Intracellular antigen determination by flow cytometry in leukaemic cells

Area of Science:

• Immunology
• Cell Biology
• Hematology

Background:

• Flow cytometry is crucial for diagnosing diseases by analyzing intracellular antigens.
• Existing fixation and permeabilization methods can be complex.
• A need exists for simple, reliable techniques for intracellular antigen detection.

Purpose of the Study:

• To evaluate a straightforward fixation and permeabilization protocol using 2% paraformaldehyde and Tween 20.
• To assess the method's reliability for analyzing intracellular antigens in leukemia.
• To compare intracellular versus surface antigen expression in specific leukemia subtypes.

Main Methods:

• A simple fixation and permeabilization technique was employed.
• The method utilized 2% paraformaldehyde and Tween 20.
• Intracellular antigens were analyzed in three leukemia cases using flow cytometry.

Main Results:

• The 2% paraformaldehyde and Tween 20 method proved reliable and easy to perform.
• Abundant intracellular kappa light chains were detected in a plasma cell leukemia case.
• Intracellular CD3 and CD22 expression exceeded surface expression in pre-T-acute lymphoblastic leukemia and pre-pre B-acute lymphoblastic leukemia, respectively.

Conclusions:

• The evaluated method is a simple and reliable approach for flow cytometric analysis of intracellular antigens.
• This technique facilitates the sensitive detection of intracellular markers relevant to leukemia diagnosis.
• The findings highlight the utility of this method for characterizing specific leukemia subtypes based on intracellular antigen profiles.