The isolation and sequence of a novel gene from a human functional T cell line.

Abstract

The study of T cell activation, proliferation, and effector function has been in vitro cultures of clonal T cell lines (1-3). Growth of such cell lines depends on periodic stimulation with growth factors, including IL-2, and with antigen that is recognized by the Ti/CD3 cell surface complex. It is believed that these IL-2-dependent, antigen-specific T cell lines, which function in vitro as helper or cytolytic cells, are normal counterparts of the T lymphocytes that perform these important effector and regulatory roles in vivo .To identify genes and gene products involved in the molecular mechanisms of T cell activation and effector function, we have begun a systematic search for cDNA clones expressed in functional cytolytic and Th cell lines but not in cell lines of other lineages . Here we report the structure of two overlapping cDNA clones that represent a gene, designated 519, that is expressed in 10 functional T cell lines but not in 14 established tumor lines, including 6 T cell tumor lines. We also show that the expression of 519 mRNA in PBL is increased more than 10-fold by mitogenic or antigenic stimulation. This increase is first detected 3-5 d after initiation of the culture. It is thus possible that the 519 gene product is involved in the activation and subsequent proliferation and/or differentiation of resting T cells. The data also show that functional T cell lines express gene products that are not readily detected in long term T cell tumor lines. Materials and MethodsCell Lines. The characterization and growth of the funct have been described (4, 5).Construction of cDNA Libraries. Constructi specific sequences was as described (6, 7) with minor modifications. Briefly, total cytoplasmic RNA was extracted from the CD8+, HLA-A2-specific functional cytolytic T cell line AH2 (4) by NP-40 lysis, and poly(A)+ mRNA was prepared by oligo(dT) chromatography (8). Single-stranded cDNA was synthesized using reverse transcriptase primed with i T cell lines and clones

High Cytotoxic and Specific Migratory Potencies of Senescent CD8+CD57+ Cells in HIV-Infected and Uninfected Individuals

“…One of the genes most overexpressed in CD8 ϩ CD57 ϩ (vs CD8 ϩ CD57 Ϫ ) cells codes for granulysin. This protein is present in the cytotoxic granules of CTL and NK cells and is a critical effector of the antimicrobial activity of CTLs against intracellular pathogens (37,38). Granulysin is also a chemoattractant and a proinflammatory activator that actively recruits immune cells to inflammation sites (39), and its presence shows that the antiviral role of CD8 ϩ CD57 ϩ cells involves mechanisms other than direct lysis of HIV-infected cells.…”

Granulysin, a T Cell Product, Kills Bacteria by Altering Membrane Permeability

“…A third mechanism involving direct T cell antimicrobial activity was suggested from studies identifying granulysin (1) in cytoplasmic granules of cytolytic T cells (2,3). Granulysin is a member of the saposin-like protein (SAPLIP) 3 family, including amoebapores, antimicrobial proteins that amoebas use to prevent growth of phagocytosed bacteria (4).…”

Analysis of serum granulysin in patients with hematopoietic stem‐cell transplantation: Its usefulness as a marker of graft‐versus‐host reaction

“…It has been reported that there are two independent granulysin secretion pathways: one is Ca-independent and another is Ca-dependent [15,23]. In an earlier paper, we reported that serum granulysin levels increase gradually after 40 years old and that an increase of serum granulysin represents recovery and maturation of immune function after cell therapy for the immune compromised patients over a long time scale.…”

“…Granulysin is a recently defined cytolytic granule protein of NK cells and cytotoxic T lymphocytes (CTLs) [15]. Granulysin is synthesized as a 15-kDa precursor form, which is then sorted to the cytolytic granules and processed there into a 9-kDa effector form [16].…”