bluuebunny/crossref_metadata_2025 huggingface dataset:
New insights into rheumatoid arthritis by whole transcriptome profiling of autoreactive B cells
Authors: Mahendra Ankit , Yang Xingyue , Qiu Peng , Park Daechan , Coarfa Cristian , Agarwal Sandeep K , Varadarajan Navin
Published: January 2023

B cells are important in the pathogenesis of rheumatoid arthritis (RA). By utilizing a flow-cytometric assay to reliably detect the low frequency of autoreactive B cells (ABC) in the blood of RA patients, we performed RNA sequencing of citrulline-specific ABCs and compared them with hemaglutinin (HA) specific B cells. To the best of our knowledge, this is the first complete transcriptome map of human ABCs in autoimmunity, and our data shows a specific up regulation of amphiregulin (AREG) (q-value = 3.1×10−12, fold-change = 250); a member of the EGF growth factor of ligands. Also, the receptor signaling pathways associated with AREG including EGF mediated signaling (FDR q-value = 0.06) and the NRG1 mediated signaling (FDR q-value = 0.03) were enriched, supporting an important role for AREG. Consistent with the well-recognized role of TNF in RA, our RNA-seq data revealed highly significant enrichment of both the TNF pathway (FDR q-value = 0.02) and genes that are targets of TNF (FDR q-value = 0.08) in ABCs from RA patients. Comparisons of our differential expression gene analyses with known risk loci of RA (SNPs) identified genes with decreased expression including TNFAIP3 (leading to inflammatory responses), and REL (anti-apoptotic); and increased expression including CCR6 (maturation and differentiation), and ETS1 (development and maturation) within ABCs than HA specific B cells. Ig sequencing of ABC show normal mutational targeting frequencies but increased somatic hymermutation. In aggregate, our results indicate that the pro-inflammatory nature of IL15 and TNF mediated signaling might promote a feed-forward loop promoting the altered differentiation of ABCs through the AREG-TNF signaling axis, which in turn promotes inflammation.
Anti-nuclear antibodies in autoimmune prone mice are protective against HIV
Authors: Schroeder Kristin M , Strauch Pamela , Harper Michael , Pelanda Roberta , Santiago Mario L , Torres Raul M
Published: January 2023

HIV broadly neutralizing antibodies (bnAbs) recognize conserved regions of the HIV envelope trimer, are capable of neutralizing up to 90% of cross-clade HIV strains in vitro, and have auto- and poly-reactive specificities. Multiple bnAbs, with different specificities for HIV envelope, have been reported to recognize autoantigens, such as membrane phospholipids, histones, and human ubiquitin ligase 3. Further, bnAb CH98 has been identified in an HIV infected individual with systemic lupus erythematosus (SLE) and was found to recognize both dsDNA and the CD4 binding site, as well as neutralize HIV strains of multiple clades in vitro. From this data, our lab hypothesized that both central and peripheral B cell tolerance checkpoints inhibit potentially neutralizing HIV-Ab responses, and that autoimmune prone individuals are capable of mounting neutralizing Ab responses. In order to further investigate this, we immunized age-matched, wild type and SLE prone strains of mice with HIV envelope proteins in adjuvant. We found that a subset of SLE prone mice neutralized multiple heterologous HIV strains 14 days post immunization, which was not observed in wild type mice. This neutralizing Ab response positively correlated with age and total serum IgM levels. SLE prone mice produced envelope-specific IgG of similar magnitude and isotypes as wild type mice, following envelope immunization. Using an array with 92 autoantigens, we found that the subset of SLE prone mice, which neutralize HIV, produce higher levels of histone H2A and dsDNA reactive IgM, as compared to the non-neutralizing SLE and wild type mice. Lastly, in vivo and in vitro studies show that activation of the autoimmune B cells by adjuvant is required for this neutralization activity.
The role of plasma cell precursors in a mouse model of lupus
Authors: Rosean Timothy R , Loo William , Erickson Loren D
Published: January 2023

Systemic lupus erythematosus (SLE) is an autoimmune disease, which occurs when terminally-differentiated B cells, called plasma cells (PC), secrete anti-nuclear antibodies (ANA). Once formed, PCs persist in the bone marrow for the lifetime of the host, but the mechanisms of this long-term persistence are poorly understood. Our lab and others have identified a novel subset of terminally-differentiated B-lineage cells. These cells, called plasma cell precursors (PCpre) are a post-germinal center cell population, which are both long-lived and capable of self-renewal. While PCpre do not actively secrete antibody, they can terminally differentiate into mature, long-lived PCs. The role of PCpre in the production of ANA has yet to be elucidated. To understand the role of PCpre in autoimmunity, our lab utilized the congenic New Zealand Black Autoimmunity 2 (NBA2) mouse strain, which develops B cell abnormalities similar to SLE. NBA2 mice develop significantly higher frequencies and numbers of PCpre compared to controls. PCpre from NBA2 mice express the transcription factors IRF-4, BLIMP-1, and XBP-1 that are known to be upregulated in PC. In addition, PCpre express high levels of B220 and MHC class II. Adoptive transfer of PCpre cells from NBA2 mice into B6 controls results in ANAs in recipient mice by day 14, demonstrating the pathogenic potential of PCpre in mouse models of SLE. Furthermore, these PCpre are capable of long-term survival as they persist in the recipient for at least 40 days. Based on these findings, we hypothesize that PCpre are involved in the renewal of the PC niche in the NBA2 model of SLE. Current research is focused on the mechanisms by which PCpre undergo proliferation and terminal differentiation.
Dnase1L3 trafficking is redirected from secretion to the nucleus in lupus-prone mice
Authors: Abbott Kennady N , Wu Wenbo , Shi Guilan , Salter Russell , Keyel Peter
Published: January 2023

Pediatric-onset Systemic Lupus Erythematosus (SLE) is linked to a deficiency of the endonuclease Dnase1L3. Dnase1L3 can function extracellularly as a barrier to transfection or localize to the nucleus of macrophages to cleave DNA during apoptosis, though the mechanism by which Dnase1L3 protects from pediatric-onset SLE is unknown. Two polygenic lupus murine models, MRL-lpr and NZB/W F1, bear the activity-reducing T89I mutation in Dnase1L3. Dnase1L3 T89I has an eight-fold decrease in barrier-to-transfection activity, while the endonuclease activity is only decreased two-fold compared to the wildtype, providing a tool with which to dissect Dnase1L3 trafficking in SLE. We hypothesized that Dnase1L3 T89I localizes to the nucleus, which decreases barrier-to-transfection activity and enhances SLE onset. We transfected HEK cells with wild type or T89I Dnase1L3, and determined the subcellular localization of Dnase1L3. We found that the majority of wild type Dnase1L3 is secreted from the cell, while Dnase1L3 T89I showed increased nuclear localization. Since Dnase1L3 alters inflammasome function, we tested whether inflammatory stimuli alter Dnase1L3 trafficking. LPS treatment of cells transfected with Dnase1L3 T89I reduced its nuclear localization. Altered Dnase1L3 trafficking may impair inflammasome function, since macrophages from NZB/W F1 mice secreted less IL-1β than wild type mice. Overall, the increase in nuclear localization of Dnase1L3 T89I suggests that barrier to transfection or extracellular nuclease activity may promote IL-1β release and protect against pediatric-onset SLE.
Chronic Interferon-gamma expression triggers a female bias in the development of primary biliary cholangitis
Authors: Young Howard A , Leung Patrick , Tsuneyama Koichi , Valencia Julio , Hodge Deborah L , Kim Seohyun , Back Tim , Karwan Megan , Merchant Anand , Feng Dechun , Gao Bin , Park Ogyi , Gershwin M Eric , Bae Rachel
Published: January 2023

An important issue in autoimmunity research has been the ability to identify the mechanisms that link genetic susceptibility to environmental influences. In most autoimmune diseases, including primary biliary cholangitis (PBC), the serologic hallmarks of disease precede clinical pathology by years, thus making early diagnosis and potential treatment difficult if not unlikely. Thus identifying animal models that present early events becomes essential to understanding disease initiation and progression. Although murine models of PBC have been identified, there are none that fully recapitulate the human disease. Herein we have utilized a novel mouse model exhibiting dysregulation of interferon-gamma (IFNg) expression, characterized by prolonged and chronic expression of IFNg as a result of the replacement of the IFNg 3′ UTR AU-rich element with random nucleotides (designated ARE-Del−/−). These C57 BL/6 ARE-Del−/− mice develop autoimmune cholangitis with a female predominance characterized by upregulation of total serum bile acids, portal duct inflammation, spontaneous production of AMA and increased skin pigmentation. RNA seq data has revealed that female ARE-Del−/− mice have >1000 genes deferentially expressed in hepatocytes compared to control mice, including genes that represent early responses in disease progression. Furthermore, the data indicates that the Type 1 IFN pathway is also involved, implying that Type II interferon interplays with Type I IFNs in the initiation of PBC. Thus persistent IFNg expression, along with Type I and Type II IFN pathway interactions, is critical for the pathogenesis of PBC and recapitulates the sex bias observed in patients.
Next generation sequencing reveals HLA and KIR susceptibility alleles for rheumatoid arthritis
Authors: Geraghty Daniel , Nakanishi Mari , Ide Takeshi , Tanaka Hidenori , Miyazaki Yuki , Saji Hiroh , Inoue Kazuya , Murata Norikazu , Ashida Tsuneo , Hatake Katsuhiko , Ishitani Akiko
Published: January 2023

Previous associations of KIR with rheumatoid arthritis (RA) have been reported in some but not all populations studied, possibly due to limited genotyping in some studies, and gene content homogeneity in some populations. In this study, HLA and KIR typing was carried out, including KIR haplotype and allele typing, using state of the art sequencing methodologies in a re-examination of the association of these gene families with RA in a Japanese cohort. An additional cohort of pollen allergy patients was examined in an effort to distinguish common genetic elements in a phenotype functionally reciprocal to RA. DNAs from 116 RA patients, 167 pollen allergy patients, and 185 healthy controls were examined for KIR haplotype, allele type and HLA class I and II allele types using next generation sequencing (NGS). Association analysis was carried out with healthy controls classified into two groups, positive and negative for allergen specific IgE antibodies, including a pollen allergy group for comparison with RA. Significant results were observed with allele types KIR2DS4*007 and KIR3DL1*00501, which strongly associated with disease, while KIR 3DL1*001 and 3DL1*02901 associated with a protective phenotype. These findings were significant when compared with the IgE positive control group while the IgE-negative group did not demonstrate significance. Given that KIR3DL1 is an inhibitory receptor and the KIR3DL1*00501 allele has been reported as a low expression allele, these findings are consistent with a model of weak suppression of NK cytotoxic activity as a contributing factor in RA. Further support for this model was observed from the reciprocity of the genetic associations between RA and pollen allergy.
Role of the TNF-family cytokine TL1A and its receptor DR3 in systemic autoimmunity and glomerulonephritis
Authors: Meylan Francoise , Hayes Erika , Ferdinand John , Farley Taylor , Richoz Nathan , Gabay Odile , Siegel Richard M
Published: January 2023

DR3 (Tnsfrsf25) is a death domain-containing tumor necrosis factor receptor found primarily on activated T cells and innate lymphocytes. Interaction with its ligand the TNF-family cytokine TL1A plays critical roles in diverse autoimmune disease models in facilitating T cell accumulation at the site of inflammation while the systemic response was unaffected. To determine whether DR3 could play a role in systemic autoimmunity, we studied the role of DR3 in the MLR/lpr mouse model of lupus. Lpr mice have a mutation in the TNF family receptor Fas (CD95, Tnfrsf6) that impair its expression, causing a systemic autoimmune disease that mimics systemic lupus erythematosus (SLE) observed in humans. DR3 deficient lpr mice developed identical degrees of lymphadenopathy, splenomegaly compared to control littermates. Accumulation of autoantibodies was also not dependent on DR3. However, DR3 deficiency significantly protected MRL/lpr mice from IgG and C3 glomerular immune complex deposition and DR3 deficient kidneys had decreased medullary T cell accumulation, decreased periglomerular and medullary macrophage infiltrates, reduced proteinuria and reduced renal pathology. In addition, DR3-deficient CD4+ T cell transfer in a chronic graft-versus-host disease (GVHD) model showed similar levels of autoantibodies and immunoglobulin class switching than with wild-type CD4+ T cells but decreased proteinuria and reduced renal pathology. These findings show that DR3 specifically promotes renal pathology in the MRL/lpr mouse model of lupus nephritis and in chronic GVHD without affecting systemic autoimmunity. Blockade of TL1A-DR3 interactions may be a novel strategy to counter nephritis and other organ damage in human SLE.
Meta-Analysis from Gene Expression Profiles of Lupus Affected Tissues Reveals Novel Immune Cell Contributions
Authors: Grammer Amrie C. , Heuer Sarah , Catalina Michelle , Robl Robert , Madamanchi Sushma , Wanjari Pankhuri , Min Junghwan , Bachali Prathyusha , Kretzler Matthias , Berthier Celine , Suarez-Farinas Mayte , Davis Laurie , Lauwerys Bernard , Houssiau Frédéric , Lipsky Peter
Published: January 2023

Immunologic mechanisms causing tissue damage in autoimmune diseases such as SLE are not fully understood. To gain additional insight, gene expression profiles obtained from lupus affected skin, synovium and kidney were obtained and compared to meta-analyzed data obtained from active lupus B, T and myeloid cells. More than 300 arrays from lupus patients and appropriate controls were analyzed to determine differentially expressed (DE) genes (8279 discoid lupus skin, 5465 synovium, 6381 glomerulus, 5587 tubulointerstitum). Notably, the majority of lupus affected tissue DE genes were detected in more than one tissue and 439 were differentially expressed in all tissues. A variety of approaches assessed the molecular pathways and cellular phenotypes accounting for these common lupus affected tissue DE genes. Curated STRING-based interaction analysis identified a number of pathways including co-stimulation of T cells, activation of B and myeloid cells, antigen presentation, TLR signaling and p38 activation; from a total of 193 IPA-documented pathways, 59 were common to all tissues including p38 signaling, TLR signaling, maturation of dendritic cells, B cell activation and ICOS-ICOSL in T cells. Novel bioinformatics approaches documented that more than 50% of the DE genes in the tissues were associated with immune cell function and 11–18% were unique to the immune system. Further analysis using LINCS and additional novel software highlighted specific molecular pathways of immune cell activation including the JAK/STAT and the IL12 pathways. These results demonstrate the value of comprehensive application of orthogonal curated bioinformatics tools in identifying the role of immune cells in lupus pathogenesis and tissue damage.
Exacerbation of MPTP induced neurodegeneration by a CD4+ effector T cell line specific for N-α-synuclein in a mouse model of Parkinson’s disease
Authors: Wilshusen Rebecca A , Mosley R. Lee
Published: January 2023

Inflammation and oxidative stress play critical roles in PD. Moreover, T cell-mediated immunity regulates innate immunity to control or exacerbate neuroinflammation. We have previously shown in the MPTP mouse model that adoptive transfer of effector T cells (Teffs) which recognize nitrated α-synuclein (N-α-syn) as non-self exacerbates microglial-mediated neuroinflammation with amplified dopaminergic neurodegeneration. The specific mechanisms by which these Teffs modulate neurodegeneration remain enigmatic. However, increasing evidence suggests neurotoxic inflammatory activities have a profound effect on the pathogenesis of PD. Herein, we report on the generation and long-term culture of a N-α-syn specific CD4+ Teff line that induces increased production of reactive nitrogen species from microglia and also intensifies the loss of tyrosine hydroxylase+ (TH+) neurons of MPTP-intoxicated mice. Flow cytometric analysis showed the Teffs line exhibited very distinct expression of CD3 and CD4 cell surface markers and intracellular IL-17a cytokine, suggestive of characteristics consistent with a Th17 cell. Co-culture of the Teffs with a dopaminergic cell line significantly increased cell death of dopaminergic cells in comparison to controls. Lastly, adoptive transfer of the Teffs line to MPTP-intoxicated mice increased the loss of TH+ neurons. Together, these results support the notion that Teffs may play a pivotal role in the progression of neurodegeneration in PD. Further investigation of the mechanisms behind the exacerbation of neurodegeneration caused by Teffs has the potential to advance therapeutic strategies that target neurotoxic mechanisms to attenuate neuroinflammation and ameliorate neuronal loss in PD.
Immunogenetic analysis of autoimmune hepatitis in Alaska Native People
Authors: Ferrante Andrea , Kennedy Stephanie N.
Published: January 2023

Autoimmune hepatitis (AIH) is a chronic inflammatory liver disease, in which a multiplicity of genes, operating alone or in concert, determines the risk that environmental factors act as a trigger of an adaptive immune response against liver self-antigens. Susceptibility to AIH is conferred by genes of the human leukocyte antigen (HLA) system, especially alleles of class II HLA molecules (HLAII). Genetic risk is also conferred by immune-related genes outside the HLA region, whereas the environmental triggers are not known. Though AIH has a global occurrence, its prevalence among Alaska Native People (AN) is the highest reported in the literature, yet efforts to understand the causes of such a high prevalence have been limited. To identify potential risk factors in this population, we have determined by Next-Generation Sequencing HLAII haplotypes and variants of CTLA-4, AIRE, TNFA*2, and VDR expressed in 48 AN AIH patients. The most common HLA alleles identified are HLA-DRB1*04:01:01, HLA-DRB1*14:01:01 and HLADQB1*03:01:01. The most common haplotype is HLA-DRB1*04:01:01, DQB1*03:01:01. DRB1*0401 and 1401 are significantly more represented in the affected cohort than in the general AN population (DRB1*0401: 0.427 vs 0.114; DRB1*1401: 0.104 vs 0.032; p<0.01), as well as DQB1*0301 (0.615 vs. 0.381, p<0.01). Significant SNPs have also been identified for CTLA-4 (+49A/G, G=52.1%, more represented in our cohort than in the general population) and VDR (FokI polymorphism, F allele frequency = 71.9% vs. general population 62.5%). Self-antigen presentation by the identified HLAII alleles in the context of reduced tolerance associated with the above polymorphisms might explain development of disease.
Prevention of LPS-induced lung inflammation by PLAG administration via blocking of IL-6-STAT3-MIP-2 pathway
Authors: Lee Ha-Reum , Shin Su-Hyun , Yoo Nina , Yoon Sun Young , Kim Joo Heon , Nah Seong-Su , Kim Myung-Hwan , Sohn Ki-Young , Kim Heung-Jae , Chong Saeho , Han Yong-Hae , Kim Jae Wha
Published: January 2023

Neutrophil plays a key role in the innate immune system through as the first leukocyte to be migrated to the acute inflammatory region. Lung inflammation is an acute respiratory failure and links closely to excessive neutrophil recruitment and severe mortality. The intranasal LPS introduced mice were shown the over-flowed neutrophils into bronchoalveolar. PLAG is a synthetic monoacetyldiacylglycerol (acetyl-DAG). DAG is a lipid second messenger for PKC activator which is associated with an important for cellular functions. The transmigrated neutrophils in alveolar by LPS stimulation were completely blocked by PLAG administration. LPS-induced cell migration associated factors such as IL-6, MIP-2 and S100A8/9 were decreased by addition of PLAG. STAT3 phosphorylation which related to neutrophil migration was significantly down regulated in PLAG dieted mice. Interestingly, the selectivity of PLAG in the ameliorating of excessive neutrophil migration was verified when compared with PLAG analogues and metabolites. Post treatment of PLAG has also therapeutic effect on LPS-induced lung inflammation. These data could be utilized as a most potential therapeutic agent for lung inflammation through regulation of neutrophil transmigration via blocking of IL-6-STAT3-MIP-2 pathway.
Research of MFG-E8, OPG and SOCS-3 on preeclampsia
Authors: Ilhan Fulya , Atabay Hatice , Ozkan Zehra Sema , Doni Nebiye Yentur , Can Sevim Tuncer
Published: January 2023

Preeclampsia is a specific disease and closely associated with systemic inflammatory response. The aim of this study is research to possible role of some inflammatory factors and the controlling factor in inflammation in the etiology of preeclampsia or reflection of this situation on placental tissue. Three parameters were included in the study regarded as potential risk factors: Milk Fat Globul Epidermal Growth Factor-8 (MFG-E8); is an endometrial epithelial proteins and regulates inflammation and apoptosis. MFG-E8 is an important in the development of endometrium and chorionic villus. Osteoprotegerin (OPG); is pro-angiogenic factor, it can be important for plasental angiogenesis. Suppressor of cytokine signaling-3 Receptor (SOCS-3) has a role on prevents the inflammation. Serum and placental tissue samples were used as materials. Control group is composed of 50 healthy women volunteers with no history of preeclampsia and 28–34th weeks of pregnancy. The experimental group consisted 40 preeclamptic women in 28–34th week of pregnancy. All of the measurement analyzed through the ELISA (Enzyme-Linked Immuno Sorbent Analysis) method and findings evaluated comparatively. MFG-E8, SOCS-3 and OPG levels analysed with ELISA in the serum and placental tissue. OPG levels in serum raised and displayed statistically significant difference with respect to control group (p<0.01). In respect to the serum MFG-E8 and SOCS-3 are not observed any difference between two groups. MFG-E8 tissue levels are statistically significantly low in preeclamptic group with respect to healthy control group and that there is no changes in the levels of SOCS-3 (p<0.01).
Pinpointing IL-23-target cells in autoimmunity and inflammation using novel mouse models
Authors: Yang Yi
Published: January 2023

Interleukin-23 is the first cytokine linked to proinflammatory Th17 cells. Although IL-23 is dispensable for the differentiation of Th17 cells in vitro, it is absolutely required for their late maturation and acquisition of pathogenicity in vivo. IL-23 signaling promotes and exacerbates autoimmunity in many disease settings. On the contrary, IL-23 protects mice from pathogen-induced infection. Key cellular mediators of IL-23 in various immune responses remain to be understood since the receptor of IL-23 (IL-23R) is expressed by multiple immune cells. To elucidate the major cellular mediators of IL-23 in both pathogenic and beneficiary immune responses, we developed IL-23R conditional knock-out and IL-23R conditional knock-in mice. These mice allow us to turn on/off IL-23R expression in desired cell types by crossing with lineage-specific Cre-recombinase mice. To understand the significance of Th17 cell-intrinsic and –extrinsic IL-23R signaling in vivo, we examined these mice in various diseases. Interestingly, we found that CD4-autonomous IL-23R signaling is necessary and sufficient for the development of EAE. In contrast, IL-23R mediated protection against Citrobacter rodentium, a mucosal pathogen, is not impaired in CD4 T cell-specific IL-23R deficient animals. Thus, our data suggests a contrasting requirement for the IL-23-Th17 axis in CNS vs. mucosal sites. These novel mice shall be valuable in studying IL-23 responses in other diseases.
Evaluation of the Inflammatory Response in Non-human Primate (NHP) Models of Poly-trauma
Authors: Vernon Philip J , Schaub Leasha , Grossman Heather , Glaser Jacob , Sheppard Forest
Published: January 2023

Background

Trauma is a leading cause of mortality, with those surviving their initial injuries remaining at risk for death due to deleterious inflammation. Preclinical investigations of immunomodulatory therapeutics are hindered by the limitations of available animal models. We sought to develop a translatable poly-trauma model in NHPs to facilitate future translational immunotherapeutic evaluation.

Methods

NHPs (n=24) were randomly assigned to: 60min hemorrhage shock alone(n=8) or in combination with laparotomy (n=8) ± femur fracture (n=8), resuscitation and 24hr survival. Blood was collected at baseline (BLSN), end of shock (EOS), end of resuscitation (EOR), T=360min and T=24hr; lung tissue was collected at T=24hr. Blood and tissue cytokines were quantified by a 23 plex multiplex. Results reported as mean±SEM, Statistics: Two-tailed T-Test, with p<0.05 significant.

Results

Circulating IL-6, IL-10, IL-15, MCP-1, TGFa and VEGF were significantly elevated in all models as early as EOS; IL-1RA, IL-15, TNFa, IL-6, IL-10, MCP-1, TGFa and IL-5 were significantly increased as early as EOS and directly correlated with injury severity. Lung tissue G-CSF, IL-1b, IL-RA, IL-6, IL-10, IL-15, IL-17A, IL-18, MCP-1, sCD40L, TGFa and VEGF were significantly elevated and directly correlated with injury severity.

Conclusion

Increases in cytokines identified in all models are consistent with those reported in human trauma studies. Cytokines strongly associated with injury severity score (ISS) in humans directly correlated with model severity (IL-6 and IL-10). These models have uncovered cytokine responses not characterized in poly-trauma (IL-15), demonstrating their utility in identifying novel targets for human investigation.
Modeling cytokines and chemokines through a rooted correlation network
Authors: Qeadan Fares , Dwivedi Sunita , Page Kimberly , Gravitt Patti
Published: January 2023

Several studies have investigated the role of cytokines/chemokines in infection and chronic and autoimmune diseases using different statistical models. Generally, only two mediators are evaluated at a time, by comparing group-specific mean concentration differences. Restricted pairwise analysis from a large array of markers which inherently operate at a systems level to regulate immune response ignores important inter-correlation structures, compromising clinical and epidemiological inference. We propose the use of a rooted correlation network as a novel statistical method which allows group-specific comparisons of a panel of soluble immune markers while adjusting for the interaction complexity.

We examined soluble immune markers in luminal fluid samples from 90 HPV infected female participants with and without soil transmitted helminth (STH) co-infection in Peru. We constructed a lower triangular correlation matrix using Spearman’s correlation with Sidak adjustment for both the STH-infected and -uninfected women. Rooted correlation network was constructed for the cytokines and chemokines that were insignificant in the non-helminth group but became significant in the helminth one. The generated cytokine network demonstrated statistically significant association of Th2 pathway cytokines with the canonical Th2 cytokine IL-4. Using this new statistical model, we demonstrate that the female genital tract of STH-infected women reveals polarization towards Th2 immune response (IL-25, IL-21, IL-5, IL-10, IL-8, and IL-31) in comparison to STH-uninfected women, which was not easily recognizable when using standard bivariate methods with multiple comparisons.
Macrophage migration inhibitory factor-induced autophagy is involved in thrombin-mediated endothelial cell hyperpermeability
Authors: Yeh Trai-Ming , Chao Chiao-Hsuan
Published: January 2023

Sepsis is a disease in which systemic inflammation-induced vascular leakage and disseminated intravascular coagulation lead to multiple organ failure. Despite a high mortality andmorbidity, there is currently no specific drug for sepsis. Thrombin, a serineprotease that mediates key sepsis-related pathologies, both activates coagulation cascades and acts as a mediator to induce vascular hyperpermeability and inflammation. It is known that thrombin can induce endothelial cells to secrete macrophage migration inhibitor factor (MIF) and MIFcan cause endothelial hyperpermeability via autophagy. However, it is unclearwhether MIF-induced autophagy is involved in thrombin-mediated vascular hyperpermeability. Herein, we show that thrombin treatment induced hyperpermeability, which was accompanied by VE-cadherin translocation in the human microvascular endothelial cell line (HMEC-1). Additionally, thrombin could induce MIF secretion and autophagy in HMEC-1 cells. Inhibition of MIF by its antibody or inhibitors could reduce thrombin-induced endothelial hyperpermeability and autophagy. However, inhibition of autophagy prevented only endothelial hyperpermeability, not MIF secretion. Finally, both MIF and autophagy inhibitors reduced thrombin- induced vascular leakage in vitro. Together, our data suggest that MIF-induced autophagy is involved in thrombin-mediated endothelial cell hyperpermeability. Further studies designed to test whether autophagy inhibitors could prevent endothelial barrier dysfunction in sepsis may provide potential therapeutic modalities to reduce sepsis-related mortality.
The role of the interleukin-12/STAT4 axis in breast cancer
Authors: Nunez Angel R
Published: January 2023

Breast cancer (BC) is one of the leading causes of cancer-related deaths among women worldwide. Interleukin-12 (IL-12) has demonstrated anti-cancer effects in vitro and in vivo. IL-12 signaling occurs mainly through STAT4, and it induces anti-tumorigenic actions such as promotion of apoptosis, differentiation of T helper 1 (Th1) cells, increasing immune cell’s cytotoxicity and preventing angiogenesis. The role of the IL-12/STAT4 axis in human breast cancer has not been studied thoroughly. Our aim was to assess if the IL-12/STAT4 pathway is differentially expressed and contribute to a pro-tumor environment in BC. We hypothesize that genetic and molecular impairments lead to a downregulation of the IL-12/STAT4 axis and are associated with breast cancer risk. mRNA and protein levels of key IL-12/STAT4 molecules were measured using peripheral mononuclear blood cells (PBMCs) from BC patients and non-BC controls. Interleukin-12 B (IL12B), STAT4 and interferon gamma (IFNG) expression was lower in BC samples when compared with the controls. These results suggest that the IL-12/STAT4 axis pathway is differentially expressed in BC, and prompts more specific analyses such as whether this is BC subtype-specific.
Low-dose gamma radiation does not mitigate inflammatory cytokine responses in a murine model of Benzo[a]pyrene-induced lung cancer
Authors: Bruce Veronica R , Gott Katherine , Scott Bobby , Wilder Julie
Published: January 2023

Low-dose radiation has been reported to modify the immune system and may lead to lung cancer suppression. In a series of short-term studies, A/J mice were injected i.p. with a carcinogen, Benzo[a]pyrene (B[a]P), and/or whole-body irradiated with low-dose gamma radiation (LDR). We previously reported that B[a]P promoted a pro-inflammatory cytokine response (IL-1β, IL-6, IL-17, TNF-α) whereas LDR promoted an anti-inflammatory response (IL-4, IL-10) and increased IL-2 in supernatants of Concanavalin A-stimulated splenocytes. Consequently, we enumerated cytokine-producing cells via intracellular flow cytometry and examined cytokine production capacity by median fluorescence intensity (MFI) analysis. In the spleen, B[a]P enhanced IL-1β and IL-17 producing Gr-1+ and CD19+ cells. Although B[a]P and LDR enhanced IL-6, or IL-2, IL-4, and IL-10 production respectively, the corresponding cell types were unidentifiable. MFI was not affected by either B[a]P or LDR. In the lung, B[a]P increased IL-1β production in CD19+ and CD4+, IL-6 in CD4+, IL-17 in Gr-1+ and CD19+, and TNF-α in CD19+ and CD8+ cells. B[a]P also increased IL-4, IL-10, and IL-13 expressing CD4+ cells, and IL-10 expressing CD8+ cells. Increased MFI was observed for several of these cells types. Neither cytokine producing cell numbers nor MFI were affected by LDR. Although we observed B[a]P-mediated anti-inflammatory cytokine production in the lung, we postulate that this is a tissue specific response to ease inflammation through feedback mechanisms. Finally, we found no evidence that LDR mitigates B[a]P-mediated inflammatory cytokine responses in either tissue. We believe it is unlikely that LDR suppresses B[a]P-mediated cancer suppression through immune modulation.
Establishment of tissue-resident macrophage core and tissue specific programs during embryogenesis
Authors: Mass Elvira , Ballesteros Ivan , Farlik Matthias , Halbritter Florian , Klughammer Johanna , Bock Christoph , Geissmann Frederic
Published: January 2023

Tissue resident macrophages develop during embryogenesis, are maintained independently of hematopoietic stem cells during adulthood, and display tissue-specific functions and phenotypes. To understand the genetic program that drives macrophage differentiation from distinct progenitors and their tissue-specific identity, we performed a systematic spatio-temporal analysis of macrophage development in mice. Transcriptional and in situ analyses revealed that a core macrophage program is established very early and within a short time window and is conserved throughout fetal development and in adult macrophages. A new genetic mouse model supports the transcriptional data as it allows to specifically label differentiating macrophages, and therefore targets adult macrophages in all tissues. Interestingly, immediately after colonization of the target tissue, we could identify tissue-specific macrophage programs that are maintained until adulthood. Additionally, we identified a novel transcriptional regulator, which is expressed in early fetal liver macrophages, and whose inactivation results in an impaired Kupffer cell development. In summary, we present a macrophage developmental atlas, identify novel tissue-specific markers and transcriptional regulators, and show that tissue specialization is mainly influenced by ontogenetic gene expression programs.
Expression and function of Olig2 in eosinophils
Authors: Chung Il Yup , Hwang Sae Mi , Uhm Tae Gi
Published: January 2023

Eosinophils are granulocytes that are associated with homeostasis and type 2 immune responses, allergy and parasitic infection. Olig2 is a basic helix loop helix transcription factor that is known to play a key role in cell fate decision and subsequent differentiation of oligodendrocytes. Herein, we report the expression and possible function of Olig2 in eosinophils. When cord blood (CB) CD34+ hematopoietic stem cells differentiated toward eosinophils during a 24 day culture period, Olig2 protein was expressed only in CB eosinophils at day 24, a time when these cells were considered fully differentiated and matured, while Olig2 protein was not detected at day 18 or at earlier time points. Olig2 protein was also abundantly expressed in human peripheral blood eosinophils, but not neutrophils, monocytes, lymphocytes or CB mast cells. RNA-seq analysis showed that numerous genes, especially those encoding eosinophil surface molecules, were highly upregulated along with OLIG2. Among the genes examined, SIGLEC-8 mRNA and protein were downregulated in parallel with OLIG2 by an Olig2 siRNA or a shRNA. In reporter gene and chromatin immunoprecipitation experiments, an E-box in the first intron was found to stimulate SIGLEC-8 gene transcription and bind Olig2. Occupancy of the E-box by Olig2 was also seen upon differentiation of immature eosinophilic AML14.3D10 cells. Hence, at least one important aspect of eosinophil differentiation is driven by Olig2, a transcription factor that has not previously been reported in normal granulocytes.
Role of Wnt-4 signaling in dendritic cells lineage and differentiation
Authors: Jain Pooja , Scharer Christopher , Kawasawa Yuka , Sreedhar Sreesha , Khan Zafar , Boss Jeremy
Published: January 2023

Dendritic cells (DCs) are important mediators of the innate and adaptive immune responses. The network of genes governing the differentiation of these cells has been a topic of discussion for a long time, mainly because of their similarity to other myeloid lineages such as monocytes and macrophages. In order to gain insights into the characterization of factors that contribute to the lineage restricted progenitors in the bone marrow that govern DC differentiation, we performed epigenetic analyses utilizing next generation sequencing of bone marrow-derived CD11c+ DCs (BMDCs). Fms-like tyrosine kinase 3 ligand (Flt3L) is a hematopoietic growth factor that is crucial to the development and commitment of DCs from the common myeloid progenitor (CMP) to the common dendritic progenitor (CDP), subsequently yielding conventional and plasmacytoid DCs. The combined analysis of the DNA methylome by Reduced Representation Bisulfite sequencing (RRBS) and the transcriptome by RNA-seq identified a hypermethylation and a reduction in gene expression of Wnt-4 signaling. Wnt-4 gene family is implicated in several developmental processes especially in determination of cell fate and embryogenesis. A comparative validation analyses at RNA and protein levels on BM cells, CMPs and BMDCs yielded confirmed changes in Wnt-4 and in another cell-cycle factor, Cyclin-D1. Further studies are being done to identify intersection of DNA methylation/RNA-seq data to see if changes in methylation are correlated with transcription, in the Wnt signaling enabling us to further understand the differentiation and maturation of DC lineages.
Characterization of a novel bone marrow resident cell population with the potential to regulate early hematopoiesis and skeletogenesis.
Authors: Root Sierra Hope , Matthews Brya , Kalajzic Ivo , Aguila H. Leonardo
Published: January 2023

Hematopoiesis and skeletogenesis are tightly regulated with participation of cells from both systems. The main hematopoietic component is the osteoclast, a cell crucial for bone remodeling. However, beyond bone homeostasis, other hematopoietic cells may exist as regulators of hematopoiesis within the bone marrow (BM) microenvironment. Histological analyses of mice expressing cre under the control of Dental Matrix Protein-1 (Dmp-1) promoter and crossed with the reporter strain Ai9, showed a fluorescent (tomato+) dendritic cell–like population interspersed within the BM space. These tomato+ cells were isolated, and showed efficient expression of RANK-L and IL-7, suggesting that they could regulate hematopoiesis. These cells also express surface markers of myeloid lineage including: CD11b, F4.80, CD169, and Ly-6C. In vitro hematopoietic progression assays failed on generating these cells from isolated hematopoietic stem cells. However, they can be generated and expanded ex vivo from CD11b+ tomato negative cells. Furthermore, using a tamoxifen inducible Dmp-1/tomato mouse, hematopoietic tomato+ cells appeared 2 days after tamoxifen treatment. In a transgenic mouse bearing the tamoxifen inducible Dmp-1/tomato plus a transgene allowing the ablation of bone formation after injecting ganciclovir; bone ablation for 16 days followed by tamoxifen two days prior to analysis, showed a compromise in the generation of tomato+ hematopoietic cells indicating that their generation is linked to bone formation. Finally, these cells exist in the BM of late gestation embryos and neonates suggesting a possible fetal origin. Currently we are addressing their phagocytic function and their ability to influence hematopoietic progression.
c-Kit inhibitors prevent respiratory viral infection-induced acute asthma via the blockade of innate lymphoid cell activation
Authors: Chang Ya-Jen , Lai Alan Chuan-Ying , Wu Han-Chih , Umetsu Dale
Published: January 2023

Asthma is a major public health problem that affects nearly 10% of the general population and is thought to be mediated by allergen-specific Th2 cells, adaptive immunity and allergic inflammation. Current treatments for asthma generally focus on minimizing the Th2-driven eosinophilic inflammation with the use of anti-inflammatory therapies such as corticosteroids. However, this approach, targeting the adaptive Th2 cell immunity with corticosteroids, has not reduced the high rates of hospitalizations for acute asthma, most often caused by acute viral infection, including influenza. Recently, type 2 innate lymphoid cells (ILC2) expressing c-Kit have been shown to expand during influenza infection and directly caused airway hyper reactivity (AHR), a cardinal feature of asthma. We now show that the treatment of influenza-infected mice with c-Kit kinase inhibitors blocked the development of AHR and airway inflammation and greatly reduced the number and function of ILC2 cells, whereas the treatment with corticosteroids, commonly used to treat acute asthma, failed to prevent the influenza-induced AHR. We also demonstrated that c-Kit function is required for normal ILC2 cell function and for influenza-induced AHR, and that c-Kit kinase inhibitor inhibits murine lung ILC2 cell proliferation and cytokine productions in vitro and in vivo. Furthermore, the treatment of c-Kit inhibitor did not affect in vivo viral clearance. Our study demonstrated the crucial role of c-Kit in modulating the ILC2 activation in the lungs, particularly in respiratory viral infection induced acute asthma.
Imbalance of Peripheral CD4+T Cell Subsets in Active Vitiligo
Authors: Zhen Yu , Li Shanshan
Published: January 2023

Accumulating studies have indicated that vitiligo is one kind of autoimmune diseases and CD4+T cells play important roles in the pathogenesis of vitiligo. However, there have been very limited papers concerning the profile of circulating CD4+T cell subsets in active vitiligo. To clarify this issue, we investigated the changes of peripheral Th1, Th2, Th17, Tfh and Tregs from 30 patients with active vitiligo. We found that compared with healthy controls, the percentages of circulating Th1 and Th17 were significantly increased, while the proportions of Th2 and Tregs showed no significant difference to healthy controls. Similar results were got in qualification of their corresponding transcription factors at mRNA level. In addition, the percentage of CD4+CXCR5+Tfh was significantly elevated in vitiligo patients. Compared with healthy controls, the expression level of IL-17A was significantly increased in sera of patients with active vitiligo, while the productions of IFN-γ, IL-4, TGF-β had no significant change. The imbalance observed between the effector and suppressor CD4+T cells in the current study suggests their involvement in the pathogenesis of vitiligo. Besides these changes of cellular immunity, increased Tfh indicates humoral immunity may also be responsible for development of vitiligo.
Autoimmune T cells binds to self antigen weakly in the thymus but greatly upregulate the binding strength in the periphery
Authors: Liu Baoyu , Blanchfield J. Lori , Zhu Cheng , Evavold Brian D.
Published: January 2023

How autoimmune TCRs progress through thymocyte selection to egress in the periphery and induce disease is unclear. Autoimmune TCRs are believed to be of low affinity for self antigens due to the process of thymocyte negative selection. Using an experimental autoimmune encephalomyelitis (EAE) mouse model, we have cloned two tetramer positive myelin oligodendrocyte glycoprotein (MOG)35-55-specific TCRs (named B8 and 1C9) from polyclonal CD4+ T cells; both TCRs have a high two dimensional (2D) affinity for MOG (similar to a pathogen-specific TCR SMARTA for an LCMV epitope gp66-77) by the micropipette adhesion frequency assay. A second critical feature of antigen recognition is the bond lifetime between the TCR and pMHC that occurs under applied force. To this end, we have produced retrogenic mice for the two TCRs and used their thymocytes on biomembrane force probe (BFP) to measure single TCR-pMHC bond lifetimes. Despite their high affinity, both TCRs had short bond lifetimes with the MOG antigen displaying a slip bond where bond lifetimes decrease with increasing force. In contrast, the same TCRs on naïve T cells in the periphery displayed much longer bond lifetimes (up to 100 fold) and strong catch bonds (a counterintuitive type of molecular bond that strengthens under force), suggesting physiological regulation of TCR kinetics within single T cell clones. In comparison, the SMARTA TCR formed indistinguishable catch bonds with gp66-77 on thymocytes and naïve T cells. Therefore, these data identify bond lifetime under force as a unique kinetic property of autoimmune TCRs where weak interactions in the thymus potentially avoid negative selection yet in the periphery the TCRs bind with sufficiently long lifetime to promote autoimmunity.
Vitamin D increases the human endosomal TLR response against Mycobacterium tuberculosis nucleic acids
Authors: Cervantes Jorge L. , Oak Esther , Garcia John , Salazar Juan C.
Published: January 2023

Mycobacterium tuberculosis (Mtb) infects macrophages where it avoids elimination by interfering with host defense mechanisms. Type I IFNs are produced by activation of endosomal TLRs after recognition of nucleic acids. TLR importance is underlined by the fact that the TLR-adaptor molecule MyD88 pathway in macrophages is important in confining Mtb within phagolysosomes, mycobacterial clearance, and an adequate innate and adaptive immune response against Mtb. Vitamin D (VD) has a positive effect in macrophage activation, phagocytosis, restriction of Mtb growth, and killing of the mycobacteria.

We aimed to characterize the endosomal TLR-signaling of human macrophages, as they are appear crucial in determining the fate of the disease, and explore if variations in Type I IFN responses correlate with differences in Mtb nucleic acids derived from different Mtb genotypes. Stimulation with DNA from Mtb of different lineages elicited differential NF-κB and IRF inflammatory responses in THP-1 derived macrophages. We also studied the effect of VD on these responses, as it is known to activate macrophages and enhance their antimycobacterial activity. Pretreatment with VD increased the response to DNA from HN878 and virulent strain H37Rv. It also enhanced the response to RNA from M. smegmatis and attenuated M. bovis strain BCG.

No previous study has evaluated Type I or Type II IFNs response differences between different Mtb strains. This is of great importance as type I IFNs appear to strongly inhibit macrophages responsiveness to IFN-γ, and are associated with active TB. Understanding VD-mediated activation occurring at the pulmonary alveoli will impact the design of novel therapeutic and successful immunization strategies.