Specific antibody immune response against the parasitic portion of a glutathione-S-transferase fusion protein.
The humoral immune response against an Entamoeba histolytica recombinant protein has been investigated. The 628 bp Bam HI-Eco RI DNA fragment (L1b) from the M11 cDNA clone, partially coding for a 220 kDa (L220) protein, was ligated in-frame into the pGEX-3X plasmid vector to produce the fusion protein GST-L1b. BALB/c mice were immunized with different doses of the GST-L1b fusion protein (10-500 microg). GST-L1b doses of 100/50/50, 300, or 500 microg induced an antibody response (IgG1>IgG3, IgG2a>IgG2b) specific for the amoebic part of the fusion protein (L1b). These antibodies were able to recognize the native protein in amoebic total extract. Anti-GST antibodies were not detected. On the other hand, doses of 10/10/10 or 200/100/100 microg induced antibodies able to recognize both GST (IgG2a>IgG1>IgG2b) and L1b (IgG1, IgG2a>IgG3>IgG2b). When mice were immunized with GST alone (100/50/50, 300 or 500 microg), antibodies against GST-L1b or GST were not detected. However, GST doses of 10/10/10 or 200/100/100 microg induced an antibody response able to recognize both GST-L1b and GST. We propose that an immunization protocol similar to the one used in this work may allow induction of high antibody titers specific against the parasite segment of a GST-fusion protein.
López-Monteon
Aracely
A
Experimental Pathology Department, CINVESTAV-IPN, México DF, México.
Ramos-Ligonio
Angel
A
Pérez-Castillo
Leticia
L
Talamás-Rohana
Patricia
P
Rosales-Encina
José Luis
JL
D000818:Animals; D000913:Antibodies, Protozoan / Q000276:immunology*; D004748:Entamoeba histolytica / Q000235:genetics* / Q000276:immunology; D005260:Female; D005982:Glutathione Transferase / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D007106:Immune Sera / Q000097:blood / Q000276:immunology; D015151:Immunoblotting; D007074:Immunoglobulin G / Q000276:immunology; D037102:Lectins / Q000235:genetics* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism
D000913:Antibodies, Protozoan; D007106:Immune Sera; D007074:Immunoglobulin G; D037102:Lectins; D015800:Protozoan Proteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase