Warning: mkdir(): Permission denied in /home/virtual/lib/view_data.php on line 81
Warning: fopen(upload/ip_log/ip_log_2024-05.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 Immunohistochemical localization of 36 and 29 kDa proteins in sparganum
Immunohistochemical localization of 36 and 29 kDa proteins in sparganum
L S Kim,Y Kong,S Y Kang and S Y Cho
Department of Parasitology, College of Medicine, Chung-Ang University, Seoul 156-756, Korea.
Abstract
Antigenic proteins of 36 and 29 kDa were localized in Spirometra mansoni plerocercoid (sparganum) immunohistochemically by avidin biotin complex (ABC) staining. When polyclonal antibodies such as BALB/c mouse serum immunized with crude saline extract of sparganum or confirmed sparganosis sera were reacted as primary antibodies, the positive chromogen (3-amino, 9-ethylcarbazole) reactions were recognized at syncytial tegument, tegumental cells, muscle and parenchymal cells and lining cells of excretory canals. A monoclonal antibody (MAb) which was reacting to 36 and 29 kDa proteins in the extract of the worm was localized at the syncytial tegument and tegumental cells. The present results suggested that the potent antigenic proteins of 36 and 29 kDa in sparganum were produced at the tegumental cells and transported to the syncytial tegument.
Figures
Fig. 1 SDS-PAGE/immunoblot finding of SP-53 monoclonal antibody. In SDS-PAGE, a linear gradient gel of 10~15% was used for protein separation in reducing condition.
Figs. 2-9 Fig. 2. Nagative control in ABC staining (×100). A 1:1000 diluted healthy human serum was used as a primary antibody. Positive reactions were not observed in any tissue.
Fig. 3. Another negative control (×100). Uninfected mouse serum was reacted at dilution of 1:100 as a primary antibody instead of human serum. No structures were stained with AEC chromogen.
Fig. 4. Positive control (×100). A hyperimmune BALB/c mouse serum was reacted as a primary antibody. Syncytial tegument, tegumental cells, parenchymal cells, excretory canals and muscle cells were stained positively.
Fig. 5. Another positive control of immunized BALB/c mouse serum (×200). Tegument, parenchymal cells and excretory canals were stained with AEC chromogen.
Fig. 6. When a 1:200 diluted patient serum was reacted, strong positive reactions were observed at surface of syncytial tegument, tegumental cells, a part of muscle cells, parenchymal cells and excretory canals (×100).
Fig. 7. Staining with another patient serum (×200). Stronger reactions were shown at surface of syncytial tegument and tegumental cells.
Fig. 8. MAb reacting to 36 and 29 kDa was used as a primary antibody (×100). Surface of syncytial tegument and tegumental cells showed strong positive reactions.
Fig. 9. Another example of ABC staining treated with 36 and 29 kDa MAb (×400). Strong positive reactions at syncytial tegument and tegumental cells were observed.
References
1.
Chappell CL, Dresden MH. Purification of cysteine proteinases from adult Schistosoma mansoni. Arch Biochem Biophys 1987;256(2):560–568.
2.
Cho SY, Kang SY, Kong Y. Purification of antigenic protein of sparganum by immunoaffinity chromatography using a monoclonal antibody. Korean J Parasitol 1990;28(3):135–142.
3.
Choi SH, Kang SY, Kong Y, Cho SY. Antigenic protein fractions reacting with sera of sparganosis patients. Korean J Parasitol 1988;26(3):163–167.
4.
Fukase T, et al. Jpn J Parasitol 1985;34:351–360.
5.
Fukase T, et al. Jpn J parasitol 1986;35:141–144.
6.
Kim CH, Yang J. Imunological characterization of antigens from cysticercus and sparganum and their application to immunodiagnosis 1. Immunological characteristics of crude antigenic components from Cysticercus cellulosae. Korean J Parasitol 1988;26(4):245–254.
7.
Kim CH, Choi WS. [Immunohistochemical observation on the antigens inducing IgG and IgM antibodies against sparganum]. Korean J Parasitol 1991;29(4):339–353.
8.
Kim H, Kim SI, Cho SY. Serological Diagnosis Of Human Sparganosis By Means Of Micro-ELISA. Korean J Parasitol 1984;22(2):222–228.
9.
Kong Y, Kang SY, Cho SY. Single step purification of potent antigenic protein from sparganum by gelatin-affinity chromatography. Korean J Parasitol 1991;29(1):1–7.
10.
Kwak KH, et al. Korean J Zool 1988;31:225–235.
11.
Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227(5259):680–685.
12.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193(1):265–275.
13.
Ohnishi Y, et al. Jpn J Parasitol 1986;35:25–33.
14.
Song CY, et al. Proc Mol Biol Genet 1990;5:141–145.
15.
Tsang VC, Peralta JM, Simons AR. Enzyme-linked immunoelectrotransfer blot techniques (EITB) for studying the specificities of antigens and antibodies separated by gel electrophoresis. Methods Enzymol 1983;92:377–391.
PDF Links
Full text via DOI
Download Citation 
