Proteomic and transcriptomic characterization of interferon-alpha-induced human primary T helper cells.
Interferon-alpha (IFN-alpha) is a multifunctional cytokine that modulates immune response. In spite of the numerous comprehensive studies on the effects of IFN-alpha on various cell types, novel characteristics of this versatile agent emerge continuously. In the present study a differential proteomic approach was used to identify new IFN-alpha-regulated proteins in human primary CD4(+) T cells. Two IFN-alpha-inducible proteins, soluble N-ethylmaleimide-sensitive factor attachment protein alpha (alpha-SNAP) and cleavage stimulation factor-64 (CstF-64) previously not described in this context, were identified. Additionally, several proteins already known as IFN-stimulated genes were observed. The results of proteomics experiments were further studied at the mRNA level using real-time reverse transcriptase-polymerase chain reaction (RT-PCR). Both peripheral blood and cord blood CD4(+) T cells were used in order to see if there are differences in IFN-alpha response between these populations. Differences were observed between the IFN-alpha-induced expression kinetics in peripheral blood and cord blood transcripts. The induction was more rapid in peripheral blood than in cord blood cells. CstF-64 expression was upregulated by IFN-alpha at the protein, but not at the mRNA level.
Rosengren
Arsi T
AT
Turku Centre for Biotechnology, University of Turku and Abo Akademi University, Turku, Finland. arsi.rosengren@btk.utu.fi
Nyman
Tuula A
TA
Syyrakki
Saija
S
Matikainen
Sampsa
S
Lahesmaa
Riitta
R
D001345:Autoradiography; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005312:Fetal Blood / Q000276:immunology; D006801:Humans; D007231:Infant, Newborn; D016898:Interferon-alpha / Q000235:genetics / Q000494:pharmacology*; D007371:Interferon-gamma / Q000096:biosynthesis; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008213:Lymphocyte Activation; D010449:Peptide Mapping; D040901:Proteomics; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016622:Silver Staining; D012995:Solubility; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins; D006377:T-Lymphocytes, Helper-Inducer / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*; D014158:Transcription, Genetic; D033921:Vesicular Transport Proteins / Q000737:chemistry / Q000378:metabolism
D016898:Interferon-alpha; D012333:RNA, Messenger; D052067:Soluble N-Ethylmaleimide-Sensitive Factor Attachment Proteins; D033921:Vesicular Transport Proteins; D007371:Interferon-gamma