| RRC ID |
68441
|
| Author |
Ohmori H, Takai T, Tanigawa T, Honma Y.
|
| Title |
Establishment of an enzyme release assay for cytotoxic T lymphocyte activity.
|
| Journal |
J Immunol Methods
|
| Abstract |
In the present report, we describe the establishment of a cell line that can be used as the target for measuring the activity of cytotoxic T lymphocytes (CTL) by an enzyme release assay. We transfected P3/NS1-Ag4-1 (NS-1), a myeloma cell line derived from BALB/c mice with Escherichia coli beta-galactosidase (beta-Gal) gene, and isolated a stable transformant designated as NS-1/Z that expressed a high level of the enzyme activity intracellularly. The effector cells showing cytotoxicity against NS-1/Z were induced when the spleen cells of AKR or C3H mice were cultured with mitomycin C-treated BALB/c spleen cells for 4 days. When 2 x 10(4) NS-1/Z cells were incubated with varying numbers of effector cells, beta-Gal activity was released from the target cells depending on the number of effector cells and the time of incubation for up to 8 h. A highly sensitive enzyme assay was performed by using a fluorescent substrate, 4-methylumbelliferyl-beta-D-galactoside. The cytotoxicity was specific for H-2 haplotype of the stimulator cells, and was abolished by treating the effector cells with anti-Lyt 2 plus complement. The sensitivity of the enzyme release assay was comparable to that of 51Cr release assay. These results indicate that NS-1/Z can be used as a target cell line for the non-radioactive measurement of CTL activity.
|
| Volume |
147(1)
|
| Pages |
119-24
|
| Published |
1992-2-14
|
| DOI |
10.1016/s0022-1759(12)80036-6
|
| PII |
S0022-1759(12)80036-6
|
| PMID |
1541835
|
| MeSH |
Animals
Cell Line
Chromium Radioisotopes
Male
Mice
Mice, Inbred Strains
T-Lymphocytes, Cytotoxic / immunology*
Transfection
beta-Galactosidase / genetics
beta-Galactosidase / metabolism*
|
| IF |
1.901
|
| Resource |
| Human and Animal Cells |
P3/NSI/1-AG4-1(RCB0095) |
