Isolation and Enumeration of Isotype Specific Plaque Forming Cells from the Murine Intestine to Study the Development of the Intestinal B Cell Background Response
Quantification of (specific) IgA in faeces using the ELISA technique (1,2) and localization of antibody containing cells (ACC) by immunohistology (3,4) are important methods to investigate the in vivo expression of the enteric humoral immune response. These methods have improved our knowledge of the expression of the mucosal immune system, but they have several restrictions. Immunohistological studies give a direct insight in the in vivo expression of the mucosal immune system itself, but quantitative data are very difficult to obtain and the number of ACC does not reflect the number of B cells secreting antibodies (5) contributing to the intestinal IgA response. Moreover, IgA measured in faeces is not merely a reflection of locally (intestinally) produced and secreted IgA, since systemically produced IgA can also contribute to the IgA levels in faeces (6). Knowledge about the quantitative expression of the enteric humoral immune response can be obtained by isolation of lymphocytes from the intestine which can be tested subsequently for their capacities to secrete (antigen-specific) IgA.
KeywordsLamina Propria Isolation Procedure Mucosal Immune System Antibody Secreting Cell Lamina Propria Lymphocyte
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