Allergy Asthma Immunol Res.  2014 Jul;6(4):341-349. 10.4168/aair.2014.6.4.341.

Perinatal Immunization With Vaccine-Grade Listeria monocytogenes Provides Protection Against Murine Th2 Airway Inflammation

• Aloyouni S
• Segeritz C
• Sherrid AM
• Gold MJ
• Loeffler D
• Blanchet M
• Cai B
• Hirota J
• McNagny KM
• Kollmann T

• KMID: 2260215
• DOI: http://doi.org/10.4168/aair.2014.6.4.341

Abstract

PURPOSE
Asthma is a chronic respiratory disorder that leads to inflammation and narrowing of the airways. Its global prevalence has attained epidemic levels and treatment options that reach beyond temporary relief of symptoms are urgently needed. Since the processes leading to clinically symptomatic asthma start early in life, we set out to systematically evaluate a neonatal immunotherapeutic based on Listeria monocytogenes (Lm) for the control of allergic sensitization.
METHODS
We modified Lm to express the model allergen, ovalbumin (OVA), and tested the ability of neonatal immunization with this strain to control allergic sensitization in a mouse model of OVA-induced asthma. Mice were immunized as newborns with live or heat killed LmOVA or live Lm, followed 6 weeks later by allergic sensitization with OVA. In order to determine whether the T(H)1-polarizing effect of this vaccine vector inadvertently may exacerbate development of certain T(H)1-driven allergic diseases, mice immunized as newborns were assessed in a model of adult hypersensitivity pneumonitis (HP).
RESULTS
Both LmOVA and Lm-control vaccines were highly effective in providing long-lasting protection from airway inflammation after only one immunization given perinatally. Serum antibody levels and lung cytokine production suggest that this prophylactic strategy is associated with an allergen specific T(H)1-dominated response. Specifically, LmOVA vaccinated mice displayed significantly elevated OVA-specific serum IgG2a, but no difference in anti-OVA IgE antibodies and only slightly decreased anti-OVA IgG1 antibodies. Importantly, Lm-based neonatal vaccination did not exacerbate Th1/Th17 driven HP, arguing against broad spectrum immune skewing.
CONCLUSIONS
Our findings highlight the promise of early life Lm-based immunomodulatory interventions as a prophylactic strategy for allergic asthma.

Keyword

Asthma; vaccines; listeria monocytogenes; immune system; newborn; Extrinsic allergic

MeSH Terms

Adult
Alveolitis, Extrinsic Allergic
Animals
Antibodies
Asthma
Hot Temperature
Humans
Immune System
Immunization*
Immunoglobulin E
Immunoglobulin G
Infant, Newborn
Inflammation*
Listeria monocytogenes*
Lung
Mice
Ovalbumin
Ovum
Prevalence
Vaccination
Vaccines
Antibodies
Immunoglobulin E
Immunoglobulin G
Ovalbumin
Vaccines

Figure

• 

  Fig. 1 Vaccination with Lm and LmOVA significantly decreases Th2 driven allergic airway inflammation. (A) Schematic of Lm vaccination and experimental allergic airway inflammation protocol. (B) Total cell counts in BALF from naïve and OVA challenged mice. Naïve=saline vaccination and no subsequent OVA challenge, Ctrl=saline vaccination and OVA challenge, Lm=Listeria vaccination and OVA challenge, LmOVA=Listeria-OVA vaccination and OVA challenge, HKLmOVA=heat-killed LmOVA vaccination followed by OVA challenge. (C) Differential cell counts in BALF. (D) Relative concentration of OVA-specific IgG2a. Serum was diluted 1:1,000 and then serial dilutions were evaluated for OVA-specific antibody reactivity by ELISA as described in Materials and Methods. Values are expressed as mean±SEM. (**P value≤0.01, ***P value≤0.001). The number of mice per group per experiment was 3-5.

• 

  Fig. 2 Inflammatory lung cell cytokine responses are shifted by neonatal prophylactic LmOVA vaccination. Single cell suspensions were prepared from lungs, placed in culture and cytokine production was examined 48 hours after restimulation using 500 µg/mL of OVA. Cytokine production was determined by the 6-Milliplex Mouse Cytokine/Chemokine Immunoassay. (A) IFN-γ production. (B) IL-5 production. (C) IL-10 production. Values are expressed as mean±SEM. (*P value ≤0.05, ***P value ≤0.001). The number of mice per group per experiment was 3-5.

• 

  Fig. 3 Neonatal prophylactic immunization with Lm and LmOVA prevents significant increase in airway hyperresponsiveness (AHR) to methacholine challenge. (A) Mice were immunized and challenged as described in Fig. 1 and administered increasing doses of MCh intravenously. Airway resistance (R % increase) was measured for each dose of MCh. (B) Data from (A) re-evaluated as area under the curve. Values are expressed as mean±SEM. The number of mice per group was 8.

• 

  Fig. 4 Neonatal Lm vaccination does not significantly alter sensitivity to Th1/17 driven Hypersensitivity Pneumonitis (HP). (A) Schematic of vaccination and Saccharopolyspora rectivirgula (SR)-induced HP induction protocol. (B) Total cell counts in BALF from naïve and SR-challenged mice were determined as shown in Fig. 1. (C) Differential cell counts in BALF in response to HP induction. (D), (E), (F) Clinical scores for inflammatory infiltrates attributed to airways, vessels, and lung parenchyma respectively. Lungs were fixed and stained with H&E prior to blind scoring as described in Materials and Methods. Values are expressed as mean±SEM, The number of mice per group was 6-14.

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