Blastogenic responses of splenic lymphocytes to <i>Naegleria fowleri</i> lysates and T-cell mitogen in mice with primary amoebic meningoencephalitis

, Park, Kwang Min; , Ryu, Jae Sook; , Im, Kyung Il

doi:1987.25.1.1

Korean J Parasito > 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.


Abstract
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.

Figures


	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 (●).

Tables


	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

References


1.	Barker LR. Experimental malaria: effects upon the immune response to different antigens. J Infect Dis 1971;123(1):99–101.

2.	Britten V, Hudson L. Immune suppression to Trypanosoma cruzi antigens is associated with infection but not immunisation. Trop Med Parasitol 1986;37(2):97–100.

3.	Carter RF. Description of a Naegleria sp. isolated from two cases of primary amoebic meningo-encephalitis, and of the experimental pathological changes induced by it. J Pathol 1970;100(4):217–244.

4.	Clinton BA, Stauber LA, Palczuk NC. Leishmania donovani: antibody response to chicken ovalbumin by infected golden hamsters. Exp Parasitol 1969;25(1):171–180.

5.	Cunningham DS, Kuhn RE. Trypanosoma cruzi-induced suppressor substance. I. cellular involvement and partial characterization. J Immunol 980 May;124(5):2122–2129.

6.	Ferrante A, Thong YH. Antibody induced capping and endocytosis of surface antigens in Naegleria fowleri. Int J Parasitol 1979;9(6):599–601.

7.	Ferrante A, Smyth C. Mitogenicity of Naegleria fowleri extract for murine T lymphocytes. Immunology 1984;51(3):461–468.

8.	Greenwood BM, Playfair JH, Torrigiani G. Immunosuppression in murine malaria. I. General characteristics. Clin Exp Immunol 1971;8(3):467–478.

9.	Holbrook TW, Boackle RJ, Parker BW, Vesely J. Activation of the alternative complement pathway by Naegleria fowleri. Infect Immun 1980;30(1):58–61.

*10.*	Kierszenbaum F. Immunologic deficiency during experimental Chagas\' disease (Trypanosoma cruzi infection): role of adherent, nonspecific esterase-positive splenic cells. J Immunol 1982;129(5):2202–2205.

*11.*	Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193(1):265–275.

*12.*	Martinez J, Duma RJ, Nelson EC, Moretta FL. Experimental naegleria meningoencephalitis in mice. Penetration of the olfactory mucosal epithelium by Naegleria and pathologic changes produced: a light and electron microscope study. Lab Invest 1973;29(2):121–133.

*13.*	Ramos C, Schädtler-Siwon I, Ortiz-Ortiz L. Suppressor cells present in the spleens of Trypanosoma cruzi-infected mice. J Immunol 1979;122(4):1243–1247.

*14.*	Salaman MH, Wedderburn N, Bruce-Chwatt LJ. The immunodepressive effect of a murine plasmodium and its interaction with murine oncogenic viruses. J Gen Microbiol 1969;59(3):383–391.

*15.*	Sengers RC, Jerusalem CR, Doesburg WH. Murine malaria IV Disturbed immunological responsiveness during Plasmodium berghei infection. Exp Parasitol 1971;30(1):41–53.

*16.*	Simjee AE, Gathiram V, Coovadia HM, Jackson TF, Kiepiela P, Pudifin DJ, Stretton M. Cell-mediated immunity in hepatic amoebiasis. Trans R Soc Trop Med Hyg 1985;79(2):165–168.

*17.*	Strickland GT, Petitt LE, Voller A. Immunodepression in mice infected with Toxoplasma gondii. Am J Trop Med Hyg 973 Jul;22(4):452–455.

*18.*	Strickland GT, Ahmed A, Sell KW. Blastogenic response of Toxoplasma-infected mouse spleen cells to T- and B-cell mitogens. Clin Exp Immunol 1975;22(1):167–176.

*19.*	Thong YH, Ferrante A, Shepherd C, Rowan-Kelly B. Resistance of mice to Naegleria meningoencephalitis transferred by immune serum. Trans R Soc Trop Med Hyg 1978;72(6):650–652.

*20.*	Thong YH, Shepherd C, Ferrante A, Rowan-Kelly B. Protective immunity to Naegleria fowleri in experimental amebic meningoencephalitis. Am J Trop Med Hyg 1978;27(2 Pt 1):238–240.

*21.*	Voller A, Bidwell DE, Bartlett A. Enzyme immunoassays in diagnostic medicine. Theory and practice. Bull World Health Organ 1976;53(1):55–65.