Molecular studies on LAK cells

We have developed a culture system for long-term growth of human LAK cells exhibiting an elevated, wide-spectrum anti-tumor cytotoxicity. The phenotypic and molecular properties of the final LAK cell populations indicated that they consist of three main types: a) NK-like lymphocytes (type I): NKH-1+, Ti alpha/beta-, Ti gamma/delta-, CD3-lymphocytes carrying the germline configuration of all TCR genes and expressing variable amount of the 1.0 beta mRNA and variably sized T delta transcripts; b) gamma/delta-like T lymphocytes (type II) NKH-1+, Ti alpha/beta-, Ti gamma/delta+, CD3+ lymphocytes carrying polyclonal rearrangements of the gamma and delta genes and expressing high levels of mature gamma and delta transcripts; c) alpha/beta-like T lymphocytes (type III): NKH-1+, Ti alpha/beta+, Ti gamma/delta-, CD3+ lymphocytes carrying rearrangements of all TCR genes and expressing high levels of mature alpha and beta transcripts. We took advantage of the high number of available LAK cells to clarify: 1) the origin of the NK-LAK delta transcripts. delta gene expression in LGL, NK clones and type I LAK cultures revealed six delta transcripts (3.5, 3.1, 2.2, 2.0, 1.5 and 1.3 kb), which varied in number and relative abundance in the different samples. None of the six known V delta was expressed and the delta locus was retained in its germline configuration suggesting that the delta expression is due to a partially rearranged or germline transcripts; 2) the origin of the NK-LAK truncated T beta transcript. We isolated two different clone types from a type I LAK cell cDNA library: a) J-C clones consisting of one of three J beta regions and the corresponding C beta 1 or C beta 2 regions; b) X-J-C and C-X clones, containing as yet unidentified (X) sequences. The presence of RSSs in J-C clones suggests that they derive from mRNAs transcribed from a promoter in the 5'J. Nucleotide analysis demonstrated that only one of the isolated clones had the potential to code a short T beta protein.