Pub Date : 2024-11-15DOI: 10.1016/j.cyto.2024.156804
Agata Kaczmarek, Anna Katarzyna Wrońska, Justyna Sobich, Mieczysława Irena Boguś
Cytokines are highly conserved between mammals and insects. The present study examines the multiple effects of interferon-gamma (IFN-γ) application on the immunological defence mechanisms of Galleria mellonella larvae, invertebrates which are gaining popularity as a replacement for mammalian research models in immunological studies. G. mellonella hemolymph is known to contain an IFN-γ homolog that shares 33 % similarity with its mammalian analogue, and its level in insect hemocytes increases during exposition to entomopathogenic fungus Conidiobolus coronatus. The present research examines the impact of IFN-γ on larval development, the effectiveness of fungal infection, and the morphology and physiology of wax moth immunocompetent cells. Treatment with IFN-γ enhanced wound healing, chemotaxis activity and hemocyte impedance, while reducing hemocyte phagocytosis and oxidative stress in cultured immunocompetent cells; it also appears to increase the levels of Jak-2- and NF-κB-like molecules in hemocytes. Our findings suggest that IFN-γ demonstrated considerable similarity between mammals and humans, thus further demonstrating the evolutionary conservatism of cytokines.
{"title":"The multifunctional role of IFN-γ in Galleria mellonella (Lepidoptera) immunocompetent cells","authors":"Agata Kaczmarek, Anna Katarzyna Wrońska, Justyna Sobich, Mieczysława Irena Boguś","doi":"10.1016/j.cyto.2024.156804","DOIUrl":"10.1016/j.cyto.2024.156804","url":null,"abstract":"<div><div>Cytokines are highly conserved between mammals and insects. The present study examines the multiple effects of interferon-gamma (IFN-γ) application on the immunological defence mechanisms of <em>Galleria mellonella</em> larvae, invertebrates which are gaining popularity as a replacement for mammalian research models in immunological studies. <em>G. mellonella</em> hemolymph is known to contain an IFN-γ homolog that shares 33 % similarity with its mammalian analogue, and its level in insect hemocytes increases during exposition to entomopathogenic fungus <em>Conidiobolus coronatus</em>. The present research examines the impact of IFN-γ on larval development, the effectiveness of fungal infection, and the morphology and physiology of wax moth immunocompetent cells. Treatment with IFN-γ enhanced wound healing, chemotaxis activity and hemocyte impedance, while reducing hemocyte phagocytosis and oxidative stress in cultured immunocompetent cells; it also appears to increase the levels of Jak-2- and NF-κB-like molecules in hemocytes. Our findings suggest that IFN-γ demonstrated considerable similarity between mammals and humans, thus further demonstrating the evolutionary conservatism of cytokines.</div></div>","PeriodicalId":297,"journal":{"name":"Cytokine","volume":"185 ","pages":"Article 156804"},"PeriodicalIF":3.7,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142637899","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Sepsis is an uncontrolled inflammatory response to infection and is closely associated with the occurrence of acute respiratory distress syndrome (ARDS). Low tidal volume lung ventilation and permissive hypercapnia is a recognized therapy for ARDS. However, whether permissive hypercapnia aggravates sepsis-associated encephalopathy (SAE) remains unclear. The present study investigated whether hypercapnia contributed to the development of SAE through the purinergic 2X7 receptor (P2X7R) by activating the Nod-like receptor protein 3 (NLRP3) inflammasome in sepsis.
Methods
The SAE model was established by intracranial injection of lipopolysaccharide (LPS) (1 μg/ml, 5 μl) in C57BL/6 mice. Hypercapnia was induced by mechanical ventilation with a high concentration of CO2 (5 % CO2, 21 % O2 and 74 % N2) in vivo. Toll-like receptor 4 (TLR4) and P2X7R knockout (KO) mice were employed in the study, while in vitro, BV2 microglial cells were treated with LPS or a high concentration of CO2 (15 % CO2 + 20 % O2). Immunofluorescence and western blot analysis were used to assess the expression levels of TLR4, NF-κB, phosphorylated (p)-NF-κB, P2X7R, pro-caspase-1, caspase-1, pro-IL-1β, IL-1β, pro-IL-18 and IL-18. ATP levels in the cell culture medium were detected by fluorometric assay.
Result
The results revealed that, compared with the sham group, the expression levels of TLR4, p-NF-κB, pro-IL-1β, pro-IL-18 and NLRP3 were significantly upregulated in the LPS and LPS + hypercapnia groups, but not in the hypercapnia group. Although the expression levels of caspase-1, IL-1β and IL-18 were increased slightly in the LPS group, their upregulation was more pronounced in the LPS + hypercapnia group, and it was suppressed when TLR4 was knocked out. Furthermore, P2X7R expression and ATP levels in the cell culture medium remained unchanged in the LPS group compared with the sham group but were remarkably increased both in the hypercapnia and LPS + hypercapnia groups. Additionally, P2X7R KO restrained the caspase-1, IL-1β and IL-18 increased induced by LPS injected intracranially and hypercapnia.
Conclusions
In conclusion, LPS induced the priming step of NLRP3 inflammasome activation, but had little effect on the activation step, while hypercapnia played an important role in the activation step through P2X7R, depending on the priming step stimulated by LPS.
{"title":"Hypercapnia promotes NLRP3 inflammasome activation in microglia by activating P2X7R after lipopolysaccharide-induced activation of the TLR4/NF-κB signaling pathway","authors":"Hongguang Ding , Shiying Zhang , Zhuo Li , Juhao Zeng , Hongke Zeng","doi":"10.1016/j.cyto.2024.156806","DOIUrl":"10.1016/j.cyto.2024.156806","url":null,"abstract":"<div><h3>Background</h3><div>Sepsis is an uncontrolled inflammatory response to infection and is closely associated with the occurrence of acute respiratory distress syndrome (ARDS). Low tidal volume lung ventilation and permissive hypercapnia is a recognized therapy for ARDS. However, whether permissive hypercapnia aggravates sepsis-associated encephalopathy (SAE) remains unclear. The present study investigated whether hypercapnia contributed to the development of SAE through the purinergic 2X7 receptor (P2X7R) by activating the Nod-like receptor protein 3 (NLRP3) inflammasome in sepsis.</div></div><div><h3>Methods</h3><div>The SAE model was established by intracranial injection of lipopolysaccharide (LPS) (1 μg/ml, 5 μl) in C57BL/6 mice. Hypercapnia was induced by mechanical ventilation with a high concentration of CO<sub>2</sub> (5 % CO<sub>2</sub>, 21 % O<sub>2</sub> and 74 % N<sub>2</sub>) <em>in vivo</em>. Toll-like receptor 4 (TLR4) and P2X7R knockout (KO) mice were employed in the study, while <em>in vitro</em>, BV2 microglial cells were treated with LPS or a high concentration of CO<sub>2</sub> (15 % CO<sub>2</sub> + 20 % O<sub>2</sub>). Immunofluorescence and western blot analysis were used to assess the expression levels of TLR4, NF-κB, phosphorylated (p)-NF-κB, P2X7R, pro-caspase-1, caspase-1, pro-IL-1β, IL-1β, pro-IL-18 and IL-18. ATP levels in the cell culture medium were detected by fluorometric assay.</div></div><div><h3>Result</h3><div>The results revealed that, compared with the sham group, the expression levels of TLR4, p-NF-κB, pro-IL-1β, pro-IL-18 and NLRP3 were significantly upregulated in the LPS and LPS + hypercapnia groups, but not in the hypercapnia group. Although the expression levels of caspase-1, IL-1β and IL-18 were increased slightly in the LPS group, their upregulation was more pronounced in the LPS + hypercapnia group, and it was suppressed when TLR4 was knocked out. Furthermore, P2X7R expression and ATP levels in the cell culture medium remained unchanged in the LPS group compared with the sham group but were remarkably increased both in the hypercapnia and LPS + hypercapnia groups. Additionally, P2X7R KO restrained the caspase-1, IL-1β and IL-18 increased induced by LPS injected intracranially and hypercapnia.</div></div><div><h3>Conclusions</h3><div>In conclusion, LPS induced the priming step of NLRP3 inflammasome activation, but had little effect on the activation step, while hypercapnia played an important role in the activation step through P2X7R, depending on the priming step stimulated by LPS.</div></div>","PeriodicalId":297,"journal":{"name":"Cytokine","volume":"185 ","pages":"Article 156806"},"PeriodicalIF":3.7,"publicationDate":"2024-11-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142643603","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-13DOI: 10.1016/j.cyto.2024.156800
Renée Duijzer , Daisy Dalloyaux , Melissa M. Boerrigter , Heidi Lemmers , Helga Dijkstra , Liesbeth van Emst , René H.M. te Morsche , Martin Jaeger , Leo A.B. Joosten , Joost P.H. Drenth
Rationale
The role of the innate immune system in polycystic liver disease (PLD) has been underexplored despite its potential importance in disease progression. This study explores the innate immune response in PLD patients by analyzing cytokine production of peripheral blood mononuclear cells (PBMCs) in response to various pathogens compared to healthy controls.
Methods
Samples were collected from patients with ADPLD or ADPKD and PLD. PBMCs were isolated and stimulated with LPS (1 ng), LPS (10 ng), E. coli, K. pneumoniae, S. aureus, and C. albicans. ELISA was used to measure TNF, IL-1β, IL-1Ra, IL-6, and IL-8 concentrations after 24 hours, and IL-17, IL-22, and IFNγ concentrations after 7 days. Control samples were matched for age and gender.
Results
104 patients and 12 controls were included. PLD patients showed consistent increased IL-6 concentrations compared to controls. Other cytokine levels varied per stimulus. Controls showed higher IL-8 and TNF concentrations in response to Gram-negative bacteria, while PLD patients showed higher IL-1β and IL-1Ra levels in response to S. aureus and C. albicans. No clear differences were found in IL-17, IL-22, and IFN-γ concentrations after 7 days. These observed differences were independent of demographic and clinical parameters.
Conclusion
Compared to healthy controls, the PLD patients innate immune system shows an altered response when stimulated by various pathogens. These findings underscore the importance of further investigation into the underlying mechanisms as this might help our understanding disease progression and be a potential target for new therapies.
{"title":"Exploring the innate immune response in polycystic liver disease","authors":"Renée Duijzer , Daisy Dalloyaux , Melissa M. Boerrigter , Heidi Lemmers , Helga Dijkstra , Liesbeth van Emst , René H.M. te Morsche , Martin Jaeger , Leo A.B. Joosten , Joost P.H. Drenth","doi":"10.1016/j.cyto.2024.156800","DOIUrl":"10.1016/j.cyto.2024.156800","url":null,"abstract":"<div><h3>Rationale</h3><div>The role of the innate immune system in polycystic liver disease (PLD) has been underexplored despite its potential importance in disease progression. This study explores the innate immune response in PLD patients by analyzing cytokine production of peripheral blood mononuclear cells (PBMCs) in response to various pathogens compared to healthy controls.</div></div><div><h3>Methods</h3><div>Samples were collected from patients with ADPLD or ADPKD and PLD. PBMCs were isolated and stimulated with LPS (1 ng), LPS (10 ng), <em>E. coli</em>, <em>K. pneumoniae</em>, <em>S. aureus</em>, and <em>C. albicans</em>. ELISA was used to measure TNF, IL-1β, IL-1Ra, IL-6, and IL-8 concentrations after 24 hours, and IL-17, IL-22, and IFNγ concentrations after 7 days. Control samples were matched for age and gender.</div></div><div><h3>Results</h3><div>104 patients and 12 controls were included. PLD patients showed consistent increased IL-6 concentrations compared to controls. Other cytokine levels varied per stimulus. Controls showed higher IL-8 and TNF concentrations in response to Gram-negative bacteria, while PLD patients showed higher IL-1β and IL-1Ra levels in response to <em>S. aureus</em> and <em>C. albicans</em>. No clear differences were found in IL-17, IL-22, and IFN-γ concentrations after 7 days. These observed differences were independent of demographic and clinical parameters.</div></div><div><h3>Conclusion</h3><div>Compared to healthy controls, the PLD patients innate immune system shows an altered response when stimulated by various pathogens. These findings underscore the importance of further investigation into the underlying mechanisms as this might help our understanding disease progression and be a potential target for new therapies.</div></div>","PeriodicalId":297,"journal":{"name":"Cytokine","volume":"184 ","pages":"Article 156800"},"PeriodicalIF":3.7,"publicationDate":"2024-11-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142610820","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-10DOI: 10.1016/j.cyto.2024.156801
Zhengda Pei , Mengxiang Tian
The Cyclic GMP-AMP synthase (cGAS)-Stimulator of interferon genes [1] signaling pathway has emerged as a pivotal immune response mechanism, activating immune defenses upon detection of both exogenous and endogenous DNA within cells. Its activation is intricately linked to various diseases and inflammatory processes, spanning autoimmune disorders, infectious ailments, and malignancies. Among pancreatic diseases, encompassing acute pancreatitis, chronic pancreatitis, and pancreatic cancer, current clinical treatment efficacy remains suboptimal. Here, we elucidate the molecular intricacies of the cGAS-STING signaling pathway and delineate its therapeutic potential in acute pancreatitis, chronic pancreatitis, and pancreatic cancer. Additionally, we offer an overview of recent advancements in STING agonists and antagonists, assessing their therapeutic potential in pancreatic-related disorders. In summary, by exploring the multifaceted roles of the cGAS-STING signaling pathway and its implications in pancreatic diseases, we aim to shed light on potential avenues for therapeutic intervention and management in these challenging clinical contexts.
{"title":"The cGAS-STING pathway as a novel therapeutic strategy for pancreatic diseases","authors":"Zhengda Pei , Mengxiang Tian","doi":"10.1016/j.cyto.2024.156801","DOIUrl":"10.1016/j.cyto.2024.156801","url":null,"abstract":"<div><div>The Cyclic GMP-AMP synthase (cGAS)-Stimulator of interferon genes <span><span>[1]</span></span> signaling pathway has emerged as a pivotal immune response mechanism, activating immune defenses upon detection of both exogenous and endogenous DNA within cells. Its activation is intricately linked to various diseases and inflammatory processes, spanning autoimmune disorders, infectious ailments, and malignancies. Among pancreatic diseases, encompassing acute pancreatitis, chronic pancreatitis, and pancreatic cancer, current clinical treatment efficacy remains suboptimal. Here, we elucidate the molecular intricacies of the cGAS-STING signaling pathway and delineate its therapeutic potential in acute pancreatitis, chronic pancreatitis, and pancreatic cancer. Additionally, we offer an overview of recent advancements in STING agonists and antagonists, assessing their therapeutic potential in pancreatic-related disorders. In summary, by exploring the multifaceted roles of the cGAS-STING signaling pathway and its implications in pancreatic diseases, we aim to shed light on potential avenues for therapeutic intervention and management in these challenging clinical contexts.</div></div>","PeriodicalId":297,"journal":{"name":"Cytokine","volume":"184 ","pages":"Article 156801"},"PeriodicalIF":3.7,"publicationDate":"2024-11-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142610821","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-03DOI: 10.1016/j.cyto.2024.156795
Chiara Della Bella , Chiara Medici , Sofia D’Elios , Marisa Benagiano , Alessandra Ludovisi , Maria Angeles Gomez-Morales , Mario M. D’Elios , Fabrizio Bruschi
Introduction
We studied the cellular immune response in a patient infected since 10 months (along with other 51 people) during a trichinellosis outbreak caused by Trichinella spp.
Methods
A 46 years old female resulted serologically positive for trichinellosis. We isolated peripheral blood mononuclear cells (PBMCs) and incubated them with excretory/secretory antigens (ESA) of Trichinella spiralis (T1) or Trichinella pseudospiralis (T4) to produce antigen specific T cell lines and clones, analysed for the phenotype (T helper or cytotoxic cells), for their T4 or T1 antigens specificity and for their cytokine profile (IFNγ, IL-17A, IL-4) by flow cytometry, thymidine incorporation assay and ELISpot.
Results
The test performed using ESA from T1 or T4 has identified the species responsible for infection as T. pseudospiralis since the proliferative responses (evaluated by CFSE, Carboxyfluorescein succinimidyl ester, FACS analysis) was higher for T4 (72,8%) than T1 (23.6 %) antigen. The cell lines produced significant levels of IFNγ, IL-4 and IL-17A after stimulation. From the T cell line obtained in response to T1 ESA, as regards CD4 + cells, 12 % Th2, 22.8 % Th1, 6.6 % Th17, 6 % Th0, 2.2 % Th1/Th17 and 0.7 % Th2/Th17, were obtained. From the T1-specific TCL we generated 15 clones. From the TCL specific for T4 ESA, as regards CD4+, 15.2 % Th2, 27.1 % Th1, 3 % Th17, 10.3 %Th0, 1.9 % Th1/Th17 and 1 % Th2/ Th17 were obtained. From such TCL 4 clones were isolated, 1Th2, 1 Th1, 1 Th17, 1 Th1/Th17 and no Th0 nor Th2/Th17.
Conclusions
By cellular immunology techniques the species responsible of the infection resulted T. pseudospiralis, confirming the results previously obtained by serology. For the first time it was revealed in a human chronic infection the presence of Th17 cells.
{"title":"Interleukin 17 producing T cell responses in human chronic trichinellosis-insight from a case study","authors":"Chiara Della Bella , Chiara Medici , Sofia D’Elios , Marisa Benagiano , Alessandra Ludovisi , Maria Angeles Gomez-Morales , Mario M. D’Elios , Fabrizio Bruschi","doi":"10.1016/j.cyto.2024.156795","DOIUrl":"10.1016/j.cyto.2024.156795","url":null,"abstract":"<div><h3>Introduction</h3><div>We studied the cellular immune response in a patient infected since 10 months (along with other 51 people) during a trichinellosis outbreak caused by <em>Trichinella</em> spp.</div></div><div><h3>Methods</h3><div>A 46 years old female resulted serologically positive for trichinellosis. We isolated peripheral blood mononuclear cells (PBMCs) and incubated them with excretory/secretory antigens (ESA) of <em>Trichinella spiralis</em> (T1) or <em>Trichinella pseudospiralis</em> (T4) to produce antigen specific T cell lines and clones, analysed for the phenotype (T helper or cytotoxic cells), for their T4 or T1 antigens specificity and for their cytokine profile (IFNγ, IL-17A, IL-4) by flow cytometry, thymidine incorporation assay and ELISpot.</div></div><div><h3>Results</h3><div>The test performed using ESA from T1 or T4 has identified the species responsible for infection as <em>T. pseudospiralis</em> since the proliferative responses (evaluated by CFSE, Carboxyfluorescein succinimidyl ester, FACS analysis) was higher for T4 (72,8%) than T1 (23.6 %) antigen. The cell lines produced significant levels of IFNγ, IL-4 and IL-17A after stimulation. From the T cell line obtained in response to T1 ESA, as regards CD4 + cells, 12 % Th2, 22.8 % Th1, 6.6 % Th17, 6 % Th0, 2.2 % Th1/Th17 and 0.7 % Th2/Th17, were obtained. From the T1-specific TCL we generated 15 clones. From the TCL specific for T4 ESA, as regards CD4+, 15.2 % Th2, 27.1 % Th1, 3 % Th17, 10.3 %Th0, 1.9 % Th1/Th17 and 1 % Th2/ Th17 were obtained. From such TCL 4 clones were isolated, 1Th2, 1 Th1, 1 Th17, 1 Th1/Th17 and no Th0 nor Th2/Th17.</div></div><div><h3>Conclusions</h3><div>By cellular immunology techniques the species responsible of the infection resulted <em>T. pseudospiralis</em>, confirming the results previously obtained by serology. For the first time it was revealed in a human chronic infection the presence of Th17 cells.</div></div>","PeriodicalId":297,"journal":{"name":"Cytokine","volume":"184 ","pages":"Article 156795"},"PeriodicalIF":3.7,"publicationDate":"2024-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142566849","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The exact role of different immune cells and cytokines in control or promotion of intracellular growth of leishmania has still remained a controversial topic. The aim of the present study was to study effects of cellular changes and relevant cytokines in cell mediated immunity by diphencyprone (DCP) in pathogenicity of acute L.major infection in BALB/c mice.
Methods
45 healthy female BALB/c mice were injected with L. major promastigotes under the base of tail. The mice were randomly divided to three groups of 15 mice: (1) control group without any treatment. (2) acetone group: Acetone was applied topically on the cutaneous lesions weekly and (3) DCP group: DCP was applied topically on the cutaneous lesions with increasing concentrations to induce local allergy. The mice were followed by the end of eighth week, and then macroscopic changes, histopathology, immunology studies, and organ parasite burden were determined.
Results
In DCP group, in comparison to other groups the ulcer size and parasite burden in ulcer site and spleen increased, significantly. There was a deep lymphohistiocytic infiltration in the ulcer site. Total IgG, IgG1, and IgG2a levels as well as IgG2a/IgG1 ratio and intracellular IFN-gamma in CD8+ lymphocytes were significantly higher. IL4 and T CD8+ lymphocytes were significantly lower in DCP group. The IgG2a/IgG1 ratio was more than 1 in all groups.
Conclusion
Our findings demonstrated that DCP reduced the CD8+ lymphocytes and IL-4 production. In spite of increased IgG2a/IgG1 ratio, the parasite burden and inflammation severity increased in infected mice. The results can show the pivotal role of CD8+ lymphocytes in conjunction with Th1 lymphocytes in the control of acute leishmania infection in mice.
{"title":"Diphencyprone reduces the CD8+ lymphocytes and IL-4 and enhences IgG2a/IgG1 ratio in pathogenicity of acute leishmania major infection in BALB/c mice","authors":"Pourandokht Mousavian , Vahid Mashayekhi Goyonlo , Mohammad Javanbakht , Mahmoud Reza Jafari , Hamidreza Moosavian , Monovar Afzal Aghaei , Mohammadreza Malekzadeh","doi":"10.1016/j.cyto.2024.156792","DOIUrl":"10.1016/j.cyto.2024.156792","url":null,"abstract":"<div><h3>Background</h3><div>The exact role of different immune cells and cytokines in control or promotion of intracellular growth of <em>leishmania</em> has still remained a controversial topic. The aim of the present study was to study effects of cellular changes and relevant cytokines in cell mediated immunity by diphencyprone (DCP) in pathogenicity of acute <em>L.</em>major infection in BALB/c mice.</div></div><div><h3>Methods</h3><div>45 healthy female BALB/c mice were injected with <em>L. major</em> promastigotes under the base of tail. The mice were randomly divided to three groups of 15 mice: (1) control group without any treatment. (2) acetone group: Acetone was applied topically on the cutaneous lesions weekly and (3) DCP group: DCP was applied topically on the cutaneous lesions with increasing concentrations to induce local allergy. The mice were followed by the end of eighth week, and then macroscopic changes, histopathology, immunology studies, and organ parasite burden were determined.</div></div><div><h3>Results</h3><div>In DCP group, in comparison to other groups the ulcer size and parasite burden in ulcer site and spleen increased, significantly. There was a deep lymphohistiocytic infiltration in the ulcer site. Total IgG, IgG1, and IgG2a levels as well as IgG2a/IgG1 ratio and intracellular IFN-gamma in CD8<sup>+</sup> lymphocytes were significantly higher. IL4 and T CD8<sup>+</sup> lymphocytes were significantly lower in DCP group. The IgG2a/IgG1 ratio was more than 1 in all groups.</div></div><div><h3>Conclusion</h3><div>Our findings demonstrated that DCP reduced the CD8<sup>+</sup> lymphocytes and IL-4 production. In spite of increased IgG2a/IgG1 ratio, the parasite burden and inflammation severity increased in infected mice. The results can show the pivotal role of CD8<sup>+</sup> lymphocytes in conjunction with Th1 lymphocytes in the control of acute <em>leishmania</em> infection in mice.</div></div>","PeriodicalId":297,"journal":{"name":"Cytokine","volume":"184 ","pages":"Article 156792"},"PeriodicalIF":3.7,"publicationDate":"2024-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142566848","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-01DOI: 10.1016/j.cyto.2024.156798
Li Yuan , Yongshuang Zhou , Ruiyu Wang , Xin Huang , Ruilin Tang , Fang Yan
Background
Acute kidney injury (AKI) is common in septic patients and strongly associated with adverse outcomes. The pathophysiology of AKI in septic patients remains elusive, and detection of patients at risk of AKI or at risk of progression to severe and persistent AKI is critical for timely and adequate support measures, including mitigating further renal damage. Therefore, identification of biomarkers associated with septic-associated AKI that contribute to improve septic AKI is an area of intensive research.
Methods
A total of 116 consecutive patients with sepsis were categorized into two groups (AKI and non-AKI) based on the occurrence of AKI within 24 h of admission to the intensive care unit (ICU). Serum levels of soluble TLR2 (sTLR2), as well as biomarkers such as interleukin(IL)-6, IL-22, IL-10, creatinine, urea, procalcitonin, hypersensitive C-reactive protein (hs-CRP), and D-Dimer (D2), were measured within 24 h after ICU admission. Demographic and clinical characteristics including sequential organ failure assessment (SOFA) scores and acute physiology and chronic health evaluation II (APACHE II) scores were recorded. Logistic regression analysis was conducted to identify potential predictive biomarkers. Receiver operating characteristic (ROC) curve analysis was employed to determine the optimal model for predicting septic-associated AKI.
Results
Patients in the AKI group exhibited significantly higher serum concentrations of IL-6, IL-10, sTLR2, creatinine, urea, hs-CRP, procalcitonin, D2 and lower serum albumin concentrations as well as higher APACHE II scores compared to those in the non-AKI group. Logistic regression analysis revealed that APACHE II scores, log10-transformed sTLR2 concentration, creatinine and D2 concentration were valuable predictors of AKI among septic patients. ROC curves demonstrated that log10-transformed sTLR2 concentration exhibited comparable predictive value to creatinine in determining the incidence of sepsis-associated AKI. The model with variables of APACHE II score, Log10-transformed serum TLR2 concentration, creatinine and D2 concentration yielded the greatest area under the curve of 0.863.
Conclusion
Elevated levels of sTLR2 in early-stage of septic patients may serve as a promising novel biomarker for predicting sepsis-associated AKI.
背景：急性肾损伤（AKI）是脓毒症患者的常见病，与不良预后密切相关。脓毒症患者急性肾损伤的病理生理学仍然难以捉摸，而检测有急性肾损伤风险或有进展为严重和持续性急性肾损伤风险的患者对于及时采取适当的支持措施（包括减轻进一步的肾损伤）至关重要。因此，鉴定与脓毒症相关性 AKI 有关的生物标志物以改善脓毒症 AKI 是一个需要深入研究的领域：方法：根据脓毒症患者入住重症监护室（ICU）后 24 小时内发生 AKI 的情况，将 116 名脓毒症患者分为两组（AKI 组和非 AKI 组）。研究人员在患者入住重症监护室 24 小时内测量了血清中可溶性 TLR2（sTLR2）的水平，以及白细胞介素（IL）-6、IL-22、IL-10、肌酐、尿素、降钙素原、超敏 C 反应蛋白（hs-CRP）和 D-二聚体（D2）等生物标志物。记录了人口统计学和临床特征，包括序贯器官衰竭评估（SOFA）评分和急性生理学和慢性健康评估 II（APACHE II）评分。进行逻辑回归分析以确定潜在的预测性生物标记物。采用接收者操作特征（ROC）曲线分析来确定预测脓毒症相关性 AKI 的最佳模型：结果：与非 AKI 组患者相比，AKI 组患者血清中 IL-6、IL-10、sTLR2、肌酐、尿素、hs-CRP、降钙素原、D2 的浓度明显升高，血清白蛋白浓度降低，APACHE II 评分升高。逻辑回归分析显示，APACHE II 评分、经 log10 变形的 sTLR2 浓度、肌酐和 D2 浓度是脓毒症患者发生 AKI 的重要预测指标。ROC 曲线显示，在确定脓毒症相关性 AKI 的发生率方面，对数 10 变形的 sTLR2 浓度与肌酐具有相当的预测价值。包含 APACHE II 评分、血清 TLR2 浓度对数 10 变形值、肌酐和 D2 浓度等变量的模型得出的曲线下面积最大，为 0.863：脓毒症患者早期的 sTLR2 水平升高可作为预测脓毒症相关性 AKI 的新型生物标记物。
{"title":"Unveiling the role of sTLR2: A novel biomarker for predicting septic-associated AKI","authors":"Li Yuan , Yongshuang Zhou , Ruiyu Wang , Xin Huang , Ruilin Tang , Fang Yan","doi":"10.1016/j.cyto.2024.156798","DOIUrl":"10.1016/j.cyto.2024.156798","url":null,"abstract":"<div><h3>Background</h3><div>Acute kidney injury (AKI) is common in septic patients and strongly associated with adverse outcomes. The pathophysiology of AKI in septic patients remains elusive, and detection of patients at risk of AKI or at risk of progression to severe and persistent AKI is critical for timely and adequate support measures, including mitigating further renal damage. Therefore, identification of biomarkers associated with septic-associated AKI that contribute to improve septic AKI is an area of intensive research.</div></div><div><h3>Methods</h3><div>A total of 116 consecutive patients with sepsis were categorized into two groups (AKI and non-AKI) based on the occurrence of AKI within 24 h of admission to the intensive care unit (ICU). Serum levels of soluble TLR2 (sTLR2), as well as biomarkers such as interleukin(IL)-6, IL-22, IL-10, creatinine, urea, procalcitonin, hypersensitive C-reactive protein (hs-CRP), and D-Dimer (D2), were measured within 24 h after ICU admission. Demographic and clinical characteristics including sequential organ failure assessment (SOFA) scores and acute physiology and chronic health evaluation II (APACHE II) scores were recorded. Logistic regression analysis was conducted to identify potential predictive biomarkers. Receiver operating characteristic (ROC) curve analysis was employed to determine the optimal model for predicting septic-associated AKI.</div></div><div><h3>Results</h3><div>Patients in the AKI group exhibited significantly higher serum concentrations of IL-6, IL-10, sTLR2, creatinine, urea, hs-CRP, procalcitonin, D2 and lower serum albumin concentrations as well as higher APACHE II scores compared to those in the non-AKI group. Logistic regression analysis revealed that APACHE II scores, log10-transformed sTLR2 concentration, creatinine and D2 concentration were valuable predictors of AKI among septic patients. ROC curves demonstrated that log10-transformed sTLR2 concentration exhibited comparable predictive value to creatinine in determining the incidence of sepsis-associated AKI. The model with variables of APACHE II score, Log10-transformed serum TLR2 concentration, creatinine and D2 concentration yielded the greatest area under the curve of 0.863.</div></div><div><h3>Conclusion</h3><div>Elevated levels of sTLR2 in early-stage of septic patients may serve as a promising novel biomarker for predicting sepsis-associated AKI.</div></div>","PeriodicalId":297,"journal":{"name":"Cytokine","volume":"184 ","pages":"Article 156798"},"PeriodicalIF":3.7,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142563537","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-11-01DOI: 10.1016/j.cyto.2024.156797
Alessandro Cannavo , Monica Gelzo , Caterina Vinciguerra , Graziamaria Corbi , Marco Maglione , Vincenzo Tipo , Antonietta Giannattasio , Giuseppe Castaldo
Endothelial-cell-specific molecule-1 (ESM-1) also called endocan is a well-known biomarker for detecting inflammation, endothelial dysfunction (ED), and cardiovascular (CV) risk in COVID-19 patients. Upon SARS-CoV-2 infection, a small percentage of children develop Multisystem Inflammatory Syndrome in children (MIS-C). Whether endocan can be used as a biomarker of MIS-C is unknown. In this study, we assessed ESM-1 levels in MIS-C (n = 19) and healthy controls (HC; n = 17). We observed a significant increase in serum ESM-1 levels in MIS-C vs HC (p = 0.0074). In addition, ROC curve analysis demonstrated that this factor has a reasonable discriminatory power between MIS-C patients and HC (AUC of 0.7585). Notably, after one week of hospitalization and care, ESM-1 levels decreased, and this reduction was observed also for other inflammatory and pro-thrombotic markers like C-reactive protein, procalcitonin, fibrinogen, D-dimer, and ferritin, suggesting a general recovery trend in MIS-C patients. In fact, we observed that serum ESM-1 levels positively correlated with procalcitonin (PCT) (r = 0.468; p = 0.043). Finally, logistic regression analysis demonstrated an association between endocan levels and cardiac complications like myocarditis. Therefore, this study suggests that ESM-1 is a valuable diagnostic and prognostic biomarker in patients with MIS-C that may help identify those MIS-C patients at higher risk for cardiovascular complications and guide treatment strategies.
{"title":"Serum endocan (ESM-1) as diagnostic and prognostic biomarker in Multisystem inflammatory syndrome in children (MIS-C)","authors":"Alessandro Cannavo , Monica Gelzo , Caterina Vinciguerra , Graziamaria Corbi , Marco Maglione , Vincenzo Tipo , Antonietta Giannattasio , Giuseppe Castaldo","doi":"10.1016/j.cyto.2024.156797","DOIUrl":"10.1016/j.cyto.2024.156797","url":null,"abstract":"<div><div>Endothelial-cell-specific molecule-1 (ESM-1) also called endocan is a well-known biomarker for detecting inflammation, endothelial dysfunction (ED), and cardiovascular (CV) risk in COVID-19 patients. Upon SARS-CoV-2 infection, a small percentage of children develop Multisystem Inflammatory Syndrome in children (MIS-C). Whether endocan can be used as a biomarker of MIS-C is unknown. In this study, we assessed ESM-1 levels in MIS-C (n = 19) and healthy controls (HC; n = 17). We observed a significant increase in serum ESM-1 levels in MIS-C vs HC (p = 0.0074). In addition, ROC curve analysis demonstrated that this factor has a reasonable discriminatory power between MIS-C patients and HC (AUC of 0.7585). Notably, after one week of hospitalization and care, ESM-1 levels decreased, and this reduction was observed also for other inflammatory and pro-thrombotic markers like C-reactive protein, procalcitonin, fibrinogen, D-dimer, and ferritin, suggesting a general recovery trend in MIS-C patients. In fact, we observed that serum ESM-1 levels positively correlated with procalcitonin (PCT) (r = 0.468; p = 0.043). Finally, logistic regression analysis demonstrated an association between endocan levels and cardiac complications like myocarditis. Therefore, this study suggests that ESM-1 is a valuable diagnostic and prognostic biomarker in patients with MIS-C that may help identify those MIS-C patients at higher risk for cardiovascular complications and guide treatment strategies.</div></div>","PeriodicalId":297,"journal":{"name":"Cytokine","volume":"184 ","pages":"Article 156797"},"PeriodicalIF":3.7,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142563535","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-31DOI: 10.1016/j.cyto.2024.156796
Jin Cao , Zhaowen Yang
Background
Rheumatoid arthritis (RA) remains a devastating autoimmune disease characterized by joint damage, inflammation, and disability. This study investigates the function of lymphocyte antigen 96 (LY96) in the inflammatory response in RA and explores its regulatory mechanism.
Methods
A mouse model of RA was developed using type II collagen, and the LY96 expression in the ankle joint tissue was determined. Upstream regulators targeting LY96 were investigated using bioinformatics, followed by chromatin immunoprecipitation and luciferase reporter assays for validation. Gain- or loss-of-functions of LY96 and forkhead box A2 (FOXA2) were performed to analyze their roles in arthritis score, pathological changes, and inflammatory responses in mice. The effects of FOXA2 and LY96 on pro-inflammatory activation of macrophages were additionally investigated in vitro using a mouse RAW264.7 macrophage model with lipopolysaccharide treatment.
Results
LY96 mRNA and protein (MD-2) levels were increased in the RA mice. Knockdown of LY96 alleviated arthritis severity, joint deformities, inflammation, and cartilage destruction in mice. In vitro, the LY96 knockdown reduced the pro-inflammatory activation of RAW264.7 macrophages by inhibiting the TLR4/NF-κB inflammatory signaling transduction. FOXA2 was identified as a transcriptional repressor of LP96 poorly expressed in RA. Overexpression of FOXA2 similarly alleviated inflammation and reduced M1-type macrophages in vivo and in vitro. However, these changes were reversed by the additional LY96 upregulation.
Conclusion
This study suggests that FOXA2 represses LY96 transcription to inhibit the TLR4/NF-κB signaling transduction, thus reducing pro-inflammatory activation of macrophages in the context of RA.
背景类风湿性关节炎（RA）仍然是一种以关节损伤、炎症和残疾为特征的破坏性自身免疫性疾病。本研究调查了淋巴细胞抗原 96（LY96）在 RA 炎症反应中的功能，并探讨了其调控机制。方法利用 II 型胶原蛋白建立了 RA 小鼠模型，并测定了 LY96 在踝关节组织中的表达。采用生物信息学方法研究了以 LY96 为靶标的上游调控因子，并通过染色质免疫沉淀和荧光素酶报告实验进行了验证。对 LY96 和叉头框 A2（FOXA2）的功能增益或缺失进行了研究，以分析它们在小鼠关节炎评分、病理变化和炎症反应中的作用。此外，研究人员还利用小鼠 RAW264.7 巨噬细胞模型和脂多糖处理，在体外研究了 FOXA2 和 LY96 对巨噬细胞促炎激活的影响。敲除 LY96 可减轻小鼠关节炎的严重程度、关节畸形、炎症和软骨破坏。在体外，LY96基因敲除通过抑制TLR4/NF-κB炎症信号转导，减少了RAW264.7巨噬细胞的促炎激活。FOXA2被鉴定为LP96的转录抑制因子，在RA中表达较差。FOXA2 的过表达同样减轻了炎症，并减少了体内和体外的 M1 型巨噬细胞。结论这项研究表明，FOXA2抑制LY96转录以抑制TLR4/NF-κB信号转导，从而减少RA情况下巨噬细胞的促炎激活。
{"title":"FOXA2 inhibits the TLR4/NF-κB signaling pathway and alleviates inflammatory activation of macrophages in rheumatoid arthritis by repressing LY96 transcription","authors":"Jin Cao , Zhaowen Yang","doi":"10.1016/j.cyto.2024.156796","DOIUrl":"10.1016/j.cyto.2024.156796","url":null,"abstract":"<div><h3>Background</h3><div>Rheumatoid arthritis (RA) remains a devastating autoimmune disease characterized by joint damage, inflammation, and disability. This study investigates the function of lymphocyte antigen 96 (LY96) in the inflammatory response in RA and explores its regulatory mechanism.</div></div><div><h3>Methods</h3><div>A mouse model of RA was developed using type II collagen, and the LY96 expression in the ankle joint tissue was determined. Upstream regulators targeting LY96 were investigated using bioinformatics, followed by chromatin immunoprecipitation and luciferase reporter assays for validation. Gain- or loss-of-functions of LY96 and forkhead box A2 (FOXA2) were performed to analyze their roles in arthritis score, pathological changes, and inflammatory responses in mice. The effects of FOXA2 and LY96 on pro-inflammatory activation of macrophages were additionally investigated <em>in vitro</em> using a mouse RAW264.7 macrophage model with lipopolysaccharide treatment.</div></div><div><h3>Results</h3><div>LY96 mRNA and protein (MD-2) levels were increased in the RA mice. Knockdown of LY96 alleviated arthritis severity, joint deformities, inflammation, and cartilage destruction in mice. <em>In vitro</em>, the LY96 knockdown reduced the pro-inflammatory activation of RAW264.7 macrophages by inhibiting the TLR4/NF-κB inflammatory signaling transduction. FOXA2 was identified as a transcriptional repressor of LP96 poorly expressed in RA. Overexpression of FOXA2 similarly alleviated inflammation and reduced M1-type macrophages <em>in vivo</em> and <em>in vitro</em>. However, these changes were reversed by the additional LY96 upregulation.</div></div><div><h3>Conclusion</h3><div>This study suggests that FOXA2 represses LY96 transcription to inhibit the TLR4/NF-κB signaling transduction, thus reducing pro-inflammatory activation of macrophages in the context of RA.</div></div>","PeriodicalId":297,"journal":{"name":"Cytokine","volume":"184 ","pages":"Article 156796"},"PeriodicalIF":3.7,"publicationDate":"2024-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142560781","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-10-29DOI: 10.1016/j.cyto.2024.156786
Stephanie A. Goldstein , Melissa Winder , Camille Carter , J. Bair Diamond , Eric Bowles , Thomas B. Martins , Harry R. Hill , David K. Bailly
Background
Chylothorax after pediatric cardiac surgery is associated with increased morbidity and mortality. Poor understanding exists regarding inflammation within the pleural fluid. Our aim was to determine the relationship between proinflammatory markers and chylothorax.
Methods
This is a single-center prospective observational cohort study. Pediatric patients after cardiac surgery with >20 mL/kg/day of chest tube output were included from January 2022 through January 2023. The pleural fluid was tested for 13 cytokine concentrations using a multiplexed immunoassay and for albumin and C-Reactive Protein (CRP) levels. Bivariable comparisons and Spearman’s rank correlations were made.
Results
Out of 63 patients, chylothorax occurred in 20 (32 %), of which 10 (50 %) were neonates. Cytokine concentrations, CRP, and albumin levels were not different between chylothorax and non-chylothorax patients. Higher levels of four proinflammatory cytokines (IL-1β, IL-5, IL-8, IL-13) correlated with longer chest tube drainage (r = 0.29, 0.43, 0.28, 0.39 respectively). There were higher concentrations of IL-1β, IL-5, IL-8 and IL-13 in each progressively longer quartile of days to resolution. The longest quartile of days to resolution (12–50 days) showed the highest median cytokine levels.
Conclusions
The 13 cytokines tested were not associated with the diagnosis of chylothorax. However, higher IL-1β, IL-5, IL-8, and IL-13 concentrations were correlated with longer days to resolution. These findings demonstrate an inflammatory component to effusion resolution and may indicate an inflammatory phenotype that is distinct from chylothorax.
{"title":"Cytokines in chest tube drainage after pediatric cardiac surgery – Is chylothorax the only phenotype?","authors":"Stephanie A. Goldstein , Melissa Winder , Camille Carter , J. Bair Diamond , Eric Bowles , Thomas B. Martins , Harry R. Hill , David K. Bailly","doi":"10.1016/j.cyto.2024.156786","DOIUrl":"10.1016/j.cyto.2024.156786","url":null,"abstract":"<div><h3>Background</h3><div>Chylothorax after pediatric cardiac surgery is associated with increased morbidity and mortality. Poor understanding exists regarding inflammation within the pleural fluid. Our aim was to determine the relationship between proinflammatory markers and chylothorax.</div></div><div><h3>Methods</h3><div>This is a single-center prospective observational cohort study. Pediatric patients after cardiac surgery with >20 mL/kg/day of chest tube output were included from January 2022 through January 2023. The pleural fluid was tested for 13 cytokine concentrations using a multiplexed immunoassay and for albumin and C-Reactive Protein (CRP) levels. Bivariable comparisons and Spearman’s rank correlations were made.</div></div><div><h3>Results</h3><div>Out of 63 patients, chylothorax occurred in 20 (32 %), of which 10 (50 %) were neonates. Cytokine concentrations, CRP, and albumin levels were not different between chylothorax and non-chylothorax patients. Higher levels of four proinflammatory cytokines (IL-1β, IL-5, IL-8, IL-13) correlated with longer chest tube drainage (<em>r</em> = 0.29, 0.43, 0.28, 0.39 respectively). There were higher concentrations of IL-1β, IL-5, IL-8 and IL-13 in each progressively longer quartile of days to resolution. The longest quartile of days to resolution (12–50 days) showed the highest median cytokine levels.</div></div><div><h3>Conclusions</h3><div>The 13 cytokines tested were not associated with the diagnosis of chylothorax. However, higher IL-1β, IL-5, IL-8, and IL-13 concentrations were correlated with longer days to resolution. These findings demonstrate an inflammatory component to effusion resolution and may indicate an inflammatory phenotype that is distinct from chylothorax.</div></div>","PeriodicalId":297,"journal":{"name":"Cytokine","volume":"184 ","pages":"Article 156786"},"PeriodicalIF":3.7,"publicationDate":"2024-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142537873","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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