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In vivo mucosal delivery of bioactive human interleukin 1 receptor antagonist produced by Streptococcus gordonii.

Ricci S, Macchia G, Ruggiero P, Maggi T, Bossù P, Xu L, Medaglini D, Tagliabue A, Hammarström L, Pozzi G, Boraschi D - BMC Biotechnol. (2003)

Bottom Line: RFVP/IL-1ra displayed full biological activity in vitro in assays of inhibition of IL-1beta-induced lymphocyte proliferation and was released by recombinant S. gordonii in vivo both at the vaginal and the gastrointestinal mucosa of mice.RFVP/IL-1ra appeared beneficial in the model of ulcerative colitis represented by IL-2-/- mice (knock-out for the interleukin-2 gene), as shown by the body weight increase of IL-2-/- mice locally treated with S. gordonii producing RFVP/IL-1ra.These results indicate that recombinant S. gordonii can be successfully used as a delivery system for the selective targeting of mucosal surfaces with therapeutic proteins.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Molecular Microbiology and Biotechnology, Department of Molecular Biology, University of Siena, Policlinico Le Scotte, Viale Bracci, 53100 Siena, Italy. riccisus@unisi.it

ABSTRACT

Background: Interleukin-1 (IL-1) is a cytokine involved in the initiation and amplification of the defence response in infectious and inflammatory diseases. IL-1 receptor antagonist (IL-1ra) is an inactive member of the IL-1 family and represents one of the most potent mechanisms for controlling IL-1-dependent inflammation. IL-1ra has proven effective in the therapy of acute and chronic inflammatory diseases in experimental animal models and also in preliminary clinical trials. However, optimisation of therapeutic schedules is still needed. For instance, the use of drug delivery systems targeting specific mucosal sites may be useful to improve topical bioavailability and avoid side effects associated with systemic administration.

Results: In order to develop systems for the delivery of IL-1ra to mucosal target sites, a Streptococcus gordonii strain secreting human IL-1ra was constructed. The recombinant IL-1ra produced by S. gordonii was composed of the four amino acid residues RVFP of the fusion partner at the N-terminus, followed by the mature human IL-1ra protein. RFVP/IL-1ra displayed full biological activity in vitro in assays of inhibition of IL-1beta-induced lymphocyte proliferation and was released by recombinant S. gordonii in vivo both at the vaginal and the gastrointestinal mucosa of mice. RFVP/IL-1ra appeared beneficial in the model of ulcerative colitis represented by IL-2-/- mice (knock-out for the interleukin-2 gene), as shown by the body weight increase of IL-2-/- mice locally treated with S. gordonii producing RFVP/IL-1ra.

Conclusions: These results indicate that recombinant S. gordonii can be successfully used as a delivery system for the selective targeting of mucosal surfaces with therapeutic proteins.

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Body weight of IBD-affected IL-2-deficient mice treated with RVFP/IL-1ra producing S. gordonii. Two groups of six IL-2-/- mice were inoculated intragastrically with 1 × 1010 CFU of either S. gordonii strain GP1294 (RVFP/IL-1; solid triangles) or GP204 (control; open triangles). One untreated mouse (open circles) served as a control of disease progression. Starting at 4 weeks of age, bacteria were administered (indicated by arrows) to mice twice per week for a total of 7 consecutive weeks. Body weight (in grams) of individual mice was recorded twice per week up to 15 weeks of age, and expressed as mean ± SD. Statistically significant (t test) differences between mice treated with S. gordonii GP1294 and those receiving GP204 are represented by asterisks (*, P < 0.05; **, P < 0.01).
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Figure 3: Body weight of IBD-affected IL-2-deficient mice treated with RVFP/IL-1ra producing S. gordonii. Two groups of six IL-2-/- mice were inoculated intragastrically with 1 × 1010 CFU of either S. gordonii strain GP1294 (RVFP/IL-1; solid triangles) or GP204 (control; open triangles). One untreated mouse (open circles) served as a control of disease progression. Starting at 4 weeks of age, bacteria were administered (indicated by arrows) to mice twice per week for a total of 7 consecutive weeks. Body weight (in grams) of individual mice was recorded twice per week up to 15 weeks of age, and expressed as mean ± SD. Statistically significant (t test) differences between mice treated with S. gordonii GP1294 and those receiving GP204 are represented by asterisks (*, P < 0.05; **, P < 0.01).

Mentions: Mice deficient in the interleukin-2 gene (IL-2-/-) spontaneously develop a progressive IBD similar to human ulcerative colitis between 4 to 15 weeks of age [35,36]. The disease is associated with increased levels of IL-1 [37,38], and treatment with systemic IL-1ra diminishes colitis severity [37]. As we showed that S. gordonii GP1294 was able to release IL-1ra in the GI tract of mice (Table 1), it was thus examined whether production of IL-1ra by S. gordonii in the GI tract of IL-2-/- mice could be effective in decreasing the IBD-like pathology. Starting at 4 weeks of age, animals were treated for 7 consecutive weeks with either S. gordonii GP1294 or the GP204 control strain. As a measure of disease progression, body weight was recorded up to the 15th week of age. A significant weight gain was observed in the first week of treatment (week 4 to 5) in the animal group inoculated with the GP1294 strain (Figure 3). A more consistent weight increase was achieved from week 8 to week 10.5 in mice treated with S. gordonii GP1294, with statistically significant differences compared to animals inoculated with the control strain (Figure 3). After treatment interruption at week 10, differences in body weight between the two animal groups could no longer be detected. These results suggest that recombinant S. gordonii, that is able to release bioactive IL-1ra in the GI tract, can improve the symptoms of ulcerative colitis. It may be hypothesized that such improvement is due to the inhibition of IL-1-dependent inflammation by IL-1ra, although further investigation is needed to confirm this hypothesis.


In vivo mucosal delivery of bioactive human interleukin 1 receptor antagonist produced by Streptococcus gordonii.

Ricci S, Macchia G, Ruggiero P, Maggi T, Bossù P, Xu L, Medaglini D, Tagliabue A, Hammarström L, Pozzi G, Boraschi D - BMC Biotechnol. (2003)

Body weight of IBD-affected IL-2-deficient mice treated with RVFP/IL-1ra producing S. gordonii. Two groups of six IL-2-/- mice were inoculated intragastrically with 1 × 1010 CFU of either S. gordonii strain GP1294 (RVFP/IL-1; solid triangles) or GP204 (control; open triangles). One untreated mouse (open circles) served as a control of disease progression. Starting at 4 weeks of age, bacteria were administered (indicated by arrows) to mice twice per week for a total of 7 consecutive weeks. Body weight (in grams) of individual mice was recorded twice per week up to 15 weeks of age, and expressed as mean ± SD. Statistically significant (t test) differences between mice treated with S. gordonii GP1294 and those receiving GP204 are represented by asterisks (*, P < 0.05; **, P < 0.01).
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Figure 3: Body weight of IBD-affected IL-2-deficient mice treated with RVFP/IL-1ra producing S. gordonii. Two groups of six IL-2-/- mice were inoculated intragastrically with 1 × 1010 CFU of either S. gordonii strain GP1294 (RVFP/IL-1; solid triangles) or GP204 (control; open triangles). One untreated mouse (open circles) served as a control of disease progression. Starting at 4 weeks of age, bacteria were administered (indicated by arrows) to mice twice per week for a total of 7 consecutive weeks. Body weight (in grams) of individual mice was recorded twice per week up to 15 weeks of age, and expressed as mean ± SD. Statistically significant (t test) differences between mice treated with S. gordonii GP1294 and those receiving GP204 are represented by asterisks (*, P < 0.05; **, P < 0.01).
Mentions: Mice deficient in the interleukin-2 gene (IL-2-/-) spontaneously develop a progressive IBD similar to human ulcerative colitis between 4 to 15 weeks of age [35,36]. The disease is associated with increased levels of IL-1 [37,38], and treatment with systemic IL-1ra diminishes colitis severity [37]. As we showed that S. gordonii GP1294 was able to release IL-1ra in the GI tract of mice (Table 1), it was thus examined whether production of IL-1ra by S. gordonii in the GI tract of IL-2-/- mice could be effective in decreasing the IBD-like pathology. Starting at 4 weeks of age, animals were treated for 7 consecutive weeks with either S. gordonii GP1294 or the GP204 control strain. As a measure of disease progression, body weight was recorded up to the 15th week of age. A significant weight gain was observed in the first week of treatment (week 4 to 5) in the animal group inoculated with the GP1294 strain (Figure 3). A more consistent weight increase was achieved from week 8 to week 10.5 in mice treated with S. gordonii GP1294, with statistically significant differences compared to animals inoculated with the control strain (Figure 3). After treatment interruption at week 10, differences in body weight between the two animal groups could no longer be detected. These results suggest that recombinant S. gordonii, that is able to release bioactive IL-1ra in the GI tract, can improve the symptoms of ulcerative colitis. It may be hypothesized that such improvement is due to the inhibition of IL-1-dependent inflammation by IL-1ra, although further investigation is needed to confirm this hypothesis.

Bottom Line: RFVP/IL-1ra displayed full biological activity in vitro in assays of inhibition of IL-1beta-induced lymphocyte proliferation and was released by recombinant S. gordonii in vivo both at the vaginal and the gastrointestinal mucosa of mice.RFVP/IL-1ra appeared beneficial in the model of ulcerative colitis represented by IL-2-/- mice (knock-out for the interleukin-2 gene), as shown by the body weight increase of IL-2-/- mice locally treated with S. gordonii producing RFVP/IL-1ra.These results indicate that recombinant S. gordonii can be successfully used as a delivery system for the selective targeting of mucosal surfaces with therapeutic proteins.

View Article: PubMed Central - HTML - PubMed

Affiliation: Laboratory of Molecular Microbiology and Biotechnology, Department of Molecular Biology, University of Siena, Policlinico Le Scotte, Viale Bracci, 53100 Siena, Italy. riccisus@unisi.it

ABSTRACT

Background: Interleukin-1 (IL-1) is a cytokine involved in the initiation and amplification of the defence response in infectious and inflammatory diseases. IL-1 receptor antagonist (IL-1ra) is an inactive member of the IL-1 family and represents one of the most potent mechanisms for controlling IL-1-dependent inflammation. IL-1ra has proven effective in the therapy of acute and chronic inflammatory diseases in experimental animal models and also in preliminary clinical trials. However, optimisation of therapeutic schedules is still needed. For instance, the use of drug delivery systems targeting specific mucosal sites may be useful to improve topical bioavailability and avoid side effects associated with systemic administration.

Results: In order to develop systems for the delivery of IL-1ra to mucosal target sites, a Streptococcus gordonii strain secreting human IL-1ra was constructed. The recombinant IL-1ra produced by S. gordonii was composed of the four amino acid residues RVFP of the fusion partner at the N-terminus, followed by the mature human IL-1ra protein. RFVP/IL-1ra displayed full biological activity in vitro in assays of inhibition of IL-1beta-induced lymphocyte proliferation and was released by recombinant S. gordonii in vivo both at the vaginal and the gastrointestinal mucosa of mice. RFVP/IL-1ra appeared beneficial in the model of ulcerative colitis represented by IL-2-/- mice (knock-out for the interleukin-2 gene), as shown by the body weight increase of IL-2-/- mice locally treated with S. gordonii producing RFVP/IL-1ra.

Conclusions: These results indicate that recombinant S. gordonii can be successfully used as a delivery system for the selective targeting of mucosal surfaces with therapeutic proteins.

Show MeSH
Related in: MedlinePlus