Flash detection of anti-H-2 antibodies using chemiluminescence, without complement

Within the first minute following their exposure to a specific anti-H-2 serum and in the absence of complement, murine spleen cells generate a chemiluminescence phenomenon which is precisely measurable by photometry in the presence of luminol. The reaction lasts approximately 10 to 20 minutes, it also generated by bone marrow and, although weakly, by peritoneal cells. In contrast, thymus cells remain totally unresponsive. The striking inhibition of the phenomenon in the presence of superoxide dismutase confirmed that it is indeed due to O2(-). This rapid, simple, inexpensive, extremely sensitive and reproducible technique offers the advantage of focusing on the histocompatibility antigens borne by monocytes, macrophages and granulocytes. Lastly, this O2(-) production triggered by anti-H-2 antibodies should be kept in mind when discussing the mechanism of vascular damage in allograft rejection.