bluuebunny/crossref_metadata_2025_split huggingface dataset:
Thromboxane (TXA2) Attenuates Lung Inflammatory Responses and Th9 Cell Differentiation and Function During Allergic Lung Inflammation
Authors: Li Hong , Bradbury Alyce , Edin Matthew L , Gruzdev Artiom , Sparks Matthew , Hoopes Samantha L , DeGraff Laura M , Bortner Carl D , Coffman Thomas , Zeldin Darryl C 
Published: January 2023

Abstract
Thromboxane(TXA2) is a potent bronchial smooth muscle spasmogen implicated in the pathogenesis of asthma. T helper type 9 (Th9) cells, a subpopulation of CD4+ T cells, play an important role in the pathogenesis of asthma;however, it is unknown whether TXA2 regulates Th9 cell differentiation or function during allergic lung inflammation. Hence, we used an in vivo ovalbumin (OVA)-induced allergic inflammation model to study the role of TXA2 and its receptor in Th9 cell differentiation during allergic lung inflammation. Following OVA sensitization/exposure, the percentage of IL‑9+/CD4+ T cells was significantly decreased in lungs, blood and spleen of mice that were implanted with mini pumps containing thromboxane (TXA2). Also, TP−/− mice had a significantly higher percentage of Th9 cells after OVA sensitization/exposure compared to WT mice, and also exhibited increased allergic lung inflammation. In vitro experiments showed that differentiation of naïve CD4+ T cells to Th9 cells was significantly inhibited by TXA2. TXA2 also inhibited Th9 cell differentiation of OVA-sensitized CD4+ memory T cells. CD4+ T cells did not produce significant amounts of TXA2, but expressed the TP receptor during Th9 cell differentiation. TXA2 activation of TP receptors increased cAMP production and enhanced phosphorylation of p38 MAPK, ERK and PI3K in CD4+ T cells during Th9 differentiation in vitro. These effects were markedly attenuated in TP−/− cells. Importantly, TXA2 signaling suppressed activation of the Il9 promoter via a mechanism that involved binding of PBX1 and NFE2 transcription factors to their corresponding response elements. Thus, TXA2 acts through TP receptors to suppress Il9 promoter activation and Th9 cell differentiation.
DC-independent TSLP-mediated acute allergic airway inflammation via CD11c+ Interstitial Macrophages
Authors: Lai Jen-Feng , Thompson Lucas J , Ziegler Steven F 
Published: January 2023

Abstract
Thymic stromal lymphopoietin (TSLP) is an epithelial-derived cytokine important for the development of type-2 inflammatory responses at mucosal surfaces. In humans, TSLP has been found to be elevated in the lungs of asthmatics, and in mouse models TSLP can promote type-2 airway inflammation. DC have been shown to respond directly to TSLP and subsequently promote the differentiation of Th2 cells. In addition, other lung-resident MHCII-expressing cells, such as Alveolar Macrophages (AM) and Interstitial Macrophages (IM), have been shown to regulate the activation of T cells. We have identified a subset of CD11c-expressing Interstitial Macrophages (CD11cIM) addition to AM and IM, which can directly induce proliferation and differentiation of CD4 T cells in a TSLP-dependent manner. Following exposure to TSLP, these cells are more potent then DC for expansion and Th2 cytokine production of CD4 T cells. More importantly, CD11cIM are necessary and sufficient for the induction of Th2 responses and downstream eosinophilic inflammation in the lungs. These findings indicate a novel unique and critical role for TSLP-signaling on CD11cIM in the development of acute Th2-dependent allergic airway inflammation in pulmonary microenvironment.
Smith-Lemli-Opitz syndrome reveals requirement for sterol biosynthesis in the innate immune response
Authors: Gabor Kristin , Madenspacher Jennifer H , Wassif Christopher A , Bushel Pierre R , Porter Forbes D , Fessler Michael B 
Published: January 2023

Abstract
Smith-Lemli-Opitz Syndrome (SLOS) is a rare neurodevelopmental disorder caused by mutation of 7-dehydrocholesterol (7DHC) reductase (DHCR7), a terminal enzyme in cholesterol biosynthesis. SLOS patients have increased incidence of infections, but no mechanism of immunosuppression has been identified. As cholesterol is a key component of membrane raft microdomains and rafts have been implicated in Toll-like receptor (TLR) signaling, we examined whether TLR responsiveness is altered in SLOS. Primary fibroblasts from SLOS patients with varying degrees of deficit in DHCR7 activity were used to assess the TLR4 response. Upon stimulation, significantly lower NF-kB activation and IL-6 and IL-8 production were observed in SLOS cells than in controls. Across patients, a direct relation between residual DHCR7 enzyme activity and IL-6 production was observed, together with an inverse relation between clinical severity scores and IL-8 production. Microarray analysis revealed that several pro-inflammatory chemokines are less responsive to LPS in SLOS. Deficient LPS-induced cytokines were also observed in macrophages from Dhcr7 mutant mice and in RAW264.7 macrophages treated with DHCR7 inhibitors. Supporting cholesterol deficit as the cause of the TLR4 impairment, the LPS response of SLOS macrophages was partially rescued by cholesterol repletion. Mechanistically, confocal microscopy revealed that TLR4 localization to rafts is decreased in SLOS macrophages. Taken together, these findings indicate that the SLOS mutation confers an abnormal TLR4 response. Deficient innate immunity may be a clinically relevant contributor to SLOS pathogenesis, and SLOS highlights the importance of cholesterol biosynthesis to innate immunity.
Autoimmune variant PTPN22 is associated with impaired responses to influenza virus vaccination
Authors: Crabtree Juliet Nicole , He Wenqian , Guan Weihua , Miller Matthew S , Peterson Erik J 
Published: January 2023

Abstract
High-affinity antibody production, T cell activation, and Interferon upregulation all contribute to protective immunity that occurs in humans following influenza immunization. Hematopoietic cell-specific PTPN22 encodes Lymphoid Phosphatase (Lyp), which regulates lymphocyte antigen receptor and Pattern Recognition Receptor (PRR) signaling. A PTPN22 variant R620W (LypW) predisposes to autoimmune and infectious disease, and confers altered signaling through antigen receptors and PRRs. We tested the hypothesis that LypW-bearing humans would have diminished immune response to trivalent influenza vaccine (TIV). LypW carriers exhibited decreased induction of influenza-specific CD4 T cells expressing effector cytokines, and failed to increase antibody affinity following TIV. No differences between LypW carriers and non-carriers were observed in virus-specific CD8 T cell responses, early interferon transcriptional responses, or myeloid APC costimulatory molecule upregulation. LypW association with defects in TIV-induced CD4 T cell expansion and antibody affinity maturation suggests that LypW may predispose to diminished capacity to generate protective immunity against influenza.
Investigating mechanisms restricting responsiveness of B cells specific for foreign antigens
Authors: Steach Holly R , Taylor Justin J. 
Published: January 2023

Abstract
In order to prevent antibody-mediated autoimmunity, B cells reactive against self-antigens are subjected to tolerance mechanisms including induction of a state of restricted functionality termed anergy. We hypothesize that anergy also restricts responsiveness of B cells specific for foreign antigens due to B cell receptors (BCR) cross-reactivity with endogenously expressed molecules. Using a previously published antigen-specific enrichment strategy, we have shown that 60–80% of antigen-specific B cells fail to respond to antigen immunization. While some of this poor responsiveness is likely a result of low affinity, many B cells binding high levels of antigen are found within the non-responding population. A broadly-inclusive marker for self-reactivity in mice with an intact polyclonal B cell repertoire has been identified using the orphan nuclear receptor Nur77, known to be up-regulated downstream of BCR signaling. Using Nur77-eGFP reporter mice, we have found that populations of B cells specific for non-self antigens display a range of baseline Nur77 expression similar to that of total B cells, perhaps indicating a similar extent of self-antigen recognition. Coordinately, we have found that antigen-specific naive B cells expressing low levels of Nur77 are more responsive to immunization than their counterparts expressing high levels of Nur77. These data suggest that responsiveness of naive B cells specific for foreign antigens is restricted by cross-reactivity with self-antigens. Our results could lend a unique perspective to the paradigmatic definition of B cell anergy and offer an accession to immunogen design as the principle factor governing protective humoral immune responses following vaccination.
GATA2 is critical for mast cell differentiation and maintenance
Authors: Li Yapeng , Liu Bing , Lukin Kara , Finkelman Fred , Hagman James , Roers Axel , Huang Hua 
Published: January 2023

Abstract
The GATA binding protein 2 (GATA2) plays critical roles in hematopoietic stem cell survival and proliferation, granulocyte–monocyte progenitor differentiation, and basophil and mast cell differentiation. However, precise roles of GATA2 in mast cell differentiation and maintenance have not been delineated. We identified GATA2 is essential for pre-basophil and mast cell progenitors (pre-BMPs) to differentiate into mast cells. We found that GATA2 is critical in maintaining the expression of a wide range of genes that are important for performing mast cell functions. To determine the in vivo role of GATA2 in mast cell development, we generated connective tissue mast cell-specific Gata2 deficient mice. We found that Gata2 was required for connective tissue mast cell development. These mice failed to develop IgE-mediated anaphylaxis. GATA2 regulated the expression of Hdc gene, which is critical for IgE-mediated anaphylaxis, by binding to regulatory regions of the Hdc gene to promote its transcription. Our study reveals GATA2 is critical for mast cell differentiation and maintenance.
Defining the role of CCR8 in promoting thymocyte development and thymic epithelial homeostasis
Authors: Thyagarajan Hiran Maye , Hu Zicheng , Ehrlich Lauren I R 
Published: January 2023

Abstract
Generation of functional and self-tolerant T cells in the thymus is crucial to preventing autoimmune diseases. Medullary thymic epithelial cells (mTECs) and dendritic cells (DCs) present diverse auto-antigens to developing thymocytes; strong TCR reactivity in response to self-antigens causes thymocytes to undergo negative selection, eliminating autoreactive cells from the repertoire. Chemokine receptors expressed on the surface of developing thymocytes have been shown to promote accumulation of post-positive selection thymocytes in the medulla, where they may encounter mTECs and DCs presenting auto-antigens. CCR8 is highly expressed by post-positive selection CD4 thymocytes while its ligands, CCL1 and CCL8 are expressed by mTECs. Thus, CCR8 signaling may promote medullary entry of post-positive selection thymocytes and efficient interactions with mTECs and enforcing central tolerance. Our preliminary data indicate that CCR8−/− thymi have a reduced medullary area, implicating impaired thymocyte:mTEC interactions that impacts normal proliferation and differentiation of mTECs. Furthermore, 2-photon imaging reveals that CCR8 deficiency results in a significant increase in the path straightness of CD4 SP thymocytes, which could impact the efficiency of interactions with medullary stromal cells. Strikingly, CCR8 deficiency results in increased cellularity of CD4SP OTII TCR transgenic thymocytes in a wild-type host, suggesting CCR8 is critical for efficient clonal deletion to endogenous auto-antigens. Thus, our preliminary data indicate that CCR8 contributes to the migratory properties and stromal interactions of post-positive selection thymocytes that promote efficient negative selection.
IRF8 represses GM-CSF expression in tumor cells to mediate myeloid-derived suppressor cell differentiation.
Authors: Paschall Amy V , Redd Priscilla , Zhang Ruihua , Xiong Huabao , Abrams Scott I , Liu Kebin 
Published: January 2023

Abstract
Myeloid-derived suppressive cells (MDSCs) are immature myeloid cells that are induced by inflammatory mediators. Massive accumulation of MDSCs is a hallmark of cancer in mice and human patients. Proinflammatory factors are believed to induce MDSC differentiation. In an early study, we showed that T cell-produced GM-CSF, a proinflammatory cytokine, induces MDSC accumulation under physiological conditions. We further determined that IRF8 represses MDSC accumulation in vivo. Tumor cells are known to produce abundant GM-CSF; we therefore sought to test the hypothesis that IRF8 regulates GM-CSF expression to mediate MDSC accumulation in the tumor-bearing host. We observed that mouse sarcoma cells rapidly induce MDSC accumulation, which is positively correlated with tumor growth kinetics. Analysis of purified MDSCs from tumor-bearing mice indicated that GM-CSF mRNA levels increase and IRF8 expression decreases as tumor sizes increase. GM-CSF and IRF8 expression kinetics are therefore inversely correlated in MDSCs in vivo. Analysis of the csf2 gene promoter DNA sequence identified an IRF8-binding consensus element, and ChIP analysis determined that IRF8 binds to this DNA element in the csf2 promoter. Analysis of human sarcoma specimens revealed that the IRF8 promoter DNA is hypermethylated. Silencing IRF8 expression using IRF8-specific shRNA increased GM-CSF expression. In summary, we determine that tumor cells use GM-CSF to induce MDSC differentiation, and IRF8 functions as a transcriptional repressor of GM-CSF expression to suppress MDSC accumulation. Human sarcoma cells may use DNA methylation to silence IRF8 and increase GM-CSF expression to promote MDSC accumulation, thereby suppressing the anti-tumor immune response.
Prognostic significance of tumor infiltrating immune cells in oral squamous cell carcinoma
Authors: wang zhi , Fang Juan , Song Jingjing , Chen yichen , Ma Da , Liu Xiangqi , cheng bin 
Published: January 2023

Abstract

Background
Oral squamous cell carcinoma (OSCC) is the most common malignancy in oral maxillofacial region. Although modest improvements in the survival rates have been reported, the prognosis remains unsatisfactory. Tumor infiltrating immune cells have been proved playing critical roles in tumor prognosis. We conducted this study to evaluate the predictive value of CD4+ T cells, T-bet+ Th 1 cells, CD8+ T cells, CD57+ NK cells and CD68+ macrophages for OSCC outcome.

Methods
78 OSCC patients were included and followed up for at least 3 years. Clinical data and Paraffin-embedded samples were collected. CD4, CD8, T-bet, CD57 and CD68 expressions were examined by immunohistochemistry. The Kaplan –Meier method, log-rank test and Cox proportional hazards model were used to estimate the prognostic factors for Overall survival. ROC and AUC were used to evaluate the prognostic value.

RESULTS
Multivariate analysis revealed that no lymph node involvement, lower CD4 (+) T cells and high CD57 (+) NK cells infiltration were all significantly related with longer survival (P &lt; 0.01). The respective predictive accuracy of CD4+ T cells, CD8+ T cells and CD57+ NK cells were all superior to TNM staging. CD57 expression provided the highest accuracy (AUC=0.759; 95% CI, 0.652–0.886).

Conclusion
Our study highlighted the predominant role of innate immune system in tumor immunity. Adaptive immunity seemed to be the major victim of immune tolerance. When compared to TNM staging, CD4, CD8 and CD57 expression significantly improves mortality prediction for OSCC patients. This study will provide new strategies to patient management of OSCC.
Modeling immune system-tumor interactions using humanized mice
Authors: Aryee Ken-Edwin , Brehm Michael A , Shultz Leonard D , Jurczyk Agata 
Published: January 2023

Abstract
The interplay between tumor cells, the non-malignant stoma and hematolymphoid cells (of both innate and adoptive lineages) determines the kinetics of cancer progression. However, the study of tumor-immune system interactions has primarily been restricted to mouse models. The implant of tumors into humanized mice that are engrafted with human immune systems provides a unique opportunity to interrogate the interactions between human immune cells and malignancies, and to test novel anti-tumor immunotherapies. Here we show the use of the BLT (bone marrow/liver/thymus) humanized mouse model to study the interactions between the human immune system and melanoma. The BLT mouse supports high levels of human immune system development including HLA-restricted T cells, B cells and innate immune cells. For these studies, we used NOD-scid IL2rnull (NSG) transgenically expressing SCF, GM-CSF and IL-3 (NSG-SGM3), which enhances human innate immune cell development and function. Following implant of patient derived melanomas into NSG-SGM3 BLT mice, we observed the infiltration of human T cells, mast cells, NK cells and macrophages into the tumor microenviroment. Moreover the recovery of high levels of FoxP3+ regulatory T cells, M2 macrophage and conventional CD4 and CD8 T cells expressing PD-1 and CTLA4 from the tumor suggests that the melanoma growing within NSG-SGM3 BLT mice creates an immune regulatory environment similar to that seen in patients. These results indicate that tumors implanted into NSG-SGM3 BLT mice can be used as an effective model for understanding tumor-immune system interactions.
Neuromodulatory activities of CD4+CD25+Foxp3+ regulatory T cells in 3xTg-AD Alzheimer’s disease model
Authors: Bae Hyunsu , Baek Hyunjung , Ye Minsook , Kang Geun-Hyung , Lee Chanju , Lee Gihyun , Lee Sujin 
Published: January 2023

Abstract
Alzheimer’s disease (AD) is the most prevalent cause of dementia marked by progressive cognitive and functional impairment and memory loss. AD patients display neuropathological lesions including deposition of extracellular amyloid-beta (Aβ) peptide and the formation of neurofibrillary tangles. Although the mechanisms causing brain atrophy and neuronal death are largely unknown, increasing evidence suggests that immunity plays a critical role in the pathogenesis of AD. In the present study, we investigated the effect of regulatory T cells (Tregs) on AD mouse model. First, we transiently depleted Treg population using anti-CD25 antibody. Depletion of Tregs for 4 months resulted in a marked aggravation of the spatial learning deficits of 6-month-old 3xTg-AD mice. Importantly, deposition of Aβ plaques and CD11b+ microglia/macrophage were significantly increased in the hippocampal CA1 and CA3 regions of the Treg depleted 3xTg-AD compared to the vehicle-treated 3xTg-AD group. Next, we explored the effect of Teffs (effector T cells; CD4+CD25−) and Treg (CD4+CD25+) in adoptive transfer model. Systemic transplantation of purified Tregs into 3xTg-AD mice improved cognitive function and reduced deposition of Aβ plaques. In contrast, adoptive transfer of Teffs showed increased Aβ plaques burden and diminished behavioral function. Our finding strongly suggested that systemic Treg administration ameliorates disease progression and could be an effective AD treatment.
Internalization of T-Cell Receptor Mimic Antibody RL6A by Brain Derived Endothelial Cells Analyzed by Flow Cytometry
Authors: Park Jee Hyun , Hawkins Oriana E. , Chintala Navin K. , Weidanz Jon A. , Bickel Ulrich 
Published: January 2023

Abstract
T-cell receptor mimic monoclonal antibodies (TCRm) recognize peptide-MHC class I complexes. They combine the specificity of T-cell receptors with high binding affinities in the low nanomolar to picomolar range characteristic of monoclonal antibodies. The TCRm RL6A targets YLLPAIVHI peptide-HLA-A2 complexes, where the peptide is derived from the p68 RNA helicase protein. Previous work showed that this target is expressed by brain endothelial cells forming the blood-brain barrier (BBB). Upon binding, RL6A crosses the BBB via a receptor-mediated transcytosis mechanism. Confocal microscopy indicated that, after internalization, RL6A co-localized with the early endosomal marker (EEA1). Here we used conventional flow cytometry and imaging flow cytometry to quantitatively analyze the internalization process. Endothelial cell monolayers formed by hCMEC/D3 cells were incubated with unlabeled RL6A for time periods up to 120 min. Surface bound RL6A was detected by a secondary, PE-labeled antibody. RL6A disappeared in a biphasic pattern from the surface of hCMEC/D3 cells, with an initial half-life (t1/2) of internalization of about 3 min and a second, much slower phase (half life about 3 h) likely reflecting recycling to the membrane. In a complementary approach using imaging flow cytometry (Amnis ImagestreamX) and IDEAS analysis software, the internalization of AlexaFluor488-RL6A could be directly visualized and evaluated. The results confirmed that RL6A internalizes rapidly into hCMEC/D3 cells. In conclusion, flow cytometry is a suitable tool to analyze trafficking of specific peptide-HLA complexes in endothelial cells.
Enhancing CD4+ T cell function as a strategy for improving antibiotic therapy of Mycobacterium tuberculosis infection
Authors: Costa Diego L , Mayer-Barber Katrin D , Andrade Bruno B , Amaral Eduardo P , Maiga Mamoudou , Namasivayam Sivaranjani , Mittereder Lara , Orland Sandra , Barber Daniel L , Sher Alan 
Published: January 2023

Abstract
In order to develop appropriate combined immuno/chemotherapeutic interventions to treat Mtb infection, it is important to understand the interaction between the host immune response and antibiotic therapy of this pathogen. We and others have observed that CD4 T+ cell responses to Mtb decline after conventional chemotherapy. In an attempt to sustain CD4+ T cell responsiveness during antibiotic treatment, we repeatedly immunized mice with the immunodominant peptides (ESAT-64-17 or Ag85B280-294) from the major Mtb antigens ESAT-6 and Ag85B beginning at the start of drug treatment 28days post aerosol infection. When bacillary loads were assayed after 4 weeks of treatment, we found that peptide administration failed to enhance the bacterial clearance observed with chemotherapy alone. Similarly, weekly adoptive transfer of in vitro differentiated Th1 cells specific for ESAT-6 (C7) or Ag85B (P25) during antibiotic treatment failed to improve bacterial clearance despite the successful migration and proliferation of these cells in the lung parenchyma. These findings suggest that therapeutic vaccination by boosting CD4+ T cell responses during the initial phase of Mtb chemotherapy is unlikely to improve the efficacy of treatment. Nevertheless, additional experiments have suggested a major role for endogenous T lymphocytes in drug-mediated control of Mtb infection. Thus, we have observed that antibiotic treatment is ineffective in T cell deficient (TCRα−/−) mice that have received higher dose infections. We are currently evaluating the nature of this T cell requirement and whether it can be targeted to improve the outcome of Mtb therapy.
This work was supported by the intramural research program of the NIAID.