Measurement of intracellular ATP content by the luciferin-luciferase photometric reaction appears to be an extremely sensitive method for detecting complement dependent cytotoxicity (DCD) mediated by antibodies directed against H-2 antigens. Within the 15 min following the addition of complement, a considerable loss of ATP is observed in the antibody-coated target cells. The reaction is detectable using normal spleen lymphocytes as target cells, completely specific and much more sensitive than the classical dye exclusion test. Preliminary findings from our current study indicate that this simple and very rapid CDC method should be considered for the purpose of HLA ABC and D typing in humans.

Adenosine Triphosphate↗

PubMedDate not supplied

[Antibody-dependent cell-mediated cytotoxicity of ADCC].

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Antibody-Dependent Cell Cytotoxicity↗

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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.

Animals↗