Characterization of a non-granule associated pore-forming protein in agranular lymphocytes

Recently, two populations of small lymphocytes (SL) have exhibited non‐major histocompatibility complex (MHC) restricted lysis. Recent studies by numerous laboratories have demonstrated that resting T cells triggered through CD3 and CD28 costimulations can result in immediate, non‐MHC restricted kil...

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Bibliographic Details
Published in:Journal of leukocyte biology 1995-06, Vol.57 (6), p.897-903
Main Authors: Ortaldo, J.R., Wiltrout, T.A., Sayers, T.J., Yagita, H., Winkler‐Pickett, R.T.
Format: Article
Language:English
Subjects:
Cells, Cultured
Cytotoxicity, Immunologic
Humans
Killer Cells, Natural - chemistry
Lymphocyte Subsets - chemistry
lymphocytes
Lymphocytes - chemistry
membrane
Membrane Glycoproteins - analysis
NK cells
Perforin
Pore Forming Cytotoxic Proteins
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Summary:Recently, two populations of small lymphocytes (SL) have exhibited non‐major histocompatibility complex (MHC) restricted lysis. Recent studies by numerous laboratories have demonstrated that resting T cells triggered through CD3 and CD28 costimulations can result in immediate, non‐MHC restricted killing. Our recent studies with CD3‐, CD56+ SL demonstrated that although these cells contained no cytoplasmic granules detected with electron microscopy, they mediated potent NK and ADCC activity. In the present study, we have used a Western blotting technique that allows for the detection and quantitation of total cellular levels of pore‐forming protein (PFP). We have found that freshly isolated peripheral non‐granulated lymphocytes (both CD3+ and CD3‐) contain PFP. In addition, CD3‐ CD56+ SL contain levels of PFP similar to those of the highly granular CD3‐ LGL. In search of non‐granule PFP, we exploited the rat NK (RNK) cell lines as a source of other potential cytotoxic factors. A membrane associated PFP, based on Western blotting, was isolated from rat RNK cells. Unlike PFP isolated from granules, this PFP was active after culture in Ca2+‐containing medium. However, the lytic activity isolated from the non‐granule PFP of RNK cells was inhibited by monoclonal antibodies to PFP. Collectively, these studies indicate that PFP is present in small agranular lymphocytes (both NK and T cells) and that it is not stored in large cytoplasmic granules. The implication of our results for the acquisition and activation of lytic ability in NK and T cells will be discussed. J. Leukoc. Biol. 57: 897–903; 1995.
ISSN:0741-5400
1938-3673
DOI:10.1002/jlb.57.6.897