Production of monoclonal antibodies against the 8 kDa subunit of Echinococcus granulosus Antigen B (EgAgB8/2) using DNA immunization.
Cystic echinococcosis (CE), an endemic cosmopolitan zoonotic helminthic disease caused by the larval stage of Echinococcus granulosus, lacks reliable diagnostic tools that fulfill the criteria of high sensitivity and specificity. Antigen B (AgB), a thermostable lipoprotein that constitutes a considerable fraction of the cystic hydatid fluid (HF), is being considered as a suitable source for vaccination and immunodiagnosis of CE due to its high specificity. Genetic immunization was used to immunize BALB/c mice with the second subunit of antigen B (EgAgB8/2) for the production of monoclonal antibodies (MAb). Fusion products between the spleen cells and myeloma cells produced six MAbs of the following isotypes: IgG2a (two clones), IgG2b (three clones), and IgM (one clone). The MAbs were tested for their specificity to crude sheep hydatid fluid (CSHF) versus other antigens prepared from other helminthic parasites including Toxocara canis, Acanthocheilonema viteae, Fasciola hepatica, Schistosoma mansoni, and Taenia. Five MAbs reacted with E. granulosus antigens, one showed cross reactivity with S. mansonia antigens, and one showed a high reactivity with E. granulosus but was cross reactive with all helminthic antigens tested. Using SDS-PAGE and immunoblotting under reducing conditions, all MAbs identified the four AgB subunits with molecular weights of 8, 16, 24, and 36 kDa. Further work on the specificity and sensitivity of these MAbs as well as their use in detecting circulating parasite antigens and in antigen purification will be assessed in future studies.
Al-Qaoud
Khaled M
KM
Department of Biological Sciences, Yarmouk University, Irbid, Jordan. akhaled@yu.edu.jo
Al-Omari
Mariam M
MM
Al-Aghbar
Mohammad
M
Abdel-Hafez
Sami K
SK
D000818:Animals; D000909:Antibodies, Helminth / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D000947:Antigens, Helminth / Q000276:immunology*; D001483:Base Sequence; D004247:DNA / Q000737:chemistry*; D004446:Echinococcus / Q000378:metabolism*; D005260:Female; D006825:Hybridomas / Q000378:metabolism*; D008074:Lipoproteins / Q000737:chemistry*; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D012689:Sequence Homology, Nucleic Acid; D012756:Sheep; D013154:Spleen / Q000378:metabolism
D000909:Antibodies, Helminth; D000911:Antibodies, Monoclonal; D000947:Antigens, Helminth; D008074:Lipoproteins; C514022:antigen B, Echinococcus granulosus; D004247:DNA