title: In Vivo Killing Capacity of Cytotoxic T Cells Is Limited and Involves Dynamic Interactions and T Cell Cooperativity.
abstract: According to in vitro assays, T cells are thought to kill rapidly and efficiently, but the efficacy and dynamics of cytotoxic T lymphocyte (CTL)-mediated killing of virus-infected cells in vivo remains elusive. We used two-photon microscopy to quantify CTL-mediated killing in mice infected with herpesviruses or poxviruses. On average, one CTL killed 2-16 virus-infected cells per day as determined by real-time imaging and by mathematical modeling. In contrast, upon virus-induced MHC class I downmodulation, CTLs failed to destroy their targets. During killing, CTLs remained migratory and formed motile kinapses rather than static synapses with targets. Viruses encoding the calcium sensor GCaMP6s revealed strong heterogeneity in individual CTL functional capacity. Furthermore, the probability of death of infected cells increased for those contacted by more than two CTLs, indicative of CTL cooperation. Thus, direct visualization of CTLs during killing of virus-infected cells reveals crucial parameters of CD8(+) T cell immunity.
authors.affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany. Electronic address: halle.stephan@mh-hannover.de.
authors.forename: Stephan
authors.initials: S
authors.lastname: Halle
authors.affiliation: Institute of Virology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Kirsten Anja
authors.initials: KA
authors.lastname: Keyser
authors.affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Felix Rolf
authors.initials: FR
authors.lastname: Stahl
authors.affiliation: Institute of Virology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Busche
authors.affiliation: Institute of Virology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Anja
authors.initials: A
authors.lastname: Marquardt
authors.affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Xiang
authors.initials: X
authors.lastname: Zheng
authors.affiliation: Institute of Experimental Hematology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Galla
authors.affiliation: Institute for Immunology, Ludwig-Maximilians-Universität München, 80336 München, Germany; Institute of Molecular Immunology, Helmholtz Zentrum München, 81377 München, Germany.
authors.forename: Vigo
authors.initials: V
authors.lastname: Heissmeyer
authors.affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Heller
authors.affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Jasmin
authors.initials: J
authors.lastname: Boelter
authors.affiliation: Institute of Virology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Karen
authors.initials: K
authors.lastname: Wagner
authors.affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Bischoff
authors.affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Rieke
authors.initials: R
authors.lastname: Martens
authors.affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Asolina
authors.initials: A
authors.lastname: Braun
authors.affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Kathrin
authors.initials: K
authors.lastname: Werth
authors.affiliation: Department of Systems Immunology and Braunschweig Integrated Centre of Systems Biology, Helmholtz Centre for Infection Research, 38124 Braunschweig, Germany.
authors.forename: Alexey
authors.initials: A
authors.lastname: Uvarovskii
authors.affiliation: Department of Systems Immunology and Braunschweig Integrated Centre of Systems Biology, Helmholtz Centre for Infection Research, 38124 Braunschweig, Germany.
authors.forename: Harald
authors.initials: H
authors.lastname: Kempf
authors.affiliation: Department of Systems Immunology and Braunschweig Integrated Centre of Systems Biology, Helmholtz Centre for Infection Research, 38124 Braunschweig, Germany; Institute for Biochemistry, Biotechnology, and Bioinformatics, Technische Universität Braunschweig, 38124 Braunschweig, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Meyer-Hermann
authors.affiliation: Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, 2333 ZA Leiden, the Netherlands.
authors.forename: Ramon
authors.initials: R
authors.lastname: Arens
authors.affiliation: Institute for Infectious Diseases and Zoonoses, Ludwig-Maximilians-Universität München, 80539 München, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Kremer
authors.affiliation: Institute for Infectious Diseases and Zoonoses, Ludwig-Maximilians-Universität München, 80539 München, Germany.
authors.forename: Gerd
authors.initials: G
authors.lastname: Sutter
authors.affiliation: Institute of Virology, Hannover Medical School, 30625 Hannover, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Messerle
authors.affiliation: Institute of Immunology, Hannover Medical School, 30625 Hannover, Germany. Electronic address: foerster.reinhold@mh-hannover.de.
authors.forename: Reinhold
authors.initials: R
authors.lastname: Förster
mesh_terms: D000818:Animals; D020013:Calcium Signaling; D002450:Cell Communication; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D006566:Herpesviridae Infections / Q000276:immunology*; D006801:Humans; D057131:Immune Evasion; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D036641:Microscopy, Fluorescence, Multiphoton; D018146:Muromegalovirus / Q000276:immunology*; D054353:Perforin / Q000235:genetics / Q000378:metabolism*; D016176:T-Lymphocyte Subsets / Q000276:immunology* / Q000821:virology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology* / Q000821:virology; D014615:Vaccinia / Q000276:immunology*; D014616:Vaccinia virus / Q000276:immunology*
chemical_list: D054353:Perforin