bluuebunny/crossref_metadata_2025_split huggingface dataset:
The NOD-like receptor NLRP12 plays a critical role in hepatic inflammation and cancer
Authors: Zaki Hasan , Udden SM Nashir 
Published: January 2023

Abstract
Hepatocellular carcinoma (HCC) is a deadly human cancer associated with chronic inflammation. The cytosolic NOD-like receptor NLRP12 has emerged as a negative regulator of inflammation, but its role in HCC is unknown. Cancer genomics database analysis showed that NLRP12 expression is significantly reduced in human HCC and around 2% HCC patients carry mutations in NLRP12. We investigated the role of NLRP12 in HCC using mouse models of HCC induced by carcinogen diethylnitrosamine (DEN). Nlrp12−/− mice were highly susceptible to DEN-induced HCC with increased inflammation, hepatocyte proliferation, and tumor burden. Consistently, Nlrp12−/− tumors showed higher expression of proto-oncogenes cJun and cMyc and downregulation of tumor suppressor p21. Interestingly, antibiotics treatment dramatically diminished tumorigenesis in Nlrp12−/− mouse livers. Signaling pathway analyses demonstrated higher JNK activation in Nlrp12−/− HCC and cultured hepatocytes during stimulation with microbial pattern molecules. Inhibition of JNK activation or NLRP12 overexpression reduced proliferative and inflammatory responses of Nlrp12−/− hepatocytes. In summary, this study demonstrated for the first time that NLRP12 negatively regulates HCC pathogenesis via downregulation of JNK-dependent inflammation and proliferation of hepatocytes.
Humanized PD-1/PD-L1 (HuPD) mice facilitate the direct functional comparison of immune checkpoint inhibitors in vivo
Authors: Barham Whitney Jo , Harrington Susan M. , Liu Xin , Dong Haidong 
Published: January 2023

Abstract
Though anti-PD-1/L1 antibody therapies have generated unprecedented responses in a limited number of patients, the biological and immunological parameters that define their effective use are still largely unknown. The absence of appropriate pre-clinical models has limited our ability to compare the mechanistic differences between the approved PD-1/L1 antibodies in vivo and to determine the context in which each might be most effective. To address this critical need, we have produced humanized PD-1 and PD-L1 mice in which the extracellular domains (antibody binding sites) of mouse PD-1 and PD-L1 are replaced with the sequences of human PD-1 and PD-L1. The PD-1 and PD-L1 in the resulting “HuPD” mice can be recognized by FDA-approved antibodies, while the intracellular domains are kept intact for normal downstream signals. Following characterization of the new mouse model, in our first in vivo treatment studies, HuPD mice were immunized with OVA/Poly I:C in the context of FDA-approved PD-1/L1 antibodies and antigen-specific T cell production was quantified. Nivolumab, Pembrolizumab, and Atezolizumab all increased effector CD8+T cell production against the OVA antigen as expected given their immune stimulating function. Surprisingly, OVA-tetramer staining was unable to detect antigen-specific CD8+T cells in the spleens of immunized mice treated with Avelumab. This may reflect the unique ability of Avelumab to induce ADCC, and indicates that our model is capable of distinguishing key differences among the antibodies in vivo. Ongoing studies will identify how these differences might be exploited to render the most effective results for patients in a variety of tumor types and immune environments.
A novel TNFR2 antibody induces T cell co-stimulation and promotes durable anti-tumor immunity
Authors: Sampson James F , Fulton Ross B , Kurella Vinodh B , Richards Jennifer M , Camblin Adam J , Wong Christina S , Koshkaryev Alexander , Lulo James E , Qiu James A , Kumar Sandeep , Luus Lia , Jiao Yang , Suchy James , Grabow Stephanie , Paragas Violette , Razlog Maja , Xi Lihui , Muda Marco , Drummond Daryl C , Tam Eric M , Raue Andreas 
Published: January 2023

Abstract
TNFR2, an emerging therapeutic target in immuno-oncology, is upregulated upon T cell activation and is expressed by tumor-infiltrating effector and regulatory T (Treg) cells. We generated a novel monoclonal antibody, Y9, specific to murine TNFR2 and investigated its mechanism of action. In vitro, Y9 stimulation of purified T cells increased proliferation and effector function, indicating that Y9 acts as an agonist and can provide co-stimulation.
In vivo, Y9 treatment of mice with established tumors resulted in complete tumor clearance across a variety of models. The activity of Y9 on immune cells was confirmed by its decreased activity in mice depleted of NK or CD8+ T cells. Unlike the proposed Treg depletion mechanism of other therapeutic antibodies, depletion of Treg cells is not the primary mechanism of action of Y9 treatment. Instead, decreased TNFR2 and other co-inhibitory receptor surface expression was observed following treatment. Y9 activity depended on FcγR binding, which facilitated enhanced agonist activity.
Based on this evidence, we developed MM-401, a human antibody targeting TNFR2. MM-401 has agonistic activity; upon incubation of MM-401 with human CD4+ and CD8+ T cells, we observed upregulation of activation markers and cytokine production comparable to MEDI6469 (anti-OX40). We also observed that MM-401 promotes antibody-dependent cellular cytotoxicity (ADCC) in an NK cell-mediated in vitro assay and a reduction in the number of Treg cells in human ovarian cancer ascites. These data suggest that MM-401 could also promote anti-tumor immunity by mediating ADCC, as well as by direct co-stimulation of T cell responses, and justifies the continued development of MM-401 as a novel cancer immunotherapy.
The TNFR1/mTNFα signaling axis initiates tumor-promoting inflammation and is associated with poor outcome in neuroblastoma
Authors: Tomolonis Julie Anne , Ngai Ho , Courtney Amy , Metelitsa Leonid S 
Published: January 2023

Abstract
Neuroblastoma (NB) is a poorly differentiated, aggressive pediatric solid tumor for which half of high-risk cases have no identified genetic alteration. While such high-risk tumors are often infiltrated by macrophages that generate an inflammatory gene signature predictive of poor outcome, the initiation mechanism of this pro-tumorigenic inflammation remains unknown.
In our analysis of clinical outcome-linked NB gene expression data, high tumor expression of TNFR1 was strongly correlated with poor event-free survival (5-year EFS 15% vs 70%, bonf p=9.3e-07), highlighting the potential importance of TNFα signaling in NB. Using TNFα knockout human NB cell lines, we determined that monocytes, but not NB cells, produce TNFα during in vitro co-culture. Furthermore, using a TNFα converting enzyme inhibitor to prevent secretion of soluble TNFα, we found that the membrane-bound form of TNFα (mTNFα) expressed on monocytes was critical for NF-κB pathway activation and downstream production of IL-6, a known NB growth factor. Neutralization of TNFα or its receptor TNFR1, but not TNFR2, in a monocyte-NB co-culture prevented NF-κB activation in monocytes, reducing IL-6 production by 50% compared to controls (p &lt;0.01).
Our results demonstrate that TNFR1-expressing NB cells activate monocytes via contact-dependent mTNFα signaling, leading to NF-κB activation and production of IL-6, a known driver of NB progression and therapy resistance. These findings reveal a novel, druggable mechanism of tumor-promoting inflammation in NB.
Regulation of PD-L1 expression in stimulated antigen specific CD8+ T cells by immunoglobulin 4 (IgG4)
Authors: Daftarian Pirouz M , Ouerkaxi Wushouer 
Published: January 2023

Abstract

Background
Many studies have focused on finding a correlation between the responsiveness to immune checkpoint (IC) therapies and the expression level of IC markers. Here, we report the studies on the expression of co-inhibitory molecules on healthy donor peripheral mononuclear cells (PBMC). We co-cultured PBMCs from different donors with recall antigen peptide to analyze the responses to PD-1 blockade antibody treatment. We have used our previously reported optimized recall antigen assay in which the expanded antigen specific CD8+ T cells (TET+) were measured as a readout after the peptide stimulation. Interestingly, the IgG4 appears to reduce the TET+ cell population. To further study this potential effect of IgG4, we conducted a series of immune-profiling studies by comparing TET+ gated and TET− CD8+ gated T cells.

Results and conclusion
Healthy donor PBMCs were stimulated with recall peptide alone, or peptide plus either PD-1 blockade, or IgG4 and evaluated the PD-1, PD-L1, Tim-3, and Lag-3 expressions in either T cells or in monocytes. We then compared such expressions in TET+ and TET− CD8+ T cells. Unexpectedly, IgG4 isotype control treatment was associated with upregulated PD-L1 expression only in the TET+ CD8+ T cells. To confirm whether the decrease of TET+ population in our assay was caused by the up regulation of PD-L1, we blocked the PD-1 signal transduction pathway by treating the cells with a validated PD-1 blocking antibody. As a result of PD-1 blockade, the expansion of TET+ cell population was recovered by anti-PD-1 blocking treatment. Taken together, these results suggest that IgG4 treatment with recall peptide up regulates PD-L1 expression and therefore reduces the expansion of antigen specific CD8+ T cells in some donors.
Effects of NLRP1 risk polymorphisms on IL-18 levels and gene expression
Authors: Poole Brian D , Westhoff Laura Judge , Gill Erin , Nishiguchi Michelle 
Published: January 2023

Abstract

Background
NLRP1 is a central component of the inflammasome. Several polymorphisms in the gene are linked to increased risk for autoimmune and autoinflammatory diseases. We examined the effects of these risk polymorphisms on cytokine levels and NLRP1 expression levels in human volunteers.

Materials and Methods
NLRP1 was genotyped at the SNPs rs2670660 and rs12150220 using TaqMan on peripheral blood cells. Homozygous individuals were selected for further study.
Serum IL-18 and IL-1B levels were quantified using ELISA. NLRP1 gene expression was measured semiquantitatively by SybrGreen real-time PCR.

Results
The risk variants at rs2670660 and rs12150220 were almost never found separately. There was no case in which a volunteer with homozygous risk SNP variant also had a homozygous protective variant at the other, indicating very strong linkage (p&lt;2.33 × 10-13).
Volunteers with the risk haplotype (AAAA) had significantly higher levels of serum IL-18 than volunteers with the protective haplotype (GGTT) (0.439 ng/μL compared to 0.152 ng/μL, p=0.024)
A trend towards increased NLRP1 expression was seen in the risk haplotype compared to the protective (4.4-fold increase, p=0.101).

Discussion
The risk haplotype showed increased production of IL-18 even in non-autoimmune individuals, with a trend towards higher NLRP1 expression. This suggests that the risk variants lead to higher NLRP1 activity, and therefore a pro-inflammatory phenotype. Because of the strong linkage of the AAAA and GGTT genotypes, it was impossible to differentiate which SNP was responsible for these effects. Further studies with increased numbers of volunteers should allow an increased understanding of the contribution of NLRP1 risk variants to autoimmune disease
The role of factor Y in primary biliary cirrhosis
Authors: Cho Min Jeing , Dho So Hee , Shin Saeam , Lee Ji Yeon , Lee Yeong Un , Lee Kyung-a , Kim Lark Kyun 
Published: January 2023

Abstract
Primary biliary cirrhosis (PBC) is an autoimmune disease which is characterized by bile duct injuries result from excessive immune responses. In female over the age of 40, the incidence of PBC has been estimated to be one out of 1000 people. Liver transplantation is required for patients who progress to end-stage PBC. However, risk factors and causes of PBC remain elusive.
We performed whole-exome sequencing in a family with four sisters diagnosed with PBC and one healthy half-sister. Additionally, Sanger sequencing was conducted with 62 unrelated PBC patient specimens. Here, we identified a potential PBC causative gene factor Y, a regulator of cell death and inflammation. We investigated whether factor Y plays a role in PBC using factor Y null human macrophage cell line generated by CRIPSR/Cas9 technology. We found ablation of factor Y induces inflammatory cytokines compared with control cells upon treatment with LPS or TNF. Surprisingly, molecules secreted by factor Y null cell line induced fibrosis in human hepatic stellate cell, LX-2. These results indicate that factor Y is a potential target for treatment of PBC.
Critical role of macrophages in checkpoint blockade induced autoimmune diabetes in the NOD mouse
Authors: Hu Hao , Zakharov Pavel , Unanue Emil R 
Published: January 2023

Abstract
We confirmed that the PD-1/PD-L1 regulatory pathway is one control mechanism of autoimmune diabetes in NOD mice: administration of anti-PD-1 but not anti-CTLA-4 antibodies rapidly induced diabetes in young mice showing a control phase established very early in the autoimmune process. We confirmed that both CD4 and CD8 T cells and resident macrophages were essential for the PD-1 regulation to take effect.
We performed an unbiased single-cell RNA sequencing analysis of islets after anti-PD-1 treatment. We identified a subpopulation of highly activated macrophage showing an IFNγ signature with high level of CXCL9 and PD-L1 and a monocyte signature distinguishable by the markers Ly6c and CCR2. Flow cytometry analysis of islets upon anti-PD-1 treatment identified a predominance of the monocyte derived macrophages compared to the islet resident macrophages. Depleting blood monocytes by clodronate liposomes inhibited the recruitment of the monocyte derived macrophages and prevented the development of acute diabetes induced by anti-PD-1 treatment. To analyze the pathogenicity of this monocyte-derived macrophages, we performed cytotoxicity assays by co-culturing macrophages stimulated with LPS&IFNγ and a β-cell line Min6: the highly activated macrophage killed β-cells in a manner dependent on IFNγ and inducible nitric oxide.
In sum, our study shows highly activated macrophage derived from monocytes are cytotoxic for beta cells and promoting diabetes progression. Our findings also indicate that a consequence of checkpoint blockade is the recruitment and activation of cytocidal macrophages derived from monocytes. Targeting monocyte-derived macrophages may prevent some of the checkpoint blockade related immune-adverse effects.
NOX2-deficient neutrophils facilitate joint inflammation in serum-induced arthritis
Authors: Liao Yi-Chu , Shieh Chi-Chang K 
Published: January 2023

Abstract
Immune-mediated inflammatory arthritis, including rheumatoid arthritis (RA), is a family of chronic inflammatory diseases of joints. NADPH oxidases 2 (NOX2) complex in phagocytes generates high concentrations of ROS to kill the engulfed microbes. Inherited mutations in the subunits of NOX2 result in chronic granulomatous disease (CGD). Subjects with defective NOX2 are susceptible to recurrent infections and a variety of autoimmune conditions. According to our previous studies, NOX2-deficient mice developed more severe immune-mediated arthritis, but depletion of neutrophils suppressed the development of serum-induced arthritis in these mice. Based on the hypothesis that NOX2-deficient neutrophils play a pro-inflammatory role in immune-mediated arthritis, we investigated the activity of neutrophil from joints to facilitate inflammation. We found that there are more neutrophils accumulations in NOX2-deficient joints, compared to WT joints. Both NOX2-deficient /WT neutrophils affected the anti-CD3 and anti-CD28 induced T lymphocyte proliferation, however, NOX2-deficient neutrophils were less suppressive. Moreover, NETs formation and high level of citrullinated histone H3 were detected in joints of NOX2-deficient mice at day 3 of arthritis induction. Furthermore, neutrophils from RA joints were found to suppress the T cell proliferation. Finally, carbonylated protein and total GSH level were lower in synovial fluid of RA patients when compared with those in osteoarthritis joints. Our results implicate that neutrophils may be an important cellular factor that lead to more severe arthritis in the NOX2-deficient subjects.
Kindlin-3 organizes a ring of clustered high affinity β2 integrins during human neutrophil spreading under flow
Authors: Fan Zhichao , Wen Lai , Yeh Yi-Ting , Kiosses William B , Gutierrez Edgar , Groisman Alex , Francois Joshua , Lasheras Juan C , del Alamo Juan C , Chien Shu , Ginsberg Mark H , Petrich Brian G , Ley Klaus 
Published: January 2023

Abstract
Neutrophils are vital for inflammation and immune defense. Dependent on β2 integrins, spherical neutrophils spread on vascular endothelium after arrest, which is critical for their recruitment from circulation to resist high shear flow and to initiate intravascular crawling. Here, we use high-resolution quantitative dynamic footprinting microscopy to monitor neutrophil spreading on a substrate of recombinant ICAM-1 and P-selectin under flow. A homogenous binding assay using the conformation-reporter antibodies mAb24 (reporting high-affinity β2, H+) and KIM127 (reporting extended β2, E+) showed three conformations of activated β2 integrins. E−H+ β2 integrins increased before E+H− and E+H+ conformations at the beginning of neutrophil spreading. Integrin extension depended on Syk-mediated integrin outside-in signaling. The ring of E−H+ and E+H+, but not E+H− β2 integrins was fully formed during late neutrophil spreading just before migration. Using kindlin-3-GFP fusion proteins, a ring of kindlin-3 was observed before the ring of H+ integrins appeared. These findings show spatially coordinated integrin activation during spreading. The previously unrecognized E−H+ conformation is the pioneer integrin for neutrophil spreading and appears to be organized by kindlin-3.
The pathogenic role of choroid plexus infiltrating T cells in Neuropsychiatric Lupus
Authors: Moore Erica , Huang Michelle , Reynolds Cara , Putterman Chaim 
Published: January 2023

Abstract
Neuropsychiatric systemic lupus erythematosus (NPSLE) manifests as a range of symptoms, including cognitive abnormalities, mood disorders, and psychosis. The pathogenesis is complex and not yet completely understood. We recently proposed that the choroid plexus (CP) could be an alternative entry site to the blood brain barrier for immune mediators to enter the central nervous system. In the MRL/lpr lupus mouse model, the CP is heavily infiltrated with T and B cells, which is reminiscent of human NPSLE patients whose autopsies revealed infiltrates in the CP. Nevertheless, the role of CP infiltrating T cells in the pathogenesis is not known and is the objective for this study. In two cohorts, we evaluated whether intracerebroventricularly (ICV) delivered MRL/lpr CP T cells accelerate neuropsychiatric disease in young female MRL/lpr mice. Six week old female MRL/lpr mice were injected with MRL/lpr CP CD3+ T cells, MRL/lpr splenic CD3+ T cells, or PBS. The T cells were pooled from the CP and spleen of 16 week old MRL/lpr mice and expanded with anti-CD3/CD28 antibodies (1 ug/mL) and mIL-2 (10 U/mL) for 4 days. About ~200,000 cells were injected into the lateral ventricle from the bregma at AP −0.34 mm, ML −1.0 mm, and depth −2.0 mm. After 4 weeks post-injection, we found that CP T cell recipient mice had increased cognitive deficits compared to PBS controls. Histological assessment revealed an increase in lymphocytic infiltration, the number of proliferating cells, and IgG and complement deposition, in the choroid plexus of CP T recipient mice. Our result indicate that T cells can migrate to and proliferate in the CP following ICV injection, and that brain infiltrating MRL/lpr T cells may be a key contributor to the pathogenesis of NPSLE.
Oxysterols induce proinflammatory response via Focal Adhesion Kinase (FAK) signaling
Authors: Casimiro Kim Roxana Chiok , Pokharel Swechha Mainali , Bose Santanu 
Published: January 2023

Abstract
We recently reported that the oxysterol 25-hydroxycholesterol (25HC), which is an oxygenated form of cholesterol, promotes proinflammatory response via Integrin-Focal Adhesion Kinase (FAK) signaling. However, it is unknown whether other non-25HC oxysterols possess similar proinflammatory activity induced by FAK signaling pathway. Thus, we tested two non-25HC oxysterols, 22-hydroxycholesterol (22HC) and 7-alphahydroxycholesterol (7-alpha-HC), for their ability to induce proinflammatory response in macrophages. Both of these oxysterols induced a proinflammatory response since we observed production of the proinflammatory cytokine tumor necrosis factor alpha (TNF) from macrophages treated with 22HC and 7-alpha-HC. Furthermore, this response was mediated via FAK signaling as TNF production from 22HC treated macrophages was abrogated in the presence of a FAK-inhibitor. Taken together, this study suggests that non-25HC oxysterols can utilize a similar signaling pathway as 25HC to induce proinflammatory response. Studies are currently in progress to dissect the molecular mechanisms involved in triggering proinflammatory response by 22HC and 7-alpha-HC.
Follicular dendritic cell migration is facilitated by the expression of matrix metalloproteinase 3 through TNFα-ERK1/2-AP-1 signaling
Authors: Pak Hyo-Kyung , Kim Yong-Woo , Lee A-Neum , Roh Jin , Chung Yoo-Sam , Park Chan-Sik 
Published: January 2023

Abstract
Folliculardendritic cells (FDCs) are the crucial stromal components in the germinal center and have the pivotal roles in high-affinity antibody production. FDCs are developed and maintained in the germinal centers by lymphotoxin α1/β2 and tumor necrosis factor-α (TNFα). However, many of the features of FDCs are still not identified, particularly its migration. Here, we demonstrate for the first time that the migration of human FDCs is enhanced by TNFα-mediated metalloproteinase 3 (MMP3) expression. MMP3 was highly expressed in the normal human germinal centers. MMP3 expression was markedly increased in human primary FDCs by TNFα. TNFα induced the ERK1/2 phosphorylation and the promoter activity of the AP-1 binding sites in the Mmp3 promoter region. TNFα treatment increased FDC migration and the knockdown of MMP3 significantly reduced the TNFα-induced migration of FDCs. Overall, we identified a novel control mechanism for the expression of MMP3 in FDCs to modulate the migration, which may play an important role in lymphoid follicle formation. Since lymphoid follicles are observed in the lesions of autoimmune diseases and lymphomas, controlling the MMP3/TNFα-mediated migration of FDCs can provide novel insight into therapeutic modalities for aberrant germinal center-associated disorders.