Genotoxicity of genetic recombinant human erythropoietin in a novel test system.
The nude (athymic) mouse was used as a novel test system for the evaluation of genotoxicity of genetic recombinant human erythropoietin (rhEPO). A fibroblast cell line derived from the kidneys of baby hamsters, BHK-21, or a subclone of BHK-21 transfected with an expression vector containing the human EPO gene, named BXE cells, were implanted in nude mice. The concentration of EPO in the plasma of mice bearing BXE increased in relation to the increase in the weight of the tumor formed from growth of BXE cells. Increased values of hematocrit (Ht), ratio of reticulocytes to erythrocytes (RET ratio) and the number of red blood cells in mice bearing BXE indicated that excessive hematopoiesis was occurring in the host. However, the concentrations of EPO in the plasma of the mice bearing BHK-21 did not increase in relation to the cell mass and consequently the Ht values and RET ratios in these mice were not affected. Marked increases in the frequencies of micronucleated polychromatic erythrocytes (MNPCE) and micronucleated RET (MNRET) were noted in the mice bearing BXE, although no chromosomal aberrations were found in the spleen and marrow cells of the same mice. The increased levels of RET, MNRET and MNPCE seemed to result from acceleration of erythroblastic maturation and proliferation by rhEPO. It is, therefore, concluded that errors in the processes of enucleation or differentiation of erythrocytes should be equally considered as possible mechanisms alongside errors in genetic repair processes for the increased frequencies of MNPCE and MNRET.