1. In vivo imaging of CD8 + T cell-mediated elimination of malaria liver stages
- Author
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Vitaly V. Ganusov, Andrea J. Radtke, Reka K. Kelemen, Fidel Zavala, Rogerio Amino, Sze Wah Tse, Scot C. Kuo, Ian A. Cockburn, Robert Ménard, Laura Mac-Daniel, Johns Hopkins Malaria Research Institute [Baltimore], Johns Hopkins Bloomberg School of Public Health [Baltimore], Johns Hopkins University (JHU)-Johns Hopkins University (JHU), W. Harry Feinstone Department of Molecular Microbiology and Immunology [Baltimore, MD, États-Unis] (MMI), Biologie et Génétique du Paludisme, Institut Pasteur [Paris], Genome Science and Technology, The University of Tennessee [Knoxville], Johns Hopkins University School of Medicine [Baltimore], R.M. is supported by Grant ANR-10-LABX-62-IBEID from the French Government’s Investissement d’Avenir program, and R.A. by the French National Research Agency Grant ANR-10-JCJC-1302-PlasmoPEP. Work in the F.Z. laboratory was supported by National Institutes of Health Grant AI44375 (to F.Z.) and an European Molecular Biology Organization short-term fellowship (to I.A.C.). F.Z. and I.A.C. receive support from the Bloomberg Family Foundation., We thank Dr. Yun-Chi Chen for providing recombinant vaccinia virus, Dr. Pascale Gueirard for animal breeding, Center for Production and Infection of Anopheles for providing mosquitoes, and IMAGOPOLE for the imaging platform., ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), ANR-10-JCJC-1302,PlasmoPEP,Imagerie in vivo de la phase pre-erythrocitaire du paludisme(2010), and Institut Pasteur [Paris] (IP)
- Subjects
lymphocytes ,Plasmodium ,Epitopes, T-Lymphocyte ,CD8-Positive T-Lymphocytes ,MESH: CD8-Positive T-Lymphocytes/immunology ,Parasite Load ,Mice ,Interleukin 21 ,0302 clinical medicine ,MESH: Reverse Transcriptase Polymerase Chain Reaction ,MESH: Receptors, Antigen, T-Cell/metabolism ,Cytotoxic T cell ,MESH: Animals ,IL-2 receptor ,Mice, Inbred BALB C ,0303 health sciences ,Microscopy, Confocal ,Multidisciplinary ,Reverse Transcriptase Polymerase Chain Reaction ,MESH: Malaria/parasitology ,CD28 ,Biological Sciences ,Natural killer T cell ,Adoptive Transfer ,MESH: Microscopy, Confocal/methods ,MESH: Liver/parasitology ,3. Good health ,Cell biology ,MESH: CD8-Positive T-Lymphocytes/ultrastructure ,Liver ,[SDV.IMM.IA]Life Sciences [q-bio]/Immunology/Adaptive immunology ,MESH: Malaria/immunology ,MESH: Epitopes, T-Lymphocyte/metabolism ,MESH: Mice, Transgenic ,MESH: Models, Immunological ,Green Fluorescent Proteins ,Antigen presentation ,Receptors, Antigen, T-Cell ,MESH: Mice, Inbred BALB C ,Mice, Transgenic ,MESH: Green Fluorescent Proteins/metabolism ,Streptamer ,Biology ,Time-Lapse Imaging ,Cell Line ,MESH: Plasmodium/immunology ,03 medical and health sciences ,Animals ,Antigen-presenting cell ,MESH: Mice ,030304 developmental biology ,MESH: Liver/immunology ,Models, Immunological ,MESH: Time-Lapse Imaging/methods ,immunity ,Malaria ,MESH: Cell Line ,MESH: Parasite Load ,MESH: Adoptive Transfer ,030215 immunology - Abstract
CD8 + T cells are specialized cells of the adaptive immune system capable of finding and eliminating pathogen-infected cells. To date it has not been possible to observe the destruction of any pathogen by CD8 + T cells in vivo. Here we demonstrate a technique for imaging the killing of liver-stage malaria parasites by CD8 + T cells bearing a transgenic T cell receptor specific for a parasite epitope. We report several features that have not been described by in vitro analysis of the process, chiefly the formation of large clusters of effector CD8 + T cells around infected hepatocytes. The formation of clusters requires antigen-specific CD8 + T cells and signaling by G protein-coupled receptors, although CD8 + T cells of unrelated specificity are also recruited to clusters. By combining mathematical modeling and data analysis, we suggest that formation of clusters is mainly driven by enhanced recruitment of T cells into larger clusters. We further show various death phenotypes of the parasite, which typically follow prolonged interactions between infected hepatocytes and CD8 + T cells. These findings stress the need for intravital imaging for dissecting the fine mechanisms of pathogen recognition and killing by CD8 + T cells.
- Published
- 2013