title: Comprehensive comparative and semiquantitative proteome of a very low number of native and matched epstein-barr-virus-transformed B lymphocytes infiltrating human melanoma.
abstract: Melanoma, the deadliest form of skin cancer, is highly immunogenic and frequently infiltrated with immune cells including B cells. The role of tumor-infiltrating B cells (TIBCs) in melanoma is as yet unresolved, possibly due to technical challenges in obtaining TIBCs in sufficient quantity for extensive studies and due to the limited life span of B cells in vitro. A comprehensive workflow has thus been developed for successful isolation and proteomic analysis of a low number of TIBCs from fresh, human melanoma tissue. In addition, we generated in vitro-proliferating TIBC cultures using simultaneous stimulation with Epstein-Barr virus (EBV) and the TLR9 ligand CpG-oligodesoxynucleotide (CpG ODN). The FASP method and iTRAQ labeling were utilized to obtain a comparative, semiquantitative proteome to assess EBV-induced changes in TIBCs. By using as few as 100 000 B cells (∼5 μg protein)/sample for our proteomic study, a total number of 6507 proteins were identified. EBV-induced changes in TIBCs are similar to those already reported for peripheral B cells and largely involve changes in cell cycle proliferation, apoptosis, and interferon response, while most of the proteins were not significantly altered. This study provides an essential, further step toward detailed characterization of TIBCs including functional in vitro analysis.
authors.affiliation: Division of Immunology, Allergy and Infectious Diseases, Department of Dermatology, Medical University of Vienna , Waehringer Guertel 18-20, A-1090 Vienna, Austria.
authors.forename: Margarita
authors.initials: M
authors.lastname: Maurer
authors.forename: André C
authors.initials: AC
authors.lastname: Müller
authors.forename: Katja
authors.initials: K
authors.lastname: Parapatics
authors.forename: Winfried F
authors.initials: WF
authors.lastname: Pickl
authors.forename: Christine
authors.initials: C
authors.lastname: Wagner
authors.forename: Elena L
authors.initials: EL
authors.lastname: Rudashevskaya
authors.forename: Florian P
authors.initials: FP
authors.lastname: Breitwieser
authors.forename: Jacques
authors.initials: J
authors.lastname: Colinge
authors.forename: Kanika
authors.initials: K
authors.lastname: Garg
authors.forename: Johannes
authors.initials: J
authors.lastname: Griss
authors.forename: Keiryn L
authors.initials: KL
authors.lastname: Bennett
authors.forename: Stephan N
authors.initials: SN
authors.lastname: Wagner
mesh_terms: D001402:B-Lymphocytes / Q000378:metabolism* / Q000821:virology; D049109:Cell Proliferation; D002469:Cell Separation; D015870:Gene Expression; D015972:Gene Expression Regulation, Neoplastic; D063990:Gene Ontology; D004854:Herpesvirus 4, Human / Q000502:physiology*; D006801:Humans; D008545:Melanoma / Q000276:immunology* / Q000473:pathology / Q000556:secondary; D058977:Molecular Sequence Annotation; D020543:Proteome / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D020543:Proteome; D012333:RNA, Messenger