Platforms

Research focus / Technologies

(in collaboration with the Institute of Molecular Immunology and the Institute of Medical Microbiology and Immunology at the Technische Universität München)

To evaluate the impact of immunotherapies it is necessary to have reliable monitoring tools available to assess immune responses of patients participating in clinical trials.

The central aim of the immune monitoring platform at the Institute of Virology is the development, establishment and standardization of different state-of-the-art methods for the analysis of immune responses against different infections.

The immune monitoring group is escorting several clinical trials with the aim to evaluate novel vaccines and therapeutic interventions for human immunodeficiency virus (HIV), hepatitis B and hepatitis C virus (HBV/HCV).

In addition to the assessment of immunologic effects in interventional studies, immune responses towards various viral infections (Epstein-Barr virus (EBV), Cytomegalovirus (CMV), HBV, HCV and HIV) are monitored in different cohorts.

To assess and define immunological correlates of viral control and protection against infection, complex analysis of multiple parameters is required. Therefore we develop, standardize, validate and perform technologies for multi-parametric functional and phenotypic analysis of T- (CD4+ and CD8+) and B-cell responses. Furthermore we focus on the development of systems to analyze, organize and store data for high throughput experimental procedures.


Available technologies:

     

  • Multiparameter flow cytometry-based assays (up to 20 parameter)
    - Intracellular cytokine staining
    - Phenotypic T-cell analysis (T-cell subset analysis, T-cell exhaustion)
    - Proliferation assay (CFSE-based)
    - Cytotoxicity / apoptose assay (CD107a, Annexin V, Granzyme B / Perforin)

     

  • ELISpot and FluoroSpot assay for quantification of antigen specific T- and B- cell responses
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  • MHC class I multimer staining
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  • Database storage of multi-color flow cytometry data
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  • Software application to analyze large data sets from polychromatic flow cytometry
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Research topics

Current research topics

     

  • Monitoring  hepatitis B virus (HBV) and hepatitis C virus (HCV)-specific T- and B-cell responses
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  • Identification of mechanisms responsible for T-cell failure in chronically HBV-infected patients and to develop therapeutic strategies to restore T-cell function
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  • Characterization of HIV-specific immune responses in different cohorts of HIV-1 infected subjects.
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  • Monitoring EBV-specific T-cell responses in a cohort of young competitive athletes to determine the impact of intensive physical training on antiviral immunity
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  • Development of a pharmacodynamic T-cell monitoring to allow individualized immunosuppressive therapy 
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Group

Immunomonitoring group at the Institute of Virology:

  • Dr. Tanja Bauer, PhD 
  • Dr. Claudia Dembek, PhD (currently on maternity leave)
  • Nina Körber, PhD student
  • Carolina Russo, PhD student
  • Mirjana Lekic, technician

 

Former group members

  • Dr. Antonio Cosma, PhD
  • Dr. Sarah Kutscher, PhD
  • Simone Allgayer, technician
  • Kim Dietrich, technician

Available instrumentation / Booking

  • BD LSRFortessa™ cell analyzer
    5 laser, 20 parameter SORP instrument (355 nm (UV), 405 nm (violet), 488nm (blue), 532nm (green), 640 nm (red)), equipped with HTS option. 
    Configuration
  • BD LSRII™ cell analyzer
    3 laser, 12 parameter instruments (408 nm (violet), 488nm (blue), 633 nm (red)), equipped with HTS option.
    Configuration
  • ImmunoSpot S6 Ultra-V Analyzer
    Visible light assays (enzymatic ELISpot) and fluorescence--based FluoroSpot assays (combinations of two fluorochromes FITC and PE)

Booking

All analyzers are available for use by external users.
Before external users can book and use the equipment, they must attend a basic training session in order to obtain log-in credentials to the instruments.

For more details please contact

Tanja.BauernoSp@m@helmholtz-muenchen.de

Training / Workshops


In cooperation with the CIMT immunoguiding program (CIP; http://cimt.eu/) we offer different workshops with the main objective to address practical aspects of T-cell assays and their analysis (ELISPOT, flow cytometry-based assays). The workshops aim to support researchers to establish, improve or validate T-cell assays in their own laboratories.

Completed workshop

Practical workshop dedicated to the analysis of multiparametric flow cytometry data.
Workshop schedule

Upcoming workshop

Workshop on Fluorospot assays
Tentative date: June 2013

Workshop programs are published at the CIMT homepage http://cimt.eu/ and via the CIMT newsletter.

Publications

     

  • Buchmann P, Dembek C, Kuklick L, Jäger C, Tedjokusumo R, von Freyend MJ, Drebber U, Janowicz Z, Melber K, Protzer U.: A novel therapeutic hepatitis B vaccine induces cellular and humoral immune responses and breaks tolerance in hepatitis B virus (HBV) transgenic mice. Vaccine. 2013 Jan 7. [Epub ahead of print]
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  • Dembek CJ, Kutscher S, Allgayer S, Russo C, Bauer T, Hoffmann D, Goebel FD, Bogner JR, Erfle V, Protzer U, Cosma A.: Longitudinal changes in HIV-1-specific T-cell quality associated with viral load dynamic. J Clin Virol. 2012 Oct;55(2):114-20.
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  • Braeckman T, Van Herck K, Jilg W, Bauer T, Van Damme P.: Two decades of hepatitis B vaccination in mentally retarded patients: effectiveness, antibody persistence and duration of immune memory. Vaccine. 2012 Jul 6;30(32):4757-61.
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  • Kutscher S, Bauer T, Dembek C, Sprinzl M, Protzer U.: Design of therapeutic vaccines: hepatitis B as an example. Microb Biotechnol. 2012 Mar;5(2):270-82.
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  • Malnati MS, Heltai S, Cosma A, Reitmeir P, Allgayer S, Glashoff RH, Liebrich W, Vardas E, Imami N, Westrop S, Nozza S, Tambussi G, Buttò S, Fanales-Belasio E, Ensoli B, Ensoli F, Tripiciano A, Fortis C, Lusso P, Poli G, Erfle V, Holmes H.: A new antigen scanning strategy for monitoring HIV-1 specific T-cell immune responses. J Immunol Methods. 2012 Jan 31;375(1-2):46-56.
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  • Bauer T, Sprinzl M, Protzer U.: Immune control of hepatitis B virus. Dig Dis. 2011;29(4):423-33.
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  • Kutscher S, Allgayer S, Dembek CJ, Bogner JR, Protzer U, Goebel FD, Erfle V, Cosma A.: MVA-nef induces HIV-1-specific polyfunctional and proliferative T-cell responses revealed by the combination of short- and long-term immune assays. Gene Ther. 2010 Nov;17(11):1372-83.
  • Dembek CJ, Kutscher S, Heltai S, Allgayer S, Biswas P, Ghezzi S, Vicenzi E, Hoffmann D, Reitmeir P, Tambussi G, Bogner JR, Lusso P, Stellbrink HJ, Santagostino E, Vollbrecht T, Goebel FD, Protzer U, Draenert R, Tinelli M, Poli G, Erfle V, Malnati M, Cosma A.: Nef-specific CD45RA+ CD8+ T cells secreting MIP-1beta but not IFN-gamma are associated with nonprogressive HIV-1 infection. AIDS Res Ther. 2010 Jul 2; 7:20.
  • Kutscher S, Allgayer S, Dembek CJ, Bogner JR, Protzer U, Goebel FD, Erfle V, Cosma A. MVA-nef induces HIV-1-specific polyfunctional and proliferative T-cell responses revealed by the combination of short- and long-term immune assays. . 2010 Jun 10.Gene Ther. 2010 Nov;17(11):1372-83. Epub 2010 Jun 10.
  • Di Bonito P, Grasso F, Mochi S, Petrone L, Fanales-Belasio E, Mei A, Cesolini A, Laconi G, Conrad H, Bernhard H, Dembek CJ, Cosma A, Santini SM, Lapenta C, Donati S, Muratori C, Giorgi C, Federico M. Anti-tumor CD8+ T cell immunity elicited by HIV-1-based virus-like particles incorporating HPV-16 E7 protein. Virology. 2009 Dec 5;395(1):45-55. Epub 2009 Oct 2.
  • Kutscher S, Dembek CJ, Allgayer S, Heltai S, Stadlbauer B, Biswas P, Nozza S, Tambussi G, Bogner JR, Stellbrink HJ, Goebel FD, Lusso P, Tinelli M, Poli G., Erfle V.,.Pohla H., Malnati.M., Cosma A;The intracellular detection of MIP-1beta enhances the capacity to detect IFN-gamma mediated HIV-1-specific CD8 T-cell responses in a flow cytometric setting providing a sensitive alternative to the ELISPOT. AIDS Res Ther. 2008 Oct 6;5:22."
  • Mamani-Matsuda M, Cosma A, Weller S, Faili A, Staib C, Garçon L, Hermine O, Adoue D, Fieschi C, Pers JO, Arakelyan N, Varet B, Sauvanet A, Berger A, Paye F, Andrieu JM, Michel M, Godeau B, Buffet P, Reynaud CA, Weill JC:The human spleen is the main reservoir for life long memory B-cells.Blood 2008, Mar 3 [Epub ahead of print]
  • Hoffmann D, Seebach J, Cosma A, Goebel FD, Strimmer K, Schätzl HM, Erfle V: Therapeutic vaccination reduces HIV sequence variability. FASEB J 2008 Jan16 [Epub ahead of print]
  • Brave A, Gudmundsdotter L, Gasteiger G, Hallermalm K, Kastenmüller W, Rollman E, Boberg A, Engstrom G, Reiland S, Cosma A, Drexler I, Hinkula J, Wahren B, Erfle V: Immunization of mice with the nef gene from Human Immunodeficiency Virus type 1: Study of immunological memory and long-term toxicology. Infect Agent Cancer 2007 Jul 10; 2:14
  • Cosma A, Nagaraj R, Staib C, Diemer C, Wopfner F, Schätzl H, Busch DH, Sutter G, Goebel FD, Erfle, V: Evaluation of modified vaccinia virus Ankara as an alternative vaccine against Smallpox in chronically HIV type 1-infected individuals undergoing HAART. AIDS Research and Human Retroviruses 2007, 23/6, 782-793. ultström et al. Immunol Lett. 2004
  • Cosma A, Bühler S, Nagaraj R, Staib C, Hammarin AL, Wahren B, Goebel FD, Erfle V, Sutter G:> Clin Diagn Lab Immun 2004, 11/2: 406-410
  • Cosma A, Nagaraj R, Bühler S, Hinkula J, Busch DH, Sutter G, Goebel FD, Erfle V: Therapeutic vaccination with MVA-HIV-1 nef elicits Nef-specific T-helper cell responses in chronically HIV-1 infected individuals. Vaccine 2003, 22: 21-29