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Václav Horejsí

Name of the laboratory

Members of the laboratory

Head

Václav Horejsí vaclav.horejsi@img.cas.cz

Research Fellows

Pavla Angelisová pavla.angelisova@img.cas.cz

Tomáš Brdicka tomas.brdicka@img.cas.cz

Pavel Otáhal pavel.otahal@img.cas.cz

Technicians

Jana Pokorná jana.pokorna@img.cas.cz

Ladislava Sobotková ladislava.sobotkova@img.cas.cz

Martkéta Voborská marketa.voborska@img.cas.cz

Ph.D. Student

Tereza Havlová tereza.havlova@img.cas.cz

Matous Hrdinka matous.hrdinka@img.cas.cz

Ondrej Stepánek ondrej.stapanek@img.cas.cz

Peter Dráber pdj@img.cas.cz

Research activities

A major topic of our laboratory has been for many years structure and function of human leukocyte surface molecules. Many membrane (glyco)proteins and glycolipids are of essential importance for all aspects of leukocyte biology and functioning of the immune system. Important functional features of a number of them and their mutual interactions remain poorly understood. We have been dealing mainly with proteins present in membrane microdomains called membrane rafts. These minute membrane structures are distinguished from the rest of the membrane by their high content of glycosphingolipids, cholesterol and glycolipid (GPI)-anchored proteins, while transmembrane proteins are mostly excluded from them. Importantly, they accumulate cytoplasmic signaling molecules such as heterotrimeric G-proteins and Src-family protein tyrosine kinases. These membrane „signaling islets“ (which were actually co-discovered by our laboratory (Stefanova et al., Science,254:1016,1991) appear to be of crucial importance for immunoreceptor signaling. In the past we discovered or co-discovered  a number of their protein components, such as GPI-anchored proteins CD14, CD48, CD59, CD108, and namely transmembrane adaptor proteins (Horejsi et al., Nat Rev Immunol 4:603,2004) such as PAG (Brdicka et al, J.Exp.Med. 191: 1591, 2000), NTAL (Brdicka et al. 196: 1617, 2002), LIME (Brdickova et al, J Exp Med. 198:1453, 2003).

In the recent period 2005-2010 we continued, mostly in collaboration with others, our studies on signaling molecules present in membrane rafts, namely several transmembrane adaptor proteins PAG, NTAL and LIME and their involvement in immunoreceptor signaling; we also characterized a few novel transmembrane adaptor proteins (LST1A, PRR7, Nvl) (manuscripts currently in preparation).  Furthermore, we discovered an apparently novel type of “heavy” rafts, differing from the “classical” ones by higher protein-lipid ratio and containing a number of transmembrane proteins (Otahal et al., J.Immunol. 2010 184: 3689, 2010).

Furthermore, we produced a number of monoclonal antibodies recognizing many CD molecules, intracellular signaling proteins etc., as valuable research tools (http://www.img.cas.cz/mi/Horejsi-Table.html). Most of these mAbs are commercialized by institute-linked spin-off biotech companies (e.g. www.exbio.cz).

Techniques available

• Monoclonal antibody production in mice
• Polyclonal antibody production
• Cloning technique
• Antibody characterization
• Mycoplasma testing
• Immunochemical, cytofluorometric, microscopic methods

Selected publications (2000-present)

• Otáhal, P., Angelisová, P., Hrdinka, M., Brdicka, T., Novák, P.,  Drbal, K. Horejsi, V. A new type of membrane raft-like microdomains and their possible involvement in TCR signaling. J. Immunol. 2010;184:3689-96.
• Wu W, Slaastad HS, de la Rosa Carrillo D, Frey T, Tjonnfjord GE, Boretti E, Aasheim HC, Horejsi V, Lund-Johansen F. Antibody array analysis with label-based detection and resolution of protein size. Mol Cell Proteomics. 2009;8:245-57.
• Veracini L, Simon V, Richard V, Schraven B, Horejsi V, Roche S, Benistant C. The Csk-binding protein PAG regulates PDGF-induced Src mitogenic signaling via GM1. J Cell Biol. 2008;182:603-14.
• Lo Monaco E, Sibilio L, Melucci E, Tremante E, Suchànek M, Horejsi V, Martayan A, Giacomini P.  HLA-E: strong association with beta2-microglobulin and surface expression in the absence of HLA class I signal sequence-derived peptides. J Immunol. 2008;181:5442-50.
• Khunkaewla P, Schiller HB, Paster W, Leksa V, Cermák L, Anděra L, Hořejší V, Stockinger H. LFA-1-mediated leukocyte adhesion regulated by interaction of CD43 with LFA-1 and CD147. Mol Immunol. 2008; 45:1703-11.  
• Iwaki S, Spicka J, Tkaczyk C, Jensen BM, Furumoto Y, Charles N, Kovarova M, Rivera J, Horejsi V, Metcalfe DD, Gilfillan AM. Kit- and FceRI-induced differential phosphorylation of the transmembrane adaptor molecule NTAL/LAB/LAT2 allows flexibility in its scaffolding function in mast cells. Cell Signal. 2008; 20, 195-205.
• Gregoire C, Simova S, Wang Y, Sansoni A, Richelme S, Schmidt-Giese A, Simeoni L, Angelisova P, Reinhold D, Schraven B, Horejsi V, Malissen B, Malissen M. Deletion of the LIME adaptor protein minimally affects T and B cell development and function. Eur. J. Immunol. 2007;37, 3259-69.
• Meraner P, Horejsi V, Wolpl A, Fischer GF, Stingl G, Maurer D. Dendritic cells sensitize TCRs through self-MHC-mediated Src family kinase activation. J Immunol. 2007;178:2262-71.
• Tessarz AS, Weiler S, Zanzinger K, Angelisova P, Horejsi V, Cerwenka A. Non-T cell activation linker (NTAL) negatively regulates TREM-1/DAP12-induced inflammatory cytokine production in myeloid cells. J Immunol. 2007;178:1991-9.
• Tedoldi S, Paterson JC, Hansmann ML, Natkunam Y, Ruediger T, Angelisova P, Du MQ, Roberton H, Roncador G, Sanchez L, Pozzobon M, Masir N, Barry R, Pileri S, Mason DY, Marafioti T, Horejsi V. Transmembrane adaptor molecules: A new category of lymphoid cell markers. Blood. 2005; 107:213-21.
• Wang Y, Horvath O, Hamm-Baarke A, Richelme M, Gregoire C, Guinamard R, Horejsi V, Angelisova P, Spicka J, Schraven B, Malissen B, Malissen M. Single and combined deletions of the NTAL/LAB and LAT adaptors minimally affect B-cell development and function. Mol Cell Biol. 2005;25:4455-65.
• Stork B, Engelke M, Frey J, Horejsi V, Hamm-Baarke A, Schraven B, Kurosaki T, Wienands J. Grb2 and the Non-T Cell Activation Linker NTAL Constitute a Ca(2+)-Regulating Signal Circuit in B Lymphocytes. Immunity. 21:681-91, 2004.
• Volna P, Lebduska P, Draberova L, Simova S, Heneberg P, Boubelik M, Bugajev V, Malissen B, Wilson BS, Horejsi V, Malissen M, Draber P.  Negative regulation of mast cell signaling and function by the adaptor LAB/NTAL. J Exp Med. 200:1001-13, 2004.
• Tkaczyk C, Horejsi V, Iwaki S, Draber P, Samelson LE, Satterthwaite AB, Nahm DH, Metcalfe DD, Gilfillan AM. NTAL phosphorylation is a pivotal link between the signaling cascades leading to human mast cell degranulation following Kit activation and Fc{epsilon}RI aggregation. Blood. 104:207-14, 2004.
• Carven GJ, Chitta S, Hilgert I, Rushe MM, Baggio RF, Palmer M, Arenas JE, Strominger JL, Horejsi V, Santambrogio L, Stern LJ. Monoclonal antibodies specific for the empty conformation of HLA-DR1 reveal the conformational change associated with peptide binding. J Biol Chem. 279:16561-70, 2004.
• Brdicková N, Brdicka T, Angelisová P, Horváth O, Spicka J, Hilgert I, Paces J, Simeoni L, Kliche S, Merten C, Schraven B, Horejsí V. LIME: a new membrane raft-associated adaptor protein involved in CD4 and CD8 coreceptor signaling. J Exp Med. 198:1453-1462, 2003.
• Gonen-Gross T, Gazit R, Achdout H, Hanna J, Mizrahi S, Markel G, Horejsi V, Mandelboim O.  Special organization of the HLA-G protein on the cell surface. Hum Immunol. 64: 1011-1016, 2003. 
• Vang T, Abrahamsen H, Myklebust S, Horejsi V, Tasken K. Combined spatial and enzymatic regulation of Csk by cAMP and protein kinase A inhibits TCR signaling. J Biol Chem.  16: 17597-600, 2003
• Davidson D, Bakinowski M, Thomas ML, Horejsi V, Veillette A. Phosphorylation-dependent regulation of T-cell activation by PAG/Cbp, a lipid raft-associated transmembrane adaptor. Mol Cell Biol. 23: 2017-28, 2003
• Baumgartner M, Angelisova P, Setterblad N, Mooney N, Werling D, Horejsi V, Langsley G. Constitutive exclusion of Csk from Hck-positive membrane microdomains permits Src kinase-dependent proliferation of Theileria-transformed B-lymphocytes. Blood 101: 1874-81, 2003
• Brdicka T, Imrich M, Angelisová P, Brdicková N, Horváth O, Spicka J, Hilgert I, Lusková P, Dráber P, Novák P, Engels N, Wienands J, Simeoni L,  Osterreicher J, Aguado E, Malissen M, Schraven B, Horejsi V. NTAL (Non-T cell activation linker): a transmembrane adaptor protein involved in immunoreceptor signaling. J.Exp.Med.  196: 1617-26, 2002
• Boyson JE, Erskine R, Whitman MC, Chiu M, Lau JM, Koopman LA, Valter M, Angelisova P, Horejsi V, Strominger JL. Disulfide bond-mediated dimerization of HLA-G on the cell surface. Proc. Natl. Acad. Sci. U.S.A. 99: 16180-85, 2002
• Cermák, L., Símová, S., Pintzas, A., Horejsí, V., and Andera, L: Apoptosis of TF-1 cells induced by engagement of CD43: roles of transcription, expression of Daxx and adhesion molecules. J. Biol.Chem. 277: 7955-7961, 2002
• Kropshofer, H., Spindeldreher, S., Röhn, T.A., Platania, N., Grygar, C., Wölpl, A., Langen, H., Horejsi, V., and Vogt, A.B.: Peptide-selective enrichment of MHC II-peptide complexes in tetraspan microdomains distinct from lipid rafts. Nature Immunol. 3: 61-68, 2002.
• Brdicková N, Brdicka T, Andera L, Spicka J, Angelisová P, Milgram SL, Horejsí V.: Interaction between two adapter proteins, PAG and EBP50: a possible link between membrane rafts and actin cytoskeleton. FEBS Lett. 507:133-6, 2001
• K.Drbal, P.Angelisová, I.Hilgert, J.Cerny, P.Novák, V.Horejsí: A proteolytically truncated form of free CD18, the common chain of leukocyte integrins, as a novel marker of activated myeloid cell. Blood 98: 1561-1566, 2001
• T.Brdicka, D.Pavlistová, A.Leo, E.Bruyns, V.Korínek, P.Angelisová, J.Scherer, A.Shevchenko, A.Shevchenko, I.Hilgert, J.Cerny, K.Drbal, Y.Kuramitsu, B.Kornacker, V.Horejsí, B.Schraven: Phosphoprotein associated with glycosphingolipid-enriched microdomains (PAG), a novel ubiquitously expressed transmembrane adaptor protein, binds the protein tyrosine kinase Csk and is involved in regulation of T cell activation. J. Exp. Med. 191: 1591-1604, 2000.

Reviews

• Horejsi V. Lipid rafts and their roles in T-cell activation. Microbes Infect. 2005;7:310-6
• Horejsi V, Zhang W, Schraven B.  Transmembrane adaptor proteins: organizers of immunoreceptor signalling. Nat Rev Immunol. 4:603-16, 2004.
• Horejsí V.: The roles of membrane microdomains (rafts) in T cell activation. Immunol. Revs. 191: 148-163, 2003
• Leo, A., Wienands, J., Baier, G., Horejsí, V., and Schraven, B.: Adapters in lymphocyte signaling. J. Clin. Invest. 109: 301-309, 2002