| Home | E-Submission | Sitemap | Contact us |  
Korean J Parasitol > Volume 29(3):1991 > Article

Original Article
Korean J Parasitol. 1991 Sep;29(3):267-277. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1991.29.3.267
Copyright © 1991 by The Korean Society for Parasitology
Natural killer cell activity in Naegleria fowleri infected mice
K R Lee,C O Shin and K Im
Department of Parasitology, College of Medicine and Institute of Tropical Medicine, Yonsei University, Seoul 120-752, Korea.

The natural killer (NK) cell activity of splenocytes and recycling capacity of NK cells were observed by combining the 51Cr-release cytotoxicity assay and single cell cytotoxicity assay against YAC-1.

The ICR mice were infected intranasally with Naegleria fowleri, that is a pathogenic free-living amoeba. The mice infected with 1 × 105 trophozoites showed mortality rate of 76.7% and mean survival time of 12.9 days.

The cytotoxic activity of NK cells in infected mice was significantly higher than that of non-infected mice during the period between 12 hours and day 3 after infection, and highest on day 1. The target-binding capacity of NK cells in infected mice was not different from that of non-infected ones. Maximal killing potential and maximal recycling capacity were remarkably increased in infected mice as compared with the control.

The results obtained in this observation indicated that elevated NK cell activity in mice infected with N. fowleri was not due to target-binding capacity of NK cells but due to the increased activity of NK cells and increased recycling capacity of individual NK cells.


Fig. 1
Data from Table 1 are used to exemplify the calculation of Vmax. Initial number of target cells(T) is poltted against number of dead target cells(V) at the end of the test. The points fit well to the theoretical Michaelis-Menten saturation curve. A Line-weaver-Burk polt is used to calculate Vmax and Km. 1/T=X plotted against 1/V=Y. Data are taken from Table 1 and Fig. 1. Regression analysis is used to obtain a straight line (r=0.990). Vmax equals the reciprocal of the Y intercept and Km equals the negative reciprocal of the X intercept.

Fig. 2
Natural killer cell cytotoxic activity against YAC-1 target cells in mice infected with Naegleria fowleri.

Fig. 3
Single cell cytotoxic activity against YAC-1 target cells in mice infected with Naegleria fowleri.

Fig. 4
Vmax against YAC-1 target cell in mice infected with Naegleria fowleri.

Fig. 5
Maximal recycling capacity against YAC-1 target cells in mice infected with Naegleria fowleri.


Table 1
A representative experiment showing data used from calculations of Vmax and Km

Table 2
Cumulative number of death in mice inoculated intranasally with Naegleria fowleri

Table 3
NK activity, Vmax, TBC, fraction of dead conjugates, and estimation of active NK & their maximal recycling capacity*

Table 4
Correlation matrix between Vmax, TBC, active NK, MRC and NK activity

1. Abo T, Balch CM. A differentiation antigen of human NK and K cells identified by a monoclonal antibody (HNK-1). J Immunol 1981;127(3):1024–1029.
2. Arai S, et al. J Immunol 1983;138:651–658.
3. Berke G, Gabison D. Energy requirements of the binding and lytic steps of T lymphocyte-mediated cytolysis of leukemic cells in vitro. Eur J Immunol 1975;5(10):671–675.
4. Bhagwandeen SB, Carter RF, Naik KG, Levitt D. A case of hartmannellid amebic meningoencephalitis in Zambia. Am J Clin Pathol 1975;63(4):483–492.
5. Bloom BR. Natural killers to rescue immune surveillance?. Nature 1982;300(5889):214–215.
6. Bradley TP, Bonavida B. Mechanism of cell-mediated cytotoxicity at the single cell level. IV. Natural killing and antibody-dependent cellular cytotoxicity can be mediated by the same human effector cell as determined by the two-target conjugate assay. J Immunol 1982;129(5):2260–2265.
7. Burton RC, et al. J Immunol 1981;126:1985–1991.
8. Carter RF. Primary amoebic meningo-encephalitis: clinical, pathological and epidemiological features of six fatal cases. J Pathol Bacteriol 1968;96(1):1–25.
9. Cikes M, Friberg S Jr, Klein G. Progressive loss of H-2 antigens with concomitant increase of cell-surface antigen(s) determined by Moloney leukemia virus in cultured murine lymphomas. J Natl Cancer Inst 1973;50(2):347–362.
10. Clark IA, et al. Nature 1974;252:318–325.
11. Cursons RTM, et al. NIJ Mar Freshwater Res 1976;10:245–262.
12. 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.
13. 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.
14. Eldridge A, et al. Br Med J 1967;1:299–303.
15. Steinhauer EH, Doyle AT, Reed J, Kadish AS. Defective natural cytotoxicity in patients with cancer: normal number of effector cells but decreased recycling capacity in patients with advanced disease. J Immunol 1982;129(5):2255–2259.
16. Ferrante A, Thong YH. Antibody induced capping and endocytosis of surface antigens in Naegleria fowleri. Int J Parasitol 1979;9(6):599–601.
17. Ferrante A, Smyth C. Mitogenicity of Naegleria fowleri extract for murine T lymphocytes. Immunology 1984;51(3):461–468.
18. Fowler M, Carter RF. Acute pyogenic meningitis probably due to Acanthamoeba sp.: a preliminary report. Br Med J 1965;2(5464):740–742.
19. Galli SJ, Dvorak AM, Ishizaka T, Nabel G, Der Simonian H, Cantor H, Dvorak HF. A cloned cell with NK function resembles basophils by ultrastructure and expresses IgE receptors. Nature 1982;298(5871):288–290.
20. Gatti G, Cavallo R, Sartori ML, Marinone C, Angeli A. Cortisol at physiological concentrations and prostaglandin E2 are additive inhibitors of human natural killer cell activity. Immunopharmacology 1986;11(2):119–128.
21. Gidlund M, Orn A, Wigzell H, Senik A, Gresser I. Enhanced NK cell activity in mice injected with interferon and interferon inducers. Nature 1978;273(5665):759–761.
22. Grimm EA, Bonavida B. Studies of the induction and expression of T cell mediated immunity. VI. Heterogeneity of lytic efficiency exhibited by isolated cytotoxic T lymphocytes prepared from highly enriched populations of effector-target conjugates. J Immunol 1977;119(3):1041–1047.
23. Grimm E, Bonavida B. Mechanism of cell-mediated cytotoxicity at the single cell level. I. Estimation of cytotoxic T lymphocyte frequency and relative lytic efficiency. J Immunol 1979;123(6):2861–2869.
24. Hatcher FM, Kuhn RE. Destruction of Trypanosoma cruzi by Natural killer cells. Science 1982;218(4569):295–296.
25. Herberman RB, Nunn ME, Lavrin DH. Natural cytotoxic reactivity of mouse lymphoid cells against syngeneic acid allogeneic tumors. I. Distribution of reactivity and specificity. Int J Cancer 1975;16(2):216–229.
26. Herberman RB, Holden HT. Natural cell-mediated immunity. Adv Cancer Res 1978;27:305–377.
27. Herberman RB, Ortaldo JR. Natural killer cells: their roles in defenses against disease. Science 1981;214(4516):24–30.
28. Henney CS, Kuribayashi K, Kern DE, Gillis S. Interleukin-2 augments natural killer cell activity. Nature 1981;291(5813):335–338.
29. Hiserodt JC, et al. J Immnunol 1982;135:1782–1786.
30. Hiserodt JC, et al. J Immunol 1982;136:2272–2278.
31. Holbrook NJ, Cox WI, Horner HC. Direct suppression of natural killer activity in human peripheral blood leukocyte cultures by glucocorticoids and its modulation by interferon. Cancer Res 1983;43(9):4019–4025.
32. Jondal M, Pross H. Surface markers on human b and t lymphocytes. VI. Cytotoxicity against cell lines as a functional marker for lymphocyte subpopulations. Int J Cancer 1975;15(4):596–605.
33. Visvesvara GS, Jones DB, Robinson NM. Isolation, identification, and biological characterization of Acanthamoeba polyphaga from a human eye. Am J Trop Med Hyg 1975;24(5):784–790.
34. Kasai M, Leclerc JC, McVay-Boudreau L, Shen FW, Cantor H. Direct evidence that natural killer cells in nonimmune spleen cell populations prevent tumor growth in vivo. J Exp Med 1979;149(5):1260–1264.
35. Kenny M. Health Lab Sci 1971;8:5–10.
36. Kiessling R, Klein E, Wigzell H. "Natural" killer cells in the mouse. I. Cytotoxic cells with specificity for mouse Moloney leukemia cells. Specificity and distribution according to genotype. Eur J Immunol 1975;5(2):112–117.
37. Kingston D, Warhurst DC. Isolation of amoebae from the air. J Med Microbiol 1969;2(1):27–36.
38. Lallinger GJ, et al. Immunology 1987;55:1289–1293.
39. Lang NP, Ortaldo JR, Bonnard GD, Herberman RB. Interferon and prostaglandins: effects on human natural and lectin-induced cytotoxicity. J Natl Cancer Inst 1982;69(2):339–343.
40. Lopez C. Genetics of natural resistance to herpesvirus infections in mice. Nature 1975;258(5531):152–153.
41. 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.
42. Naginton J, Watson PG, Playfair TJ, McGill J, Jones BR, Steele AD. Amoebic infection of the eye. Lancet 1974;2(7896):1537–1540.
43. Onsrud M, Thorsby E. Influence of in vivo hydrocortisone on some human blood lymphocyte subpopulations. I. Effect on natural killer cell activity. Scand J Immunol 1981;13(6):573–579.
44. Ortaldo JR, et al. Ann Rev Immunol 1984;2:355–359.
45. Petranyi G, et al. Immunogenetics 1976;3:15–22.
46. Quan PC, Ishizaka T, Bloom BR. Studies on the mechanism of NK cell lysis. J Immunol 1982;128(4):1786–1791.
47. Reynolds CW, Timonen T, Herberman RB. Natural killer (NK) cell activity in the rat. I. Isolation and characterization of the effector cells. J Immunol 1981;127(1):282–287.
48. Richards CS. Two new species of Hartmannella amebae infecting freshwater mollusks. J Protozool 1968;15(4):651–656.
49. Roder JC, Kiessling R, Biberfeld P, Andersson B. Target-effector interaction in the natural killer (NK) cell system. II. The isolation of NK cells and studies on the mechanism of killing. J Immunol 1978;121(6):2509–2517.
50. Rowan-Kelly B, Ferrante A, Thong YH. Activation of complement by Naegleria. Trans R Soc Trop Med Hyg 1980;74(3):333–336.
51. Rumpold H, Kraft D, Obexer G, Bock G, Gebhart W. A monoclonal antibody against a surface antigen shared by human large granular lymphocytes and granulocytes. J Immunol 1982;129(4):1458–1464.
52. Ryser JE, Sordat B, Cerottini JC, Brunner KT. Mechanism of target cell lysis of cytolytic T lymphocytes. I. Characterization of specific lymphocyte-target cell conjugates separated by velocity sedimentation. Eur J Immunol 1977;7(2):110–117.
53. Schmoller H. [Description Of Some Cultivated Amoebae Of Marine Origin]. J Protozool 1964;11:497–502.
54. Talmadge JE, Meyers KM, Prieur DJ, Starkey JR. Role of NK cells in tumour growth and metastasis in beige mice. Nature 1980;284(5757):622–624.
55. Timonen T, Reynolds CW, Ortaldo JR, Herberman RB. Isolation of human and rat natural killer cells. J Immunol Methods 1982;51(3):269–277.
56. Thong YH, Ferrante A, Rowan-Kelly B, OKeefe DE. Immunization with culture supernatant in experimental amoebic meningoencephalitis. Trans R Soc Trop Med Hyg 1979;73(6):684–685.
57. Trischmann T, Tanowitz H, Wittner M, Bloom B. Trypanosoma cruzi: role of the immune response in the natural resistance of inbred strains of mice. Exp Parasitol 1978;45(2):160–168.
58. Ullberg M, Jondal M. Recycling and target binding capacity of human natural killer cells. J Exp Med 1981;153(3):615–628.
59. Warner JF, Dennert G. Effects of a cloned cell line with NK activity on bone marrow transplants, tumour development and metastasis in vivo. Nature 1982;300(5887):31–34.
60. Wright SC, et al. J Immunol 1981;125:1561–1567.
61. Wright SC, et al. J Immunol 1983;130:2479–2485.
62. Young JD, Podack ER, Cohn ZA. Properties of a purified pore-forming protein (perforin 1) isolated from H-2-restricted cytotoxic T cell granules. J Exp Med 1986;164(1):144–155.
63. Davies M. Biological membranes. Nature 1970;228(5269):394.
Editorial Office
c/o Department of Medical Environmental Biology
Chung-AngUniversity College of Medicine, Dongjak-gu, Seoul 06974, Korea
Tel: +82-2-820-5683   Fax: +82-2-826-1123   E-mail: kjp.editor@gmail.com
About |  Browse Articles |  Current Issue |  For Authors and Reviewers
Copyright © 2021 by The Korean Society for Parasitology and Tropical Medicine.     Developed in M2PI