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Synthesis of complement component C5 by human B and T lymphoblastoid cell lines
W Reed1, R A Roubey, J G Dalzell
1Department of Medicine, University of North Carolina, Chapel Hill 27599.
Immunogenetics
|January 1, 1990
Summary
Human B and T lymphoblastoid cell lines synthesize complement component 5 (C5). This immune protein was detected in cell lysates and supernatants, with C5 mRNA levels comparable to hepatoma cells.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- Complement component 5 (C5) is a crucial protein in the complement system.
- The synthesis and expression of C5 in human lymphoid cell lines are not well understood.
Purpose of the Study:
- To investigate the synthesis of C5 in human B and T lymphoblastoid cell lines.
- To characterize the expression and molecular properties of C5 produced by these cell lines.
Main Methods:
- Enzyme-linked immunosorbent assay (ELISA) using C5-specific monoclonal antibodies (mAbs).
- Detection of C5 in cell culture supernatants and whole cell lysates.
- Analysis of C5 messenger RNA (mRNA) size and abundance.
- Immunoaffinity chromatography for C5 isolation and subunit analysis.
Main Results:
- C5 synthesis was detected in all eight examined B and T lymphoblastoid cell lines.
- C5 was found in culture supernatants and/or cell lysates.
- Lymphoblastoid cells expressed C5 mRNA similar in size to HepG2 hepatoma cells.
- C5 mRNA levels in lymphoblastoid cells were approximately one-tenth of those in HepG2 cells.
- Isolated lymphoblastoid C5 exhibited similar subunit composition to plasma-derived C5, with a slightly smaller alpha subunit.
Conclusions:
- Human B and T lymphoblastoid cell lines are capable of synthesizing C5.
- The expression of C5 in these lymphoid cells involves both mRNA production and protein secretion/intracellular presence.
- Lymphoblastoid-derived C5 shares structural similarities with plasma C5, suggesting functional relevance.