SFB796 - Sub project B2


Control of HSV-1 specific replication and transmission mechanisms in human dendritic cells


Project summary

Ralph Steinman called the dendritic cells (DC) "nature's adjuvant" to stress their importance in eliciting the immune response and to underline the fact that they are the only known antigen-presenting cells so far that can activate naïve T-cells. Of course they also play a fundamental role in triggering an antiviral response. This study aims to further explore the interplay between this important cell type and two members of the herpesviridae, namely HSV-1 and hCMV. While we were able to show that HSV-1 replicates in mature DC (mDC), we are now interested in the mechanisms that cause the low number of progeny particles. As we have hints that the nuclear egress might be hampered, we will start with investigating the disruption of the nuclear lamina. Another focus of our research will lie on the transfer of progeny HSV-1 from infected mDC to T-cells. We reported the transfer of HSV-1 from infected mDC to Vero cells or primary keratinocytes during the first funding period. Preliminary data suggest that transfer is also possible to CD8+ and CD4+ T-cells while infection of those with free virus is not possible. It would represent a very elegant way of immune evasion if infiltrating activated T-cells can be infected and eventually inactivated by HSV-1 transfer from infected mDC. We will address the question if the transfer is also gE-dependent as we showed for the transfer to Vero cells and keratinocytes or if trogocytosis or the virological synapse plays a role. In parallel, the phenotype of the infected T-cells will be analyzed.

We and others reported earlier that the loss of CD83 from the surface of mDC is a major immune evasion mechanism. In this part of the proposed project we want to deepen our insights in the mode of action underlying the loss of CD83 after (i) HSV-1 and (ii) hCMV infection. Finally, as during hCMV infection a soluble form of CD83 (sCD83) seems to contribute to the suppression of T-cell activation, we will determine the 3-D structure by (i) crystallography and (ii) by NMR. As sCD83 has been shown to be active in the prevention of the experimental autoimmune encephalomyelitis and of transplant rejections by reducing T-cell responses there might be a common mechanism. The structure of the protein will yield new insights in this mechanism.

Figure:   Viruses specifically target CD83 to evade Immune Responses.
Herpes simplex Virus Type 1 (HSV-1) as well as Human Cytomegalovirus (HCMV) target CD83 to influence T-cell stimulation by mDCs, but use completely different mechanisms. (1) The infection of mDCs with HSV-1 results in a rapid intracellular degradation of surface associated CD83. (2) This results in "mDCs", with low levels of membrane-bound CD83 (CD83- mDCs) and strongly reduced capacity to stimulate T-cells. (3) In strong contrast, the infection of mDCs with HCMV does not lead to degradation, but to a proteolytic cleavage of the surface CD83. (4) Thus, a form of soluble CD83 is generated that again inhibits T-cell stimulation. (5) Both – shedding of sCD83 as well as degradation of mCD83 – result in strongly reduced T-cell activation, leading to strongly reduced immune responses. mCD83 = membrane-bound CD83; sCD83 = soluble CD83; HSV-1 = Herpes simplex Virus Type 1; HCMV = Human Cytomegalovirus.


Project relevant publications

  • Heilingloh CS, Grosche L, Kummer M, Mühl-Zürbes P, Kamm L, Scherer M, Latzko M, Stamminger T, Steinkasserer A.   (2017).   The Major Immediate-Early Protein IE2 of Human Cytomegalovirus Is Sufficient to Induce Proteasomal Degradation of CD83 on Mature Dendritic Cells.   Front Microbiol. 2017 Feb 1;8:119. doi: 10.3389/fmicb.2017.00119

  • Heilingloh CS, Kummer M, Mühl-Zürbes P, Draßner C, Daniel C, Klewer M, Steinkasserer A.   (2015).   L-particles transmit viral proteins from HSV-1-infected mDCs to uninfected bystander cells inducing CD83 down-modulation.   Journal of Virology 89(21):11046-11055

  • Stein MF, Blume K, Heilingloh CS, Kummer M, Biesinger B, Sticht H, Steinkasserer A.   (2015).   CD83 and GRASP55 interact in human dendritic cells.   Biochem. Biophys. Res. Comm. 459:42-48

  • Heilingloh CS, Mühl-Zürbes P, Steinkasserer A, Kummer M.   (2014).   Herpes simplex virus type 1 ICP0 induces CD83 degradation in mature dendritic cells independent of its E3 ubiquitin ligase function.   J. Gen. Virology 95(Pt 6):1366-1375

  • Theodoridis AA, Eich C, Figdor FG, Steinkasserer A.   (2011).   Infection of dendritic cells with herpes simplex virus type 1 induces rapid degradation of CYTIP, thereby modulating adhesion and migration.   Blood 118(1): 107-115

  • Goldwich A, Prechtel AT, Mühl-Zürbes P, Pangratz NM, Stössel H, Romani N, Steinkasserer A, Kummer M.   (2011).   Herpes simplex virus type I (HSV-1) replicates in mature dendritic cells but can only be transferred in a cell-cell contact-dependent manner.   J Leukoc Biol. 2011 Jun;89(6): 973-979

  • Staab C, Mühl-Zürbes P, Steinkasserer A, Kummer M.   (2010).   Eukaryotic expression of functionally active recombinant soluble CD83 from HEK 293T cells.   Immunobiology 215: 849-854.

  • Xu X, Husseiny MI, Goldwich A, Hensel M.   (2010).   Efficacy of intracellular activated promoters for generation of Salmonella-based vaccines.   Infect Immun. 78(11): 4828-4838.

  • Kummer M, Prechtel AT, Mühl-Zürbes P, Turza NM, Steinkasserer A.   (2009).   HSV-1 upregulates the ARE-binding protein tristetraprolin in a STAT1- and p38-dependent manner in mature dendritic cells.   Immunobiology 214(9-10): 852-860.

  • Eisemann J, Prechtel AT, Mühl-Zürbes P, Steinkasserer A, Kummer M.   (2009).   Herpes simplex virus type I infection of mature dendritic cells leads to reduced LMP7-mRNA-expression levels.   Immunobiology 214(9-10): 861-867.

  • Kummer, M., Turza, N. M., Boutell, C., Everett, R. D., Steinkasserer A., Prechtel, A. T.   (2007).   Herpes Simplex Virus Type 1 (HSV-1) induces CD83 degradation in mature dendritic cells with immediate early kinetics via the cellular proteasome.   J Virology 81(12): 6326-6338.

  • Prechtel, A. T., Turza, N. M., Theodoridis, A. and Steinkasserer A.   (2007).   CD83 knockdown in monocyte-derived dendritic cells by small interfering RNA (siRNA) leads to a diminished T-cell-stimulation.   J Immunology 178: 5454-5464.

  • Prechtel, A., Turza, N. M., Kobelt, D. J., Eisemann, J. I., Coffin, R. S., McGrath, Y., Hacker, C., Ju, X., Zenke, M. and Steinkasserer A.   (2005).   Infection of mature dendritic cells with herpes simplex virus type 1 dramatically reduces lymphoid chemokine-mediated migration.   J Gen Virol 86: 1645-1657.

  • Zinser, E., Lechmann, M., Golka, A., Lutz, M. B. and Steinkasserer A.   (2004).   Prevention and treatment of experimental autoimmune encephalomyelitis (EAE) by soluble CD83.   J Exp Med 200: 345-351.

  • Lechmann, M., Krooshoop, D. J. E. B., Dudziak, D., Kremmer, E., Kuhnt, C., Figdor, C. G., Schuler, G. and A. Steinkasserer   (2001).   The extracellular domain of CD83 inhibits dendritic cell-mediated T-cell stimulation and binds to a ligand on dendritic cells.   J Exp Med 194: 1813-1821.