caa a22 4500 378874241 CHVBK 20180305123416.0 cr unu---uuuuu 161128e20030220xx s 000 0 eng 10.1515/BC.2003.030 doi (NATIONALLICENCE)gruyter-10.1515/BC.2003.030 Interaction of Heat Shock Protein 70 Peptidewith NK Cells Involves the NK Receptor CD94 [Elektronische Daten] [C. Gross, D. Hansch, R. Gastpar, G. Multhoff] Full-length Hsp70 protein (Hsp70) and the C-terminal domain of Hsp70 (Hsp70C) both stimulate the cytolytic activity of naive natural killer (NK) cells against Hsp70-positive tumor target cells. Here, we describe the characterization of Hsp70-NK cell interaction with binding studies using the human NK cell line YT. Binding of recombinant Hsp70 protein (Hsp70) and the C-terminal domain of Hsp70 (Hsp70C) to YT cells is demonstrated by immunofluorescence studies. A phenotypic characterization revealed that none of the recently described HSP-receptors (α 2-macroglobulin receptor CD91, Toll-like receptors 2, 4, 9, CD14) are expressed on YT cells. Only the C-type lectin receptor CD94 is commonly expressed by YT cells and Hsp70 reactive NK cells. A correlation of the cell density-dependent, variable CD94 expression and the binding capacity of Hsp70 was detected. Furthermore, Hsp70 binding could be completely abrogated by preincubation of YT cells with a CD94-specific antibody. Competition assays using either unlabeled Hsp70 protein or an unrelated protein (GST) in 20-fold excess and binding studies with escalating doses of Hsp70 protein provide evidence for a specific and concentration-dependent interaction of Hsp70 with YT cells. In addition to Hsp70 and Hsp70C, a 14-mer Hsp70 peptide termed TKD is known to exhibit comparable stimulatory properties on NK cells. Similar to fulllength Hsp70 protein (10 ug/ml-50 ug/ml), a specific binding of this peptide to YT cells was observed at 4C, at equivalent concentrations (2.0 ug/ml-8.0 ug/ml). Following a 30 min incubation period at 37C, membrane-bound Hsp70 protein and Hsp70 peptide TKD were completely taken up into the cytoplasm. Copyright © 2003 by Walter de Gruyter GmbH & Co. KG Biochemistry nationallicence Molecular biology nationallicence Cellular biology nationallicence Gross C. aut Hansch D. aut Gastpar R. aut Multhoff G. aut Biological Chemistry Walter de Gruyter 384/2(2003-02-20), 267-279 1431-6730 384:2<267 2003 384 bchm https://doi.org/10.1515/BC.2003.030 text/html Onlinezugriff via DOI 1 research article jats NATIONALLICENCE 856 40 https://doi.org/10.1515/BC.2003.030 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Gross C. aut NATIONALLICENCE 700 1- Hansch D. aut NATIONALLICENCE 700 1- Gastpar R. aut NATIONALLICENCE 700 1- Multhoff G. aut NATIONALLICENCE 773 0- Biological Chemistry Walter de Gruyter 384/2(2003-02-20), 267-279 1431-6730 384:2<267 2003 384 bchm CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-gruyter