Complement-Mediated Cytolysis of Cell-Wall Mutants of Chlamydomonas reinhardtii

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Treatment of the cell wall-less mutant CW 15 of Chlamydomonas reinhardtii with human serum leads to a marked increase of the cell volume, followed by an irreversible cytolysis. Heat-inactivated serum as a control reveals no cytotoxic effects on CW 15. Experiments with C4-, properdin-, C3-, and factor H-depleted sera indicate the alternative pathway of complement as being responsible for the serum-mediated lysis. After immunofluorescence marking as well as electromicroscopically after negative staining the membrane attacking complex of complement, C5b-9, could be demonstrated on the surface of CW 15. These results together with the observation that cells of the wild-type strain 11-32c of C. reinhardtii are not lysed by active serum suggest that only protoplasts of Chlamydomonas carry surface structures capable to activate the alternative pathway of complement. In order to find out whether other cell wall mutants of C. reinhardtii, besides CW 15, can also activate the human complement system, we tested three strains each of the three known mutant categories. Strains CW 4, CW 9, and CW 19, representing category A, and strains CW 3, CW 10, and CW 92, representing category C, and CW 8 and CW 18, accounting for category B, were cytolysed by normal human serum. Only one type used in our experiments, CW 20 of category B, resisted serum treatment, suggesting the need to redefine this category.
日本植物生理学会の論文