Recombinant Norwalk virus–like particles given orally to volunteers: Phase I study

doi:10.1016/S0016-5085(99)70548-2

Gastroenterology

Volume 117, Issue 1, July 1999, Pages 40-48

https://doi.org/10.1016/S0016-5085(99)70548-2 Get rights and content

Abstract

Background & Aims: Norwalk virus (NV) is a major cause of epidemic gastroenteritis. The NV capsid is composed of a single protein that forms recombinant (rNV) virus-like particles (VLPs). In mice, these VLPs are immunogenic when administered orally without adjuvant, and they elicit serum immunoglobulin (Ig) G and intestinal IgA responses. The aim of this study was to evaluate the safety and immunogenicity of rNV VLPs in healthy volunteers. Methods: Twenty antibody-positive adults were orally administered rNV VLPs in sterile Milli-Q water on days 1 and 21. Vaccine safety and serum rNV-specific total and subclass IgG and IgA antibody responses were monitored. The immune response induced by the VLPs was compared with the response elicited by replicating virus. Results: No side effects were observed or reported by the volunteers. Serum IgG responses to rNV VLPs were dose-dependent, and all vaccinees given 250 μg of rNV VLPs responded with ≥4-fold increases in serum IgG titers. Most of the volunteers (83%; 15 of 18) responded after the first rNV VLP dose and showed no increase in serum IgG titer after the second dose. Conclusions: Orally administered rNV VLPs are safe and immunogenic in healthy adults when administered without adjuvant and are useful to test the mucosal delivery of immunogens.

GASTROENTEROLOGY 1999;117:40-48

Section snippets

Vaccination studies

Informed consent was obtained from 24 adult volunteers (12 women and 12 men; age, 18–46 years) of various ethnic background (14 white, 2 black, and 6 Asian). This study was approved by the Baylor Affiliate Human Study Review Board. For this phase I vaccination study, volunteers with serum antibody titers of ≤1280 to rNV VLPs by ELISA were selected. Two doses of rNV VLPs (100 or 250 μg) in sterile Milli-Q water (Milli-Q Water System; Millipore, Bedford, MA) or Milli-Q water alone were

rNV particles

The rNV vaccine was sterile, and endotoxin levels were consistently negative (i.e., below 0.02 endotoxin units per milligram of particles). Electron microscopy analyses showed numerous VLPs that appeared structurally similar to native NV virions (see Figure 1 in Ball et al.¹⁴). Sodium dodecyl sulfate–polyacrylamide gel electrophoresis analyses revealed a major 58K band and minor bands at approximately 50K and 30K corresponding to rNV cleavage products (data not shown). No toxicity was observed

Discussion

Many pathogens initiate infection at a mucosal surface, but most vaccines are administered by a parenteral route. The potential advantages of oral administration of antigen include ease of delivery, reduced side effects, and the production of secretory IgA (sIgA) at mucosal surfaces.23, 24 Although most current oral vaccine formulations are attenuated live bacteria or viruses, there is interest in new formulations, and many adjuvants and encapsulation methods are being developed to enhance the

Acknowledgements

The authors thank D. Dimitrov and T. McPherson for technical assistance and R. Atmar, M. Ciarlet, and M. Conner for critical comments regarding the manuscript.

References (40)

MS Ho et al.
Viral gastroenteritis aboard a cruise ship
Lancet
(1989)
H Spiegelberg
Biological activities of immunoglobulins of different class and subclasses
Adv Immunol
(1974)
J Mestecky et al.
The human IgA system: a reassessment
Clin Immunol Immunopathol
(1986)
O Molberg et al.
CD4⁺ T cells with specific reactivity against astrovirus isolated from normal human small intestine
Gastroenterology
(1998)
B Jiang et al.
Heterotypic protection from rotavirus infection in mice vaccinated with virus-like particles
Vaccine
(1999)
AZ Kapikian et al.
Norwalk group of viruses
MK Estes et al.
Norwalk virus and other enteric caliciviruses
CW Hedberg et al.
Outbreaks of food-borne and waterborne viral gastroenteritis
Clin Microbiol Rev
(1993)
ME Hardy et al.
Specific proteolytic cleavage of recombinant Norwalk virus capsid protein
J Virol
(1995)
JE Kaplan et al.
The frequency of a Norwalk-like pattern of illness in outbreaks of acute gastroenteritis
Am J Public Health
(1982)

RL Fankhauser et al.

Molecular epidemiology of “Norwalk-like viruses” in outbreaks of gastroenteritis in the United States

J Infect Dis

(1998)

J Vinjé et al.

The incidence and genetic variability of small round structured viruses in outbreaks of gastroenteritis in The Netherlands

J Infect Dis

(1997)

G Cukor et al.

Human viral gastroenteritis

Microbiol Rev

(1984)

TW Sharp et al.

Epidemiology of Norwalk virus during an outbreak of acute gastroenteritis aboard a US aircraft carrier

J Med Virol

(1995)

X Jiang et al.

Expression, self-assembly, and antigenicity of the Norwalk virus capsid protein

J Virol

(1992)

BVV Prasad et al.

Three-dimensional structure of baculovirus-expressed Norwalk virus capsids

J Virol

(1994)

KY Green et al.

Comparison of the reactivities of baculovirus-expressed recombinant Norwalk virus capsid antigen with those of the native Norwalk virus antigen in serologic assays and some epidemiologic observations

J Clin Microbiol

(1993)

JM Ball et al.

Oral immunization with recombinant Norwalk virus–like particles induces a systemic and mucosal immune response in mice

J Virol

(1998)

DY Graham et al.

Norwalk virus infection of volunteers: new insights based on improved assays

J Infect Dis

(1994)

JJ Gray et al.

Detection of immunoglobulin M (IgM), IgA, IgG Norwalk virus–specific antibodies by indirect enzyme-linked immunosorbent assay with baculovirus-expressed Norwalk virus capsid antigen in adult volunteers challenged with Norwalk virus

J Clin Microbiol

(1994)

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^☆: Supported by Applied Technology program grants 04949-033 and 004949-055 from the Texas Higher Education Coordinating Board, National Institutes of Health training grant T32-DK07664, and General Clinical Research Center grant MO1 RR00188.

^☆☆: Address requests for reprints to: Mary Estes, Ph.D., Division of Molecular Virology, Baylor College of Medicine, One Baylor Plaza, Houston, Texas 77030. e-mail: [email protected]; fax: (713) 798-3586.

^★: Dr. Ball's current address is: Department of Pathobiology, Texas A&M University, Texas Veterinary Medical Center, College Station, Texas.

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Alimentary TractRecombinant Norwalk virus–like particles given orally to volunteers: Phase I study☆,☆☆,★

Abstract

Section snippets

Vaccination studies

rNV particles

Discussion

Acknowledgements

Lancet

Adv Immunol

Clin Immunol Immunopathol

Gastroenterology

Vaccine

Norwalk group of viruses

Norwalk virus and other enteric caliciviruses

Outbreaks of food-borne and waterborne viral gastroenteritis

Clin Microbiol Rev

Specific proteolytic cleavage of recombinant Norwalk virus capsid protein

J Virol

The frequency of a Norwalk-like pattern of illness in outbreaks of acute gastroenteritis

Am J Public Health

Molecular epidemiology of “Norwalk-like viruses” in outbreaks of gastroenteritis in the United States

J Infect Dis

The incidence and genetic variability of small round structured viruses in outbreaks of gastroenteritis in The Netherlands

J Infect Dis

Human viral gastroenteritis

Microbiol Rev

Epidemiology of Norwalk virus during an outbreak of acute gastroenteritis aboard a US aircraft carrier

J Med Virol

Expression, self-assembly, and antigenicity of the Norwalk virus capsid protein

J Virol

Three-dimensional structure of baculovirus-expressed Norwalk virus capsids

J Virol

Comparison of the reactivities of baculovirus-expressed recombinant Norwalk virus capsid antigen with those of the native Norwalk virus antigen in serologic assays and some epidemiologic observations

J Clin Microbiol

Oral immunization with recombinant Norwalk virus–like particles induces a systemic and mucosal immune response in mice

J Virol

Norwalk virus infection of volunteers: new insights based on improved assays

J Infect Dis

Detection of immunoglobulin M (IgM), IgA, IgG Norwalk virus–specific antibodies by indirect enzyme-linked immunosorbent assay with baculovirus-expressed Norwalk virus capsid antigen in adult volunteers challenged with Norwalk virus

J Clin Microbiol

Alimentary Tract
Recombinant Norwalk virus–like particles given orally to volunteers: Phase I study☆,☆☆,★