Immunological tests by anti-free-living amoebas serum produced in experimental animals I. Immobilization of free-living amoebas <i>in vitro</i> by rabbit antiserum

, Im, Kyung Il; , Oh, Hyea Sook

doi:1978.16.1.41

Korean J Parasito > Volume 16(1):1978 > Article

Original Article


Korean J Parasitol. 1978 Jun;16(1):41-46. English. Published online Mar 20, 1994. http://dx.doi.org/10.3347/kjp.1978.16.1.41
Copyright © 1978 by The Korean Society for Parasitology


Immunological tests by anti-free-living amoebas serum produced in experimental animals I. Immobilization of free-living amoebas in vitro by rabbit antiserum
Kyung-Il Im and Hyea-Sook Oh
Institute of Tropical Medicine and Department of Parasitology, College of Medicine, Yonsei University, Korea.


Abstract
Rabbits were immunized with free-living amoebas by intravenous injections. The amoebas were Acanthamoeba culbertsoni and Naegleria fowleri and obtained by axenic cultivation in CGVS medium. Each rabbit received 10(6) of Acanthamoeba culbertsoni and 10⁵ of Naegleria fowleri trophozoites respectively every other day in three doses and finally one booster dose at 1 week later. Antiserum was collected from thc following day of the booster injection up to 2 months period, and stored at -30℃ until use. The immobilization test was performed. One drop of amoeba suspension was mixed with the test serum on slide and observed the mobile state under microscope. 1. Maximal immobilizing phenomenon observed in 30 minutes and, then gradually recovered to normal state. 2. Inactivation of antiserum at 56℃ for 30 minutes did not affect the immobilization phenomenon. 3. The immobilization rates decreased by the serial dilution of antiserum. At dilution more than 1:8, the immobilization was almost the same as in the normal serum. 4. The immobilizing antibody in anti-Acanthamoeba culbertsoni rabbit serum showed highest titre in 3rd day after booster immunization and from first to 6th week in anti-Naegleria fowleri rabbit serum. 5. Cross matching of Acanthamoeba culbertsoni and Naegleria fowleri showed antigenic difference of the two species. It is suggested that the immobilization reaction may be of value as a supplementary test in the diagnosis of primary amoebic meningoencephalitis.

Figures


	Fig. 1 Immobilization of free-living amoebas (A; Acanthamoeba culbertsoni, N; Naegleria fowleri) in respective immune serum by time


	Fig. 2 Immobilization of Naegleria fowleri in inactivated serum and non-inactivated serum.


	Fig. 3 Immobilization of free-living amoebas in the immune serum by dilution.


	Fig. 4 Immobilization titer of Acanthamoeba culbertsoni (A) immune serum and Naegleria fowleri (N) immune serum by duration.


	EXPLANATION OF PLATES Fig. 1.Acanthamoeba culbertsoni, mobile trophozoites Fig. 2.Acanthamoeba culbertsoni, immobilized one Fig. 3.Acanthamoeba culbertsoni, agglomerated trophozoites Fig. 4.Naegleria fowleri, mobile trophozoite Fig. 5.Naegleria fowleri, immobilized one

Tables


	Table 1 CGVS Medium for Free-living Amoeba


	Table 2 Antigenic relationship between Acanthamoeba culbertsoni and Naegleria fowleri exposure in each antiserum for 30 minutes

References


1.	Biagi F, Buentello L. Immobilization reaction for the diagnosis of amebiasis. Exp Parasitol 1961;11:188–190.

2.	Biagi F, Beltran F, Ortega PS. Remobilization of Entamoeba histolytica after exposure to immobilizing antibodies. Exp Parasitol 1966;18(1):87–91.

3.	Cho KM, et al. Yonsei Medical Journal 1969;10(2):173–178.

4.	Chung PR. [Immobilization-Agglomeration Reaction For The Diagnosis Of Trichomonas Vaginalis Infection]. Korean J Parasitol 1970;8(1):13–21.

5.	Cole BA, Kent JF. Immobilization of Endamoeba histolytica in vitro by antiserum produced in the rabbit. Proc Soc Exp Biol Med 1953;83(4):811–814.

6.	Nelson RA Jr, Mayer MM. Immobilization of Treponema pallidum in vitro by antibody produced in syphilitic infection. J Exp Med 1949;89(4):369–393.

7.	Zaman V. An immobilization reaction against Balantidium coli. Nature 1962;194:404–405.

8.	Zaman V. Studies on the Immobilization Reaction in the Genus Balantidium. Trans R Soc Trop Med Hyg 1964;58:255–259.