Expression, processing and transcriptional regulation of granulysin in short-term activated human lymphocytes

• Published in: BMC immunology 2007-06, Vol.8 (1), p.9-9, Article 9
• Main Authors: Latinovic-Golic, Sonja, Walch, Michael, Sundstrom, Hanna, Dumrese, Claudia, Groscurth, Peter, Ziegler, Urs
• Format: Article
• Language: English
• Subjects: Antigens, Differentiation, T-Lymphocyte - genetics; Antigens, Differentiation, T-Lymphocyte - metabolism; Epitopes - immunology; Gene expression; Gene Expression Regulation; Genetic aspects; Humans; Killer Cells, Lymphokine-Activated - cytology; Killer Cells, Lymphokine-Activated - immunology; Kinetics; Listeria; Listeria - immunology; Lymphocyte Activation; Lymphocytes; Microscopy, Confocal; Physiological aspects; Properties; Protein Processing, Post-Translational; RNA, Messenger - genetics; RNA, Messenger - metabolism; T cells; T-Lymphocytes - cytology; T-Lymphocytes - immunology; Time Factors; Transcription, Genetic
• ISSN: 1471-2172; 1471-2172
• DOI: 10.1186/1471-2172-8-9

Granulysin, a cytotoxic protein expressed in human natural killer cells and activated T lymphocytes, exhibits cytolytic activity against a variety of intracellular microbes. Expression and transcription have been partially characterised in vitro and four transcripts (NKG5, 519, 520, and 522) were identified. However, only a single protein product of 15 kDa was found, which is subsequently processed to an active 9 kDa protein. In this study we investigated generation of granulysin in lymphokine activated killer (LAK) cells and antigen (Listeria) specific T-cells. Semiquantitative RT-PCR revealed NKG5 to be the most prominent transcript. It was found to be up-regulated in a time-dependent manner in LAK cells and antigen specific T-cells and their subsets. Two isoforms of 519 mRNA were up-regulated under IL-2 and antigen stimulation. Moreover, two novel transcripts, without any known function, comprising solely parts of the 5 prime region of the primary transcript, were detected. A significant increase of granulysin expressing LAK cells as well as antigen specific T-cells was shown by fluorescence microscopy. On the subset level, increase in CD4+ granulysin expressing cells was found only under antigen stimulation. Immunoblotting showed the 15 kDa form of granulysin to be present in the first week of stimulation either with IL-2 or with bacterial antigen. Substantial processing to the 9 kDa form was detected during the first week in LAK cells and in the second week in antigen specific T-cells. This first comprehensive study of granulysin gene regulation in primary cultured human lymphocytes shows that the regulation of granulysin synthesis in response to IL-2 or bacterial antigen stimulation occurs at several levels: RNA expression, extensive alternative splicing and posttranslational processing.