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Published on: March 17, 2013
CD5+ B cells predominate in peripheral tissues of rabbit
1Department of Microbiology and Immunology, Loyola University Chicago, Stritch School of Medicine, Maywood, IL 60153.
Insights
Nearly all adult rabbit B lymphocytes express CD5, a stark contrast to mice and humans. This suggests early development and self-renewal maintain the rabbit B cell population throughout life.
Area of Science:
- Immunology
- Cell Biology
Background:
- Restricted usage of VH genes is noted in rabbit B lymphocytes and human/murine CD5 B lymphocytes.
- This observation suggests a potential high prevalence of CD5+ B lymphocytes in rabbits.
Purpose of the Study:
- To investigate the expression of CD5 on rabbit B lymphocytes.
- To determine the frequency of CD5+ B lymphocytes in peripheral lymphoid tissues of adult rabbits.
Main Methods:
- Cloned the rabbit CD5 gene using human CD5 cDNA probe.
- Developed anti-rabbit CD5 monoclonal antibodies (mAb) via transfection and immunization.
- Analyzed CD5 expression on rabbit lymphocytes using two-color immunofluorescence and Western blot.
Main Results:
- Successfully cloned the rabbit CD5 gene and developed specific mAb.
- Western blot confirmed a 67-kDa CD5 protein in rabbit lymphoid tissues.
- Two-color immunofluorescence revealed that essentially all peripheral B lymphocytes in adult rabbits express CD5.
Conclusions:
- Rabbit peripheral B lymphocytes predominantly express CD5, unlike in mice and humans.
- This unique expression pattern may indicate early ontogeny and self-renewal as the primary maintenance mechanism for rabbit B cells.
Abstract:
Restricted usage of VH genes is observed in rabbit B lymphocytes and in human and murine CD5 B lymphocytes. This observation raised the possibility that most rabbit B lymphocytes were CD5+. To investigate this we cloned the CD5 gene from a rabbit cosmid library, using a probe derived from human CD5 cDNA. The rabbit CD5 gene was transfected into a murine T cell line and then we used the transfectants to develop anti-rabbit CD5 mAb. By Western blot analysis, the mAb reacted with a 67-kDa protein in lysates prepared from mesenteric lymph node and spleen cells. We determined the frequency of CD5+ B lymphocytes in peripheral lymphoid tissues of adult rabbits by two-color immunofluorescence analysis using anti-CD5 mAb and anti-L chain antibodies. The analysis showed that essentially all peripheral B lymphocytes in adult rabbits express CD5. The observation that CD5 is expressed on nearly all rabbit B lymphocytes contrasts markedly to mouse and human, where only a small number of B lymphocytes express CD5. We propose that most peripheral B lymphocytes in rabbit, as in chicken, develop early in ontogeny and are maintained throughout life by a self-renewing process.
