Changes of splenocyte IFN-gamma mRNA synthesis in rats infected with Paragonimus westermani

Abstract

Changes in the expression level of splenocyte IFN-gamma mRNA of Sprague-Dawley (SD) rats infected with Paragonimus westermani were analyzed by competitive reverse transcription-polymerase chain reaction (RT-PCR) followed by southern blot. The template RNA was extracted from the splenocytes of rats infected with 20 metacercariae of P. westermani. The products of competitive RT-PCR were subjected to southern blot and enhanced chemiluminescence (ECL), and analyzed with a densitometer. In comparison with that of uninfected control rat splenocytes (value of 1), the levels of mRNA expression of IFN-gamma had changed to 0.747 at 1 week post infection (PI), 0.00175 at 2 week PI, 0.0217 at 3 week PI, 0.194 at 4 week PI and then to 0.537 at 5 week PI. The level at 7 week PI had returned to 1.25, comparable with that of uninfected rats. These results show that, when infected with P. westermani, the levels of IFN-gamma mRNA of SD rat splenocytes were remarkably reduced by more than 500 times at 2 week PI and restored to normal level at 7 week PI.

Fig. 1

Analysis of the levels of rat IFN-γ mRNA with competitive RT-PCR and enhanced chemiluminescence. Splenocytes were collected each week from 1 week PI to 7 week PI, excluding 6 week PI. A. Autoradiography of hybridized southern blot. B. Graph of the density ratio.