STUDIES ON SPECIFIC SEROLOGICAL ANTIGENS IN METACERCARIAE AND JUVENILES OF Paragoniums westermani AND ITS MONOCLONAL ANTIBODIES

The polyclonal antibodies to juveniles of Paragoniums westermani (PwJ-PcAbs) from sera of Wistar rats infected with Paragoniums westerrnani (P. w.)were purified by Sephadex G 200 chromatography. Next the shared serological antigens of P. w. metacercaria and juveniles (PwMJ-SAg) from the crude antigens of the metacercariae (M-NS-Ag) were purified with immuno-affinity chromatography on cyanogen bromide-activated cross-linked Sepharose 4B beads coupled withPwJ-PcAbs. PwMJ-SAg, agroup of glycoprotein molecules shown by the staining test, were specific serological antigens of P. w. metacercariae and juveniles, identified by the immunoabsorb test and immunoelectrophoresis. By SDSPAGE, PwMJ-SAg were fractionated to seven bands, including major bands A(27.SK) and Bi(19.5 K), the two major serological antigen molecules. 20 sera samples from the patients with the nonpulmonary type of P. w. paragonimiasis were detected using PwMJ-SAg and M-NS-Ag by Dot-ELISA, and the difference of sensitivity between two antigens was highly statistically significant (P<0.001). BALB/c mice, in the early stage of infection with P. w. metacercaria, were immunized with PwMJ-SAg. The spleen cells of the mice were isolated and fused with SP_2/o, a murine myeloma cell line. After three subclonal cultures, eight cell lines secreting monoclonal antibodies (McAbs) to PwMJ-SAg were prepared from 384 wells of hybridoma cells. All McAbs were IgG_1 subclass.

The polyclonal antibodies to juveniles of Paragoniums westermani (PwJ-PcAbs) from sera of Wistar rats infected with Paragoniums westerrnani (P. w.)were purified by Sephadex G 200 chromatography. Next the shared serological antigens of P. w. metacercaria and juveniles (PwMJ-SAg) from the crude antigens of the metacercariae (M-NS-Ag) were purified with immuno-affinity chromatography on cyanogen bromide-activated cross-linked Sepharose 4B beads coupled withPwJ-PcAbs. PwMJ-SAg, agroup of glycoprotein molecules shown by the staining test, were specific serological antigens of P. w. metacercariae and juveniles, identified by the immunoabsorb test and immunoelectrophoresis. By SDSPAGE, PwMJ-SAg were fractionated to seven bands, including major bands A(27.SK) and Bi(19.5 K), the two major serological antigen molecules. 20 sera samples from the patients with the nonpulmonary type of P. w. paragonimiasis were detected using PwMJ-SAg and M-NS-Ag by Dot-ELISA, and the difference of sensitivity between two