Immunology and Flow Cytometry

The aim of our immunological analyses is to investigate the complex processes of the immune response to pathogens. In addition to immunological detection methods, such as ELIspot and ELISA, a major part of our analyses is done with flow cytometry.

For flow cytometrical investigations, individual cells or particles are analysed and, if necessary, sorted on the basis of their physical and molecular properties. This enables us to record certain properties of cells, cell populations or cell lines, either by means of staining with fluorescent conjugated antibodies or based on the expression of fluorescent proteins. The cells are guided past one or more lasers individually, the characteristic scattered and fluorescent light is recorded separately by detectors and subsequently evaluated by a computer. The FACS (Fluorescence-Activated Cell Sorting) method additionally isolates individual cells physically according to their fluorescent properties. The separation of specific cell populations permits further investigations or analyses.

A special feature of flow cytometric analyses of non-human primates are the few commercially available NHP-specific antibodies. Therefore, mainly anti-human antibodies are used in this research area, but not all of them cross-react with NHP.  Within the scope of our investigations, we continuously expand the list or cross-reactive antibodies with different NHP species (Rhesus monkeys, long-tailed macaques, marmosets, baboons) to establish extensive dyeing panels to identify and characterize various immune cell populations, including activation, differentiation, cytokine-expression etc., both among healthy animals and within the scope of miscellaneous infection trials.

Selected publications:
 
 Neumann B, Shi T, Gan LL, Klippert A, Daskalaki M, Stolte-Leeb N, Stahl-Hennig C. Comprehensive panel of cross-reacting monoclonal antibodies for analysis of different immune cells and their distribution in the common marmoset (Callithrix jacchus). J Med Primatol. 2016 Jun;45(3):139-46. doi: 10.1111/jmp.12216.

Mietsch M, Paqué K, Drummer C, Stahl-Hennig C, Roshani B. The aging common marmoset's immune system: From junior to senior. Am J Primatol. 2020 Jun;82(6):e23128. doi: 10.1002/ajp.23128. Epub 2020 Apr 4.

Neumann B, Sopper S, Stahl-Hennig C. OMIP-026: Phenotypic analysis of B and plasma cells in rhesus macaques; Cytometry A. 2015 Sep;87(9):800-2.doi: 10.1002/cyto.a.22712.

Neumann B, Klippert A, Raue K, Sopper S, Stahl-Hennig C. Comparative phenotypical analysis of B cells in fresh and cryopreserved mononuclear cells from blood and tissue of rhesus macaques. J Leukoc Biol. 2015 Jan;97(1):19-30. doi: 10.1189/jlb.1HI0514-243R.

Joas S, Sauermann U, Roshani B, Klippert A, Daskalaki M, Mätz-Rensing K, Stolte-Leeb N, Heigele A, Tharp GK, Gupta PM, Nelson S, Bosinger S, Parodi L, Giavedoni L, Silvestri G, Sauter D, Stahl-Hennig C, Kirchhoff F. Nef-Mediated CD3-TCR Downmodulation Dampens Acute Inflammation and Promotes SIV Immune Evasion. Cell Rep. 2020 Feb 18;30(7):2261-2274.e7. doi: 10.1016/j.celrep.2020.01.069.

Denner J, Längin M, Reichart B, Krüger L, Fiebig U, Mokelke M, Radan J, Mayr T, Milusev A, Luther F, Sorvillo N, Rieben R, Brenner P, Walz C, Wolf E, Roshani B, Stahl‐Hennig C & Abicht JM. Impact of porcine cytomegalovirus on long-term orthotopic cardiac xenotransplant survival. SciRep. 2020; 10:17531. doi: 10.1038/s41598-020-73150-9