Abstract
This work aimed to study compare of extraction, purification and study gene expression of perforin from ALL, CLL and healthy, to chive this goal, blood samples were collected from 10 Leukemia cases (5 ALL and 5 CLL) and 5 healthy. Comparison of the molecular weight of purified perforin from these groups shows no detectable differences as compared with that of standard perforin which specified as 70KDa using SDS-PAGE. However, gene expression by using western blotting, it was found that the level of perforin in these groups were different due to differential gene expression between patients, and healthy control. Hence high levels were found in healthy individuals and CLL patient as compared with ALL patients as expressed by the presence of intense band.
Highlights
NTRODUCTION A 65-kDa protein with sequence homology to complement components C6 to C9, is stored in cytoplasmic granules of CTL and plays a major role in the secretary pathway (1)
When perforin is released from the granules of activated CTLs or NK cells, it forms a complex with granzymes and proteoglycans that binds to the target cell plasma membrane and promotes entry of the granzymes into target cell and serves as a channel for the influx of enzyme derived from the CTL granules (3)
Perforin are derived from cytoplasmic granules present in large granular lymphocytes, NK cells or CTL cells, since isolated granules are cytotoxic in the presence of Ca+ and form poly perforin (5)
Summary
NTRODUCTION A 65-kDa protein with sequence homology to complement components C6 to C9, is stored in cytoplasmic granules of CTL and plays a major role in the secretary pathway (1). Upon binding of CTL to the target cell and appropriate engagement of the T-cell receptor (TCR), the cytoplasmic granules containing perforin and granzymes are released vectorially onto the target cell (2). When perforin is released from the granules of activated CTLs or NK cells, it forms a complex with granzymes and proteoglycans that binds to the target cell plasma membrane and promotes entry of the granzymes into target cell and serves as a channel for the influx of enzyme derived from the CTL granules (3). Perforin are derived from cytoplasmic granules present in large granular lymphocytes, NK cells or CTL cells, since isolated granules are cytotoxic in the presence of Ca+ and form poly perforin (5). Antibodies raised against poly perforin recognize granules and SDS PAGE analysis of isolated granules revealed the presence of three major proteins with approximate molecular weights of 27, 29, and 66 KDa (7)
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