Recombinant Norwalk virus-like particles administered intranasally to mice induce systemic and mucosal (fecal and vaginal) immune responses - PubMed (original) (raw)

Comparative Study

Recombinant Norwalk virus-like particles administered intranasally to mice induce systemic and mucosal (fecal and vaginal) immune responses

R A Guerrero et al. J Virol. 2001 Oct.

Abstract

Recombinant Norwalk virus-like particles (rNV VLPs) were administered to BALB/c mice by the intranasal (i.n.) route to evaluate the induction of mucosal antibody responses. The results were compared to systemic and mucosal responses observed in new and previous studies (J. M. Ball, M. E. Hardy, R. L. Atmar, M. E. Connor, and M. K. Estes, J. Virol. 72:1345-1353, 1998) after oral administration of rNV VLPs. Immunizations were given in the presence or absence of a mucosal adjuvant, mutant Escherichia coli heat-labile toxin LT(R192G). rNV-specific immunoglobulin G (IgG) and fecal IgA were evaluated by enzyme-linked immunosorbent assay. The i.n. delivery of rNV VLPs was more effective than the oral route at inducing serum IgG and fecal IgA responses to low doses of rNV particles. Vaginal responses of female mice given VLPs by the i.n. and oral routes were also examined. All mice that received two immunizations with low doses i.n. (10 or 25 microg) of rNV VLPs and the majority of mice that received two high doses orally (200 microg) in the absence of adjuvant had rNV-specific serum IgG, fecal, and vaginal responses. Additional experiments evaluated whether rNV VLPs can function as a mucosal adjuvant by evaluating the immune responses to two soluble proteins, keyhole limpet hemocyanin and chicken egg albumin. Under the conditions tested, rNV VLPs did not enhance the serum IgG or fecal IgA response to these soluble proteins when coadministered by the i.n. or oral route. Low doses of nonreplicating rNV VLPs are immunogenic when administered i.n. in the absence of adjuvant, and addition of adjuvant enhanced the magnitude and duration of these responses. Recombinant NV VLPs represent a candidate mucosal vaccine for NV infections in humans.

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Figures

FIG. 1

Serum IgG responses after in and oral administration to BALB/c mice of rNV VLPs in the presence or absence of LT(R192G). (A and B) Serum IgG responses at 36 dpi (A) and 417 dpi (B). The x axis shows the dose of VLPs administered on days 1 and 21, as well as the presence or absence of adjuvant. The GMT was calculated, including responders and nonresponders. The error bars show the standard error of the mean. Above each column is the number of responders over the total number of mice tested, and the GMT of only the responders is shown in parentheses. Statistical analysis was done with the Mann-Whitney U test. For each panel, identical symbols above two different columns indicate that these two groups were significantly different.

FIG. 2

Fecal antibody responses after i.n. and oral administration to BALB/c mice of rNV VLPs in the presence or absence of LT(R192G). (A and B) Fecal IgA responses at 36 dpi (A) and 417 dpi (B). The x axis shows the dose of VLPs administered on days 1 and 21, as well as the presence or absence of adjuvant. The left panel y axis shows the GMT of antibody for each group of mice, and the right panel y axis represents the response expressed as mean of the ratio of NV-specific IgA in nanograms per milliliter to total IgA in micrograms per milliliter. The GMT of only the responders is shown in parentheses. Statistical analysis was done with the Mann-Whitney U test. For each panel, identical symbols above different columns indicate that these two groups were significantly different. There were no statistical differences between the groups in panel B.

FIG. 3

Vaginal antibody responses after i.n. and oral administration to BALB/c mice of rNV VLPs in the presence or absence of LT(R192G). (A and B) Vaginal responses at 40 dpi (A) and 365 dpi (B). The GMT was calculated, including responders and nonresponders. The error bars show the standard error of the mean. Above each column is the number of responders over the total number of mice tested, and the GMT of only the responders is shown in parentheses. The y axis on the left panel shows the GMT of antibody for each group of mice, and the y axis on the right panel represents the response expressed as mean of the ratio of NV-specific IgA in nanograms per milliliter to total IgA in micrograms per milliliter. Statistical analysis was done with the Mann-Whitney U test. Identical symbols above two different columns indicate that these two groups were significantly different.

FIG. 4

BALB/c mouse antibody responses to the oral administration of OVA in the presence of rNV VLPs or LT(R192G). (A) Serum IgG responses. (B) Fecal IgA responses. The x axis shows the dose of OVA, rNV VLPs, or LT(R192G) administered by the oral route. For panel A, the y axis represents the GMT of antibody for each group of mice. For panel B, the y axis represents the response expressed as the mean of the ratio of OVA-specific IgA in nanograms per milliliter to total IgA in micrograms per milliliter. The GMT of only the responders is shown in parentheses. Statistical analysis was done with the Mann-Whitney U test. There were no statistically significant differences between any of the groups.

FIG. 5

BALB/c mouse antibody responses after i.n. administration with OVA in the presence of rNV VLPs or LT(R192G). (A and B) Serum IgG responses (A) and fecal IgA responses (B) 28 dpi for mice immunized with a two week interval. (C and D) Serum IgG responses (C) and fecal IgA responses (D) 35 dpi for mice immunized with a 3-week interval. The x axis shows the dose of OVA, rNV VLPs, or LT(R192G) administered by the i.n. route on days 1 and 14 (A and B) or on days 1 and 21 (C and D). The lightly shaded column received OVA with 30 μg of rNV VLPs on days 1 and 14. For panels A and C, the y axis represents the GMT of antibody for each group of mice. For panels B and D, the y axis represents the response expressed as the mean of the ratio of OVA-specific IgA in nanograms per milliliter to total IgA in micrograms per milliliter. The GMT of only the responders is shown in parentheses. Statistical analysis was done with the Mann-Whitney U test. For each panel, identical symbols above two different columns indicate that these two groups were significantly different.

FIG. 6

BALB/c mouse antibody responses after i.n. administration with KLH. (A and B) Serum IgG responses (A) and fecal IgA responses (B) 35 dpi for mice immunized with a 3-week interval. The x axis shows the dose of KLH, rNV VLPs, or LT administered on days 1 and 21. For panel A, the y axis shows the GMT of antibody for each group of mice. The coadministration of rNV VLPs at different doses did not enhance the response, but the response was enhanced with coadministration of 10 μg of LT (P = 0.009; Mann-Whitney test). For panel B, the y axis represents the response expressed as mean of the ratio of KLH-specific IgA in nanograms per milliliter to total IgA in micrograms per milliliter. The coadministration of rNV VLPs did not enhance the response, but the coadministration of LT enhanced the response to KLH (P = 0.011). The GMT of only the responders is shown in parentheses. Statistical analysis was done with the Mann-Whitney U test. For each panel, identical symbols above two different columns indicate that these two groups were significantly different.

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Recombinant Norwalk virus-like particles administered intranasally to mice induce systemic and mucosal (fecal and vaginal) immune responses - PubMed (original) (raw)