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An alternative method of panning for rat B lymphocytes
C D Severson1, D L Burg, D E Lafrenz
1Veterans Administration Medical Center, Iowa City, IA.
Immunology Letters
|August 1, 1987
Summary
A novel panning technique uses nonspecific proteins to isolate surface immunoglobulin-positive (sIg+) B lymphocytes. This method is cost-effective, avoids B cell activation, and yields highly purified, functional B cells.
Area of Science:
- Immunology
- Cell Biology
Background:
- Isolating pure B lymphocytes is crucial for immunological research.
- Conventional panning methods using anti-immunoglobulin antibodies can activate B cells.
Purpose of the Study:
- To develop a new, cost-effective method for purifying B lymphocytes.
- To avoid B cell activation during the isolation process.
Main Methods:
- Rat lymph node cells were incubated with bovine serum albumin to promote adherence.
- Adherent cells were analyzed for surface immunoglobulin (sIg) and MHC class II expression.
- Further purification was achieved using anti-T cell antibodies and complement.
Main Results:
- The novel panning method yielded >90% surface immunoglobulin-positive (sIg+) and MHC class II-positive cells.
- The isolated B cell population contained minimal T cells.
- Purified sIg+ B cells demonstrated functional activity in BCGF and BCDF assays.
Conclusions:
- This new method offers an easy, inexpensive alternative to anti-immunoglobulin panning.
- The technique effectively isolates highly purified, functional B lymphocytes without pre-activation.
- This approach enhances the reliability of B cell studies.