Techniques for the fractionation of C′ 1 and C′ 2, and for the specific inactivation of C′ 4 of horse complement are described, and shown to be satisfactory. These three components of complement and also conglutinin are found to be essential to the process of conglutination.
The experiments reported do not exclude the possibility of additional or unidentified fractions of horse serum playing a part in the reaction. Whether or not C′ 3 is essential to the process of conglutination could not be determined from the evidence available.
It was found that by the procedures employed the C′ 4 fraction may be supplied either from the horse complement or from the heated bovine serum added as a source of conglutinin or from both.
When horse complement and bovine antibody against sheep cells are used it is found that conglutination will only result when the complement components are absorbed on to the immune complex in a particular sequence. The sequence is that C′ 1 is absorbed first on to the sensitized cells, and then C′ 2 and C′ 4 are absorbed together. Both these components must be presented together to the sensitized cells carrying C′ 1 if conglutination is to result. Finally, conglutinin acts on the sensitized cells which have absorbed the three complement components, and conglutination results.
The significance of these findings, together with matters of a more general nature, is discussed.