Warning: mkdir(): Permission denied in /home/virtual/lib/view_data.php on line 81

Warning: fopen(upload/ip_log/ip_log_2024-03.txt): failed to open stream: No such file or directory in /home/virtual/lib/view_data.php on line 83

Warning: fwrite() expects parameter 1 to be resource, boolean given in /home/virtual/lib/view_data.php on line 84
Membrane proteins and their antigenicity of Toxoplasma gondii
| Home | E-Submission | Sitemap | Contact us |  
top_img
Korean J Parasito Search

CLOSE

Korean J Parasito > Volume 26(3):1988 > Article

Original Article
Korean J Parasitol. 1988 Sep;26(3):155-162. English.
Published online Mar 20, 1994.  http://dx.doi.org/10.3347/kjp.1988.26.3.155
Copyright © 1988 by The Korean Society for Parasitology
Membrane proteins and their antigenicity of Toxoplasma gondii
Won Young Choi,Ho Woo Nam and Jae Eul Yoo
Catholic Institute of Parasitic Diseases, Catholic University Medical College, Seoul 135-040, Korea.
Abstract

Surface membrane proteins of virulent RH strain and tissue cyst-forming Fukaya strain of Toxoplasma gondii were analyzed by SDS-polyacrylamide gel electrophoresis after LPO-catalyzed surface iodination and lectin blotting, then identified the zoite-specific antigens. Prior to the analyses, purification of RH tachyzoites from mouse peritoneal exudate and of Fukaya bradyzoites from mouse brain tissues were performed by centrifugation on the discontinuous Percoll density-gradient. Tachyzoites were obtained at the interface of 50 per cent and 60 per cent Percoll solution and brain cysts were harvested at the interfaces of 40-50 per cent and 50-60 per cent, then bradyzoites were obtained by treating the cysts with hypertonic solution. The LPO-catalyzed iodination detected 15 KDa and 14 KDa proteins of bradyzoites and 30 KDa protein of tachyzoites as major bands with several other minor bands. But Con A blotting revealed some bands of 200 K-50 KDa glycoproteins of bradyzoites and 52 KDa band as major and minor bands of 33 K-20 KDa of tachyzoites. Phytohemagglutinin did not detect any band in the two forms. EITB with anti-Fukaya antibody and anti-RH antibody revealed cross-reactivities between the two forms. Despite the cross-reactivity, anti-Fukaya antibody reacted with 15 KDa band of bradyzoites specifically and, anti-RH antibody with 52 KDa, 30 KDa, and 25 KDa bands of tachyzoites, respectively. It was identified that 15 KDa protein in bradyzoite, which was not a glycoprotein, was a major membrane protein with sufficient antigenicity, and in the case of tachyzoite, 52 KDa surface glycoprotein (gp52) with specific antigenicity might be added to the major surface protein, p30.

Figures


Fig. 1
Photomicrograph of a purified T. gondii brain cyst of Fukaya strain. Cysts were purified by Percoll density-gradient centrifugation of mouse brain infected 1 month previously.


Fig. 2
Coomassie-stained pattern (CM) and lactoperoxidase(LPO)-catalyzed surface iodination pattern of bradyzoites of Fukaya strain (F) and tachyzoites of RH strain (R). Numerals stand for molecular weight (×10-8).


Fig. 3
Western blot analysis of bradyzoites (F) and tachyzoites (R) probed with 125I-Con A (Con A) and 125I-PHA (PHA). Coomassie-stained pattern was also shown as CM. Numerals stand for molecular weight (×10-3).


Fig. 4
EITB patterns of brayzoites (F) and tachyzoites (R). Reactions with anti-Fukaya antibody were designated as Anti-F and those with anti-RH antibody were described as Anti-R. Numerals stand for molecular weight (×10-3).

References
1. Araujo FG, Dubey JP, Remington JS. Antigenic similarity between the coccidian parasites Toxoplasma gondii and Hammondia hammondi. J Protozool 1984;31(1):145–147.
 
2. Balber AE, Frommel TO. Trypanosoma brucei gambiense and T. b. rhodesiense: concanavalin A binding to the membrane and flagellar pocket of bloodstream and procyclic forms. J Protozool 1988;35(2):214–219.
 
4. Choi WY, Yoo JE, Nam HW, Oh CY, Kim SW, Katakura K, Kobayashi A. Toxoplasma antibodies by indirect latex agglutination tests in zoo animals. Korean J Parasitol 1987;25(1):13–23.
 
5. Cornelissen AW, Overdulve JP, Hoenderboom JM. Separation of Isospora (Toxoplasma) gondii cysts and cystozoites from mouse brain tissue by continuous density-gradient centrifugation. Parasitology 1981;83(Pt 1):103–108.
  
6. Couvreur G, Sadak A, Fortier B, Dubremetz JF. Surface antigens of Toxoplasma gondii. Parasitology 1988;97(Pt 1):1–10.
  
7. Dempster RP. Toxoplasma gondii: purification of zoites from peritoneal exudates by eight methods. Exp Parasitol 1984;57(2):195–207.
  
8. Dubey JP, Frenkel JK. Feline toxoplasmosis from acutely infected mice and the development of Toxoplasma cysts. J Protozool 1976;23(4):537–546.
 
9. Dubremetz JF, Rodriguez C, Ferreira E. Toxoplasma gondii: redistribution of monoclonal antibodies on tachyzoites during host cell invasion. Exp Parasitol 1985;59(1):24–32.
  
10. Glass WF 2nd, Briggs RC, Hnilica LS. Use of lectins for detection of electrophoretically separated glycoproteins transferred onto nitrocellulose sheets. Anal Biochem 1981;115(1):219–224.
  
11. Grimwood BG, Hechemy K, Stevens RW. Toxoplasma gondii: purification of trophozoites propagated in cell culture. Exp Parasitol 1979;48(2):282–286.
  
12. Handman E, Goding JW, Remington JS. Detection and characterization of membrane antigens of Toxoplasma gondii. J Immunol 1980;124(6):2578–2583.
 
13. Jaffe CL, McMahon-Pratt D. The identification of membrane glycoconjugates in Leishmania species. J Parasitol 1988;74(4):548–561.
  
14. Johnson AM, McDonald PJ, Neoh SH. Monoclonal antibodies to Toxoplasma cell membrane surface antigens protect mice from toxoplasmosis. J Protozool 1983;30(2):351–356.
 
15. Kasper LH, Crabb JH, Pfefferkorn ER. Isolation and characterization of a monoclonal antibody-resistant antigenic mutant of Toxoplasma gondii. J Immunol 1982;129(4):1694–1699.
 
16. Kasper LH, Crabb JH, Pfefferkorn ER. Purification of a major membrane protein of Toxoplasma gondii by immunoabsorption with a monoclonal antibody. J Immunol 1983;130(5):2407–2412.
 
17. Kasper LH, Bradley MS, Pfefferkorn ER. Identification of stage-specific sporozoite antigens of Toxoplasma gondii by monoclonal antibodies. J Immunol 1984;132(1):443–449.
 
18. Kasper LH, Currie KM, Bradley MS. An unexpected response to vaccination with a purified major membrane tachyzoite antigen (P30) of Toxoplasma gondii. J Immunol 1985;134(5):3426–3431.
 
19. Kasper LH. Isolation and characterization of a monoclonal anti-P30 antibody resistant mutant of Toxoplasma gondii. Parasite Immunol 1987;9(4):433–445.
  
20. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227(5259):680–685.
  
21. Levine ND. Tazonomy of Toxoplasma. J Protozool 1977;24(1):36–41.
 
23. Ofek I, Sharon N. Lectinophagocytosis: a molecular mechanism of recognition between cell surface sugars and lectins in the phagocytosis of bacteria. Infect Immun 1988;56(3):539–547.
 
24. Petri WA Jr, Joyce MP, Broman J, Smith RD, Murphy CF, Ravdin JI. Recognition of the galactose- or N-acetylgalactosamine-binding lectin of Entamoeba histolytica by human immune sera. Infect Immun 1987;55(10):2327–2331.
 
25. Pettersen EK. Acta Pathol Microbiol Scan 1979;87:217–220.
26. Rodriguez C, Afchain D, Capron A, Dissous C, Santoro F. Major surface protein of Toxoplasma gondii (p30) contains an immunodominant region with repetitive epitopes. Eur J Immunol 1985;15(7):747–749.
  
27. Rodrigues MM, Xavier MT, Mendon. a-Previato L, Barcinski MA. Novel 17-kilodalton Leishmania antigen revealed by immunochemical studies of a purified glycoprotein fraction recognized by murine T lymphocytes. Infect Immun 1988;56(7):1766–1770.
 
28. Sabin AB. JAMA 1941;116:801–807.
29. Santoro F, Afchain D, Pierce R, Cesbron JY, Ovlaque G, Capron A. Serodiagnosis of toxoplasma infection using a purified parasite protein (P30). Clin Exp Immunol 1985;62(2):262–269.
 
30. Thorell JI, Johansson BG. Enzymatic iodination of polypeptides with 125I to high specific activity. Biochim Biophys Acta 1971;251(3):363–369.
 
31. Towbin H, Staehelin T, Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A 1979;76(9):4350–4354.
  
32. Tryon JC, Weidner E, Larson AD, Hart LT. A rapid method for isolating purified Toxoplasma tachyzoites from peritoneal exudate and cell culture. J Parasitol 1978;64(6):1127–1129.
  
Editorial Office
Department of Molecular Parasitology, Samsung Medical Center, School of Medicine, Sungkyunkwan University,
2066 Seobu-ro, Jangan-gu, Suwon 16419, Gyeonggi-do, Korea.
Tel: +82-31-299-6251   FAX: +82-1-299-6269   E-mail: kjp.editor@gmail.com
About |  Browse Articles |  Current Issue |  For Authors and Reviewers
Copyright © 2024 by The Korean Society for Parasitology and Tropical Medicine.     Developed in M2PI