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Large intestine-targeted, nanoparticle-releasing oral vaccine to control genitorectal viral infection.

Zhu Q, Talton J, Zhang G, Cunningham T, Wang Z, Waters RC, Kirk J, Eppler B, Klinman DM, Sui Y, Gagnon S, Belyakov IM, Mumper RJ, Berzofsky JA - Nat. Med. (2012)

Bottom Line: Oral vaccine delivery seems preferable but runs the risk of the vaccine's destruction in the upper gastrointestinal tract.Conversely, vaccine targeted to the small intestine induced only small intestinal immunity and provided no rectal or vaginal protection, demonstrating functional compartmentalization within the gut mucosal immune system.Therefore, using this oral vaccine delivery system to target the large intestine, but not the small intestine, may represent a feasible new strategy for immune protection of rectal and vaginal mucosa.

View Article: PubMed Central - PubMed

Affiliation: Vaccine Branch, Center for Cancer Research, National Cancer Institute, US National Institutes of Health, Bethesda, Maryland, USA; Department of Oncology, Air Force General Hospital, Beijing, China.

ABSTRACT
Both rectal and vaginal mucosal surfaces serve as transmission routes for pathogenic microorganisms. Vaccination through large intestinal mucosa, previously proven protective for both of these mucosal sites in animal studies, can be achieved successfully by direct intracolorectal (i.c.r.) administration, but this route is clinically impractical. Oral vaccine delivery seems preferable but runs the risk of the vaccine's destruction in the upper gastrointestinal tract. Therefore, we designed a large intestine-targeted oral delivery with pH-dependent microparticles containing vaccine nanoparticles, which induced colorectal immunity in mice comparably to colorectal vaccination and protected against rectal and vaginal viral challenge. Conversely, vaccine targeted to the small intestine induced only small intestinal immunity and provided no rectal or vaginal protection, demonstrating functional compartmentalization within the gut mucosal immune system. Therefore, using this oral vaccine delivery system to target the large intestine, but not the small intestine, may represent a feasible new strategy for immune protection of rectal and vaginal mucosa.

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I.C.R. delivered nanoparticles enter the large intestinal mucosa and orally delivered FS30D/PLGA nanoparticle-releasing microparticles selectively targets the large intestinal mucosa for uptake. (a) Colorectal mucosal uptake of PLGA nanoparticles after i.c.r. delivery of PLGA/FITC-BSA nanoparticles. Cells were isolated from the colorectum 3 days after administration and measured for fluorescence-positive cells. (P < 0.01 between PLGA/FITC-BSA and PBS treated. Representative of three independent experiments, n = 12 15 per group). (b) Induction of antigen-specific colorectal mucosal T cells after i.c.r. delivery of PLGA nanoparticles encapsulating PCLUS3-18IIB and MALP2+poly(I:C)+CpG vaccine (PLGA/PeptAg+TLRL). Three weeks after one immunization, colorectal cells were isolated and measured for P18-I10 specific CD8+ T cells by tetramer staining. P < 0.01 between PLGA/PeptAg+TLRL and PLGA alone. Representative of two independent experiments. n = 10 per group. (c) Gut mucosal uptake of PLGA particles after oral delivery of FS30D/PLGA or L100-55/PLGA. Cells were isolated from the small and large intestine at day 2 for measurement of fluorescence-positive cells. **P < 0.02 on white bar indicates the difference from small intestine. ***P < 0.001 on black bar indicates the difference from the small intestine. n = 9 12 per group. (d) Representative examples of flow cytometry from experiments shown in c.
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Figure 1: I.C.R. delivered nanoparticles enter the large intestinal mucosa and orally delivered FS30D/PLGA nanoparticle-releasing microparticles selectively targets the large intestinal mucosa for uptake. (a) Colorectal mucosal uptake of PLGA nanoparticles after i.c.r. delivery of PLGA/FITC-BSA nanoparticles. Cells were isolated from the colorectum 3 days after administration and measured for fluorescence-positive cells. (P < 0.01 between PLGA/FITC-BSA and PBS treated. Representative of three independent experiments, n = 12 15 per group). (b) Induction of antigen-specific colorectal mucosal T cells after i.c.r. delivery of PLGA nanoparticles encapsulating PCLUS3-18IIB and MALP2+poly(I:C)+CpG vaccine (PLGA/PeptAg+TLRL). Three weeks after one immunization, colorectal cells were isolated and measured for P18-I10 specific CD8+ T cells by tetramer staining. P < 0.01 between PLGA/PeptAg+TLRL and PLGA alone. Representative of two independent experiments. n = 10 per group. (c) Gut mucosal uptake of PLGA particles after oral delivery of FS30D/PLGA or L100-55/PLGA. Cells were isolated from the small and large intestine at day 2 for measurement of fluorescence-positive cells. **P < 0.02 on white bar indicates the difference from small intestine. ***P < 0.001 on black bar indicates the difference from the small intestine. n = 9 12 per group. (d) Representative examples of flow cytometry from experiments shown in c.

Mentions: Uncoated PLGA nanoparticles were manufactured in the range from 300 500 nm (418 nm ± 88 SD) (Supplementary Fig. 2a), with 90% encapsulation efficacy (Supplementary Fig. 2b). We first conducted a proof of principle experiment in which fluorescent fluorescein-containing PLGA/FITC-BSA nanoparticles were delivered directly to the colon of mice by i.c.r. administration. At day 3, a proportion of the cells isolated from the large intestinal lamina propria were detected positive for fluorescence expression (Fig. 1a), indicating an uptake of the nanoparticles. Nanoparticle uptake was primarily found in CD11b+B220int macrophages and secondarily in CD11c+CD11b+ dendritic cells (DCs) in the lamina propria of the large intestine (Supplementary Fig. 3a). Transmission electron microscopy indicates PLGA nanoparticles in the cytoplasm (Supplementary Fig. 3b). Uptake of PLGA nanoparticles by DCs (derived from bone marrow) was also confirmed in vitro by flow cytometry (Supplementary Fig. 4a), fluorescence microscopy (Supplementary Fig. 4b) and transmission electron microscopy (Supplementary Fig. 4c).


Large intestine-targeted, nanoparticle-releasing oral vaccine to control genitorectal viral infection.

Zhu Q, Talton J, Zhang G, Cunningham T, Wang Z, Waters RC, Kirk J, Eppler B, Klinman DM, Sui Y, Gagnon S, Belyakov IM, Mumper RJ, Berzofsky JA - Nat. Med. (2012)

I.C.R. delivered nanoparticles enter the large intestinal mucosa and orally delivered FS30D/PLGA nanoparticle-releasing microparticles selectively targets the large intestinal mucosa for uptake. (a) Colorectal mucosal uptake of PLGA nanoparticles after i.c.r. delivery of PLGA/FITC-BSA nanoparticles. Cells were isolated from the colorectum 3 days after administration and measured for fluorescence-positive cells. (P < 0.01 between PLGA/FITC-BSA and PBS treated. Representative of three independent experiments, n = 12 15 per group). (b) Induction of antigen-specific colorectal mucosal T cells after i.c.r. delivery of PLGA nanoparticles encapsulating PCLUS3-18IIB and MALP2+poly(I:C)+CpG vaccine (PLGA/PeptAg+TLRL). Three weeks after one immunization, colorectal cells were isolated and measured for P18-I10 specific CD8+ T cells by tetramer staining. P < 0.01 between PLGA/PeptAg+TLRL and PLGA alone. Representative of two independent experiments. n = 10 per group. (c) Gut mucosal uptake of PLGA particles after oral delivery of FS30D/PLGA or L100-55/PLGA. Cells were isolated from the small and large intestine at day 2 for measurement of fluorescence-positive cells. **P < 0.02 on white bar indicates the difference from small intestine. ***P < 0.001 on black bar indicates the difference from the small intestine. n = 9 12 per group. (d) Representative examples of flow cytometry from experiments shown in c.
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Figure 1: I.C.R. delivered nanoparticles enter the large intestinal mucosa and orally delivered FS30D/PLGA nanoparticle-releasing microparticles selectively targets the large intestinal mucosa for uptake. (a) Colorectal mucosal uptake of PLGA nanoparticles after i.c.r. delivery of PLGA/FITC-BSA nanoparticles. Cells were isolated from the colorectum 3 days after administration and measured for fluorescence-positive cells. (P < 0.01 between PLGA/FITC-BSA and PBS treated. Representative of three independent experiments, n = 12 15 per group). (b) Induction of antigen-specific colorectal mucosal T cells after i.c.r. delivery of PLGA nanoparticles encapsulating PCLUS3-18IIB and MALP2+poly(I:C)+CpG vaccine (PLGA/PeptAg+TLRL). Three weeks after one immunization, colorectal cells were isolated and measured for P18-I10 specific CD8+ T cells by tetramer staining. P < 0.01 between PLGA/PeptAg+TLRL and PLGA alone. Representative of two independent experiments. n = 10 per group. (c) Gut mucosal uptake of PLGA particles after oral delivery of FS30D/PLGA or L100-55/PLGA. Cells were isolated from the small and large intestine at day 2 for measurement of fluorescence-positive cells. **P < 0.02 on white bar indicates the difference from small intestine. ***P < 0.001 on black bar indicates the difference from the small intestine. n = 9 12 per group. (d) Representative examples of flow cytometry from experiments shown in c.
Mentions: Uncoated PLGA nanoparticles were manufactured in the range from 300 500 nm (418 nm ± 88 SD) (Supplementary Fig. 2a), with 90% encapsulation efficacy (Supplementary Fig. 2b). We first conducted a proof of principle experiment in which fluorescent fluorescein-containing PLGA/FITC-BSA nanoparticles were delivered directly to the colon of mice by i.c.r. administration. At day 3, a proportion of the cells isolated from the large intestinal lamina propria were detected positive for fluorescence expression (Fig. 1a), indicating an uptake of the nanoparticles. Nanoparticle uptake was primarily found in CD11b+B220int macrophages and secondarily in CD11c+CD11b+ dendritic cells (DCs) in the lamina propria of the large intestine (Supplementary Fig. 3a). Transmission electron microscopy indicates PLGA nanoparticles in the cytoplasm (Supplementary Fig. 3b). Uptake of PLGA nanoparticles by DCs (derived from bone marrow) was also confirmed in vitro by flow cytometry (Supplementary Fig. 4a), fluorescence microscopy (Supplementary Fig. 4b) and transmission electron microscopy (Supplementary Fig. 4c).

Bottom Line: Oral vaccine delivery seems preferable but runs the risk of the vaccine's destruction in the upper gastrointestinal tract.Conversely, vaccine targeted to the small intestine induced only small intestinal immunity and provided no rectal or vaginal protection, demonstrating functional compartmentalization within the gut mucosal immune system.Therefore, using this oral vaccine delivery system to target the large intestine, but not the small intestine, may represent a feasible new strategy for immune protection of rectal and vaginal mucosa.

View Article: PubMed Central - PubMed

Affiliation: Vaccine Branch, Center for Cancer Research, National Cancer Institute, US National Institutes of Health, Bethesda, Maryland, USA; Department of Oncology, Air Force General Hospital, Beijing, China.

ABSTRACT
Both rectal and vaginal mucosal surfaces serve as transmission routes for pathogenic microorganisms. Vaccination through large intestinal mucosa, previously proven protective for both of these mucosal sites in animal studies, can be achieved successfully by direct intracolorectal (i.c.r.) administration, but this route is clinically impractical. Oral vaccine delivery seems preferable but runs the risk of the vaccine's destruction in the upper gastrointestinal tract. Therefore, we designed a large intestine-targeted oral delivery with pH-dependent microparticles containing vaccine nanoparticles, which induced colorectal immunity in mice comparably to colorectal vaccination and protected against rectal and vaginal viral challenge. Conversely, vaccine targeted to the small intestine induced only small intestinal immunity and provided no rectal or vaginal protection, demonstrating functional compartmentalization within the gut mucosal immune system. Therefore, using this oral vaccine delivery system to target the large intestine, but not the small intestine, may represent a feasible new strategy for immune protection of rectal and vaginal mucosa.

Show MeSH
Related in: MedlinePlus