Visualization of herpes virus infections and immune response as well as their modulation

Immune cells (green) are visualized with 2-Photonen microscopy. Virus infected cells (red) are intact. The picture was taken from a 24h-infected popliteal lymph node of a mouse producing GFP-expressed immune cells

How does the immune system protect us against chronic viral infections? Beta-herpesviruses can chronically infect humans and mice, but only immunocompromised individuals suffer from disease. Thus, the intact immune response does play an important role in controlling a beta-herpesvirus infection.
In this project, we apply a combination of the in vivo 2-photon microscopy technique and infections with beta-herpesvirus reporter viruses, in order to directly investigate, in real time, how immune cells interact with virus-infected cells and keep the infection in check.

Scientific work programme

In this project, we are investigating the in vivo infection with the mouse cytomegalovirus (MCMV, murine herpesvirus-1) and the induction of CMV-specific immunity. In close collaboration between the MHH departments of immunology and virology, we have been able to unravel how different types of immune cells attack virus-infected cells in vivo, applying different mouse models. By visualizing individual cells we discovered that single cytotoxic T cells were unable to detect infected cells when the viral immune evasion mechanisms were intact. Only upon deletion of two important MCMV immune evasion genes the infected target cells became visible to the immune cells and could be destroyed (Halle et al., Immunity 2016).

We also explore how MCMV infection spreads in the lungs of infected new-born mice (Stahl et al., Mucosal Immunology 2015, Stahl et al., PLoS Pathogens 2013, Marquardt et al., and Journal of General Virology 2011).

Taken together, by applying high-resolution techniques we can closely monitor MCMV infection on the single cell level in different organs to better understand the protective immune response and particularly how it prevents chronic persistence of the infection in immunocompetent hosts.

Reinhold Förster is talking about his research at the CRC 900

Reinhold Förster has been part of the CRC since it was founded in 2010. His project B1, in cooperation with Martin Messerle, is concerned with the visualisation of the immune response during herpesvirus infections, using 2-photon-microscopy.

Publications of the project B1

Imaging dendritic cell functions. Bošnjak, B., Do, K.T.H., Förster, R., and Hammerschmidt, S.I. Immunol Rev 2022 306, 137-163. DOI: 10.1111/imr.13050
NK cell dysfunction in severe COVID-19: TGF-β-induced downregulation of integrin beta-2 restricts NK cell cytotoxicity. Barros-Martins, J., Förster, R., and Bošnjak, B. Signal Transduct Target Ther 2022 7, 32. DOI: 10.1038/s41392-022-00892-5
Challenges of CRISPR-based gene editing in primary T cells. Rezalotfi A, Fritz L, Förster R, Bošnjak B. Int J Mol Sci. 2022; 23(3). Accessed Mar 23, 2022.
Immune responses against SARS-CoV-2 variants after heterologous and homologous ChAdOx1 nCoV-19/BNT162b2 vaccination. Barros-Martins J, Hammerschmidt SI, Cossmann A, Odak I, Stankov MV, Morillas Ramos G, Dopfer-Jablonka A, Heidemann A, Ritter C, Friedrichsen M, Schultze-Florey C, Ravens I, Willenzon S, Bubke A, Ristenpart J, Janssen A, Ssebyatika G, Bernhardt G, Münch J, Hoffmann M, Pöhlmann S, Krey T, Bosnjak B, Förster R, Behrens GMN. Nat Med 2021 ; 27; 9
Immune responses against SARS-CoV-2 variants after heterologous and homologous ChAdOx1 nCoV-19/BNT162b2 vaccination. Barros-Martins J, Hammerschmidt SI, Cossmann A, Odak I, Stankov MV, Morillas Ramos G, Dopfer-Jablonka A, Heidemann A, Ritter C, Friedrichsen M, Schultze-Florey C, Ravens I, Willenzon S, Bubke A, Ristenpart J, Janssen A, Ssebyatika G, Bernhardt G, Münch J, Hoffmann M, Pöhlmann S, Krey T, Bosnjak B, Förster R, Behrens GMN. Nat Med 2021 ; 27; 9
Case Report: Convalescent Plasma Therapy Induced Anti-SARSCoV-2 T Cell Expansion, NK Cell Maturation and Virus Clearance in a B Cell Defcient Patient After CD19 CAR T Cell Therapy. Bosnjak B, Odak I, Ritter C, Stahl K, Graalmann T, Steinbrück L, Blasczyk R, Falk CS, Schulz TF, Wedemeyer HH, Cornberg M, Ganser A, Förster R and Koenecke C Front. Immunol. 2021 Aug 12; 12:721738.
Intranasal Delivery of MVA Vector Vaccine Induces Effective Pulmonary Immunity Against SARS-CoV-2 in Rodents. Bosnjak B, Odak I, Barros-Martins J, Sandrock I, Hammerschmidt SI, Permanyer M, Patzer GE, Greorgiev H, Gutierrez Jauregui R, Tscherne A, Schwarz JH, Kalodimou G, Ssebyatika G, Ciurkiewicz M, Willenzon S, Bubke A, Ristenpart J, Ritter C, Tuchel T, Meyer Zu Natrup C, Shin D-L, Clever S, Limpinsel L, Baumgärtner W, Krey T, Volz A, Sutter G, Förster R. Front Immunol 2021 ; 12
Fucosylated lipid nanocarriers loaded with antibiotics efficiently inhibit mycobacterial propagation in human myeloid cells. Durán V, Grabski E, Hozsa C, Becker J, Yasar H, Monteiro JT, Costa B, Koller N, Lueder Y, Wiegmann B, Brandes G, Kaever V, Lehr C-M, Lepenies B, Tampé R, Förster R, Bosnjak B, Furch M, Graalmann T, Kalinke U. J Control Release 2021 ; 334
MyD88 signaling by neurons induces chemokines that recruit protective leukocytes to the virus-infected CNS. Ghita L, Spanier J, Chhatbar C, Mulenge F, Pavlou A, Larsen P-K, Waltl I, Lueder Y, Kohls M, Jung K, Best SM, Förster R, Stangel M, Schreiner D, Kalinke U. Sci. Immunol. 6, 2021 Jun 25, eabc9165
Neutralization of the SARS-CoV-2 Delta variant after heterologous and homologous BNT162b2 or ChAdOx1 nCoV-19 vaccination. Hammerschmidt SI, Bosnjak B, Bernhardt G, Friedrichsen M, Ravens I, Dopfer-Jablonka A, Hoffmann M, Pöhlmann S, Behrens GMN, Förster R. Cell Mol Immunol 2021; 18; 10
Distribution of major lymphocyte subsets and memory T-cell subpopulations in healthy adults employing GLP-conforming multicolor flow cytometry. Schultze-Florey CR, Chukhno E, Goudeva L, Blasczyk R, Ganser A, Prinz I, Förster R, Koenecke C, Odak I. Leukemia 2021; 35; 10
Immunogenicity and efficacy of the COVID-19 candidate vector vaccine MVA-SARS-2-S in preclinical vaccination. Tscherne A, Schwarz KH, Rohde C, Kupke A, Kalodimou G, Limpinsel L, Okba NMA, Bosnjak B, Sandrock I, Odak I, Halwe A, Sauerhering L, Brosinski K, Liangliang N, Duell E, Jany S, Freudenstein A, Schmidt J, Werner A, Gellhorn Serra M, Klüver M, Guggemos W, Seilmaier M, Wendtner, C-M, Förster R, Haagmans BL, Becker S, Sutter G, Volz A. Proc Natl Acad Sci U S A 2021; 118; 28
Expression of ACKR4 demarcates the “peri-marginal sinus,” a specialized vascular compartment of the splenic red pulp. Werth K, Hub E, Gutjahr JC, Bosjnak B, Zheng X, Bubke A, Russo S, Rot A, Förster R. Cell Rep 2021; 36; 2
Generation of hiPSC-derived low threshold mechanoreceptors containing axonal termini resembling bulbous sensory nerve endings and expressing Piezo1 and Piezo2. Zhu S, Stanslowsky N, Fernandez-Trillo J, Mamo TM, Yu P, Kalmbach N, Ritter B, Eggenschwiler R, Ouwendijk WJD, Mzinza D, Tan L, Leffler A, Spohn M, Brown RJP, Kropp KA, Kaever V, Ha, T-C, Narayanan P. Grundhoff A, Förster R, Schambach A, Verjans GMGM, Schmidt M, Kispert A, Cantz T, Gomis A, Wegner F, Viejo-Borbolla A. Stem Cell Res 2021; 56
Targeted delivery of regulatory macrophages to lymph nodes interferes with T cell priming by preventing the formation of stable immune synapses. Yu K, Hammerschmidt SI, Permanyer M, Galla M, Rothe M, Zheng X, Werth K, Martens R, Lueder Y, Janssen A, Friedrichsen M, Bernhardt G, Förster R. Cell Rep. 2021 Jun 22 ;35(12):109273.
Strategic Anti-SARS-CoV-2 Serology Testing in a Low Prevalence Setting: The COVID-19 Contact (CoCo) Study in Healthcare Professionals. Behrens GMN, Cossmann A, Stankov MV, Schulte B, Streeck H, Förster R, Bosnjak b, Willenzon S, Boeck A-L, Tran AT, Thiele T, Graalmann T, Kayser MZ, Zychlinsky Scharff A, Dopfer C, Horke A, Pink I, Witte T, Wetzke M, Ernst D, Jablonka A, Happle S. Infect Dis Ther. 2020 Dec 9(4):837-849.
Combating COVID-19: MVA Vector Vaccines Applied to the Respiratory Tract as Promising Approach Toward Protective Immunity in the Lung. Förster R, Fleige H, Sutter G. Front Immunol. 2020 Aug 7 11: 1959.
Differential retention of lymph-borne CD8 memory T cell subsets in the subcapsular sinus of resting and inflamed lymph nodes. Nikolova G, Weiss S, Bosnjak B, Förster R. Cell Mol Immunol. 2020 May 12.
Efficient homing of T cells via afferent lymphatics requires mechanical arrest and integrin-supported chemokine guidance. Martens R, Permanyer M, Werth K, Yu K, Braun A, Halle O, Halle S, Patzer GE, Bošnjak B, Kiefer F, Janssen A, Friedrichsen M, Poetzsch J, Kohli K, Lueder Y, Gutierrez Jauregui R, Eckert N, Worbs T, Galla M, Förster R. Nat Commun. 2020 Feb 28 11(1):1114.
Chemokines and other mediators in the development and functional organization of lymph nodes. Eckert N, Permanyer M, Yu K, Werth K, Förster R. Immunol Rev. 2019 May 289(1):62-83.
Control of primary mouse cytomegalovirus infection in lung nodular inflammatory 1 foci by cooperation of Interferon-gamma expressing CD4 and CD8 T cells. Lueder Y, Heller K, Ritter C, Keyser KA, Wagner K, Liu X, Messerle M, Stahl FR, Halle S, Förster R PLoS Pathog. 2018 Aug 28;14(8):e1007252.
Repulsive behavior in germinal Centers. Moschovakis GL, Förster R. Science. 2017 May 19;356(6339):703-704.
Dendritic cell migration in health and disease. Worbs T, Hammerschmidt SI, Förster R. Nat Rev Immunol. 2016 Nov 28.
Chemokines and Chemokine Receptors in Lymphoid Tissue Dynamics. Schulz O, Hammerschmidt SI, Moschovakis GL, Förster R. Annu Rev Immunol. 2016 May 20;34:203-42.
In Vivo Killing Capacity of Cytotoxic T Cells Is Limited and Involves Dynamic Interactions and T Cell Cooperativity. Halle S, Keyser KA, Stahl FR, Busche A, Marquardt A, Zheng X, Galla M, Heissmeyer V, Heller K, Boelter J, Wagner K, Bischoff Y, Martens R, Braun A, Werth K, Uvarovskii A, Kempf H, Meyer-Hermann M, Arens R, Kremer M, Sutter G, Messerle M, Förster R. Immunity. 2016 Feb 16;44(2):233-45.
Mck2-dependent infection of alveolar macrophages promotes replication of MCMV in nodular inflammatory foci of the neonatal lung. Stahl FR, Keyser KA, Heller K, Bischoff Y, Halle S, Wagner K, Messerle M, Förster R. Mucosal Immunol. 2015 Jan;8(1):57-67.
IL-17-induced CXCL12 recruits B cells and induces follicle formation in BALT in the absence of differentiated FDCs. Fleige H, Ravens S, Moschovakis GL, Bölter J, Willenzon S, Sutter G, Häussler S, Kalinke U, Prinz I, Förster R. J Exp Med. 2014 Apr 7;211(4):643-51.
Nodular inflammatory foci are sites of T cell priming and control of murine cytomegalovirus infection in the neonatal lung. Stahl FR, Heller K, Halle S, Keyser KA, Busche A, Marquardt A, Wagner K, Boelter J, Bischoff Y, Kremmer E, Arens R, Messerle M, Förster R. PLoS Pathog. 2013;9(12):e1003828.
Absence of Siglec-H in MCMV infection elevates interferon alpha production but does not enhance viral clearance. Puttur F, Arnold-Schrauf C, Lahl K, Solmaz G, Lindenberg M, Mayer CT, Gohmert M, Swallow M, van Helt C, Schmitt H, Nitschke L, Lambrecht BN, Lang R, Messerle M, Sparwasser T. PLoS Pathog. 2013;9(9):e1003648.
Priming of CD8+ T cells against cytomegalovirus-encoded antigens is dominated by cross-presentation. Busche A, Jirmo AC, Welten SP, Zischke J, Noack J, Constabel H, Gatzke AK, Keyser KA, Arens R, Behrens GM, Messerle M. J Immunol. 2013 Mar 15;190(6):2767-77.
The human cytomegalovirus UL51 protein is essential for viral genome cleavage-packaging and interacts with the terminase subunits pUL56 and pUL89. Borst EM, Kleine-Albers J, Gabaev I, Babic M, Wagner K, Binz A, Degenhardt I, Kalesse M, Jonjic S, Bauerfeind R, Messerle M. J Virol. 2013 Feb;87(3):1720-32.