Transfection efficiency and cytotoxicity of cationic liposomes in primary cultures of rainbow trout (Oncorhynchus mykiss) gill cells.
Immunisation of fish by immersion has been applied for inactivated, whole cell bacterins, where the gill epithelial cells are considered as one of the prime uptake sites. Antigen entry is a critical factor for delivery of vaccine antigens through the immersion route, also for DNA vaccines, and delivery systems like cationic liposomes may enhance uptake. In this study, the aim was to examine the efficiency of cationic liposomes as a means to transfect primary cultures of rainbow trout gill cells with plasmids encoding viral or reporter proteins. Furthermore, the effects of the concentration and composition of liposomes/lipoplex on the viability of the cells were evaluated. Transfection of the gill cells was possible with both plasmids following transfection with lipoplexes of a neutral charge. Low concentrations and neutral/negatively charged formulations were favourable with respect to the toxicity of the formulations. Given that the mucous barrier covering the gills is overcome, this system might be useful for the priming of the local immunity in the fish gills.
Romøren
Kristine
K
Department of Basic Sciences and Aquatic Medicine, Norwegian School of Veterinary Science, Oslo.
Fjeld
Xuan T L
XT
Poléo
Antonio B S
AB
Smistad
Gro
G
Thu
Beate J
BJ
Evensen
Øystein
Ø
D000818:Animals; D002412:Cations; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D017930:Genes, Reporter; D005880:Gills / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D008081:Liposomes / Q000378:metabolism* / Q000633:toxicity*; D017686:Oncorhynchus mykiss; D010957:Plasmids / Q000235:genetics; D014162:Transfection / Q000379:methods*
D002412:Cations; D008081:Liposomes