Extension of the 4-week safety study for detecting immune system impairment appears not necessary: example of cyclosporin A in rats☆

Abstract A 4-week study was conducted to shed light on the question of whether compounds impairing immune homeostasis may escape the standard safety testing. Wistar rats were orally treated with cyclosporin A at dosages of 0 (control: olive oil), 1, 5 or 25 mg/kg/day. Ten rats/sex/group (study segment 1) were not immunized while six other rats/sex/group (study segment 2) were immunized 4 days before killing to perform a plaque forming cell (PFC) assay. All rats were subjected to routine safety evaluations (OECD guideline 407) and determination of IgM and IgG serum levels. Other immune parameters were evaluated using cells from spleen and mesenteric lymph nodes (segment 1). Effects on safety parameters were similar for immunized and non-immunized rats. A slight decrease of body weight gain (males, 25 mg/kg) accompanied slight clinical chemical and histomorphologic evidence of renal tubulotoxicity. Changes in safety parameters indicative of immune system alterations were: increased thymic corticomedullary ratio (≥5 mg/kg) and (25 mg/kg) minimal lymphopenia, low thymus weight, thymic cortical lymphocytolysis and low lymphoid cellularity of spleen and lymph nodes. They were associated with (males at ≥ 1 mg/kg) dose-related decreases of T-cell receptor + and CD4 + cells and increases of CDS + cells, and decreased PFC (≥5 mg/kg) and lymphoproliferative responses to mitogens and alloantigens (25 mg/kg). There were no changes in natural killer activity. The conventional assay identified the drug as a potential immunomodulator. Specific immune assays (phenotyping, PFC) improved the threshold of detection. These results did not support the incorporation of specific immune tests in the standard 4-week study protocol.