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Characterization of resting and phorbol ester or concanavalin A activated bovine lymph node cells with leukocyte
D J Hurley1, R A Wilson, C L Baldwin
1Department of Molecular and Cell Biology, Pennsylvania State University, University Park 16802.
Insights
This study characterizes bovine lymph node cells (LNC) and their surface antigen changes after activation. Phorbol dibutyrate (PDBU) and concanavalin A (Con A) differentially alter T cell markers, offering insights into immune cell activation.
Area of Science:
- Immunology
- Cell Biology
- Veterinary Science
Background:
- Bovine lymph node cells (LNC) are a model for studying immune cell activation and proliferation.
- Characterization of LNC subpopulations and their antigen expression changes upon activation was previously limited due to a lack of specific monoclonal antibodies.
- Understanding these changes is crucial for advancing bovine immunology research.
Purpose of the Study:
- To characterize bovine LNC subpopulations and their surface antigen expression.
- To investigate how activation with concanavalin A (Con A) or phorbol dibutyrate (PDBU) affects LNC surface antigen profiles.
- To compare the distinct effects of Con A and PDBU on LNC activation markers.
Main Methods:
- Utilized flow cytometry and a panel of leukocyte-specific monoclonal antibodies to analyze LNC.
- Compared antigen expression in unstimulated LNC with those treated with the mitogen Con A or the phorbol ester PDBU.
- Quantified changes in the expression and density of various cell-surface differentiation antigens, including T cell markers (CD2, CD4, CD5, CD8) and activation markers (J5).
Main Results:
- Unstimulated LNC showed proportions of T cells, B cells, and MHC Class II positive cells similar to peripheral blood leukocytes.
- PDBU treatment decreased CD4 expression while increasing CD2, CD5, CD6, MHC Class II, J5, and CD8 expression on LNC.
- Con A treatment significantly increased CD8, CD6, MHC Class II, and J5 expression but did not affect CD2, CD4, or CD5 percentages, suggesting subpopulation expansion, unlike PDBU's direct surface modification.
Conclusions:
- Both Con A and PDBU induce changes in bovine LNC surface antigen expression, mimicking patterns seen in human and mouse cells.
- PDBU and Con A exert distinct effects on LNC, with PDBU modulating surface antigen expression directly and Con A potentially driving subpopulation expansion.
- Neither activation method significantly altered the B cell subpopulation.
Abstract:
Bovine lymph node cells (LNC) have been used as a model to study cell activation and proliferation. Because monoclonal antibodies to bovine lymphoid-specific surface antigens have only recently become available, these cells have not been previously characterized in regard to subpopulations. Furthermore, it was not known how expression of lymphoid differentiation antigens and subset proportionalities might change following different modes of activation of LNC. Therefore, the distribution of cell-surface differentiation antigens in unstimulated LNC as well as in LNC incubated with the mitogen concanavalin A (Con A) or the phorbol ester, phorbol dibutyrate (PDBU), was measured using a series of leukocyte-specific monoclonal antibodies and flow cytometry. Unstimulated LNC were found to have similar proportions of T cells, B cells (sIgM positive), and MHC Class II positive cells similar to bovine peripheral blood mononuclear leukocytes (reviewed by Baldwin et al., 1988a). Treatment of the LNC with PDBU or mitogenic doses of Con A induced changes in the expression of surface antigens consistent with the changes observed with human and mouse cells after similar activation. However, these two compounds did not cause identical effects. After treatment with PDBU, the percentage of cells expressing CD4 as well as the density of surface expression decreased. An increase in the percentage of cells expressing and/or density of surface expression of the pan T cell antigens CD2, CD5, CD6, MHC Class II and J5, a T cell activation antigen, also occurred. PDBU treatment also increased the percentage of CD8 positive cells. The change in CD6 following PDBU treatment has not been reported previously. Con A treatment led to a significant increase in the percentage of cells bearing CD8, CD6, MHC Class II and J5, but it had no effect on the percentages of cells positive for the other T cell markers CD5, CD4, or CD2. Because Con A is a complete mitogen and PDBU is not, the changes observed following Con A stimulation probably reflected an expansion of a particular subpopulation. In contrast, PDBU most likely modifies surface antigen expression directly. Neither treatment affected the B cell subpopulation.
