Citation

  • Authors: Smitha, S., Raina, O. K., Singh, B. P., Samanta, S., Velusamy, R., Dangoudoubiyam, S., Tripathi, A., Gupta, P. K., Sharma, B., Saxena, M.
  • Year: 2010
  • Journal: J Helminthol 84 149-55
  • Applications: in vivo / DNA / in vivo-jetPEI-Man

Method

50 µg DNA at N/P=5 were injected IP for immunisation studies. Each mouse was administered DNA twice at a 3-week interval before a final booster was given, with either recombinant fatty acid binding protein or plasmid DNA. Mice in each group were divided into two subgroups of five mice after three DNA immunizations.

Abstract

Fasciola gigantica fatty acid binding protein (FABP) was evaluated for evoking an immune response in mice, by delivering the gene coding for this protein with mannosylated-polyethylenimine (PEI) to peritoneal cells. Mice were immunized with 50 microg recombinant plasmid DNA (Group I) or DNA-PEI-mannose (a 22 kDa linear cationic polymer with mannose ligand) (Group II) via the intraperitoneal route. Antibody studies showed no significant humoral immune response evoked to this DNA immunization with either PEI-mannose-delivered or naked DNA. However, on protein boosting of these DNA-primed mice there was a significant enhancement of antibody titre. Flow cytometric bead array was used to measure quantities of interleukin (IL)-2, IL-4, IL-5, interferon-gamma (IFN-gamma) and tumour necrosis factor (TNF) cytokines. Overexpression of T-helper 1 (Th1) cytokines such as IFN-gamma and TNF, with a lower but significant expression of the T-helper 2 (Th2) cytokine IL-5 was detected. Gene delivery using polyethylenimine-mannose ligand showed significant expression of IFN-gamma and TNF (P < 0.05), but no significant difference in IL-2, IL-4 and IL-5 (P>0.05) cytokine expression was observed between naked-DNA- and mannosylated PEI-DNA-delivered mice. Naked- or PEI-delivered-DNA immunization produced insignificant levels of IL-2 and IL-4 (P>0.05) cytokines in both groups of mice.

Pubmed