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KMID : 0380219960290020127
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Journal of Biochemistry and Molecular Biology
1996 Volume.29 No. 2 p.127 ~ p.132
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Expression of IgG1 Germline Transcripts in Germinal Center B Cells of Mouse Popliteal Lymph Nodes
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Lee In-Woo
Kim Jin-Ho
Chung Gook-Hyun
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Abstract
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Germinal centers (GCs) are formed in peripheral lymphoid tissues in response to protein antigens. In order to see if immunoglobulin isotype switching takes place in GC B-cells, we isolated GC B-cells (PNA positive cells) from mouse popliteal lymph nodes by a flow cytometer after the staining of lymph node cells with PNA-FITC and anti-B220-PE, and determined the expression of γ1 germline transcript and γ1 mRNA by RT-PCR. γ1 germline transcript and γ1 mRNA were amplified specifically in cDNAs from hybridoma expressing IgG1 or splenocytes stimulated LPS plus IL-4. Germinal center B-cells formed in popliteal lymph nodes of mice immunized with chicken ovalbumin were isolated 7 days after immunization. We sorted GC B-cells five times. Immunoglobulin γ1 germline transcripts were expressed in germinal center B-cells in three out of five sorts whereas two out of five sorts did not express γ1 germline transcripts in GC B-cells. The contents of GC B-cells ranged from 5 to 7% of total lymph node cells in most flow cytometric analyses but those of two sorted cells which did not express γ1 germline transcripts were out of normal range. These results imply that isotype switching of immunoglobulins may take place in GCs.
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KEYWORD
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flow cytometric analysis, germinal center, germline transcript, isotype switching
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