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.
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
Rakhymzhan, A, Leben R, Zimmermann H, Günther R, Mex P, Reismann D, Ulbricht C, Acs A, Brandt AU, Lindquist RL, Winkler TH, Hauser AE, Niesner RA. (2017) Synergistic Strategy for Multicolor Two-photon Microscopy: Application to the Analysis of Germinal Center Reactions In Vivo.Sci Rep. 7(1), 7101.
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.
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.
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. *equally contributing
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.
Zehentmeier, S., K. Roth, Z. Cseresnyes, Ö. Sercan, K. Horn, R. Niesner,H.D. Chang, A. Radbruch, and A.E. Hauser. (2014) Static and dynamic components synergize to form a stable survival niche for bone marrow plasma cells. Eur J Immunol 44(8), 2306-2317.
Roth K, Oehme L, Zehentmeier S, Zhang Y, Niesner R, Hauser AE. (2014) Tracking plasma cell differentiation and survival.Cytometry A.85(1),15-24.
Rinnenthal, J.L., C. Börnchen, H. Radbruch, V. Andresen, V. Siffrin, H. Spiecker, T. Seelemann, I. Moll, J. Herz, A.E. Hauser, R. Glumm, O. Griesbeck, F. Zipp, M. Behne, R. Niesner. (2013) Parallelized TCSPC for dynamic intravital fluorescence lifetime imaging: quantifying neuronal function at the molecular level. PLoS ONE. 8(4), e60100.
Niesner RA, Hauser AE. (2011) Recent advances in dynamic intravital multi-photon microscopy.Cytometry A 79(10), 789-98.
Herz, J., V. Siffrin, A.E. Hauser, A.U. Brandt, T. Leuenberger, H. Radbruch, F. Zipp, R. Niesner. (2010) Expanding two-photon intravital microscopy to the infrared by means of OPO. Biophys. J. 74(9), 715-723.
Hauser, A.E., T. Junt, T.R. Mempel, M.W. Sneddon, S.H. Kleinstein, S.E. Henrikson, U.H. von Andrian, M.J. Schlomchik, A.M. Haberman. (2007) Definition of germinal-center B cell migration in vivo reveals predominant intrazonal circulation patterns. Immunity 26(5), 655-667.
Hauser, A.E., Debes GF, Arce S, Cassese G, Hamann A, Radbruch A, Manz RA. (2002) Chemotactic responsiveness toward ligands for CXCR3 and CXCR4 is regulated on plasma blasts during the time course of a memory immune response. J Immunol. 169(3), 1277-82.