Preformulation studies with the Escherichia coli double mutant heat-labile toxin adjuvant for use in an oral vaccine

White, Jessica A.; Haghighi, Candace; Brunner, Johanna; Estrada, Marcus; Lal, Manjari; Chen, Dexiang

doi:10.1016/j.jim.2017.09.003

Cited by 13 publications

(11 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…coli strain JM83(pLC326) at Walter Reed Army Institute of Research (WRAIR) Pilot Bioproduction Facility (BPR 466–00 lot 0816, Forest Glen Annex, Silver Spring MD). The Escherichia coli double-mutant heat-labile toxin [LT(R192G/L211A), hereafter dmLT] was produced at WRAIR Pilot Bioproduction Facility (BPR-1037-00, Lot #1735), following previously described affinity-chromatography methods [26], and obtained through PATH (Washington D.C.). Since we and others [26] have previously observed that low concentrations of mLT and dmLT tend to get absorbed to polypropylene and borosilicate vials, doses were prepared in Daikyo Crystal Zenith® vials (West Pharmaceutical Services, Exton, PA).…”

Section: Methodsmentioning

confidence: 99%

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

et al. 2019

View full text Add to dashboard Cite

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.

show abstract

Section: Methodsmentioning

confidence: 99%

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

et al. 2019

View full text Add to dashboard Cite

show abstract

“…The consensus of these studies is that dmLT is uniquely able to induce systemic and mucosal responses after parenteral or mucosal immunization. The success of these studies has also led to recent preformulation studies evaluating buffers to promote stability and freeze-dried formulations to maximize dmLT adjuvant and/or antigen responses ( 66 , 67 ). dmLT has also been used in immunologic studies for mucosal vaccination in order to evaluate how immunity is generated or impacted by infections ( 68 , 69 ).…”

Section: Preclinical Studiesmentioning

confidence: 99%

The Mucosal Vaccine Adjuvant LT(R192G/L211A) or dmLT

Clements

Norton

2018

mSphere

136

128

View full text Add to dashboard Cite

show abstract

“…Besides, it has been shown to be safe in mice and humans, at least when administered orally [ 58 , 59 ]. This modified toxin has been used successfully as an adjuvant in numerous studies and with different antigens and immunization routes [ 60 – 67 ].…”

Section: Discussionmentioning

confidence: 99%

Rotavirus VP6 protein mucosally delivered by cell wall-derived particles from Lactococcus lactis induces protection against infection in a murine model

Temprana¹,

Argüelles²,

Gutierrez³

et al. 2018

PLoS ONE

View full text Add to dashboard Cite

Rotaviruses are the primary cause of acute gastroenteritis in children worldwide. Although the implementation of live attenuated vaccines has reduced the number of rotavirus-associated deaths, variance in their effectiveness has been reported in different countries. This fact, among other concerns, leads to continuous efforts for the development of new generation of vaccines against rotavirus.In this work, we describe the obtention of cell wall-derived particles from a recombinant Lactococcus lactis expressing a cell wall-anchored version of the rotavirus VP6 protein. After confirming by SDS-PAGE, Western blot, flow cytometry and electronic immunomicroscopy that these particles were carrying the VP6 protein, their immunogenic potential was evaluated in adult BALB/c mice. For that, mucosal immunizations (oral or intranasal), with or without the dmLT [(double mutant Escherichia coli heat labile toxin LT(R192G/L211A)] adjuvant were performed. The results showed that these cell wall-derived particles were able to generate anti-rotavirus IgG and IgA antibodies only when administered intranasally, whether the adjuvant was present or not. However, the presence of dmLT was necessary to confer protection against rotavirus infection, which was evidenced by a 79.5 percent viral shedding reduction.In summary, this work describes the production of cell wall-derived particles which were able to induce a protective immune response after intranasal immunization. Further studies are needed to characterize the immune response elicited by these particles as well as to determine their potential as an alternative to the use of live L. lactis for mucosal antigen delivery.

show abstract

Preformulation studies with the Escherichia coli double mutant heat-labile toxin adjuvant for use in an oral vaccine

Cited by 13 publications

References 33 publications

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

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

The Mucosal Vaccine Adjuvant LT(R192G/L211A) or dmLT

Rotavirus VP6 protein mucosally delivered by cell wall-derived particles from Lactococcus lactis induces protection against infection in a murine model

Contact Info

Product

Resources

About