Studies on the cell-mediated immunity in experimental <i>Naegleria</i> spp. infections

, Lee, S G; , Shin, H J; , Im, K I

doi:1989.27.3.177

Korean J Parasito > Volume 27(3):1989 > Article

Original Article


Korean J Parasitol. 1989 Sep;27(3):177-186. English. Published online Mar 20, 1994. http://dx.doi.org/10.3347/kjp.1989.27.3.177
Copyright © 1989 by The Korean Society for Parasitology


Studies on the cell-mediated immunity in experimental Naegleria spp. infections
S G Lee,H J Shin and K I Im
Department of Parasitology, College of Medicine and Institute of Tropical Medicine, Yonsei University, Seoul 120-752, Korea.


Abstract
Observations were made on the differences in cell-mediated immune responses in the mice infected with strongly pathogenic Naegleria fowleri ITMAP 359, weakly pathogenic Naegleria jadini 0400, or non-pathogenic Naegleria gruberi EGB, respectively. Variations in cell-mediated responses and changes in antibody titers according to the duration after infection were noted. Infections were done by dropping 5 µls saline suspension containing 10 × 10(4) trophozoites cultured axenically in the CGVS medium into the right nasal cavity of ICR mice aging about 6-7 weeks, under the anesthesia by intraperitoneal injection of secobarbital. Following infection, delayed type hypersensitivity(DTH) responses in the footpad and blastogenic responses of the mouse spleen cells using [3H]-thymidine were observed on the day 1, 4, 7, 10 and 14 after infection. For the preparation of amoeba lysates, each of cultured trophozoites were homogenized with an ultrasonicator, and centrifugated at 20,000 g. The supernatants of amoeba lysates were used as the mitogen and antigen for ELISA. Concanavalin A(Con. A) and lipopolysaccharide(LPS) were also used as mitogens in the blastogenic response. 1. The mice infected with N. fowleri showed the mortality rate of 75.7%. The rate was 6.2% for the N. jadini infected group, while no dead mouse was observed for N. gruberi infections. 2. In regard to DTH responses in the N. fowleri infected mice, the level increased in comparison to the control group but declined after 7 days. An increase was also noted for the N. jadini group after 1 day, but gradual decreases were observed through the infection period. In addition, no difference was noted between the N. gruberi infected and control groups. 3. Concerning the blastogenic response of the splenocytes, it increased after 10 days in the experimental group of N. fowleri infection, but the differences were not statistically significant compared with control group. It was evident that N. jadini group was not different from control group either, while there was a tendency of decrease in N. gruberi infected group. In regard to the blastogenic response of the splenocytes by LPS, it was found that the N. fowleri, N. jadini and N. gruberi infected groups had no differences from the control group. 4. The serum antibody titer of N. fowleri and N. jadini infected mice increased from the day 7 and 14 after infection respectively, while the N. gruberi infected mice showed no increase.

Figures


	Fig. 1 Survival tate in the mice inoculated intranasally with N. fowleri, N. jadini, or N. gruberi.


	Fig. 2 DTH levels obtained in response to subcutaneous injection with each amoeba lysate in the mice infected with N. fowleri, N. jadini, or N. gruberi (Inf.-BSA; infection-bovine serum albumin, Cont.-BSA; Control-bovine serum albumin).


	Fig. 3 Stimulation index of mitogen-treated splenocytes from non-infected (control) and N. fowleri infected mice


	Fig. 4 Stimulation index of mitogen-treated splenocytes from non-infected (control) and N. jadini infected mice.


	Fig. 5 Stimulation index of mitogen-treated splenocytes from non-infected (control) and N. gruberi infected mice.


	Fig. 6 IgG antibody level of sera in the mice infected intranasally with N. fowleri, N. jadini, or N. gruberi.

Tables


	Table 1 Mortality of the mice inoculated intranasally with N. fowleri, N. jadini, or N. gruberi


	Table 2 DTH levels obtained in response to subcutaneous injection with each amoeba lysate in the mice infected with N. fowleri, N. jadini, or N. gruberi


	Table 3 Stimulation index^* (blastogenic response) of mitogen-treated splenocytes from non-infected (control) and N. fowleri infected mice


	Table 4 Stimulation index^* (blastogenic response) of mitogen-treated splenocytes from non-infected (control) and N. jadini infected mice


	Table 5 Stimulation index^* (blastogenic response) of mitogen-treated splenocytes from non-infected (control) and N. gruberi infected mice

References


1.	Barbier D, Goyot P, Dedet JP. Leishmania braziliensis guyanensis dermal leishmaniasis: cell-mediated immunity related to clinical features. Trans R Soc Trop Med Hyg 1985;79(1):47–50.

2.	Barral-Netto M, Hofstetter M, Cheever AW, Ottesen EA. Specificity of antibody and cellular immune responses in human schistosomiasis. Am J Trop Med Hyg 1983;32(1):106–113.

3.	Britten V, Hudson L. Tropenmed parasit 1986;37:97–100.

4.	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.

5.	Culbertson CG. The pathogenicity of soil amebas. Annu Rev Microbiol 1971;25:231–254.

6.	Cursons RT, Brown TJ, Keys EA, Moriarty KM, Till D. Immunity to pathogenic free-living amoebae: role of cell-mediated immunity. Infect Immun 1980;29(2):408–410.

7.	Derrick E. Trans Roy Soc Trop Med Hyg 1948;42:191–198.

8.	Diamantstein T, Klos M, Gold D, Hahn H. Interaction between Entamoeba histolytica and the immune system. I. Mitogenicity of Entamoeba histolytica extracts for human peripheral T lymphocytes. J Immunol 1981;126(6):2084–2086.

9.	Diffley P, Skeels MR, Sogandares-Bernal F. Delayed type hypersensitivity in guinea pig infected subcutaneously with Naegleria fowleri Carter. Z Parasitenkd 1976;49(2):133–137.

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

*11.*	Haggerty RM, John DT. Serum agglutination and immunoglobulin levels of mice infected with Naegleria fowleri. J Protozool 1982;29(1):117–122.

*12.*	Hay J, Dutton GN, Hair DM. Blastogenic responses of splenic lymphocytes to toxoplasmal and retinal antigens and T- and B-cell mitogens in mice with congenital ocular toxoplasmosis. Ann Trop Med Parasitol 1985;79(1):113–115.

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

*15.*	Jacobson LM, et al. J Protozool 1987;34:83–56.

*16.*	John DT. Primary amebic meningoencephalitis and the biology of Naegleria fowleri. Annu Rev Microbiol 1982;36:101–123.

*17.*	John DT, Weik RR, Adams AC. Immunization of mice against Naegleria fowleri infection. Infect Immun 1977;16(3):817–820.

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

*20.*	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.

*21.*	Mason PR, Patterson BA. Proliferative response of human lymphocytes to secretory and cellular antigens of Trichomonas vaginalis. J Parasitol 1985;71(3):265–268.

*22.*	Rowan-Kelly B, Ferrante A, Thong YH. Activation of complement by Naegleria. Trans R Soc Trop Med Hyg 1980;74(3):333–336.

*23.*	Schurr E, et al. Tropenmed Parasit 1986;37:403–408.

*24.*	Selkirk ME, Ogilvie BM, Platts-Mills TA. Activation of human peripheral blood lymphocytes by a trypanosome-derived mitogen. Clin Exp Immunol 1981;45(3):615–620.

*25.*	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.

*26.*	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.

*27.*	Thong YH, Carter RF, Ferrante A, Rowan-Kelly B. Site of expression of immunity to Naegleria fowleri in immunized mice. Parasite Immunol 1983;5(1):67–76.

*28.*	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.

*29.*	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.

*30.*	Valenzuela G, López-Corella E, De Jonckheere JF. Primary amoebic meningoencephalitis in a young male from northwestern Mexico. Trans R Soc Trop Med Hyg 1984;78(4):558–559.

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

*32.*	Willaert E. [Isolation and in vitro culture of the amoeba of the genus Naegleria]. Ann Soc Belges Med Trop Parasitol Mycol 1971;51(6):701–708.

*33.*	Willaert E, et al. Protistologica 1973;9:147–426.