Dr. Raluca Niesner
Deutsches Rheuma-Forschungszentrum Berlin
Prof. Dr. Anja Erika Hauser
Professor for Immunodynamics
and Intravital Microscopy
Charité University Medicine Berlin
Longitudinal multi-photon microscopy and microendoscopy: Quantifying and controlling communication and function of B lymphocytes in vivo
Intravital multi-photon microscopy is the most versatile technology enabling us to understand cellular dynamics and communication involving B lymphocytes in living organisms, both under physiologic and pathologic conditions. In the first funding period, we closely collaborated with partners in the CRC to answer key questions referring to B lymphocytes. Thereby, we established new evaluation algorithms, (P07, P16) and solved central technical challenges of the intavital multi-photon microscopy. We developed a permanent micro-endoscopic implant that allowed us to repeatedly image tissue at depths of more than 500 µm within the murine femoral bone marrow, at sub-cellular resolution, over months, helping us to monitor long-term dynamics of components of the long-lived plasma cell survival niche (P16). We achieved intravital spectral multiplexing (up to 8 chromophores) enabling us to study the orchestration of various immune subsets during germinal center reactions (C03, P03). As far as the tissue response to inflammation is concerned, we developed intravital marker-free and FRET-based fluorescence lifetime imaging approaches for functional tissue imaging with P17. This method revealed important links between antibody-induced oxidative stress memory and neuronal dysfunction in chronic neuroinflammation. Still, the limited imaging time-window of only several hours in other organs than the bone marrow and the difficulty to quantify tissue function and dysfunction prevent us from taking full advantage of the multi-photon technology in investigating the biology of B lymphocytes in vivo. In this respect, we currently develop approaches for non-invasive tissue access to monitor various immune responses in one and the same individual over months. They will allow us to monitor the inflamed kidney in a lupus model (P12, P15), the inflamed lung (P23), and lymph nodes transplanted to the ear to study germinal center development (P03, C03). Having established longitudinal imaging, we will adapt our marker-free and FRET-based functional imaging to gain additional information on the B lymphocytes involvement during chronic inflammation of the kidney (P12, P15) and in immunodeficiency (P07) as well as on B cell activation during germinal center (P03, P04) and germinal-center-like reactions (P12, P15, P17, P23) by monitoring tissue-related oxidative stress generation and intracellular calcium signaling in vivo.
Reimer, D., Meyer-Hermann, M., Rakhymzhan, A., Steinmetz, T., Tripal, P., Thomas, J., Boettcher, M., Mougiakakos, D., Schulz, S.R., Urbanczyk, S., Hauser, A.E., Niesner, R.A.*, Mielenz, D.* (2020). B cell speed and B-FDC contacts in germinal centers determine plasma cell output via Swiprosin-1/EFhd2. Cell Reports 32, 108030.
Riedel, R., Addo, R., Ferreira-Gomes, M., Heinz, G. A., Heinrich, F., Kummer, J., Greiff, V., Schulz, D., Klaeden, C., Cornelis, R., Menzel, U., Kroger, S., Stervbo, U., Kohler, R., Haftmann, C., Kuhnel, S., Lehmann, K., Maschmeyer, P., McGrath, M., Naundorf, S., Hahne, S., Sercan-Alp, O., Siracusa, F., Stefanowski, J., Weber, M., Westendorf, K., Zimmermann, J., Hauser, A. E., Reddy, S.T., Durek, P., Chang, H.D., Mashreghi, M.F., and Radbruch, A. (2020). Discrete populations of isotype-switched memory B lymphocytes are maintained in murine spleen and bone marrow. Nat. Commun. 11, 2570.
Mothes, R., Ulbricht, C., Leben, R., Günther, R., Hauser, A.E., Radbruch, H.*, Niesner, R.A.* (2020). Teriflunomide does not change dynamics of NADPH oxidase activation and neuronal dysfunction during neuroinflammation. Front. Mol. Biosci. 7, 62.
Jumaa, H., Caganova, M., McAllister, E.J., Hoenig, L., He, X., Saltukoglu, D., Brenker, K., Köhler, M., Leben, R., Hauser, A.E., Niesner, R., Rajewsky, K., Reth, M., Jellusova, J. (2020). Immunoglobulin Expression in the Endoplasmic Reticulum Shapes the Metabolic Fitness of B Lymphocytes. Life Sci. Alliance 3, e202000700.
Lindquist, R.L., Niesner, R., Hauser, A.E. In the right place, at the right time: spatiotemporal conditions determining plasma cell survival and function. (2019). Front. Immunol. 10,788.
Leben, R. *, Köhler, M. *, Radbruch, H., Hauser, A.E., Niesner, R. (2019). Systematic Enzyme Mapping of Cellular Metabolism by Phasor-Analyzed Label-Free NAD(P)H Fluorescence Lifetime Imaging. IJMS 10, 788.
Reismann, D.*, Stefanowski, J.*, Günther, R., Rakhymzhan, A., Matthys, R., Nützi, R., Zehentmeier, S., Schmidt-Bleek, K., Petkau, G., Chang, H.D., Naundorf, S., Winter, Y., Melchers, F., Duda, G., Hauser, A.E.*, Niesner, R.A.* (2017). Longitudinal intravital imaging of the femoral bone marrow reveals plasticity within marrow vasculature. Nat. Commun. 8, 2153.
Pollok, K., Mothes, R., Ulbricht, C., Liebheit, A., Gerken, J.D., Uhlmann, S., Paul, F., Niesner, R., Radbruch, H.*, Hauser, A.E.*. (2017). The chronically inflamed central nervous system provides niches for long-lived plasma cells. Acta Neuropathol. Commun. 5, 88.
Rakhymzhan, A., Leben, R., Zimmermann, H., Günther, R., Mex, P., Reismann, D., Ulbricht, C., Acs, A., Brandt, A.U., Lindquist, R.L., Winkler, T.H., Hauser, A.E., Niesner, R.A. (2017). Synergistic Strategy for Multicolor Two-photon Microscopy: Application to the Analysis of Germinal Center Reactions In Vivo. Sci. Rep. 7, 7101.
Radbruch, H., R. Mothes, D. Bremer, S. Seifert, R. Köhler, J. Pohlan, L. Ostendorf, R. Günther, R. Leben, W. Stenzel, R.A. Niesner, A.E. Hauser. (2017). Analyzing Nicotinamide Adenine Dinucleotide Phosphate Oxidase Activation in Aging and Vascular Amyloid Pathology.in press
Keller, B., Z. Cseresnyes, I. Stumpf, C. Wehr, M. Fliegauf, A. Bulashevska, S. Usadel, B. Grimbacher, M. Rizzi, H. Eibel, R. Niesner, and K. Warnatz. (2017). Disturbed canonical NF-kB signaling in B cells of CVID patients. J Allergy Clin Immunol. 139(1), 220-231.
Bremer, D., F. Pache, R. Günther, J. Hornow, V. Andresen, R. Leben, R. Mothes, H. Zimmermann, A.U. Brandt, F. Paul, A.E. Hauser, H. Radbruch, R. Niesner. (2016). Longitudinal imaging of the retina reveals long-term dynamics of immune infiltration and its effect on the glial network in experimental autoimmune uveoretinitis, without evident signs of neuronal dysfunction in the ganglion cell layer. Frontiers Immunol. 7, 642.
Radbruch, H., D. Bremer, R. Günther, Z. Cseresnyes, R. Lindquist, A.E. Hauser, and R. Niesner. (2016). Ongoing oxidative stress causes subclinical neuronal dysfunction in the recovery phase of EAE. Frontiers Immunol 7: 92
Mossakowski, A.A., J. Pohlan, D. Bremer, R. Lindquist, J.M. Millward, M. Bock, K. Pollok, R. Mothes, L. Viohl, M. Radbruch, J. Gerhard, J. Bellmann-Strobl, J. Behrens, C. Infante-Duarte, A. Mähler, M. Boschmann, J.L. Rinnenthal, M. Füchtemeier, J. Herz, F.C. Pache, M. Bardua, J. Priller, A.E. Hauser, F. Paul, R. Niesner*, and H. Radbruch*. (2015). Tracking CNS and systemic sources of oxidative stress during the course of chronic neuroinflammation. Acta Neuropathologica 130(6), 799-814.
Radbruch, H., D. Bremer, R. Mothes, R. Günther, J.L. Rinnenthal, J. Pohlan, C. Ulbricht, A.E. Hauser, and R. Niesner. (2015). Intravital FRET: probing cellular and tissue function in vivo. IJMS 16(5),11713-27.
* equal contribution