Purification of human basophils by negative selection.
Journal of Immunological Methods
|October 28, 1983
Summary
Researchers developed a new method to isolate human basophils, crucial immune cells. This technique significantly increases basophil purity, enabling further study of these important cells and their functions.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- Basophils are critical immune cells involved in allergic reactions and host defense.
- Obtaining highly purified basophil populations is essential for detailed functional studies.
- Existing methods for basophil isolation often result in low purity and cell viability.
Purpose of the Study:
- To develop and optimize negative selection techniques for enriching human basophil populations.
- To assess the purity and functionality of basophils isolated using the described methods.
Main Methods:
- A two-step discontinuous gradient using isotonic Percoll was employed for initial basophil separation.
- A subsequent panning technique using anti-T-cell antibodies and goat anti-mouse IgG removed contaminating mononuclear and T-cells.
- Purified basophil histamine content and release in response to anti-IgE were measured.
Main Results:
- Centrifugation through a Percoll gradient increased basophil purity from approximately 1% to 14.6%.
- The addition of a T-cell panning step further enhanced basophil purity to 34%.
- Purified basophils retained normal histamine content and demonstrated comparable histamine release to unpurified cells.
Conclusions:
- Negative selection techniques, combining Percoll gradients and antibody-based panning, effectively enrich human basophil populations.
- The developed method significantly increases basophil purity (50-100 fold) while preserving their functional integrity.
- These enriched basophil preparations are suitable for detailed immunological investigations.
