Effect of protease class-specific inhibitors on in vitro development of the third- to fourth-stage larvae of Ascaris suum.

Third-stage larvae (L3) of Ascaris suum develop and molt to fourth-stage larvae (L4) during in vitro cultivation; consistently greater than 80% of the larvae develop to L4 during 7 days in culture (DIC). To assess the role of proteases in this process, the effect of protease class-specific inhibitors was studied. The presence of either a serine protease inhibitor (AEBSF, 100 microM) or an aspartic protease inhibitor (pepstatin A, 100 microM) had no effect on the percentage of L4 after 7 DIC. However, the presence of either a cysteine protease inhibitor (Z-Phe-Ala-FMK, 100 microM) or an aminopeptidase inhibitor (amastatin, 100 microM) resulted in 77% and 34% reductions, respectively, in the percentage of L4 compared to untreated cultures; viability of the larvae was not affected. The effect of Z-Phe-Ala-FMK on molting was time and dose dependent. In contrast to Z-Phe-Ala-FMK, E-64, another specific inhibitor of cysteine proteases, had no effect on molting. The data support a role for an aminopeptidase and suggest a role for a cysteine protease in the development of the L3 to L4 stage of A. suum.