bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human natural killer cells express functional formyl peptide receptors (133.13)
Authors: Bae Yoe-Sik , Kim Sang Doo , Kim Jung Mo , Lee Ha Young , Shim Jae Woong , Kim Hak Jung , Seo Su-Kil , Yun Jeanho 
Published: January 2023

Abstract
We tested the expression of the formyl peptide receptor (FPR) family and the functional roles of the FPR family in human natural killer (NK) cells. All tested human NK cells express two members of the FPR family (FPR1 and FPR2). The expression of FPR3 was noted to occur in a donor-specific manner. The stimulation of NK cells with FPR family-selective agonists (fMLF, MMK-1, F2L, and WKYMVm) elicited cytolytic activity in resting NK cells, but not in IL-2 activated NK cells; the cytolytic activity was not inhibited by pertussis toxin (PTX). The FPR family agonists also stimulated chemotactic migration of IL-2-activated NK cells, but not resting NK cells; the chemotactic migration was completely inhibited by PTX. WKYMVm stimulates ERK, p38 MAPK, and JNK activities in both resting and IL-2-activated NK cells. WKYMVm-induced chemotactic migration was partially inhibited by PD98059; however, the inhibition of JNK by its selective inhibitor (SP600125) dramatically inhibited the WKYMVm-induced cytolytic activity. Our findings indicate that human NK cells express functional members of the FPR family, and in turn the activation of the three members of the FPR receptor family elicit cytolytic activity in NK cells. We suggest that FPR family are potentially important therapeutic targets for the modulation of NK cell-mediated immune responses.
Role of CXCL12 and its receptor, CXCR4, in diversification of the rabbit primary antibody repertoire (133.3)
Authors: Lanning Dennis , Knight Katherine 
Published: January 2023

Abstract
The antibody repertoire of early postnatal rabbits is of limited diversity due to preferential V gene usage during VDJ gene rearrangement. Early in life, rabbit B cells expand this repertoire by diversifying their VDJ genes in gut-associated lymphoid tissue. VDJ gene diversification begins around 1 week of age, in B cells located in the basolateral region of nascent follicles. Because these B cells share similarities with B cells in germinal center dark zones, we hypothesized that activated B cells localize to the basolateral region in response to the chemokine CXCL12. To test this hypothesis, we examined B cell expression of CXCR4, the CXCL12 receptor, by in situ hybridization. We found that B cells in the basolateral region, but not those located elsewhere in the follicle, expressed CXCR4. Because antibody repertoire diversification requires signals derived from intestinal commensals, we further examined CXCR4 expression in appendices that were kept sterile by surgical ligation at birth. We found that B cells in these appendices did not express CXCR4 and did not diversify their VDJ genes. By contrast, we detected CXCL12 expression in both conventional and sterile appendices, demonstrating that it is not dependent on commensal-derived signals. Our results suggest that B cells activated by microbial-derived signals upregulate CXCR4 and migrate to the basolateral region of the follicle, likely in response to CXCL12, where they proliferate and diversify their VDJ genes.
CXCR6 mediates migration and invasion of breast cancer cells in a Src, FAK, and ERK1/2 dependent fashion (133.6)
Authors: Triplett Ashley , Sharma Praveen , Singh Rajesh , Lillard James , Singh Shailesh 
Published: January 2023

Abstract
Standard treatment modalities for breast cancer has improved the outlook for women afflicted with it. However, 40% of breast cancer patients still die from the disease, primarily due to metastasis. In this regard, chemokines and their corresponding receptor interactions in metastasis is an area of intense pre-clinical and clinical research. Our lab has demonstrated higher expression of CXCR6 in breast cancer cell lines (MCF-7 and MDA-MB-231) compared to the normal mammary epithelial cell line (MCF-10A), and expression by breast cancer tissue. We investigated the role of CXCR6 in breast cancer cell migration and invasion, which are key steps used by cancer cells to achieve metastatic goals. Breast cancer cells showed higher migratory and invasive potential towards the chemotactic gradients of CXCL16, which was significantly inhibited after CXCR6 blockade. Because our antibody array data showed phosphorylation of Src, FAK, and ERK1/2 in breast cancer cells following CXCL16 treatment, we used inhibitors against these molecules after CXCL16 treatment to determine the molecular mechanisms by which CXCR6 mediates migration and invasion. The results showed a significant decrease in migration and invasion, a reduction in F-actin polymerization, and decreased phosphorylation of Vav1, ezrin, cortactin, and paxillin. Taken together, this data suggests that Src, FAK, and ERK1/2 are required for CXCR6-mediated migration and invasion.
Cytokine-mediated regulation of CX3CL1 in human lung fibroblast: Involvement of STAT-1 signaling pathways (133.9)
Authors: Isozaki Takeo , Sato Michihito , Takahashi Ryo , Wakabayashi Kuninobu , Yajima Nobuyuki , Miwa Yusuke , Kasama Tsuyoshi 
Published: January 2023

Abstract
Introduction: CX3CL1 (fractalkine), a membrane-bound chemokine that induces both the adhesion and migration of leukocytes, is involved in the recruitment of cells into tissues undergoing inflammatory responses. To explore the regulation of CX3CL1 in pulmonary inflammation and fibrosis, CX3CL1 expression of by lung fibroblast was examined. Methods: Normal human lung fibroblasts were obtained from Promocell were incubated in the presence or absence of various inflammatory stimuli. Culture supernatants were collected, and soluble CX3CL1 levels were determined with an enzyme-linked immunosorbent assay. Expression of CX3CL1 mRNA transcripts in lung fibroblast was examined using quantitative TaqMan real-time polymerase chain reaction. Results: Interleukine (IL)-1β or interferon (IFN)-γ alone induced negligible soluble CX3CL1 secretion by human lung fibroblasts after 24 h. However, the combination of IL-1β and IFN-γ induced dramatic increases in both soluble CX3CL1 protein and mRNA transcripts in dose- and time-dependent manners. The secretion and expression of lung fibroblast-derived CX3CL1 were markedly reduced by specific inhibitors of the STAT-1 transcription factor. Conclusions: These findings suggest that lung fibroblasts are an important cellular source of CX3CL1, and may play roles in pulmonary inflammation and fibrosis like interstitial pneumonia which merges to rheumatoid arthritis and other autoimmune diseases.
Performance evaluation of two multiplex technologies for the measurement of serum cytokines in HIV-infected individuals (134.17)
Authors: Dabitao Djeneba , Margolick Joseph , Bream Jay 
Published: January 2023

Abstract
Simultaneous quantification of cytokines is a powerful tool to identify associations between host immune defense and human immunodeficiency virus (HIV) pathogenesis. None of the commercially available cytokine detection methods however, have been rigorously validated in the context of HIV infection. Here we compared performance characteristics of two multiplex assays: the Meso Scale Discovery (MSD) platform and the Cytometric Bead Array (CBA) in specimens obtained from HIV-infected subjects. Overall, the MSD had a wider dynamic range and lower limits of quantification which lead to the ability to detect endogenous levels of serum cytokines that were assumed to not be present by CBA. In recombinant cytokine-spiked sera, MSD had better precision and accuracy based on intra-assay variability less than 25% and percent recovery within 25% of added concentrations. Regardless of the quantitation range, we observed variability between the analytes in both platforms with respect to the capacity to recover a known amount of cytokine. Nonetheless, the MSD platform facilitates the reliable assessment of cytokines present in low concentrations in the serum of HIV-infected individuals.
IL-10 interruption of dendritic cell (DC) differentiation during the monocyte-to-DC transition is associated with increased levels of LC3, mature autophagosome formation, and survival of monocyte-macrophage-like cells. (134.21)
Authors: Santiago-Schwarz Frances , Valentino Alisa , Martin Carla , DiMaio Christopher 
Published: January 2023

Abstract
In immature DCs, fusion of autophagosomes containing Ag with MHC class II loading compartments (MIIC) has been linked to adaptive immunity. In monocyte (mono)-mϕs, fusion of autophagosomes with lysosomes yields autolysosomes and instead, innate immunity. Degradation of cargo (including apoptotic debris, leaky mitochondria) in autolysosomes may contribute to cell survival. Previously, we noted that despite IL-10-triggered apoptosis of newly committed DCs at the critical mono/DC interface, cells surviving such effects are non-antigen presenting mono-mϕ like cells avidly engaged in uptake of dying DCs. Here, we questioned if the surviving mono-mϕs exhibit increased autophagic activity. Flow cytometric analysis of mono-mϕs derived from human mono-DC precursors with DC growth factors +IL-10 revealed increases in the autophagy-associated LC3 protein vs. DC controls. Microscopic analysis of anti-LC3 stained cells showed increased punctate and ring-like patterns with IL-10, indicating formation of mature autophagosomes. Immunoblot analysis substantiated that cytosolic LC3I (18kd) was increased with IL-10 and converted to the autophagosome-associated LC3II (16kd) form. We advance the novel idea that autophagy occurs as a cytoprotective measure in concert with apoptotic stress triggered by IL-10. Aside from suppressing the onset of adaptive immunity by killing newly differentiated DCs, IL-10 may act to enhance innate immunity via pro-survival autophagic mechanisms.
Visualization of IL-15 expression in innate immune cells during homeostasis and inflammation (134.3)
Authors: Stoklasek Thomas , Colpitts Sara , Obar Joshua , Guo Caiying , Lefrançois Leo 
Published: January 2023

Abstract
IL-15 plays a multifaceted role in immune system homeostasis. To visualize IL-15 expression we created a transgenic mouse expressing emerald-GFP (EmGFP) under IL-15 promoter control. EmGFP expression was prevalent in dendritic cells (DC) and macrophages, but differed depending on DC subtype. Basophils, neutrophils, eosinophils, and mast cells expressed high EmGFP levels. Unexpectedly, NK cells and splenic γδ T cells also expressed EmGFP. Rapid upregulation of EmGFP after virus infection was dependent on both plasmacytoid DC and type I interferon, albeit to differing degrees. Microscopy revealed basal EmGFP expression by DC in lymphoid tissues that was greatly enhanced after virus infection. These results identified IL-15 as an innate cytokine, produced by multiple cell types during the early immune response to inflammation.
Interleukin-35 subunit dimerization and stability is unique among IL-12 cytokine family members (134.9)
Authors: Jones Lindsay , Collison Lauren , Vignali Dario 
Published: January 2023

Abstract
IL-35 is a newly discovered cytokine that contributes to the suppressive capacity of regulatory T cells. IL-35 is a heterodimer consisting of an α chain (p35) and a β chain (Ebi3). It is a member of the IL-12 cytokine family, which is characterized by the sharing of three α (p19, p28, p35) and two β (p40 and Ebi3) subunits, and also includes IL-12 (p35/p40), IL-23 (p19/p40) and IL-27 (p28/Ebi3). In this study, relative pairing and secretion efficiencies and the dimerization interfaces of IL-12 family members were characterized, with emphasis on IL-35. Specific p35 residues that are known to contribute to formation of the IL-12 interface were assessed for their contribution to the IL-35 interface in mutagenesis studies. Similarly, several candidate Ebi3 residues were screened for their contribution to both IL-27 and IL-35 interfaces. Several residues were identified as critical to the IL-12 or IL-27 interfaces. Conversely, no single mutation was identified that completely disrupts p35/Ebi3 pairing, indicating that p35 and Ebi3 may pair differently to form IL-35. Preliminary data also indicate that IL-35 is poorly secreted by transfected cells, compared to IL-12 and IL-27, suggesting IL-35 formation is unfavorable or the secreted protein in unstable. Thus IL-35 appears to be different from other IL-12 family members in terms of both dimerization interface and stability. This work is supported by grants from the NIH and ALSAC.
Pathogenic or non-pathogenic subpopulations of Th17 cells are generated by stimulation of naïve CD4 cells via antigen/APC or anti-CD3/CD28 antibodies (135.10)
Authors: Shi Guangpu , Vistica Barbara , Lovaas Jenna , Tang Cuiyan , Aziz Mehak , Gery Igal 
Published: January 2023

Abstract
Th17 cells play a major role in immunopathogenic processes. McGeachy et al, (Nat. Immunol., 2007) showed that two distinct subsets of Th17 were generated from activated CD4 cells when different polarization modes were used, with only one subset being encephalitogenic. Here, we analyzed the biological features of Th17 generated by the same polarization mechanism, i.e., incubation with IL-6 and TGF-b, but by two different modes of stimulation. Naïve CD4 cells, collected from TCR transgenic mice specific against hen egg lysozyme (HEL), were activated by either HEL and APC (“HA”), or by plate-bound antibodies against CD3/CD28 (“PbAb”). PbAb Th17 produced moderately higher levels of IL-17 than HA Th17, but the latter cells released profoundly higher levels of IL-22. Interestingly, Th17 cells of the two subsets produced similar levels of IL-10, shown by both analysis of the culture supernatants and intracellular staining. Importantly, when adoptively transferred into recipients expressing HEL in their lens, only HA Th17 cells induced ocular inflammation in recipients, while no disease was detected in eyes of PbAb cell recipients. It is also of note that the two Th17 subsets differed in their profile of chemokine transcripts, resembling the two different profiles recorded by McGeachy et al with the pathogenic and non-pathogenic subsets. Thus, the pathogenic or non-pathogenic Th17 subsets are generated in our system, without pre-activation, by two stimulation modes.
Mast cells influence disease severity in a murine model of relapsing-remitting multiple sclerosis. (135.15)
Authors: Walker Margaret , Amir-Sayed Blayne , Brown Melissa 
Published: January 2023

Abstract
Although mast cells (MCs) are critical mediators of allergic and innate immune responses, they also modulate adaptive immune responses. Utilizing WBB6c-kitW/Wv mast cell-deficient mice, MCs have been implicated in the promotion of disease severity in several murine models of human autoimmune disease. WBB6c-kitW/Wv mice have two independent mutations that profoundly inhibit MC development. We previously demonstrated that MCs influence disease severity in a murine model of chronic-progressive multiple sclerosis (CP-MS), specifically by influencing cellular trafficking into the CNS and blood brain barrier permeability. However, this model does not mimic the most common form of the human disease, relapsing-remitting MS (RR-MS). Therefore, to test the effect of a MC deficiency on a RR form of disease, we transferred the c-kitW/Wv mutations onto the SJL background, a strain that is susceptible to RR disease. These animals are MC-deficient in all tissues examined, carry mild hematopoietic deficits, but have relatively normal thymic T cell compartments. SJLc-kitW/Wv mice display significantly decreased cumulative EAE disease scores and a delay in the day of peak acute disease compared to their wild type littermates. These differences in disease phenotypes are abrogated upon reconstitution of wild type bone marrow-derived MCs. Thus, MCs also influence disease severity in a RR murine model of MS, a finding with potential clinical relevance for the heterogeneous MS patient population.
Effect of Kv1.3 inhibition on T cell development, activation and signaling (135.23)
Authors: Rosenzweig Jason , Lebson Lori , Mancini Justin , Calabresi Peter , Whartenby Katharine 
Published: January 2023

Abstract
The voltage-gated potassium channel Kv1.3 is highly expressed in effector memory T cells (TEM) but not in central memory (TCM ) or naïve T cells. TEM are pathogenic in several autoimmune diseases. Acute blockade of Kv1.3 by the sea anemone toxin ShK reduces activation induced proliferation of TEM. However, in Kv1.3-/- mice, in which the gene was absent from development, no immune system defects were found, which was attributed to compensatory channels (Koni et al. JBC, 2003). The limitations of pharmacological blockade and gene-targeted deletion led us to develop a lentiviral system to co-express a dominant negative Kv1.3 and GFP. We have been using this lentivirus to query the role of Kv1.3 both in the development and activation of T cells. We have conducted studies in which we have suppressed channel expression in the development of T cells from a gene-modified bone marrow transplant and also assessed the impact of blocking channel expression during antigen-specific activation of CD4+ T cells. Using the antigen specific mouse system allows multiple advantages over previous studies including the feature that specific T cells can be tracked and monitored and compared to unmodified T cells. Initial results showed that Kv1.3-silenced CD4+ T cells skewed towards the TCM phenotype. We will report on our additional results using this lentiviral expression system, which is allowing us to study the role of Kv1.3 in T cells in mice and its effect on signaling pathways.
Memory B cells from relapsing remitting multiple sclerosis patients elicit functional responses by CD4+ T cells in response to neuro-antigens (135.24)
Authors: Monson Nancy , Harp Christopher , Ireland Sara , Davis Laurie , Cassidy Bonnie , Cravens Petra , Stuve Olaf , Lovett-Racke Amy , Eagar Todd , Greenberg Benjamin , Racke Michael , Cowell Lindsay , Karandikar Nitin , Frohman Elliot 
Published: January 2023

Abstract
Recent evidence suggests that B and T cell interactions may be paramount in relapsing remitting multiple sclerosis (RRMS) disease pathogenesis. We hypothesized that memory B cell pools from RRMS patients may specifically harbor a subset of potent neuro-antigen presenting cells that support neuro-antigen reactive T cell expansion and activation. To assess the potential influence of neuro-antigen specific memory B cells from RRMS patients on autologous CD4+ T cell activation, we compared CD80 and HLA-DR expression, IL-10 and LTα secretion, neuro-antigen binding capacity, and functional neuro-antigen presentation by memory B cells from RRMS patients and compared these parameters to naïve B cells from RRMS patients and to memory and naïve B cells from healthy donors (HDs). We identified memory B cells from a subset of RRMS patients that were functionally capable of presenting neuro-antigen to autologous T cells. Notwithstanding the fact that the phenotypic parameters that promote efficient antigen presentation were observed to be similar between RRMS and HDs memory B cells, a corresponding capability to present neuro-antigen was not observed in HD memory B cells. Our results demonstrate for the first time that the memory B cell pool in RRMS harbors neuro-antigen specific clones that can activate T cells.
Interleukin-17A is dispensable for myocarditis but essential for the progression to dilated cardiomyopathy (135.25)
Authors: Baldeviano Geral , Barin Jobert , Talor Monica , Srinivasan Sachin , Chen Ping , Bedja Djahida , Zheng Donfeng , Gabrielson Kathleen , Iwakura Yoichiro , Rose Noel , Cihakova Daniela 
Published: January 2023

Abstract
A third of myocarditis cases progress to dilated cardiomyopathy (DCM); however, the mechanisms controlling this process are largely unknown. Immunization of BALB/c mice with myocarditogenic peptide in CFA adjuvant induced the infiltration of pathogenic IL-17A-secreting T helper type-17 cells into the inflamed heart during the acute stage; and mild-to-severe DCM in the chronic stage of the disease. However, IL-17A deficiency did not abrogate the incidence or severity of myosin-induced myocarditis, nor did it ameliorate the severe disease of IFNγ-deficient mice, suggesting that IL-17A plays a minimal role during acute myocarditis. In contrast, during the chronic stage, IL-17A-deficient mice but not WT littermates were fully protected from the progression to DCM. Flow cytometry and cytokine analysis revealed an important role for IL-17A in the upregulation of IL-6, TNFα, and IL-1β in the heart tissue, and the recruitment of CD11b+ monocytes and Gr1+ granulocytes into the heart. These differences were associated with a significant decrease in myocardial fibrosis and dysregulation of matrix metalloproteinases. Furthermore, neutralization of endogenous IL-17A after the onset of myocarditis decreased myocardial fibrosis and improved systolic function. Our findings reveal a critical role for IL-17A in adverse cardiac remodeling leading to end-stage DCM, and suggest that targeting IL-17A may be an attractive therapeutic strategy for patients with inflammatory DCM.
Exaggerated IL-23-induced psoriasis-like inflammation in mice lacking CCR4 (135.32)
Authors: Hedrick Michael , Zhang Hongwei , Farber Joshua 
Published: January 2023

Abstract
Recent evidence strongly implicates the IL-23/Th17/Th22 axis in psoriasis, and intradermal injection of IL-23 produces local psoriasis-like inflammation, which depends on IL-22 and other IL-10-family cytokines. CCR4 is a chemokine receptor thought to be important for T cell trafficking to skin, and is expressed on all Th2 cells, as well as many Th17 cells and Treg. We tested CCR4 deficient mice in the IL-23 injection model and found, surprisingly, that they had an exaggerated inflammatory response, with increased swelling of the injected skin, greater acanthosis, parakeratosis, and microabscess formation, higher numbers of infiltrating neutrophils, T cells and dendritic cells, and higher numbers of pro-inflammatory cytokines, including: IL-17a, IL-17f, IL-19, IL-22 and TNFα. Remarkably, despite the greater inflammation, Foxp3+ cells were not increased in the IL-23-injected skin of the CCR4 deficient mice, producing a higher effector T cell/Treg ratio as compared with wild-type mice. Our results suggest that CCR4 is important, not for effector functions, but for recruiting cells to down-modulate Th17/Th22-mediated inflammation in skin. These findings may help in understanding how inflammation is regulated in psoriasis and related disorders. Supported by the Intramural Research Program, NIH, NIAID.
The structural mechanism of Serum Amyloid A -mediated Secondary amyloid formation (135.34)
Authors: Lu Jinghua , Du Clos Terry , Mold Carolyn , Sun Peter 
Published: January 2023

Abstract
Secondary amyloidosis is a severe complication of many chronic inflammatory diseases, such as rheumatoid arthritis and atherosclerosis, when the C-terminally cleaved fragments of acute phase protein Serum Amyloid A (SAA) are deposited in tissues as amyloid fibrils. The pathologic process of secondary amyloidosis involves a conversion of the structure of native SAA into a predominantly antiparallel β-sheet secondary structure. We have now determined the X-ray structure of SAA in a monomeric and a hexameric form at 2.2Å and 2.7Å resolution, respectively. The hexameric SAA structure revealed a dimeric architecture arranged as two trimers. Strong interactions between the C-terminal loop region and four-helix-bundle core of the SAA monomers create a stable structure to prevent secondary structure refolding during amyloidogenesis. The presence of multimeric forms in human serum was confirmed by size-exclusion experiments. Recombinant monomeric, trimeric and hexameric SAA protein were purified from Escherichia coli and characterized. Binding studies show that the monomeric and trimeric but not the hexameric SAA bind to Toll-like receptor 2 and 4. In line with these experiments, SAA may undergo a transition between hexamers and monomers, which would activate Toll-like receptors to further complicate the chronic inflammation in disease situation.