Blastogenic responses of splenic Iymphocytes to Naegleria fowleri Iysates and T-cell mitogen in mice with primary amoebie meningoencephalitis.

실험적 뇌막수염에 있어 Naegleria fowleri 항원에 대한 세포매개성 면역 반응

This study was to observe the changes of blastogenic responses of splenic Iymphocytes to T-cell mitogens, N. fcwleri Iysate and concanaualin A, and serum antibody titer during the course of experimental PAM in mice. Naegleria fcwleri, strain 0359, was cultured in the CGVS medium axenically and inoculated intranasally with $7{\times}10^4$ trophozoites for the development of experimental PAM in mice. The amoebae were subjected to ultrasonication and centrifuged at 20,000g for 60 minutes, and filtered through $0.2{\mu\textrm{m}}$ filter membrane. The supernatant, N. fcwleri Iysate, was used as T-cell mitogen, and antigen for ELISA. The serum antibody was examined by ELISA using peroxidase conjugate. Two hundred ${\mi}l$ of $10^6$ splenocytes in RPMI 1640 containing 0% fetal calf serum were added to each well of a microtiter plate. To each well was added T-cell mitogens, $100{\mu}g/ml$ of N. fowleri Iysate or $4{\mu}g/ml$ of con. A, and the plates were incubated for 42 hours at $37^{\circ}C$ in 5% $CO_2$ incubator. Cultures were pulsed with of $methyl-(^3H)-thymidine$ 6 hour before harvesting. The mean blastogenic response of the splenocytes to N. fewleri Iysate was reduced, whereas that to con. A was also reduced up to on day 11 after infection. Both of these results were statistically significant compared with those of uninfected control group. The serum antibody titers were increased gradually up to day 15. The results indicated that there was an impairment of the blastogenic response of splenocytes to N. fowleri Iysate during the acute course of experimental PAM in mice.

N. fowleri를 감염시키고 그 경과에 따른 숙주의 면역기능 변동을 알아보고자 마우스에 N. fowleri를 감염시키고 그 경과에 따른 T임파구 기능과 혈청내 항체가를 관찰하였다. T임파구 기능을 알아보기 위하여 비장세포를 배양할 때 mitogen으로 con. A와 N. fowleri lysate를 첨가하고 42시간 배양후 $methyl-[^3H]-thymidine$을 넣어 비장세포에 uptake되는 정도를 측정하였으며 혈청내 항체가는 효소표식 면역검사법(ELISA)으로 측정하였다. 실험성적을 요약하면 다음과 같다. T임파구의 기능은 사용된 mitogen con. A와 N. fowleri lysated에 관계없이 감염 후 3일부터 감소되어 11일까지 대조군에 비해 유의하게 감소하였다. 감염후 15일에는 N. fowleri lysate를 사용하였을 경우 T임파구 기능이 계속 감소되어 있었으나 con. A를 첨가하였을때 정상대조군과 차이가 없음을 알 수 있었다.