title: A Novel Polyclonal Antiserum against Toxoplasma gondii Sodium Hydrogen Exchanger 1.
abstract: The sodium hydrogen exchanger 1 (NHE1), which functions in maintaining the ratio of Na(+) and H(+) ions, is widely distributed in cell plasma membranes. It plays a prominent role in pH balancing, cell proliferation, differentiation, adhesion, and migration. However, its exact subcellular location and biological functions in Toxoplasma gondii are largely unclear. In this study, we cloned the C-terminal sequence of T. gondii NHE1 (TgNHE1) incorporating the C-terminal peptide of NHE1 (C-NHE1) into the pGEX4T-1 expression plasmid. The peptide sequence was predicted to have good antigenicity based on the information obtained from an immune epitope database. After induction of heterologous gene expression with isopropyl-b-D-thiogalactoside, the recombinant C-NHE1 protein successfully expressed in a soluble form was purified by glutathione sepharose beads as an immunogen for production of a rabbit polyclonal antiserum. The specificity of this antiserum was confirmed by western blotting and immunofluorescence. The antiserum could reduce T. gondii invasion into host cells, indicated by the decreased TgNHE1 expression in T. gondii parasites that were pre-incubated with antiserum in the process of cell entry. Furthermore, the antiserum reduced the virulence of T. gondii parasites to host cells in vitro, possibly by blocking the release of Ca(2+). In this regard, this antiserum has potential to be a valuable tool for further studies of TgNHE1.
authors.affiliation: State Key Laboratory of Organ Failure, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Biotechnology, Southern Medical University, Guangzhou 510515, People's Republic of China.
authors.forename: Bin
authors.initials: B
authors.lastname: Xiao
authors.affiliation: State Key Laboratory of Organ Failure, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Biotechnology, Southern Medical University, Guangzhou 510515, People's Republic of China.
authors.forename: Zhenzhan
authors.initials: Z
authors.lastname: Kuang
authors.affiliation: State Key Laboratory of Organ Failure, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Biotechnology, Southern Medical University, Guangzhou 510515, People's Republic of China.
authors.forename: Yanli
authors.initials: Y
authors.lastname: Zhan
authors.affiliation: State Key Laboratory of Organ Failure, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Biotechnology, Southern Medical University, Guangzhou 510515, People's Republic of China.
authors.forename: Daxiang
authors.initials: D
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Organ Failure, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Biotechnology, Southern Medical University, Guangzhou 510515, People's Republic of China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Gao
authors.affiliation: State Key Laboratory of Organ Failure, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Biotechnology, Southern Medical University, Guangzhou 510515, People's Republic of China.
authors.forename: Ming
authors.initials: M
authors.lastname: Li
authors.affiliation: State Key Laboratory of Organ Failure, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Biotechnology, Southern Medical University, Guangzhou 510515, People's Republic of China.
authors.forename: Shuhong
authors.initials: S
authors.lastname: Luo
authors.affiliation: State Key Laboratory of Organ Failure, Guangdong Provincial Key Laboratory of Tropical Disease Research, School of Biotechnology, Southern Medical University, Guangzhou 510515, People's Republic of China.
authors.forename: Wenbo
authors.initials: W
authors.lastname: Hao
mesh_terms: D000818:Animals; D002460:Cell Line; D007106:Immune Sera / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D008297:Male; D051379:Mice; D015800:Protozoan Proteins / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D011994:Recombinant Proteins / Q000276:immunology; D012756:Sheep; D017923:Sodium-Hydrogen Exchangers / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014122:Toxoplasma / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D014123:Toxoplasmosis / Q000469:parasitology / Q000517:prevention & control
chemical_list: D007106:Immune Sera; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D017923:Sodium-Hydrogen Exchangers
keywords: Ca2+ release; TgNHE1; Toxoplasma gondii; polyclonal antiserum; soluble expression