Isolation and partial characterization of an immunogenic antigen of Giardia lamblia

• Published in: Parasitology international 2017-06, Vol.66 (3), p.324-330
• Main Authors: Quintero, Jael, Valdez, Alejandra, Samaniego, Brenda, Lopez-Romero, Gloria, Astiazaran-Garcia, Humberto, Rascon, Lucila, Breci, Linda, Garibay-Escobar, Adriana, Robles-Zepeda, Ramón, Velazquez, Carlos
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Animals; Antigens, Protozoan - administration & dosage; Antigens, Protozoan - chemistry; Antigens, Protozoan - genetics; Antigens, Protozoan - immunology; Antigens, Protozoan - isolation & purification; Cellular response; Epitopes, B-Lymphocyte - chemistry; Epitopes, B-Lymphocyte - immunology; Epitopes, B-Lymphocyte - isolation & purification; Epitopes, T-Lymphocyte - chemistry; Epitopes, T-Lymphocyte - immunology; Epitopes, T-Lymphocyte - isolation & purification; Gastroenterology and Hepatology; Giardia lamblia; Giardia lamblia - immunology; Humoral response; Immunity, Cellular; Immunity, Humoral; Immunogenic protein; Infectious Disease; Membrane Proteins - genetics; Mice; Protozoan Proteins - administration & dosage; Protozoan Proteins - genetics; Protozoan Proteins - immunology; Protozoan Proteins - isolation & purification; Protozoan Proteins - metabolism; Tandem Mass Spectrometry; Trophozoites - immunology
• ISSN: 1383-5769; 1873-0329
• DOI: 10.1016/j.parint.2017.01.007

Abstract Humoral and cellular immune responses play an important role during Giardia lamblia infection. Several Giardia proteins have been identified as immunogenic antigens based on their elicited humoral immune response. Poorly is known about Giardia antigens that stimulate a cellular immune response. The main purpose of this study was to isolate and partial characterize an immunogenic antigen (5G8) of G. lamblia . The 5G8 protein was isolated from G. lamblia trophozoite lysates by affinity chromatography using moAb 5G8-coupled CNBr-Sepharose. The isolated protein was analysed by electrospray tandem mass spectrometry (ESI-MS/MS), and by diverse bioinformatics tools ( Giardia DB, BLASTn, BLASTp and ExPASy). Additionally, several biochemical and immunological characteristics of the isolated protein were analysed. By ESI-MS/MS the amino acidic 5G8 sequence was deduced. The 5G8 antigen belongs to the VSP family proteins of G. lamblia . This protein is composed by one polypeptide chain (± 71 kDa). Using the algorithm SYFPHEITI, we identified candidate CD4+ T-cell epitopes from the 5G8 antigen, which can elicit cell-mediated immune responses. In this study, we have identified a G. lamblia protein that induces a strong immune response in infected mice. The biochemical and immunological characterization of the immunogenic 5G8 antigen may contribute to the rational design of a Giardia vaccine.