Evaluation of the reactogenicity, adjuvanticity and antigenicity of LT(R192G) and LT(R192G/L211A) by intradermal immunization in mice


Autoři: Milton Maciel, Jr. aff001;  Mark Smith aff004;  Steven T. Poole aff001;  Renee M. Laird aff001;  Julianne E. Rollenhagen aff001;  Robert W. Kaminski aff004;  Heather Wenzel aff005;  A. Louis Bourgeois aff005;  Stephen J. Savarino aff003
Působiště autorů: Henry M. Jackson Foundation for the Advancement of Military Medicine, Bethesda, MD, United States of America aff001;  Department of Microbiology and Immunology, Uniformed Services University of the Health Sciences, Bethesda, MD, United States of America aff002;  Enteric Diseases Department, Naval Medical Research Center, Silver Spring, MD, United States of America aff003;  Subunit Enteric Vaccines and Immunology, Walter Reed Army Institute of Research, Silver Spring, MD, United States of America aff004;  PATH, Washington, D.C., United States of America aff005;  Department of Pediatrics, Uniformed Services University of the Health Sciences, Bethesda, MD, United States of America aff006
Vyšlo v časopise: PLoS ONE 14(11)
Kategorie: Research Article
doi: 10.1371/journal.pone.0224073

Souhrn

The development of an effective subunit vaccine is frequently complicated by the difficulty of eliciting protective immune responses, often requiring the co-administration of an adjuvant. Heat-labile toxin (LT), an enterotoxin expressed by enterotoxigenic E. coli (ETEC) with an AB5 structure similar to cholera toxin, is a strong adjuvant. While the mucosa represents the natural route of exposure to LT and related toxins, the clinical utility of LT and similar adjuvants given by mucosal routes has been limited by toxicity, as well as the association between intranasal delivery of LT and Bell’s palsy. Single and double amino acid mutants of LT, LT(R192G)/mLT and LT(R192G/L211A)/dmLT respectively, have been proposed as alternatives to reduce the toxicity associated with the holotoxin. In the present study, we compared mLT and dmLT given via a non-mucosal route (i.e. intradermally) to investigate their adjuvanticity when co-administrated with an enterotoxigenic E. coli vaccine candidate, CfaEB. Antigenicity (i.e. ability to elicit response against LT) and reactogenicity at the injection site were also evaluated. BALB/c mice were immunized by the intradermal route with CfaEB plus increasing doses of either mLT or dmLT (0.01 to 2.5 μg). Both adjuvants induced dose-dependent skin reactogenicity, with dmLT being less reactogenic than mLT. Both adjuvants significantly boosted the anti-CfaE IgG and functional hemagglutination inhibiting (HAI) antibody responses, compared to the antigen alone. In addition to inducing anti-LT responses, even at the lowest dose tested (0.01 μg), the adjuvants also prompted in vitro cytokine responses (IFN-γ, IL-4, IL-5, IL-10 and IL-17) that followed different patterns, depending on the protein used for stimulation (CfaE or LTB) and/or the dose used for immunization. The two LT mutants evaluated here, mLT and dmLT, are potent adjuvants for intradermal immunization and should be further investigated for the intradermal delivery of subunit ETEC vaccines.

Klíčová slova:

Antibodies – Antigens – Cytokines – Immune response – Immunologic adjuvants – Vaccines – Erythema


Zdroje

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2019 Číslo 11