Ampudia Jeanette , Ng Cherie , Connelly Stephen , Putterman Chaim
Published: January 2023

Lupus nephritis (LN) is a serious end organ complication of systemic lupus erythematosus (SLE) in which T cells are thought to play an essential role. CD6 is a co-stimulatory receptor on T cells, that binds to activated leukocyte cell adhesion molecule (ALCAM), a ligand expressed on antigen presentation cells and epithelial and endothelial tissues. The CD6-ALCAM pathway plays an integral role in modulating T cell activation and trafficking, and increased levels of CD6 are associated with pathogenic T cell responses. To assess the role of the CD6-ALCAM pathway in LN pathogenesis, we tested a monoclonal antibody against CD6 in a short-term, validated, inducible murine model of lupus nephritis known as nephrotoxic serum nephritis (NTN). NTN mice were treated 3× per week with an anti-CD6 mAb (10D12, 60ug/dose, n=23) or with vehicle control (n=23). Healthy mice were also included as a control (n=12). Mice treated with the anti-CD6 mAb displayed decreased levels of proteinuria (p<0.001) and significantly improved BUN levels (p<0.01) compared to vehicle control mice. Histology also significantly improved with anti-CD6 treatment (p<0.05). RT-PCR revealed significantly decreased levels of VCAM and RANTES in the kidneys of treated mice, while anti-inflammatory IL-10 was increased, compared to vehicle control mice. Flow cytometry analysis indicated decreased accumulation of both renal-infiltrating activated T cells (CD4+CD25+CD69+, p <0.01) inflammatory macrophages (p<0.05). Overall, these results indicate that the CD6-ALCAM pathway is an important driver of inflammation and pathology in LN and, thus, a promising therapeutic option that is more selective than the immunosuppressive therapies currently offered.
Gata3 regulates the pathogenicity of Tbet-expressing Th17 cells in EAE
Authors: Butcher Matthew J , Gurram Rama Krishna , Chen Xi , Zhu Jinfang (Jeff)
Published: January 2023

Tbet+RORgt+ Th17 cells are thought to be key players in the pathology of experimental autoimmune encephalomyelitis (EAE) and GM-CSF production has been proposed to be one mechanism through which Th17 are encephalitic. However, the mechanisms through which Csf2 expression is regulated are unclear. Based on prior work highlighting a regulatory role for Gata3 in NKp46+ ILC3s, we hypothesized that Gata3 might play an analogous role in Tbet+ Th17 cells during EAE. Profiling Gata3 expression in MOG-immunized mice revealed that Th17 cells dynamically express Gata3 during their differentiation and maintain low levels thereafter. Therefore to determine how Gata3 might affect the pathogenicity of Tbet+ Th17 cells, we utilized tamoxifen inducible Gata3 knockout (Cre-ERT2 Gata3Fl/Fl) mice as donors in a transfer EAE model. In this model, a post-immunization deletion of Gata3 resulted in a failure to generate EAE symptoms due to a five-fold reduction in GM-CSF production, despite a normal Th17 and Tbet+ Th17 cell responses. To assess how Gata3 might affect the expression of Csf2 in a cell-intrinsic manner, we isolated Cd45.1/Cd45.2 and Cre-ERT2 Gata3Fl/Fl CD4+ effector T cells from the CNS of tamoxifen or vehicle treated co-transfer recipient mice for RNAseq. Interestingly, Gata3-deficient CD4+ T cells displayed stark reductions in Bhlhe40, Egr2, and Csf2 expression in a cell-intrinsic manner, which was confirmed in follow-up experiments. Taken together, our data suggests that Gata3 promotes the pathogenicity of Tbet+ Th17 cells in EAE, possibly via regulation of Bhlhe40 and GM-CSF expression.

This research was supported by the Intramural Research Program of NIAID, NIH.
The gut as a potential licensing site for central nervous system (CNS)-specific autoimmune lymphocytes
Authors: Salvador Ryan S. , Horai Reiko , Jittayasothorn Yingyos , Tang Jihong , Gery Igal , Caspi Rachel R
Published: January 2023

Autoimmune diseases of the CNS, experimental autoimmune encephalomyelitis (EAE) and experimental autoimmune uveoretinitis (EAU) can be induced by adoptive transfer (AT) of activated brain- or retina-specific T cells. The transferred T cells rest for several days in the recipient’s spleen or lungs, where they proliferate and differentiate to a migratory phenotype before invading the CNS, a process termed “licensing” for pathogenicity (PMID: 27986906). Our previous data in a spontaneous uveitis model in R161H mice bearing a retina-specific TCR indicated that the gut may serve as a priming site for uveitogenic T cells through molecular mimicry of gut flora (PMID: 26238373). We now asked if the gut might also serve as a licensing site for autopathogenic T cells.

Naive B10.RIII mice received AT of allotype-marked, activated R161H T cells and were monitored for EAU. Donor cells in tissues were detected by flow cytometry. Three days after AT, donor T cells could be detected in spleen, lung and gut, including the mesenteric lymph nodes and lamina propria of the large and small intestine. Similarly to lung and spleen, R161H T cells in the gut showed signs of proliferation by CFSE dilution. This was not altered by antibiotic treatment of recipients before AT to eliminate gut flora, suggesting that microbiota is not needed to retain effector T cells in the gut.

We hypothesize that primed retina-specific T cells can receive licensing signals in the gut. Since in spontaneous uveitis they may also be primed there, this could be of relevance to a disease situation. Further characterization of phenotypic and transcriptomic changes of “licensed” R161H cells retrieved from the gut may help to elucidate the local signals important for licensing at this site.
Sjögren’s syndrome minor salivary gland CD4+ T cells associate with glandular disease features and have a germinal center T follicular helper transcriptional profile
Authors: Farris A. Darise , Joachims Michelle L , Leehan Kerry M , Dozmorov Mikhail G , Georgescu Constantin , Pan Zijian , Rasmussen Astrid , Radfar Lida , Lewis David M , Stone Donald U , Grundahl Kiely , Scofield R. Hal , Lessard Christopher J , Wren Jonathan D , Thompson Linda F , Sivils Kathy L , Moore Jacen S
Published: January 2023

Objective

To assess the types of salivary gland (SG) T cells contributing to Sjögren’s syndrome (SS), we evaluated SG T cell subtypes for association with disease features and compared the SG CD4+ memory T cell transcriptomes of primary SS (pSS) and sicca subjects not meeting SS criteria (nSS).

Methods

SG biopsies of pSS and nSS subjects were evaluated for proportions (n=51 pSS, n=69 nSS) and absolute numbers (n=35 pSS, n=57 nSS) of CD4+ and CD8+ T cells. SG memory CD4+ T cells from focus score positive pSS (n=17) and focus score negative nSS subjects (n=15) were evaluated for gene expression by microarray. Differentially-expressed genes were identified and gene set enrichment and pathways analyses were performed.

Results

CD4+CD45RA− T cell frequencies were increased in pSS compared to nSS subjects (33.2 vs. 22.2%, p<0.0001), while CD8+CD45RA− T cells were decreased (38.5 vs. 46.0%, p=0.0014). SG fibrosis positively correlated with numbers of memory T cells, regardless of phenotype. Proportions of SG CD4+CD45RA− T cells correlated with focus score (r=0.43, p<0.0001), corneal damage (r=0.43, p<0.0001), and serum Ro antibodies (r=0.40, p<0.0001). Differentially expressed genes in CD4+CD45RA− cells of pSS versus nSS subjects indicated a T follicular helper (Tfh) profile, increased homing and increased cellular interactions. Predicted upstream drivers of the Tfh signature included TCR, TNF, TGF-β1, IL-4 and IL-21.

Conclusions

The proportions and numbers of SG memory CD4+ T cells associate with key SS features, consistent with a central role in disease pathogenesis. SG memory CD4+ T cells express a germinal center Tfh profile.
Zfp36 family members redundantly protect against T cell-mediated autoinflammation and premature mortality
Authors: Cook Melissa Erin , Bradstreet Tara R , Santeford Andrea , Kim Jongshim , Webber Ashlee , Schwarzkopf Elizabeth A , Jarjour Nicholas N , Lin Chih-Chung , Stumpo Deborah J , Apte Rajendra S , Blackshear Perry J , Edelson Brian T
Published: January 2023

Cytokine production must be tightly regulated in order to prevent auto-inflammatory diseases. The zinc finger 36 (Zfp36) family of RNA-binding proteins, including Zfp36, Zfp36l1, and Zfp36l2, are known to negatively regulate mRNA stability or translation of many transcripts, including cytokines. Polymorphisms in ZFP36L1 and ZFP36L2 have been identified in GWAS studies of a variety of human autoimmune diseases, necessitating understanding the functions of these genes. While there are reports of all three family members controlling cytokine production from T cells, delineating the exact functions of these genes has been challenging due to spontaneous phenotypes or mortality upon global deletion of single genes and potential redundancy in their functions. To overcome this, we generated Cd4-Cre+ Zfp36fl/fl Zfp36l1fl/fl Zfp36l2fl/fl mice. Only upon triple deletion, but not individual or various paired deletions, do mice spontaneously develop an inflammatory disease characterized by early mortality and immune cell infiltration into various organs, including the central nervous system, kidneys, and liver. These mice have drastically elevated levels of many cytokines in their sera. Our findings demonstrate a novel redundancy of the Zfp36 family members in regulating T cell homeostasis and suppressing cytokine-driven inflammation. We are currently investigating the specific mechanisms and mRNA targets that contribute to this phenotype and whether disease is primarily driven by CD4+ or CD8+ T cells. Understanding the individual and redundant functions of the Zfp36 family members may lead to opportunities to target them for suppression of T cell-driven autoimmunity or for activating anti-tumor or anti-pathogen T cell responses.
Aged T cells shape neuronal autoimmune disease in a model of late-onset multiple sclerosis (Supported by NIH/NIAID R01AI121147)
Authors: Wang Weikan , Oh Jiyoung , Thomas Rachel , Su Dongming
Published: January 2023

Although multiple sclerosis (MS), a T cell-mediated autoimmune demyelinating disease, mainly develops in young people, late-onset has also been observed in aged people. The aged T cell immune system is different from the young. One of characteristics is disrupted thymocyte negative selection, and relatively enhanced thymic regulatory T (tTreg) cell generation and accumulated peripheral Treg (pTreg) cells. The relatively increased polyclonal Treg cells potentially shape the MS disease courses, and may even prevent remission stages of this disease. Using the mouse model experimental autoimmune encephalomyelitis (EAE) of human MS, we determined that a late-onset EAE has two types of disease courses. I: Onset of EAE in the aged group was later than the young group, but after onset, the disease became more severe than in their young counterparts; II: Alternatively, onset of EAE in the aged group never became severe until after a second antigen challenge was given, causing the symptoms to appear and progress to severe disease much faster than in their young counterparts. We also found that this late-onset of EAE was associated with a disparate distribution of Treg cells in lymph nodes versus in the central nervous system (CNS) during the disease. The aged mice had a high proportion of pTreg cells in the lymph nodes, but a low proportion in the CNS, compared to their young counterparts. We believe that accumulated pTreg cells in the elderly, which is one of hallmarks of aged T cell immune system, potentially results in their residing in the blood-brain barrier and/or choroid plexus, which hampers trafficking of monocytes and/or antigen-specific Treg cells into the CNS. Thus, it is detrimental for a CNS repairing processes in neuronal inflammation.
The Role of Aire in the selection of Regulatory T cells in Diabetes
Authors: Bridge Jennifer Anne , Balolong Jared , Dong Shen , Cramer Nathan , Mowery Cody , Crawford Frances , Ye Jimmie , Kappler John W , Bluestone Jeffrey A , Anderson Mark S
Published: January 2023

In autoimmune diseases like T1D, it is proposed that failure in central tolerance mechanisms towards pancreatic islet self-antigens leads to the escape of pathogenic clones and loss of critical immunosuppressive antigen-specific Tregs. The importance of Aire in maintaining central tolerance against self-reactive antigens is evidenced by the multiorgan autoimmune disease seen in individuals with mutation the Aire gene. When Aire-expression is specifically ablated in mouse mTECs, there is a dramatic increase in the number of self-reactive T cells that escape negative selection, in addition, a dramatic loss of Tregs. Utilizing novel high-throughput platforms and single-cell DNA barcoding technology has allowed us to assess several thousand CD4+ T-cell clones. The use of FoxP3-GFP reporter mice permits us to identify individual Tregs that have undergone Aire-dependent selection in the thymus and compare the TCR repertoire to mice that have had Aire specifically ablated from the mTECs. Identification of unique clones and gene signatures within our large datasets has allowed us to start to qualitatively assess the functional role of the identified TCRs within the model of T1D. We have begun comparisons of our extensive datasets to insulin-specific TCRs and TCRs that we have previously identified from diabetic animals. In addition, the peptide specificities of the identified TCRs are being evaluated using Baculovirus MHC-II peptide libraries. Our proposed study aims to expand our knowledge on TCR repertoire of Aire-dependent Tregs and the antigen-specificity in T1D, improving our understanding of the respective function of thymic-derived antigen-specific Treg suppression in peripheral tissues associated with autoimmune T1D.
Inflammation associated weight cycling contributes to an obesogenic memory phenotype in adipose tissue
Authors: McDonald Sierra J , Sougiannis Alexander T , Vanderveen Brandon , Cranford Taryn , Enos Reilly T , Velazquez Kandy , Chatzistamou Ioulia , Fan Daping , Murphy Angela E
Published: January 2023

Energy expenditure through physical activity and dieting promotes weight loss. However, sustaining a healthy weight can be challenging and often a pattern known as weight cycling emerges. We evaluated the inflammatory status of mice undergoing weight cycling. Male C57BL/6 mice were randomized into four groups (n=10/group): low fat diet (LFD) for 32 weeks (LFD), high fat diet (HFD) for 32 weeks (HFD), LFD for 28 weeks then changed to HFD for 4 weeks (LFD->H), HFD for 21 weeks then changed to LFD for 7 weeks and then changed to HFD for 4 weeks (HFD->L->H). LFD->H and HFD->L->H mice did not significantly differ in body weight, body fat weight, or body fat percentage, but were significantly higher than LFD mice while remaining lower than HFD mice (p<0.05). In addition, HFD->L->H mice had smaller adipocytes of the epididymal fat pad compared to HFD and LFD->H mice which both had larger adipocytes (p<0.05) compared to LFD. TNFα and IFNγ were higher (p<0.05) in the epididymal fat pad of HFD->L->H mice compared to LFD and LFD->H mice. This was consistent with more M1-type macrophages (p<0.05) and cytotoxic T-cells (p<0.05) as indicated by ITGAX and CD8a gene expression, respectively. However, MCP1 was significantly lower (p<0.05) in HFD->L->H mice compared to LFD->H. Liver histopathological and protein analysis revealed HFD->L->H mice had greater NASH and NAS (p<0.05) scores, increased total STAT3 and NF-kB (p<0.05) compared to LFD controls. Taken together, weight cycling can lead to aberrations in the storage of triglycerides when challenged with an increased caloric intake. This is further supported by an increase in adipose tissue inflammation in weight cycling mice which is likely contributing to the observed obesogenic memory phenotype.
Imaging neuroinflammation in murine endotoxemia: a dual-tracer microPET evaluation
Authors: Turecki Lauren , Palandira Santhoshi P , Liu Hui , Addorisio Meghan , Tsaava Tea , Chang Eric , Ma Yilong , Carrion Joseph , Tracey Kevin J , Pavlov Valentin A
Published: January 2023

Brain dysfunction, including neuroinflammation, metabolic changes, and cognitive impairment has been reported in sepsis and other disorders “traditionally” characterized by peripheral immune and metabolic dysregulation (Annu Rev Immunol, 2018, 36:783–812). In this context, non-invasive evaluation of neuroinflammation is important for strategizing new diagnostic and therapeutic approaches targeting the brain. We utilized Micro Positron Emission Tomography (microPET) with [18F]Fluoro-2-deoxy-2-D-glucose (18FDG) and 11C-Peripheral Benzodiazepine Receptor ([11C]PBR) to evaluate brain metabolic alterations and microglia activation (indicating neuroinflammation) during murine endotoxemia. Lipopolysaccharide (LPS, endotoxin) (2 mg/kg) or saline was injected (i.p.) in male C57Bl/6 mice 6 hrs prior to microPET imaging. [11C] PBR28 (~0.5 mCi) was injected via the tail vein followed by 60 mins dynamic imaging. Subsequently, 18FDG (0.5 mCi) was injected i.p. with a 10 mins static scan acquired following 40 mins uptake. Brain images were analyzed in a standard anatomical space using Statistical Parametric Mapping. Significantly (P<0.01) increased glucose metabolism in endotoxemic mice (vs controls) was observed in the hippocampal CA2/CA3 region and other brain areas. Increased [11C]PBR28 binding (P<0.05) was also determined in the hippocampal CA2/CA3 region. These results indicate brain hypermetabolic activity and neuroinflammation during endotoxemia specifically affecting the hippocampus – an area with a primary role in memory and cognition. These findings support further development of PET-based evaluation of neuroinflammation in preclinical and clinical settings of inflammatory and metabolic disorders.
How safe is the application of nanotitania when it can induce inflammatory response, affect liver function or body weight in animal models?
Authors: Sharma Nilesh , Leibman Hannah , Sadrinia Cyrus
Published: January 2023

Consumer products containing significant amounts of Ti-nanoparticles include paints, toothpaste, colorants, cosmetics, sunscreens, medicine, and beverages. Ti is generally considered biologically inert, but Ti-nanoparticles were observed to cause significant toxicity in recent studies. In this study, lab mice were administered intraperitoneally TiO2 rutile crystals (30–50 nm) at the rate of 333 and 1332 mg/kg (body weight) at 0, 5 and 10 day during a 15-d period. After termination of the experiment, spleen extract and circulating blood were assayed for cytokines using ELISA. Elevated levels (p<0.05) of inflammatory cytokines (TNF-α, IFN-γ, IL-1-α, IL-1β and IL-6) were detected in spleen extracts, whereas significantly reduced levels of VEGF and Rantes were obtained from both spleen extract and blood, relative to control. Liver functions and organ weights were also significantly affected under both doses. Toxicity model based on C. elegans also demonstrated significant toxicity in respect of mortality and behavior response under the dose of 25–100 ppm. Further, gene expression studies related to immune responses are continuing to determine the toxicity effects in C. elegans and mouse models.
The kinase activity of IRAK4 controls sterile autoinflammation and manifestations of Murine Lupus
Authors: Agliano Federica , Ménoret Antoine , Karlinsey Keaton S. , Ragazzi Michael , Karginov Timofey , Vella Anthony T.
Published: January 2023

Interleukin-1 receptor-associated kinase (IRAK) 4 is a serine/threonine kinase involved in TLR/IL-1R responses. Upon TLR/IL-1R activation, autophosphorylation of IRAK4 induces proinflammatory gene transcription. In this study we hypothesize that the kinase activity of IRAK4 may be a druggable target in the treatment of sterile autoinflammation and Lupus. C57BL/6 female mice injected with the hydrocarbon oil Pristane, exhibited a significant Pristane-dependent induction of splenomegaly; whereas treatment with the IRAK4 kinase inhibitor reduced this inflammatory symptom. Conversely, no significant difference was observed in males. Using mass spectrometry, we found that the urine of Pristane-injected mice contained increased levels of putative markers for several inflammatory diseases, which were reduced by IRAK4 kinase inhibition. We showed that Pristane-injected mice had increased cell free DNA in serum, which was not impacted by kinase inhibitor treatment. However, chemokine release was significantly reduced in IRAK4 kinase inhibitor-treated mice. We further analyzed the kinase activity of IRAK4 in MRL-lpr mice, which were compared with MRL-lpr/IRAK4 kinase-inactive (KDKI) mice (where IRAK4 is expressed but its kinase activity is lost). Although MRL-lpr/IRAK4KDKI and MRL-lpr mice did not show differences in CD4+ and CD8+ cells activation, we found a significantly reduced splenomegaly in MRL-lpr/IRAK4KDKI mice compared to controls, also exhibiting a gender bias. Thus, IRAK4 kinase activity might represent a new therapeutic and druggable target in the recruitment of immune cells during sterile inflammatory diseases in humans, including Systemic Lupus Erythematous (SLE).
Lymphangiogenesis near the Cribriform Plate after Cerebral Ischemia
Authors: Choi Yun Hwa , Hsu Martin , Laaker Collin , Sandor Matyas , Fabry Zsuzsanna
Published: January 2023

Recently, it has been hypothesized that antigens, antigen-presenting cells (APC), and cerebrospinal fluid (CSF) may drain from the central nervous system (CNS) into lymphatics near the cribriform plate or dura to provide fluid homeostasis and antigen drainage. However, the role of lymphatic vessels during neuroinflammation is not well understood yet. Our lab has previously shown that lymphatic vessels near cribriform plate undergo lymphangiogenesis during experimental autoimmune encephalomyelitis (EAE), a mouse model of Multiple Sclerosis. Here, we looked to see whether lymphangiogenesis also occurs in a stroke mouse model of neuroinflammation. We established the transient middle cerebral artery occlusion (tMCAO) model of stroke by placing a filament for 60 minutes into the middle cerebral artery, that was followed by reperfusion after removing the filament. We show that lymphangiogenesis occurs near the cribriform plate, peaks at day 7 and decreases after 14 days following tMCAO. To confirm that the newly formed lymphoid vessels at the cribriform plate could connect with the CSF, we injected Evans Blue dye through the cisterna magna and confirmed the presence of the dye in these lymphoid vessels. Flow cytometry analysis showed that robust leukocyte infiltration such as microglia (CD45int, CD11b+), macrophage (CD45hi, CD11b+), dendritic cells (CD45hi, CD11b+, CD11c+), CD8 T cells (CD45hi, CD8+), CD4 T cells (CD45hi, CD4+), and B cells (CD45hi, B220+) near the cribriform plate after 7 days of tMCAO correlates with lymphangiogenesis (CD45−, Podoplanin+, CD31+). The manipulation of inflammation-induced lymphangiogenesis may lead to therapeutics for neuroinflammatory diseases.
M2-polarized macrophages as a potential cell therapy to mitigate inflammation-induced preterm birth
Authors: Garcia-Flores Valeria , Romero Roberto , Xu Yi , Miller Derek , Slutsky Rebecca , Robertson Sarah A , Gomez-Lopez Nardhy
Published: January 2023

Preterm birth is the leading cause of neonatal morbidity and mortality worldwide. A substantial fraction of spontaneous preterm births are due to intra-amniotic inflammation; yet, to date, no anti-inflammatory strategies exist to treat this clinical condition. We propose that boosting homeostatic immune cells that are reduced at the maternal-fetal interface of women who underwent preterm birth can serve as a novel strategy to prevent this syndrome. First, using immunophenotyping, we showed that women who experience preterm birth have low proportions of M2-polarized macrophages at the maternal-fetal interface. Next, we tested whether adoptive transfer of M2-polarized macrophages could prevent intra-amniotic inflammation-induced preterm birth in mice. Bone marrow-derived myeloid cells (BMDM) from non-pregnant mice were differentiated and polarized towards M2 macrophages, as proven by flow cytometry. The adoptive transfer of M2-polarized macrophages, but not BMDM, prevented preterm birth and neonatal death. To increase the translational value, M2-polarized macrophages from mid and late pregnancy were also tested, and those from mid pregnancy were more effective in preventing preterm birth. Using GFP-positive M2-polarized macrophages, we showed that adoptively transferred cells migrate into the maternal (lung, liver, uterus) and fetal (placenta, lung) tissues as well as the maternal-fetal interface prior to preterm birth. The adoptive transfer of M2-polarized macrophages dampened pro-inflammatory responses in the maternal circulation and amniotic cavity. Collectively, these findings raise the prospect of a novel cellular anti-inflammatory approach to prevent preterm birth and adverse neonatal outcomes.
The ImmunoGenetic Theory of Cancer Survival: Insights from the analysis of 200+ unselected breast cancer patients followed for over 8 years
Authors: Williams R Michael , Yunis Edmond J.
Published: January 2023

The ImmunoGenetic approach to cancer survival has been out focus since we began collaborating in 1976. We both had had considerable experience studying the genetics and immunology of anti-cancer and autoimmune diseases related to thymus derived cells. RMW studied thymus derived cells with Byron Waksman and mouse MHC genetics with Baruj Benacerraf. EJY discovered the importance of T-cells in numerous mouse models, including breast cancer, with experiments he did in Robert Good’s laboratory. With Bernard Amos, EJY discovered the importance of more than one gene in the MHC of humans...HLA. Using published data we presented at AAI2019, the importance of homozygosity at the P2 site of Class I and the NG2A NK receptor. Innate and adaptive immune responses, involving NK cells and KIR IG receptors, are genetically controlled and appear substantially to involve natural killer cells. The mechanism is nonspecific (innate) in the killing process, but also specific (adaptive) through the KIR system inhibition of NK cell killing. Here we describe the facts that survival in breast cancer, regardless of stage at diagnosis is highly dependent on the number of NK cells in peripheral blood at the time of diagnosis. Survival is longer when NK cell number is high, when tested after surgical diagnosis. Treatments alter NK cells numbers in the blood. Radiation, even after an R0 surgery, results in a prompt decrease in peripheral blood NK cell number which then returns to baseline shortly after radiation stops. We conclude that NK cells are recruited to radiated sites. Baseline numbers of CD4+, CD8+, and CD19+ cells behave differently. The complex symphony of life is genetically controlled and extends cancer patients’ survival. Treatments are genetic and immunological.
8-iso prostaglandin E2 causes nasal congestion in allergic rhinitis of mice
Authors: Murata Takahisa , Tachibana Yuri , Nakamura Tatsuro
Published: January 2023

Allergic rhinitis (AR) is a complex nasal disease accompanied with mast cell and eosinophils infiltration into nasal cavity. Nasal congestion is a typical symptom of allergic rhinitis that is difficult to treat. Thromboxane A2 (TXA2)-TP receptor axis is reported to be involved in nasal congestion; however, its detailed mechanism is unknown. Here, we assessed the involvement of TP-mediated signaling in nasal congestion of mice. Intraperitoneal sensitization and intranasal administration of ovalbumin induced rhinitis with nasal congestion in mice. Pharmacological TP blockade but not TXA2 synthase inhibition abolished the nasal congestion accompanied with vascular hyper-permeability. These data suggest the presence of some other TP ligand than TXA2 in nasal cavity. LC-MS/MS based comprehensive analysis identified 15 types of lipid metabolites in nasal lavage fluid (NALF) of AR. The levels of almost all metabolites were higher in the NALF of AR than that in non-AR. Among these, we detected a possible TP ligand, 8-iso PGE2 in the AR NALF. Both in vitro and in vivo experiments showed that treatment of 8-iso PGE2 caused vascular hyper-permeability which was inhibited by pharmacological TP blockade. In addition, intranasal administration of 8-iso PGE2 induced nasal congestion which was significantly inhibited by TP blockade. We identified 8-iso PGE2 as a novel TP ligand which exacerbate nasal congestion in rhinitis mice.
Cul5 E3 ubiquitin ligase limits Th2 and Th9 differentiation by regulating IL4 receptor level in CD4 T cells
Authors: KUMAR BINOD , Field Natania , Herbert De’broski R , Oliver Paula
Published: January 2023

Allergic asthma is a chronic airway disease characterized by increased airway resistance and airway remodeling. It is well established that increased Th2 and Th9 cells is responsible for this phenotype, the mechanisms restricting these cell numbers are poorly understood. Here we show that the E3 ubiquitin ligase, Cul5, restricts Th2 and Th9 differentiation and prevents allergic asthma. We found that Cul5 activity measured by neddylation is increased following CD4 T cell stimulation. We generated mice lacking Cul5 in T cells that showed mild Th2 inflammation in lung specifically. Upon house dust mite exposure these mice exhibited increased eosinophilia, goblet cell hyperplasia, and airways remodeling. Mechanistically, we show that in activated T cells, Cul5 associated with cytokine-induced SH2 protein (CIS), a SOCS-box protein. CIS act as a substrate receptor for Cul5 E3 ubiquitin complex and limits IL-4 receptor level on the surface, that correlated with increased Th2 and Th9 differentiation and/or persistence in vivo. These data support that Cul5 works with CIS to attenuate IL-4R signaling and reduce susceptibility to allergic asthma.
CD1d-dependent NKT cells establish Th1-polarity to promote resolution of dengue virus infection
Authors: St. John Ashley L , Choi Youngjoo , Rathore Abhay
Published: January 2023

NKT cells are innate-like T cells that can rapidly produce cytokines or acquire cytotoxicity upon activation, which can shape subsequent adaptive immunity towards invading pathogens. During infection by the arboviral pathogen dengue virus (DENV), NKT cells are recruited to the skin, yet their contributions to cutaneous virus clearance are unclear. Moreover, the individual contributions of various NKT subsets to DENV immunity are poorly defined. Here, we show CD1d-dependent (CD1d-dep) NKT cells promote DENV infection resolution in skin and draining lymph nodes (dLNs) through recruitment of cytotoxic lymphocytes (CTLs), including CD8+ T cells and NKT-like cells to the skin and CD4+ and γδT cells to the dLN. CD1ddep NKT cells and NKT-like T cells are major producers of chemotactic factors for CTLs and Th1 cytokines. Impaired Th1 immunity in CD1d-KO mice was also apparent as CD4+ and CD8+ T cells produced less IFN-γ and more IL-4 and Th2-associated IgG1 levels were higher than Th1-associated IgG2a levels and promoted antibody dependent enhanced disease. These results demonstrate that CD1ddep NKT cells establish Th1 polarity during the early response to DENV, which significantly affects infection resolution and memory formation.
Peripheral blood MAIT cell response to enterotoxigenic E. coli strains H10407 and B7A after controlled human infection models (CHIMs)
Authors: Eller Michael A. , Lal Kerri G , Kim Dohoon , Vasan Sandhya , Gramzinski Robert A. , Maciel Milton
Published: January 2023

Mucosal associated invariant T (MAIT) cells are an unconventional T cell subset with a semi-invariant TCR that recognizes microbially-derived riboflavin metabolites and elicit broad antimicrobial activity. MAIT cells are implicated in the control of bacterial infections, however, their role in enterotoxigenic E. coli (ETEC) infection, a common cause of traveler’s diarrhea, is unknown. Using multi-color polychromatic flow cytometry and RNA-seq on sorted peripheral MAIT cells of volunteers orally challenged with ETEC strains H10407 (CFA/I+LT+ST+; median time to first loose stool, mtFLS=47h; n=8) or B7A (CS6+ LT+ST+; mtFLS=13h; n=12), we observed that MAIT cells express higher levels of gut-homing molecules a4b7 and CCR9 7-days post-challenge. Moreover, MAIT cells also have an increase in expression of markers of activation CD38 (p=0.003) and cell proliferation Ki-67 (p=0.01) 7-days post-challenge, despite no change in overall frequency. RNA-seq revealed distinct transcriptional signatures 7-days post challenge in both challenge models. Gene set enrichment analysis (GSEA) showed that challenge with both ETEC strains elicit an upregulation of pathways involved in MAIT cell proliferation and division. Several pathways involving IL-1 secretion and the inflammasome were also upregulated after H10406 challenge. Meanwhile, B7A challenge upregulated innate-like pathways such as Natural Killer Cell Activation. These data suggest that MAIT cells are responding during ETEC challenge with upregulation markers of gut homing, activation and proliferation, but undergo differential means of activation after CHIMs with H10407 compared to the more fact-acting B7A ETEC strain.
Nox2 deficiency in neutrophils promotes excessive and sustained inflammatory responses at local and distal sites of inflammation
Authors: Haist Kelsey Cornell , Henson Peter M , Bratton Donna L
Published: January 2023

NADPH oxidase 2 (Nox2) plays important roles both in protection from pathogens and in responses to inflammatory stimuli, and its functional loss is the cause of Chronic Granulomatous Disease. While earlier reports implicated macrophage Nox2 as the major contributor to inflammatory responses, we sought to determine the respective contributions of Nox2 in resident macrophages or neutrophils to early inflammatory responses induced by two different stimuli and routes of induction. We hypothesized that absence of Nox2 function in resident macrophages would drive excessive, prolonged inflammation. Intraperitoneal (i.p.) zymosan or intratracheal (i.t.) killed A. fumigatus in Nox2-deficient mice both drive a severe, sustained inflammatory response locally with continually elevated levels of neutrophils, monocytes, and macrophages and granulomata formation, whereas WT mice exhibit rapidly resolving responses. We employed a novel mixed bone marrow chimera approach in which neutrophils are predominantly Nox2-sufficient yet resident macrophages are largely Nox2-deficient. We treated these mice and WT control mixed chimeras with both i.p. zymosan and i.t. killed A. fumigatus and characterized the inflammatory response in both the lung and peritoneal cavity after either stimulus. Notably, the Nox2-deficient mixed chimeras responded similarly to WT control chimeras, including reduced neutrophil and monocyte recruitment and absence of granulomata, demonstrating that Nox2 deficiency specifically in neutrophils drives excessive inflammation in response to both i.p. zymosan and i.t. A. fumigatus. These data highlight the importance of neutrophil Nox2 in normal progression and resolution of these inflammatory responses.
Accumulation and activation of group 2 innate lymphoid cells in the aged brain alleviate aging associated cognitive decline
Authors: Fung Ivan Ting Hin , Sankar Poornima , Zhang Yuanyue , Robison Lisa S. , Zhao Xiuli , D’Souza Shanti , Salinero Abigail E. , Wang Yue , Qian Jiang , Kuentzel Marcy , Chittur Sridar V. , Temple Sally , Zuloaga Kristen L. , Yang Qi
Published: January 2023

Increasing evidence has challenged the traditional view about the immune privilege of the brain, but the precise roles of immune cells in regulating brain physiology and function remain poorly understood. Here we report that tissue-resident group 2 innate lymphoid cells (ILC2) accumulate in the choroid plexus of aged brains. ILC2 in the aged brain are long-lived, relatively resistant to cellular senescence and exhaustion, and are capable of switching between cell cycle dormancy and proliferation. They are functionally quiescent at homeostasis but can be activated by IL-33 to produce large amounts of type-2 cytokines and other effector molecules in vitro and in vivo. Intracerebroventricular transfer of activated ILC2 revitalized the aged brain and enhanced the cognitive function of aged mice. Administration of IL-5, a major ILC2 product, was sufficient to repress aging-associated neuroinflammation and alleviate aging-associated cognitive decline. Targeting ILC2 in the aged brain may provide new avenues to combat aging-associated neurodegenerative disorders.
C-Type Lectin Receptor Dectin-1 Suppresses the Development of Experimental Autoimmune Encephalomyelitis
Authors: Yan Chengkai , Tang Na , Guo Hui , Zhang Jian
Published: January 2023

Dectin-1 (encoded by Clec7a) is a mainly myeloid cell-expressed natural killer (NK)-cell-receptor-like C-type lectin that functions as a fungal recognition receptor by binding to b-glucan carbohydrates. However, the role of Dectin-1 in adaptive immunity is not fully understood. In this study, we surprisingly found that mice deficient for Dectin-1 develop severe experimental autoimmune encephalomyelitis (EAE), a mouse model of multiple sclerosis (MS). The heightened EAE severity observed in Clec7a−/− mice correlates with increased Th17 response in the draining lymph nodes. Further analysis showed that Clec7a−/− mice have a reduced regulatory CD8+ T cells, a newly discovered CD8+ T cell subset defined as the surface expression of CD8, CD44, CD122, and Ly49+, which have been shown to suppress EAE development. In support of a critical role of Dectin-1 in non-CD4+ T cells in the development of EAE, Tcrb−/− mice receiving wild-type and Clec7a−/− CD4+ T cells develop a comparable degree of EAE. Therefore, our data suggest that Dectin-1 regulates EAE development possibly via controlling regulatory CD8+ T cell development.
Identification of germinal center-specific lipid species
Authors: Cho Sung Hoon , Jones Marissa , Spraggins Jeffrey , Caprioli Richard M , Boothby Mark Robin
Published: January 2023

Germinal centers (GCs) are microanatomic structures that B cells proliferate, are selected for affinity maturation, undergo antibody (Ab) class-switch recombination and differentiate into Ab-secreting plasma cells and memory B cells. Nutrient availability, metabolic reprogramming process, and spatial localizations of biomolecules can impact on GC response and quality of humoral immune response. By using matrix-assisted laser desorption/ionization-imaging mass spectrometry (MALDI-IMS), we identified that the ether lipid species are preferentially enriched in germinal centers, and the perturbation of ether lipid synthesis attenuatedGC response and antigen-specific Ab production. These results suggest that molecular programing of GC lymphocytes is tied to increased ether lipid synthesis and that these species are functionally important in humoral immunity.
Circulating classical monocytes share a common transcriptional signature with skin macrophages in Systemic Sclerosis
Authors: Makinde Hadijat M , Dominguez Salina T , Cuda Carla M , Gadhvi Gaurav , Aren Kathleen , Zeng Chang , Eickelberg Garrett J , Khanna Dinesh , Assassi Shervin , Frech Tracy , Winter Deborah R , Perlman Harris , Hinchcliff Monique
Published: January 2023

The etiology and pathogenesis of systemic sclerosis (SSc) are poorly understood. Circulating monocytes likely play a critical role in SSc progression through secretion of proinflammatory molecules and as precursors of macrophages that can reorganize the extracellular matrix, thereby leading to development of end-organ fibrosis. Here, we evaluate the transcriptional similarities between circulating classical monocytes (CMo) and skin macrophages of SSc patients. Bulk RNA-seq was performed on sorted CMo from SSc blood obtained through the Prospective Registry of Early Systemic Sclerosis (PRESS) cohort, and transcriptional profiles were analyzed along with profiles from matched control samples. Additionally, sorted CD45+ cells from skin biopsies of one SSc patient and one control patient were prepared for single-cell RNA-seq. There was a numerical expansion of skin macrophages from the control to the SSc sample, but no significant difference in the quantity of circulating CMo between control and SSc patients. Of the 152 significantly upregulated genes (DESeq2, p<0.05, fold change>2) observed in circulating CMo population and the 290 upregulated genes (fold change>2) in the skin macrophage cluster compared to their respective controls, we found 23 genes in common (p<1.23×10−8, hypergeometric distribution test). These shared genes were involved in various immune and inflammatory regulation processes. These data indicate that a common transcriptional signature may exist between circulating CMo and macrophages present in the skin of SSc patients, potentially suggesting that macrophage-specific pathways at the site of fibrosis can be detected in a circulating precursor population.
MDA-5 mediated RNA sensing signaling responds to the cellular RNAs escaped from RNA surveillance causing apoptotic cell death
Authors: Wang Qingde , Gao Wentao , Guo Xinfeng , Rodrigiguez Nathaniel , Billiar Timothy R
Published: January 2023

It is well established that invaded viral RNA triggers cell death through cytosolic RNA sensing and IFN signaling pathways. Cellular RNA, however, is tolerated, mainly due to the dynamic RNA surveillance which leads to rapid degradation of the immature and abnormally processed or potentially disease-causing RNA transcripts. Here we demonstrated that inappropriately processed cellular RNA escaped from RNA surveillance causes apoptotic cell dearth through MDA-5 mediated RNA sensing signaling. We generated a unique mouse model in which aberrant cellular RNA causes massive cell death and lethality of the mice. Through characterization of this model, we found that cell type specific deletion of the RNA editing enzyme ADAR1 led to accumulation of the cellular RNAs at insufficiently edited status which escaped from RNA surveillance. The unedited RNAs transported from nucleus to cytosol activated the RNA sensing signaling and resulted in robust type I IFN and ISG expression, concurrently occurred a massive apoptotic cell death. We found that activation of innate immune signaling was almost exclusively through the MDA-5 RNA receptor, since deletion of only MDA-5 prevented type I IFN production and the downstream ISGs. Deletion of MDA-5 not only rebalanced innate immune homeostasis, but also extinguished the apoptotic pathway, and completely rescued the lethality of the ADAR1 KO mice. In addition, we defined that it was the intrinsic apoptotic pathways activated by MDA-5 RNA sensing was responsible for the cell death. This finding indicated that cellular RNA sensing signaling initiated by RNA receptor MDA-5 could be a critical mechanism responding to the RNAs escaped from the nucleic RNA surveillance in diseased conditions.
Immunoglobulin gene rearrangement and BAFF responsive maturation defines a novel B cell population undergoing extra-BM development
Authors: Khan Wasif N , Bazile Cassandra A , Clark Emily S , Magee Jennifer , Boucher Justin C , Carlesso Gianluca , Morehouse Christopher , Shrestha Akritee , Umland Oliver , Salyakina Daria , Wesemann Duane R , Kleiman Eden
Published: January 2023

Transitional type-1 (T1) cells are peripheral immature B cells known to populate the spleen (spl) after completing their BCR assembly in the bone marrow (BM). To advance the understanding of splenic T1 (CD19posCD24hiCD21neg) B cells, we addressed the heterogeneity and biology of these cells using flow cytometry combined with genetically modified mice. Most recent emigrant T1 cells were selected by excluding CD23pos and including CD93high (AA4.1) B cells termed T12123DN. Transcriptomic analysis identified RAG1 and 2 as signature genes for this B cell population. Further separation of T12123DN cells based on surface IgM expression revealed a previously undescribed cell subset with undetectable cell surface IgM (-IgMneg). The spl-IgMneg subsets expresses RAG1/2 and actively undergoes Igk gene rearrangement at levels comparable to BM pre-B cells. Upon in vitro exposure to BAFF or transplantation into immunodeficient hosts, spl-IgMneg cells can give rise to fully mature IgMposIgDpos B cells. Furthermore, BAFF-R and NF-kB pathways are required for their efficient maturation. Our findings suggest that the spl-T1 population encompasses a subset of B cells that resemble but are distinct from the developing B cells in the BM. These spl-IgMneg B cells may represent receptor editing B cells, and/or precursor B cells undergoing BCR assembly and selection in the periphery, possibly providing an opportunity for tolerance induction to tissue restricted self-antigens and microbiota-derived antigens.