Activation of Natural Killer‐Like YT‐INDY Cells by Oligodeoxynucleotides and Binding by Homologous Pattern Recognition Proteins

• Published in: Scandinavian journal of immunology 2005-10, Vol.62 (4), p.361-370
• Main Authors: Kaur, H., Jaso‐Friedmann, L., Leary, J. H., Praveen, K., Brahmi, Z., Evans, D. L.
• Format: Article
• Language: English
• Subjects: Adjuvants, Immunologic - pharmacology; Antibodies; Blotting, Southwestern; Calcium - metabolism; Cell Line, Tumor; CpG Islands - genetics; CpG Islands - immunology; DNA-Binding Proteins - metabolism; Histones - immunology; Humans; Killer Cells, Natural - drug effects; Killer Cells, Natural - immunology; Killer Cells, Natural - metabolism; Lymphocyte Activation - drug effects; Lymphocyte Activation - immunology; Oligodeoxyribonucleotides - immunology; Oligodeoxyribonucleotides - pharmacology; Protein Binding - immunology; Receptors, Pattern Recognition - metabolism; Teleostei
• ISSN: 0300-9475; 1365-3083
• DOI: 10.1111/j.1365-3083.2005.01665.x

The present study was designed to examine the binding and signalling effects of single base and CpG dinucleotide phosphodiester (Po) oligodeoxynucleotides (ODN) on the human natural killer (NK)‐like cell line (YT‐INDY). Single base Po ODN composed of 20‐mers of guanosine (dG20), adenosine (dA20), cytosine (dC20) or thymidine (dT20) as well as ‘conventional’ Po CpG ODN were examined for their ability to bind and activate YT‐INDY cells. Binding by dG20 and CpG ODN to YT‐INDY cells was saturable and specific. dG20 binding was competitively inhibited by homologous dG20 and heterologous CpG ODN but not by dC20 and dA20. Two different YT‐INDY membrane proteins (18 and 29 kDa) were identified by ligand (Southwestern) blotting with biotinylated dG20 and CpG. The specificity of the ODN‐binding protein(s) was further confirmed by ODN depletion experiments using a teleost recombinant protein orthologue [nonspecific cytotoxic cells (NCC) cationic antimicrobial protein‐1 (ncamp‐1)] known to bind CpG and dG20. Cell proliferation and activation studies showed that dG20 and CpG treatment of YT‐INDY cells induced cellular DNA synthesis (i.e. G1 to S‐phase conversion). This signalling function was accompanied in dG20‐treated cells by proliferation 10 h posttreatment. Both dG20 and CpG ODN binding induced a calcium flux in YT‐INDY cells within seconds of treatment. These experiments demonstrated that Po single base dG20 and CpG ODN bind to a (potential) new class of cell‐surface proteins that mediate the activation of YT‐INDY cells.