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Korean J Parasitol > Volume 25(1):1987 > Article

Original Article
Korean J Parasitol. 1987 Jun;25(1):1-6. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1987.25.1.1
Copyright © 1987 by The Korean Society for Parasitology
Blastogenic responses of splenic lymphocytes to Naegleria fowleri lysates and T-cell mitogen in mice with primary amoebic meningoencephalitis
Kwang Min Park,Jae Sook Ryu and Kyung Il Im
Department of Parasitology, Yonsei University, College of Medicine, Seoul, Korea.

This study was to observe the changes of blastogenic responses of splenic lymphocytes to T-cell mitogens, N. fowleri lysate and concanavalin A, and serum antibody titer during the course of experimental PAM in mice. Naegleria fowleri, strain 0359, was cultured in the CGVS medium axenically and inoculated intranasally with 7 × 10(4) trophozoites for the development of experimental PAM in mice. The amoebae were subjected to ultrasonication and centrifuged at 20,000 g for 60 minutes, and filtered through 0.2 µm filter membrane. The supernatant, N. fowleri lysate, was used as T-cell mitogen, and antigen for ELISA. The serum antibody was examined by ELISA using peroxidase conjugate. Two hundred micro-l of 10(6) splenocytes in RPMI 1640 containing 10% fetal calf serum were added to each well of a microtiter plate. To each well was added T-cell mitogens, 100 µg/ml of N. fowleri lysate or 4 µg/ml of con. A, and the plates were incubated for 42 hours at 37 C in 5% CO(2) incubator. Cultures were pulsed with 1 micro-Ci of methyl-(3H)-thymidine 6 hour before harvesting. The mean blastogenic response of the splenocytes to N. fowleri lysate 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 lysate during the acute course of experimental PAM in mice.


Fig. 1
IgG antibody level of serum in mice infected intranasally with N.fowleri.

Fig. 2
Blastogenic responses (stimulation index) of con. A-treated splenocytes from non-infected (○) and N.fowleri-infected mice(●).

Fig. 3
Blastogenic responses (stiomulation index) of N.fowleri lysate treated splenocytes from non-infected (○) and N.fowleri-infected mice (●).


Table 1
Determination of optimal concentration of N.fowleri lysate for spleen lymphocytes blastogenesis

Table 2
Blastogenic responses (stimulation index) of con. A-treated splenocytes from non-infected and N.fowleri-infected mice

Table 3
Blastogenic responses (stimulation index) of N.fowleri lysate-treated splenocytes from non-infected and N.fowleri-infected mice

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