Gene expression profiling of porcine alveolar macrophages after antibody-mediated cross-linking of Sialoadhesin (Sn, Siglec-1).
Sialoadhesin (Sn) is the prototypic member of the Siglecs, a family of receptors mainly involved in cell-cell interactions. For several Siglecs, but not for Sn, intracellular signaling functions have been described. Because antibody-mediated cross-linking of surface transmembrane proteins is a powerful technique to investigate cell-molecular events, Sn expressed on porcine alveolar macrophages (PAM) was cross-linked with the antibody 41D3, and the expression profiles were compared with mock-treated macrophages by microarray analysis. Gene ontology analysis of 479 differentially expressed transcripts identified gene categories related to membrane localization, signal transduction, receptor and communication activities. Analyses of the human KEGG pathway database identified MAP kinase signaling, regulation of actin cytoskeleton, adipocytokine signaling, and wnt signaling as significantly altered pathways, supporting a role for Sn as intracellular signaling molecule. Real-time PCR of a subset of modulated genes confirmed these results and highlighted the reliability of a short-term cross-linking treatment for transcriptomic analysis of receptor functions.
Genini
Sem
S
Parco Tecnologico Padano-CERSA, Lodi, Italy. sem.genini@tecnoparco.org
Malinverni
Roberto
R
Delputte
Peter L
PL
Fiorentini
Silvia
S
Stella
Alessandra
A
Botti
Sara
S
Nauwynck
Hans J
HJ
Giuffra
Elisabetta
E
D000818:Animals; D000906:Antibodies / Q000494:pharmacology*; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D016676:Macrophages, Alveolar / Q000187:drug effects / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D063265:Sialic Acid Binding Immunoglobulin-like Lectins; D034421:Sus scrofa / Q000235:genetics*
D000906:Antibodies; D003432:Cross-Linking Reagents; D037102:Lectins; D012333:RNA, Messenger; D063265:Sialic Acid Binding Immunoglobulin-like Lectins