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The human lymph node germinal center cell: characterization and isolation by using two-color flow cytometry.
Journal of Immunology (Baltimore, Md. : 1950)
|September 1, 1986
Summary
Germinal center cells (GCC) are difficult to study in tissues. Flow cytometry using peanut agglutinin (PNA) successfully identified GCC in cell suspensions, revealing their B cell characteristics and activation markers.
Area of Science:
- Immunology
- Cell Biology
- Hematology
Background:
- The germinal center is crucial for B cell activation and differentiation.
- Studying germinal center cells (GCC) in tissue sections is challenging.
- Flow cytometry offers a potential solution for GCC characterization.
Purpose of the Study:
- To precisely define the immunologic features of human germinal center cells (GCC).
- To utilize flow cytometry for GCC identification and characterization in suspension.
- To compare GCC features with non-GCC lymphocytes.
Main Methods:
- Two-color flow cytometry of suspended human tonsillar lymphocytes.
- Identification of GCC using peanut agglutinin (PNA) lectin binding.
- Confirmation of PNA+ cells as GCC via immunoperoxidase staining and cell sorting.
Main Results:
- PNA+ cells were predominantly B cells with specific surface immunoglobulin and antigen profiles (e.g., CALLA+).
- Differences in B1 and B2 antigen expression were observed between PNA+ and PNA- B cells.
- A subset of T cells were PNA+, and large PNA+ cells showed high DNA synthetic activity.
Conclusions:
- Peanut agglutinin (PNA) binding effectively identifies germinal center cells (GCC) in cell suspensions.
- Flow cytometry provides detailed immunologic insights into GCC not easily obtained from tissue sections.
- PNA+ cells exhibit characteristics consistent with activated and differentiating B cells within germinal centers.