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Updated: Aug 17, 2026
Retroviral Transduction of T-cell Receptors in Mouse T-cells
Published on: October 22, 2010
Cloning of a cDNA for a T cell-specific serine protease from a cytotoxic T lymphocyte
Abstract:
A new serine protease was encoded by a clone isolated from a murine cytotoxic T-lymphocyte complementary DNA library by an RNA-hybridization competition protocol. Complementary transcripts were detected in cytotoxic T lymphocytes, spleen cells from nude mice, a rat natural killer cell leukemia, and in two of eight T-helper clones (both cytotoxic), but not in normal mouse kidney, liver, spleen, or thymus, nor in several tested T- and B-cell tumors. T-cell activation with concanavalin A plus interleukin-2 induced spleen cells to express this gene with kinetics correlating with the acquisition of cytolytic capacity. The nucleotide sequence of this gene encoded an amino acid sequence of approximately 25,700 daltons, with 25 to 35 percent identity to members of the serine protease family. The active site "charge-relay" residues (His57, Asp102, and Ser195 of the chymotrypsin numbering system) are conserved, as well as the trypsin-specific Asp (position 189 in trypsin). A Southern blot analysis indicated that this gene is conserved in humans, mouse, and chicken. This serine protease may have a role in lymphocyte lysis and a "lytic cascade."
Insights
Researchers identified a novel serine protease gene in cytotoxic T-lymphocytes. This gene
Area of Science:
- Immunology
- Molecular Biology
- Biochemistry
Background:
- Cytotoxic T-lymphocytes (CTLs) are crucial for adaptive immunity.
- The molecular mechanisms underlying lymphocyte-mediated cytotoxicity are not fully elucidated.
- Serine proteases play diverse roles in biological processes, including immune responses.
Purpose of the Study:
- To identify and characterize novel genes involved in lymphocyte-mediated cytotoxicity.
- To investigate the expression patterns and functional relevance of a newly discovered serine protease.
- To determine the evolutionary conservation of this gene.
Main Methods:
- Isolation of complementary DNA clones from a murine CTL library using RNA-hybridization.
- Detection of complementary transcripts via Northern blot analysis.
- Nucleotide sequencing and amino acid analysis of the identified gene.
- Southern blot analysis for cross-species gene conservation.
Main Results:
- A novel serine protease gene was identified and its complementary transcripts were found in CTLs, NK cell leukemia, and cytotoxic T-helper clones.
- Gene expression was induced in spleen cells upon T-cell activation, correlating with cytolytic capacity.
- The deduced amino acid sequence showed homology to serine proteases, with conserved active site residues.
- The gene is conserved across human, mouse, and chicken species.
Conclusions:
- A novel serine protease, potentially involved in lymphocyte lysis, has been characterized.
- This enzyme may play a role in the "lytic cascade" during immune responses.
- The conserved nature of the gene suggests a fundamental role in vertebrate immunity.
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