Immunofluorescence to Monitor the Cellular Uptake of Human Lactoferrin and its Associated Antiviral Activity Against the Hepatitis C Virus.
Immunofluorescence is a laboratory technique commonly used to study many aspects of biology. It is typically used to visualize the distribution and/or localization of a target molecule in cells and tissues. Immunofluorescence relies on the specificity of fluorescent-labelled antibodies against their corresponding antigens within a cell. Both direct and indirect immunofluorescence approaches can be used which rely on the use of antibodies linked with a fluorochrome. Direct immunofluorescence is less frequently used because it provides lower signal, involves higher cost and less flexibility. In contrast, indirect immunofluorescence is more commonly used because of its high sensitivity and provides an amplified signal since more than one secondary antibody can attach to each primary antibody. In this manuscript, both epifluorescence microscopy and confocal microscopy were used to monitor the internalization of human lactoferrin, an important component of the immune system, into hepatic cells. Moreover, we monitored the inhibitory potential of hLF on the intracellular replication of the Hepatitis C virus using immunofluorescence. Both the advantages and disadvantages associated with these approaches are discussed.
Département de biochimie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke.
Andréa
A
Allaire
Département de biochimie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke.
Frédéric
F
Picard-Jean
Département de biochimie, Faculté de médecine et des sciences de la santé, Université de Sherbrooke; Martin.Bisaillon@usherbrooke.ca.
Martin
M
Bisaillon
D000998:Antiviral Agents / Q000493:pharmacokinetics / Q000494:pharmacology*; D045744:Cell Line, Tumor; D019085:Fluorescent Antibody Technique, Direct / Q000379:methods*; D019084:Fluorescent Antibody Technique, Indirect / Q000379:methods*; D016174:Hepacivirus / Q000187:drug effects* / Q000502:physiology; D019698:Hepatitis C, Chronic / Q000188:drug therapy / Q000378:metabolism / Q000821:virology; D022781:Hepatocytes / Q000378:metabolism; D006801:Humans; D007781:Lactoferrin / Q000493:pharmacokinetics / Q000494:pharmacology*; D008099:Liver / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods; D008856:Microscopy, Fluorescence / Q000379:methods; D014779:Virus Replication / Q000187:drug effects
D000998:Antiviral Agents; C490098:LTF protein, human; D007781:Lactoferrin