Thiernesse N, Jeannesson P, Bernard J, Zagury D, Voisin G A
J Immunol Methods. 1978;21(1-2):79-88. doi: 10.1016/0022-1759(78)90225-9.
Viable mast cells, directly isolated by micromanipulation from a mouse peritoneal cell suspension, were deposited on the bottom of microtiter-plate wells and submitted to histamine release. Conventional antigen-induced anaphylactic degranulation as well as direct allogeneic anaphylactic degranulation were strongly inhibited when these mast cells were settled on normal tissue culture plastic surfaces. Nevertheless, normal degranulation could be recovered by pretreatment of the experimental surface with a multipositive charged molecule (poly-L-lysine). Under these conditions, we demonstrate that the degranulation of one isolated mast cell is possible and consequently, as regards the direct allogeneic anaphylactic degranulation, confirm the "self-triggering mechanism" in which the recognition of histocompatibility antigens on the membrane of the mast cell itself is the trigger to the secretory response. The technique of monocellular degranulation described in this paper provides a new tool which leads us to think that the problem of detection of anaphylactic antibody-secreting cells can be solved.
通过显微操作直接从小鼠腹腔细胞悬液中分离出的活肥大细胞，被接种到微量滴定板孔底部并进行组胺释放实验。当这些肥大细胞接种在正常组织培养塑料表面时，传统的抗原诱导过敏脱颗粒以及直接的同种异体过敏脱颗粒均受到强烈抑制。然而，通过用多阳离子分子（聚-L-赖氨酸）预处理实验表面，可以恢复正常脱颗粒。在这些条件下，我们证明了单个分离的肥大细胞的脱颗粒是可能的，因此，就直接的同种异体过敏脱颗粒而言，证实了“自我触发机制”，即肥大细胞膜上组织相容性抗原的识别是分泌反应的触发因素。本文所述的单细胞脱颗粒技术提供了一种新工具，使我们认为过敏抗体分泌细胞的检测问题可以得到解决。
