Lactococcus lactis as an adjuvant and delivery vehicle of antigens against pneumococcal respiratory infections

Figures & data

Figure 1 Innate immune response in lung. Alveolar macrophages constitute the first line of phagocytic defense against infectious agents and play a prominent role in lung immunity by initiating inflammation and immune responses. In the event that invading pathogens are too virulent or represent too large a load to be contained by macrophages alone, alveolar macrophages are capable of generating mediators that orchestrate the recruitment of large numbers of neutrophils from the pulmonary vasculature into the alveolar space. These neutrophils provide auxiliary phagocytic capacities that play a critical role in host defense against pathogens.

Figure 2 Antigen uptake and migratory pattern induction in the airways. The production of specific IgA in the respiratory tract during an infectious process is very important because it prevents colonization of mucosal sites and subsequent spreading into the systemic circulation.

Figure 3 Antigen uptake and migratory pattern for immune induction in deep lung. IgG production plays an important role in the protection against pathogens that reach the alveolar space. Opsonizing IgG antibodies are important for complement fixation and for improvement in the efficiency of macrophages killing. At the systemic level, immune activation also induces production of antibodies responsible for preventing the passage of pathogens to the blood.

Figure 4 Schematic representation of the nisin-controlled gene expression (NICE) system (40) used to express the recombinant PppA pneumococcal protein in L. lactis. L. lactis NZ9000 cells containing the signal transduction genes nisK and nisR in their chromosome were transformed with a plasmid carrying the pppA gene under the control of the inducible promoter P_nisA. Expression of the PppA recombinant protein was induced by the addition of sub-inhibitory amounts of the nisin and its insertion in the cell wall was mediated by the secretion (sec) and cell wall (CW) anchoring signals.

Table 1 Lactic acid bacteria as vaccine vehicles against S. pneumoniae

Antigen	Amplified from	LAB strain (recombinant)	Expresión system	Localization	Route Immunization	Type immune response	Protection	Ref.
Proteins
PspA	T1GR4	L. lactis F17847	pTnis- inducible with nisin	Citoplasm	Nasal Live and Inactivated vaccine	Live vaccine: Th1-type (IgG1:IgG2a = 3,49) Inactivated vaccine:Th2-type 1 response (higher IgG1:IgG2a ratio)	Significant protection in sepsis and respiratory infection models	Hanniffy et al, 2007
	Strain 245/00 Serotype 14 clade 1 PspA	Lb. casei CECT5275	pT1NX-constitutive expression from lactococcal promoter P1	Citoplasm		Nasal/IgG (mainly IgG1) but not IgA detected. Anti-PspA induces complement deposition	Increase survival of PspA-immunized mice against intraperitoneal challenge	Campos et al, 200895
PsaA	Strain 472/96 serotype 6B	Lb. casei CECT5275 Lb. plantarum NCD01193 Lb. helveticus ATCC15009 L. lactis MG1363	pT1NX-constitutive expression from lactococcal promoter P1	Citoplasm and cell wall	Nasal	IgG (serum) and IgA in saliva, nasal and bronchial washes (except Lb casei)	Reduced recovery of S. pneumoniae from nasal mucose in immunized animals after S. pneumoniae challenge.	Oliveira et al, 2006
PppA	T14, serotype 14	L. lactis NZ9000	pVE5547, inducible with nisin	Cell wall	Nasal and oral/Live (L) vaccine	Local (IgA, IgG) and systemic responses (IgG)	L vaccine: Improved resistance to systemic and respiratory pneumococcal challenge, both in active and passive immunized mice. Cross protection against serotypes 3, 6b and 23F D vaccine: Improved resistance to respiratory challenge.	Medina et al, 2008; Villena et al, 2008; Vintini et al, 2009
					Nasal/Inactivated (D) vaccine
PpmA	D39	GEM (based on hot TCA-treated non-living L. lactis NZ9000)	L. lactis PA1001a(pPA32)-Nisin inducible; expressed as PpmA-PA	Cell wall	Nasal	Systemic and mucosal	NO (only in combination with IgAlp-PA and SlrA-PA)	Audouy et al, 2006; Audouy et al, 2007
SlrA	D39	GEM (based on hot TCA-treated nonliving L. lactis NZ9000)	L. lactis PA 1001 (pPA162)-Nisin inducible; expressed as Slra-PA	Cell wall	Nasal	Systemic and mucosal	NO (only with IgAlp-PA and PpmA-PA or in combination with IgAlp-PA)	Audouy et al, 2006; Audouy et al, 2007
IgAlp	TIGR4	GEM (based on hot TCA-treated nonliving L. lactis NZ9000)	L. lactis PA1001 (pPA152)-Nisin inducible; expressed as IgAlp-PA	Cell wall	Nasal	Systemic and mucosal (lower levels with regard to PpmA-Pa and SlrA-PA)	NO (only with PpmA-PA and SlrA-PA or in combination with SrlA-PA)	Audouy et al, 2006; Audouy et al, 2007
Polysaccharides
Serotype 3	S. penumoniae WU2	L. lactis MG1363	Constitutive expression from original promoter	Mainly associated with cells	Intraperitoneal	T-cell independent response (IgM, Ig1 and Ig3)	n.d.	Gilbert et al, 2000
Serotype 14	“serotype 14” strain	L. lactis NZ9000	Two-plasmids system. Plasmid with regulatory genes inducible with nisin	Mainly supernatant	n.d.	n.d	n.d.	Nierop Groot et al, 2008

a a MG1363 (acmA⁻, htrA⁻, nisRK⁺) strain, lacking the AcmA cell-wall hydrolase activity and the housekeeping protease HtrA (which has been shown to degrade the AcmA cell-wall binding domain). The NisRK genes are required for nisin induced expression. n.d., not done.

Campos IB, Darrieux M, Ferreira DM, Miyaji EN, Silva DA, Áreas PAM, et al. Nasal immunization of mice with Lactobacillus casei expressing the pneumococcal surface protein A: induction of antibodies, complement deposition and partial protection against Streptococcus pneumoniae challenge. Microbes Infect 2008; 10:481 - 488

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