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Killer Artificial Antigen Presenting Cells (KaAPC) for Efficient In Vitro Depletion of Human Antigen-specific T Cells
Published on: August 11, 2014
Natural killer cells are deficient in the surface expression of the complement regulatory protein, decay accelerating
Abstract:
Decay-accelerating factor (DAF) is a 75,000 m.w. membrane protein that inhibits autologous complement C3 activation at the cell surface. One-color direct immunofluorescence with anti-DAF antibody and cytofluorographic analysis indicates that normal human monocytes and granulocytes are uniform in expression of DAF, whereas 23% of peripheral blood lymphocytes are DAF deficient. A two-color indirect immunofluorescence method, used to define the phenotype(s) of the DAF-deficient lymphocytes, was less efficient in DAF detection and led to overestimation of the fraction of deficient cells. Nonetheless, the difference between DAF expression by natural killer cells, identified by the CD16 and HNK-1 antigens, was marked. DAF deficiency was intermediate in cells expressing the CD2, CD3, CD4, or CD8 markers. On the basis of the phenotypic definition of natural killer cells and their contribution to the lymphocyte population, it is concluded that a uniform deficiency of DAF on natural killer cells accounts for about one-half of the DAF-deficient lymphocytes in peripheral blood of normal donors. The finding of a complete DAF deficiency in the lymphocytes from a patient with a lymphoproliferative disorder with the predominant proliferation of CD2+, CD3+, CD8+, HNK-1+ large granular lymphocytes gives additional support for the association of DAF-deficiency with natural killer cells.
Insights
Decay-accelerating factor (DAF) deficiency is common in natural killer cells, a type of lymphocyte. This finding helps explain DAF deficiency in peripheral blood and links it to specific immune cell types.
Area of Science:
- Immunology
- Cell Biology
Background:
- Decay-accelerating factor (DAF) is a membrane protein that regulates complement system activation.
- DAF protects cells from autologous complement attack.
Purpose of the Study:
- To investigate DAF expression on human peripheral blood cells.
- To identify the cell types responsible for DAF deficiency in lymphocytes.
Main Methods:
- One-color direct immunofluorescence and cytofluorography were used to assess DAF expression.
- Two-color indirect immunofluorescence was employed to phenotype DAF-deficient lymphocytes.
- Flow cytometry was utilized to analyze cell surface markers.
Main Results:
- Monocytes and granulocytes showed uniform DAF expression.
- 23% of peripheral blood lymphocytes were DAF deficient.
- Natural killer cells (CD16+, HNK-1+) exhibited marked DAF deficiency.
- T cells (CD2+, CD3+, CD4+, CD8+) showed intermediate DAF deficiency.
- A patient with a lymphoproliferative disorder had complete DAF deficiency on CD2+, CD3+, CD8+, HNK-1+ large granular lymphocytes.
Conclusions:
- DAF deficiency is primarily associated with natural killer cells.
- Natural killer cells account for approximately half of DAF-deficient lymphocytes.
- DAF deficiency in lymphocytes may be linked to natural killer cell populations.
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