Limits...
Transcutaneous immunization as preventative and therapeutic regimens to protect against experimental otitis media due to nontypeable Haemophilus influenzae.

Novotny LA, Clements JD, Bakaletz LO - Mucosal Immunol (2011)

Bottom Line: Preventative immunization with NTHI outer membrane protein (OMP) P5- and type IV pilus-targeted immunogens, delivered with the adjuvant LT(R192G-L211A), induced significantly earlier clearance of NTHI from the nasopharynges and middle ears of challenged chinchillas compared with receipt of immunogen or adjuvant alone.Moreover, therapeutic immunization resulted in significant resolution of established NTHI biofilms from the middle ear space of animals compared with controls.These data advocate TCI with the adhesin-directed immunogens as an efficacious regimen for prevention and resolution of experimental NTHI-induced OM.

View Article: PubMed Central - PubMed

Affiliation: The Research Institute at Nationwide Children's Hospital, Department of Pediatrics, Center for Microbial Pathogenesis and The Ohio State University College of Medicine, Columbus, Ohio, USA.

ABSTRACT
We have developed three nontypeable Haemophilus influenzae (NTHI) adhesin-derived immunogens that are significantly efficacious against experimental otitis media (OM) due to NTHI when delivered parenterally. We now expanded our preventative immunization strategies to include transcutaneous immunization (TCI) as a less invasive, but potentially equally efficacious, regimen to prevent OM due to NTHI. Additionally, we examined the potential of TCI as a therapeutic immunization regimen to resolve ongoing experimental OM. Preventative immunization with NTHI outer membrane protein (OMP) P5- and type IV pilus-targeted immunogens, delivered with the adjuvant LT(R192G-L211A), induced significantly earlier clearance of NTHI from the nasopharynges and middle ears of challenged chinchillas compared with receipt of immunogen or adjuvant alone. Moreover, therapeutic immunization resulted in significant resolution of established NTHI biofilms from the middle ear space of animals compared with controls. These data advocate TCI with the adhesin-directed immunogens as an efficacious regimen for prevention and resolution of experimental NTHI-induced OM.

Show MeSH

Related in: MedlinePlus

Migration of dendritic cells (DCs) from the pinnae to the nontypeable Haemophilus influenzae (NALT) after transcutaneous immunization (TCI) and discrimination of cutaneous DC cell type. Presence of CD11c+CFSE+ cells in the NALT after TCI with (a) dmLT alone, (b) rsPilA+dmLT, (c) LB1+dmLT, or (d) chimV4+dmLT was detected by flow cytometry. Numbers in each box represent the percentage of CFSE+ cells within each sample. (e) CFSE+ cells were identified to be DC-SIGN+ dermal DCs. (f) Bone marrow-derived DCs activated with chimV4+dmLT ex vivo and then injected subdermally into pinnae migrated to the NALT, whereas dmLT-activated DCs did not (inset). CFSE, carboxyfluoroscein succinimidyl ester; dmLT, double mutant of E. coli heat-labile enterotoxin; FSC, forward scatter; rsPilA, recombinant soluble PilA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3118858&req=5

fig6: Migration of dendritic cells (DCs) from the pinnae to the nontypeable Haemophilus influenzae (NALT) after transcutaneous immunization (TCI) and discrimination of cutaneous DC cell type. Presence of CD11c+CFSE+ cells in the NALT after TCI with (a) dmLT alone, (b) rsPilA+dmLT, (c) LB1+dmLT, or (d) chimV4+dmLT was detected by flow cytometry. Numbers in each box represent the percentage of CFSE+ cells within each sample. (e) CFSE+ cells were identified to be DC-SIGN+ dermal DCs. (f) Bone marrow-derived DCs activated with chimV4+dmLT ex vivo and then injected subdermally into pinnae migrated to the NALT, whereas dmLT-activated DCs did not (inset). CFSE, carboxyfluoroscein succinimidyl ester; dmLT, double mutant of E. coli heat-labile enterotoxin; FSC, forward scatter; rsPilA, recombinant soluble PilA.

Mentions: Last, we examined the migration of DCs from the pinna after TCI to identify potential sites of immune induction after immunization by this route. To do so, the amine-reactive dye carboxyfluoroscein succinimidyl ester (CFSE) was applied to the pinnae along with each vaccine formulation to allow for discrimination among cells resident within lymphoid tissues and DCs that had migrated to these sites from the pinnae.25, 26 We observed that within 1 h after TCI, only 13.6% of cells isolated from the nasal-associated lymphoid tissue (NALT) of animals immunized with adjuvant alone were CFSE+CD11c+ (Figure 6a). In contrast, TCI with either rsPilA, LB1, or chimV4 each delivered with dmLT resulted in a 2.5- to 5.4-fold relative increase (37.2, 34.4, and 72.9%, respectively) in the percentage of CFSE+ DCs detected within this lymphoid aggregate (Figure 6b–d). CFSE+CD11c+ DCs were also detected in the axillary and brachial lymph nodes, although only 1.1- to 2.0-fold more CFSE+ DCs were observed in lymph nodes after TCI with any immunogen vs. the adjuvant alone (data not shown). No difference in the percentage of CFSE+CD11c+ DCs within the cervical or mediastinal lymph nodes or spleen of animals that received an immunogen and those immunized with dmLT alone was found at this time point. These data suggested that DCs within the pinnae sampled the NTHI adhesin-derived immunogens that had been applied to the surface of the pinnae and were induced to mature. DC maturation resulted in the preferential migration of these cells to primarily the NALT, and also the axillary and brachial lymph nodes.


Transcutaneous immunization as preventative and therapeutic regimens to protect against experimental otitis media due to nontypeable Haemophilus influenzae.

Novotny LA, Clements JD, Bakaletz LO - Mucosal Immunol (2011)

Migration of dendritic cells (DCs) from the pinnae to the nontypeable Haemophilus influenzae (NALT) after transcutaneous immunization (TCI) and discrimination of cutaneous DC cell type. Presence of CD11c+CFSE+ cells in the NALT after TCI with (a) dmLT alone, (b) rsPilA+dmLT, (c) LB1+dmLT, or (d) chimV4+dmLT was detected by flow cytometry. Numbers in each box represent the percentage of CFSE+ cells within each sample. (e) CFSE+ cells were identified to be DC-SIGN+ dermal DCs. (f) Bone marrow-derived DCs activated with chimV4+dmLT ex vivo and then injected subdermally into pinnae migrated to the NALT, whereas dmLT-activated DCs did not (inset). CFSE, carboxyfluoroscein succinimidyl ester; dmLT, double mutant of E. coli heat-labile enterotoxin; FSC, forward scatter; rsPilA, recombinant soluble PilA.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3118858&req=5

fig6: Migration of dendritic cells (DCs) from the pinnae to the nontypeable Haemophilus influenzae (NALT) after transcutaneous immunization (TCI) and discrimination of cutaneous DC cell type. Presence of CD11c+CFSE+ cells in the NALT after TCI with (a) dmLT alone, (b) rsPilA+dmLT, (c) LB1+dmLT, or (d) chimV4+dmLT was detected by flow cytometry. Numbers in each box represent the percentage of CFSE+ cells within each sample. (e) CFSE+ cells were identified to be DC-SIGN+ dermal DCs. (f) Bone marrow-derived DCs activated with chimV4+dmLT ex vivo and then injected subdermally into pinnae migrated to the NALT, whereas dmLT-activated DCs did not (inset). CFSE, carboxyfluoroscein succinimidyl ester; dmLT, double mutant of E. coli heat-labile enterotoxin; FSC, forward scatter; rsPilA, recombinant soluble PilA.
Mentions: Last, we examined the migration of DCs from the pinna after TCI to identify potential sites of immune induction after immunization by this route. To do so, the amine-reactive dye carboxyfluoroscein succinimidyl ester (CFSE) was applied to the pinnae along with each vaccine formulation to allow for discrimination among cells resident within lymphoid tissues and DCs that had migrated to these sites from the pinnae.25, 26 We observed that within 1 h after TCI, only 13.6% of cells isolated from the nasal-associated lymphoid tissue (NALT) of animals immunized with adjuvant alone were CFSE+CD11c+ (Figure 6a). In contrast, TCI with either rsPilA, LB1, or chimV4 each delivered with dmLT resulted in a 2.5- to 5.4-fold relative increase (37.2, 34.4, and 72.9%, respectively) in the percentage of CFSE+ DCs detected within this lymphoid aggregate (Figure 6b–d). CFSE+CD11c+ DCs were also detected in the axillary and brachial lymph nodes, although only 1.1- to 2.0-fold more CFSE+ DCs were observed in lymph nodes after TCI with any immunogen vs. the adjuvant alone (data not shown). No difference in the percentage of CFSE+CD11c+ DCs within the cervical or mediastinal lymph nodes or spleen of animals that received an immunogen and those immunized with dmLT alone was found at this time point. These data suggested that DCs within the pinnae sampled the NTHI adhesin-derived immunogens that had been applied to the surface of the pinnae and were induced to mature. DC maturation resulted in the preferential migration of these cells to primarily the NALT, and also the axillary and brachial lymph nodes.

Bottom Line: Preventative immunization with NTHI outer membrane protein (OMP) P5- and type IV pilus-targeted immunogens, delivered with the adjuvant LT(R192G-L211A), induced significantly earlier clearance of NTHI from the nasopharynges and middle ears of challenged chinchillas compared with receipt of immunogen or adjuvant alone.Moreover, therapeutic immunization resulted in significant resolution of established NTHI biofilms from the middle ear space of animals compared with controls.These data advocate TCI with the adhesin-directed immunogens as an efficacious regimen for prevention and resolution of experimental NTHI-induced OM.

View Article: PubMed Central - PubMed

Affiliation: The Research Institute at Nationwide Children's Hospital, Department of Pediatrics, Center for Microbial Pathogenesis and The Ohio State University College of Medicine, Columbus, Ohio, USA.

ABSTRACT
We have developed three nontypeable Haemophilus influenzae (NTHI) adhesin-derived immunogens that are significantly efficacious against experimental otitis media (OM) due to NTHI when delivered parenterally. We now expanded our preventative immunization strategies to include transcutaneous immunization (TCI) as a less invasive, but potentially equally efficacious, regimen to prevent OM due to NTHI. Additionally, we examined the potential of TCI as a therapeutic immunization regimen to resolve ongoing experimental OM. Preventative immunization with NTHI outer membrane protein (OMP) P5- and type IV pilus-targeted immunogens, delivered with the adjuvant LT(R192G-L211A), induced significantly earlier clearance of NTHI from the nasopharynges and middle ears of challenged chinchillas compared with receipt of immunogen or adjuvant alone. Moreover, therapeutic immunization resulted in significant resolution of established NTHI biofilms from the middle ear space of animals compared with controls. These data advocate TCI with the adhesin-directed immunogens as an efficacious regimen for prevention and resolution of experimental NTHI-induced OM.

Show MeSH
Related in: MedlinePlus