Immune responses to the expressed products of the CSP antigen gene of Plasmodium falciparum southern China isolate FCC1/HN in Hela cell
10.3760/j.issn:0366-6999.2001.08.015
- VernacularTitle:恶性疟原虫海南FCC1/HN株CSP抗CSP抗原基因在Hela细胞中表达产物的免疫应答研究
- Author:
XinBing YU
1
;
Yanwen LIU
;
SH LUO
;
Zheng FANG
Author Information
1. 南通医学院
- Keywords:
plasmodium falciparum;
circumsporozoite protein;
Hela cell;
expressed product;
immune response
- From:
Chinese Medical Journal
2001;114(8):844-847
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a eukaryotic expression system with pcDNA3-PfCSP/Hela for the Circumsporozoite protein (CSP) gene of Plasmodium falciparum (P. falciparum), to observe the immune responses in BALB/c mice induced by the expressed proteins. Methods The recombinant plasmid pcDNA3-PfCSP was transformed into the Hela cell line. The expressed protein was isolated and analyzed by using SDS-PAGE and used for immunization of BALB/c mice by subcutaneous, intravenous, and intraperitoneal adminstration. Enzyme-linked immunosorbent assay(ELISA), Dot-ELISA, Western blot, T lymphocyte proliferation test, natural killer cell(NKC) activity assay, and CD4+ and CD8+ T cell detection were used for observation of humoral and cellular immune responses. Results Immune sera strongly reacted with the expressed protein, antibody titer was up to 1∶6400 as detected by ELISA. Western blot analysis revealed a specific band at 38.3?Kda. When the spleen cells of normal and immunized BALB/c mice were specifically stimulated with expressed protein, the optical densities were 0.12±0.03 and 0.34±0.04, respectively. The latter were significantly higher than the former (P<0.01). We used the MTT colorimetric assay to measure NKC activity of mice spleen. The results showed that the NKC activity of immunized BALB/c mice was remarkably higher than that of the controls (P<0.05). CD4+ and CD8+ T cells were detected by using monoclonal antibody immunofluorescence methods. The results showed that the percentage of CD4+ and CD8+ T cells of immunized group were significantly higher than that of control group (P<0.05). Conclusions The humoral and cell-mediated immune responses and elevated NKC activity to products made with a eukaryotic expression system could be specifically detected in BALB/c mice. These findings indicate that the expressed protein could enhance the immune function in mice.
