PMID- 29866139 OWN - NLM STAT- MEDLINE DCOM- 20190410 LR - 20190711 IS - 1742-2094 (Electronic) IS - 1742-2094 (Linking) VI - 15 IP - 1 DP - 2018 Jun 4 TI - Elimination of intravascular thrombi prevents early mortality and reduces gliosis in hyper-inflammatory experimental cerebral malaria. PG - 173 LID - 10.1186/s12974-018-1207-4 [doi] LID - 173 AB - BACKGROUND: Cerebral malaria (CM) is the most lethal outcome of Plasmodium infection. There are clear correlations between expression of inflammatory cytokines, severe coagulopathies, and mortality in human CM. However, the mechanisms intertwining the coagulation and inflammation pathways, and their roles in CM, are only beginning to be understood. In mice with T cells deficient in the regulatory cytokine IL-10 (IL-10 KO), infection with Plasmodium chabaudi leads to a hyper-inflammatory response and lethal outcome that can be prevented by anti-TNF treatment. However, inflammatory T cells are adherent within the vasculature and not present in the brain parenchyma, suggesting a novel form of cerebral inflammation. We have previously documented behavioral dysfunction and microglial activation in infected IL-10 KO animals suggestive of neurological involvement driven by inflammation. In order to understand the relationship of intravascular inflammation to parenchymal dysfunction, we studied the congestion of vessels with leukocytes and fibrin(ogen) and the relationship of glial cell activation to congested vessels in the brains of P. chabaudi-infected IL-10 KO mice. METHODS: Using immunofluorescence microscopy, we describe severe thrombotic congestion in these animals. We stained for immune cell surface markers (CD45, CD11b, CD4), fibrin(ogen), microglia (Iba-1), and astrocytes (GFAP) in the brain at the peak of behavioral symptoms. Finally, we investigated the roles of inflammatory cytokine tumor necrosis factor (TNF) and coagulation on the pathology observed using neutralizing antibodies and low-molecular weight heparin to inhibit both inflammation and coagulation, respectively. RESULTS: Many blood vessels in the brain were congested with thrombi containing adherent leukocytes, including CD4 T cells and monocytes. Despite containment of the pathogen and leukocytes within the vasculature, activated microglia and astrocytes were prevalent in the parenchyma, particularly clustered near vessels with thrombi. Neutralization of TNF, or the coagulation cascade, significantly reduced both thrombus formation and gliosis in P. chabaudi-infected IL-10 KO mice. CONCLUSIONS: These findings support the contribution of cytokines, coagulation, and leukocytes within the brain vasculature to neuropathology in malaria infection. Strikingly, localization of inflammatory leukocytes within intravascular clots suggests a mechanism for interaction between the two cascades by which cytokines drive local inflammation without considerable cellular infiltration into the brain parenchyma. FAU - Wilson, Kyle D AU - Wilson KD AD - Department of Microbiology and Immunology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. FAU - Ochoa, Lorenzo F AU - Ochoa LF AD - Center for Biomedical Engineering, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. FAU - Solomon, Olivia D AU - Solomon OD AD - Center for Biomedical Engineering, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. FAU - Pal, Rahul AU - Pal R AD - Center for Biomedical Engineering, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. FAU - Cardona, Sandra M AU - Cardona SM AD - Department of Biology, One UTSA Circle, University of Texas at San Antonio, San Antonio, TX, 78249, USA. FAU - Carpio, Victor H AU - Carpio VH AD - Department of Microbiology and Immunology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. FAU - Keiser, Philip H AU - Keiser PH AD - Department of Internal Medicine, Division of Infectious Diseases, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555-0435, USA. FAU - Cardona, Astrid E AU - Cardona AE AD - Department of Biology, One UTSA Circle, University of Texas at San Antonio, San Antonio, TX, 78249, USA. FAU - Vargas, Gracie AU - Vargas G AD - Center for Biomedical Engineering, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. AD - Department of Neuroscience and Cell Biology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. AD - Institute for Human Infections and Immunity, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. FAU - Stephens, Robin AU - Stephens R AUID- ORCID: 0000-0001-8036-600X AD - Department of Microbiology and Immunology, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. rostephe@utmb.edu. AD - Department of Internal Medicine, Division of Infectious Diseases, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555-0435, USA. rostephe@utmb.edu. AD - Institute for Human Infections and Immunity, University of Texas Medical Branch, 301 University Boulevard, Galveston, TX, 77555, USA. rostephe@utmb.edu. LA - eng GR - R01 NS078501/NS/NINDS NIH HHS/United States GR - R01NS078501/National Institute of Neurological Disorders and Stroke/ GR - 363327/University of Texas System/ GR - T32AI363327/National Institute of Allergy and Infectious Diseases/ GR - R01 AI089953/AI/NIAID NIH HHS/United States GR - R01 NS106597/NS/NINDS NIH HHS/United States GR - R01AI363327/National Institute of Allergy and Infectious Diseases/ PT - Journal Article DEP - 20180604 PL - England TA - J Neuroinflammation JT - Journal of neuroinflammation JID - 101222974 RN - 0 (Antibodies) RN - 0 (Anticoagulants) RN - 0 (Cytokines) RN - 0 (Glial Fibrillary Acidic Protein) RN - 0 (IL10 protein, mouse) RN - 0 (Tumor Necrosis Factor-alpha) RN - 0 (glial fibrillary astrocytic protein, mouse) RN - 130068-27-8 (Interleukin-10) RN - 7664-41-7 (Ammonia) RN - 9001-32-5 (Fibrinogen) RN - 9005-49-6 (Heparin) SB - IM MH - Ammonia/blood MH - Animals MH - Antibodies/therapeutic use MH - Anticoagulants/therapeutic use MH - Blood Vessels/pathology MH - Cytokines/*metabolism MH - Disease Models, Animal MH - Fibrinogen/metabolism MH - Gene Expression Regulation/genetics MH - Glial Fibrillary Acidic Protein/metabolism MH - Gliosis/drug therapy/*etiology/*prevention & control MH - Heparin/therapeutic use MH - Interleukin-10/genetics/metabolism MH - Leukocytes/pathology MH - Liver/metabolism/pathology MH - Malaria, Cerebral/*complications/mortality MH - Mice MH - Mice, Inbred C57BL MH - Mice, Transgenic MH - Plasmodium chabaudi/physiology MH - Tumor Necrosis Factor-alpha/immunology/metabolism MH - Vasculitis, Central Nervous System/drug therapy/*etiology/parasitology PMC - PMC5987620 OTO - NOTNLM OT - Astrocyte OT - Brain OT - Confocal microscopy OT - Immunofluorescence OT - Inflammation OT - Malaria OT - Microglia OT - Monocyte OT - Neuropathology OT - Vascular congestion COIS- ETHICS APPROVAL: Animals were cared for according to the Guide for the Care and Use of Laboratory Animals under the Institutional Animal Care and Use Committee-approved protocol #1006031. UTMB Animal Resource Center facilities operate in compliance with the USDA Animal Welfare Act, the Guide for the Care and Use of Laboratory Animals, under OLAW accreditation, and IACUC-approved protocols. COMPETING INTERESTS: The authors declare that they have no competing interests. PUBLISHER'S NOTE: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. EDAT- 2018/06/06 06:00 MHDA- 2019/04/11 06:00 PMCR- 2018/06/04 CRDT- 2018/06/06 06:00 PHST- 2018/02/14 00:00 [received] PHST- 2018/05/17 00:00 [accepted] PHST- 2018/06/06 06:00 [entrez] PHST- 2018/06/06 06:00 [pubmed] PHST- 2019/04/11 06:00 [medline] PHST- 2018/06/04 00:00 [pmc-release] AID - 10.1186/s12974-018-1207-4 [pii] AID - 1207 [pii] AID - 10.1186/s12974-018-1207-4 [doi] PST - epublish SO - J Neuroinflammation. 2018 Jun 4;15(1):173. doi: 10.1186/s12974-018-1207-4.