Isolation of antigen-binding virgin and memory B cells

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摘要：

One major obstacle in studying the activation of antigen-specific B cells is the small number of B cells reactive with a particular antigenic epitope. In this report, we describe a method by which large numbers of highly purified antigen-binding cells can be obtained. We have shown that by varying the haptenation level of the erythrocytes used for rosetting, we can purify antigen-binding B cells which have different affinities for the antigenic epitope. Thus, memory cells (which have receptors of higher affinity) can be prepared and these cells are essentially free of contaminating virgin cells. The effects of varying the haptenation levels on the red cells used for purifying the B cells can, in turn, be related to the precursor frequency of secreting cells following their activation with T cells and antigen.

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ABC antigen-binding cells (virgin antigen-binding cells (virgin ACT hypotonic ammonium chloride hypotonic ammonium chloride BSA bovine serum albumin bovine serum albumin BSS balanced salt solution balanced salt solution BSS/5% FCS balanced salt solution supplemented with 5% fetal calf serum balanced salt solution supplemented with 5% fetal calf serum