considered the model organism and has been explored as a vehicle
for the delivery of therapeutic molecules and antigens in the
mucosa [4, 6].
DNA vaccines consist of the administration of a bacterial plasmid under the control of a eukaryotic promoter encoding the
antigen of interest, which is then synthesized by the vaccinated
individuals themselves and presented to the immune system
[7]. These proteins are capable of stimulating the immune system,
becoming a promising technique of immunization against a variety
of pathogens and tumors [7, 8]. DNA vaccines have several advantages over other vaccination strategies such as the ability to induce
humoral and cellular immunity, longer shelf life, ease of manufacture, storage, transport and rapid production, besides the gene
vaccines are highly stable at room temperature and do not require
refrigeration [7, 8].
There are many forms for delivery of DNA vaccine: bioballistic
delivery through the skin, which is a method of epidermal DNA
delivery where Langerhans and keratinocyte cells are directly transfected by bombardment of gold particles coated with DNA plasmids [9, 10]; electroporation technique (EP) based on the
application of high voltage pulses in a tissue that generates pores
in the cell membrane, allowing the DNA vaccine to enter the
cytoplasm of cells, leading to an inflammatory process [11]; and
the bacterial delivery presented above.
Intramuscular administration is the most common form used;
it consists of the optimized gene sequence of interest using naked
DNA delivery. However, due to its poor distribution in the body
and rapid degradation, it requires higher amounts of plasmid
[12, 13]. As the mucosal surfaces are vulnerable to infection by
pathogenic microorganisms, the local mucosal immune responses
are important for protection against different diseases [14]. Thus,
the mucosal route of administration has been the most widely
explored. Among the mucosal administration routes, the oral,
nasal, sublingual, anal, and vaginal routes are the most
commonly used.
This chapter will describe protocols for the preparation and
mucosal administration (nasal, intragastric, and sublingual) in mice
of a DNA vaccine using Lactococcus lactis as a delivery vehicle.
2 Materials
Prepare all solutions using distilled or sterile water. All culture
medium must be sterilized by autoclavation (121
C/15 psi/
15 min), and the glucose solution must be filtered (0.22 μm filter)
(see Note 1).
448
Viviane Lima Batista et al.
for the delivery of therapeutic molecules and antigens in the
mucosa [4, 6].
DNA vaccines consist of the administration of a bacterial plasmid under the control of a eukaryotic promoter encoding the
antigen of interest, which is then synthesized by the vaccinated
individuals themselves and presented to the immune system
[7]. These proteins are capable of stimulating the immune system,
becoming a promising technique of immunization against a variety
of pathogens and tumors [7, 8]. DNA vaccines have several advantages over other vaccination strategies such as the ability to induce
humoral and cellular immunity, longer shelf life, ease of manufacture, storage, transport and rapid production, besides the gene
vaccines are highly stable at room temperature and do not require
refrigeration [7, 8].
There are many forms for delivery of DNA vaccine: bioballistic
delivery through the skin, which is a method of epidermal DNA
delivery where Langerhans and keratinocyte cells are directly transfected by bombardment of gold particles coated with DNA plasmids [9, 10]; electroporation technique (EP) based on the
application of high voltage pulses in a tissue that generates pores
in the cell membrane, allowing the DNA vaccine to enter the
cytoplasm of cells, leading to an inflammatory process [11]; and
the bacterial delivery presented above.
Intramuscular administration is the most common form used;
it consists of the optimized gene sequence of interest using naked
DNA delivery. However, due to its poor distribution in the body
and rapid degradation, it requires higher amounts of plasmid
[12, 13]. As the mucosal surfaces are vulnerable to infection by
pathogenic microorganisms, the local mucosal immune responses
are important for protection against different diseases [14]. Thus,
the mucosal route of administration has been the most widely
explored. Among the mucosal administration routes, the oral,
nasal, sublingual, anal, and vaginal routes are the most
commonly used.
This chapter will describe protocols for the preparation and
mucosal administration (nasal, intragastric, and sublingual) in mice
of a DNA vaccine using Lactococcus lactis as a delivery vehicle.
2 Materials
Prepare all solutions using distilled or sterile water. All culture
medium must be sterilized by autoclavation (121
C/15 psi/
15 min), and the glucose solution must be filtered (0.22 μm filter)
(see Note 1).
448
Viviane Lima Batista et al.