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Alkaline phosphatase activity as a membrane marker for activated B cells.
Journal of Immunology (Baltimore, Md. : 1950)
|July 1, 1982
Summary
Alkaline phosphatase activity in mouse spleen cells was investigated. This enzyme activity was found to be a potential marker for activated B cells, particularly after stimulation with lipopolysaccharide (LPS).
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Plasma membrane-bound alkaline phosphatase (ALP) is an enzyme with diverse biological roles.
- Identifying specific cell surface markers can aid in understanding immune cell activation states.
Purpose of the Study:
- To investigate the presence and characteristics of alkaline phosphatase activity in activated mouse spleen cells.
- To determine if membrane alkaline phosphatase can serve as a marker for activated B cells.
Main Methods:
- Assay of alkaline phosphatase activity using p-nitrophenylphosphate as a substrate in a microtiter assay.
- Isolation of activated spleen cell fractions using discontinuous Percoll gradients.
- Enzyme inhibition studies using EDTA and L-phenylalanine.
Main Results:
- Alkaline phosphatase activity was detected in mouse spleen cells activated by the B cell mitogen lipopolysaccharide (LPS) and T cell-dependent B cell mitogen PWM.
- No activity was observed with T cell mitogens PHA and Con A.
- The activity was localized to a blast-enriched fraction (30-40%) from Percoll gradients.
- Enzymatic activity was inhibited by EDTA and L-phenylalanine, consistent with alkaline phosphatase.
Conclusions:
- Membrane alkaline phosphatase activity is associated with B cell activation in mouse spleen cells.
- This enzyme activity shows potential as a specific marker for activated B cells.