A flow cytometric procedure for the isolation of antigenically responsive human basophils

Area of Science:

• Immunology
• Cell Biology
• Biotechnology

Background:

• Human basophils are crucial immune cells involved in allergic responses.
• Efficient isolation of pure basophils is essential for studying their function.
• Current methods for basophil isolation can be time-consuming and yield lower purity.

Purpose of the Study:

• To develop and validate a rapid and high-purity method for isolating human basophils.
• To demonstrate the functional integrity of isolated basophils.
• To provide a reliable technique for basophil research.

Main Methods:

• A flow cytometric cell sorting procedure was employed.
• Passive sensitization of cells using mouse FITC-IgE anti-DNP was performed.
• Isolated basophils were tested for histamine release upon stimulation with DNP21-human serum albumin conjugates.

Main Results:

• The procedure successfully isolated human basophils with high purity (>=98%).
• Approximately 1-4 X 10^5 basophils were obtained from 30 ml of whole blood.
• The isolated basophils demonstrated characteristic histamine release, confirming their functional activity.
• The entire sorting procedure was completed within 2 hours.

Conclusions:

• This flow cytometric method provides an efficient and effective means for isolating pure, functional human basophils.
• The technique is suitable for obtaining sufficient quantities of basophils for various research applications.
• This advancement facilitates further investigation into basophil-mediated immune responses.