Proteomic profiling of surface proteins on Th1 and Th2 cells.
We utilized mass spectrometry to profile cell surface protein differential expression on primary human T helper (Th1 and Th2) cells with the stable isotope labeling by amino acids in cell culture (SILAC) approach. Proteomic and microarray analyses were done concurrently and results were compared for 38 different genes. Although microarray studies displayed wide variability between donors for mRNA expression, these two approaches were shown to be corroborative for most gene products with the exception of a small subset of uncorrelated protein and message levels. The greatest differing Th1 to Th2 ratios were observed for BST2 (bone marrow stromal protein 2) and TRIM (T cell receptor interacting molecule). Both showed greater Th1 expression by proteomic methods, even though mRNA levels were approximately equal for both. To validate this method, we compared protein expression levels of a recently cloned molecule, B and T cell lymphocyte attenuator (BTLA), on Th1 and Th2 cell populations and showed greater protein expression on Th1 cells, which agrees with a previous analysis of higher BTLA mRNA expression in Th1 cells.(1).
Loyet
Kelly M
KM
Protein Chemistry Department, Immunology Department, Genentech, South San Francisco, California 94080, USA. loyet@gene.com
Ouyang
Wenjun
W
Eaton
Dan L
DL
Stults
John T
JT
D002454:Cell Differentiation; D002478:Cells, Cultured; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis; D040901:Proteomics; D018417:Th1 Cells / Q000737:chemistry* / Q000166:cytology; D018418:Th2 Cells / Q000737:chemistry* / Q000166:cytology