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Fabrication of nanoadjuvant with poly-ε-caprolactone (PCL) for developing a single-shot vaccine providing prolonged immunity.

Prashant CK, Bhat M, Srivastava SK, Saxena A, Kumar M, Singh A, Samim M, Ahmad FJ, Dinda AK - Int J Nanomedicine (2014)

Bottom Line: They did not elicit efficient CD8(+) T-cell responses.This biodegradable nanoadjuvant system may help to develop single-shot immunization for prolonged immunity without booster doses.The capability of enhanced CMI response may have high translational potential for immunization against intracellular infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, All India Institute of Medical Sciences, Jamia Hamdard, New Delhi, India.

ABSTRACT

Purpose: The aim of the study was to load a model antigen, tetanus toxoid (TT), in poly-ε-caprolactone nanoparticles (PCL NPs) of two size ranges, ie, mean 61.2 nm (small) and 467.6 nm (large), and study its effect on macrophage polarization as well as antigen presentation in human monocyte-derived macrophages in vitro, along with humoral and cell-mediated immune (CMI) response generated in Swiss albino mice following immunization with the TT-loaded NPs.

Materials and methods: PCL NPs were synthesized by solvent evaporation. The antigen-loaded PCL NPs were characterized for size, zeta potential, and protein-release kinetics. Swiss albino mice were immunized with the antigen-loaded PCL NPs. Flow cytometry was used to quantify interferon-γ- and interleukin-4-secreting cluster of differentiation (CD)4(+) and CD8(+) T cells in the spleen, and enzyme-linked immunosorbent assay was used to quantify anti-TT antibody levels in the serum of immunized mice.

Results: Small PCL NPs generated an M1/M2 type polarization of human blood monocyte-derived macrophages and T helper (Th)1/Th2 polarization of autologous CD4(+) T cells. Efficient CD8(+) T-cell responses were also elicited. Large PCL NPs failed to cause any type of macrophage polarization. They did not elicit efficient CD8(+) T-cell responses.

Conclusion: TT-loaded small PCL NPs were able to generate persistent and strong CMI and humoral responses against TT 2 months after single injection in mice without booster dose. This biodegradable nanoadjuvant system may help to develop single-shot immunization for prolonged immunity without booster doses. The capability of enhanced CMI response may have high translational potential for immunization against intracellular infection.

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(A and B) Interferon (IFN)-γ and interleukin (IL)-4 levels in supernatant of cultured splenocytes. Significant secretion of IL-4 (A) and IFN-γ (B) is observed from total splenocytes from mice immunized with small poly-ε-caprolactone nanoparticles (PCL NPs) through all three routes.Abbreviations: IM, intramuscular; SC, subcutaneous; IV, intravenous; TT, tetanus toxoid; neg cont, negative control.
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f7-ijn-9-937: (A and B) Interferon (IFN)-γ and interleukin (IL)-4 levels in supernatant of cultured splenocytes. Significant secretion of IL-4 (A) and IFN-γ (B) is observed from total splenocytes from mice immunized with small poly-ε-caprolactone nanoparticles (PCL NPs) through all three routes.Abbreviations: IM, intramuscular; SC, subcutaneous; IV, intravenous; TT, tetanus toxoid; neg cont, negative control.

Mentions: The flow-cytometry data of isolated splenocytes indicated that antigen-loaded small NPs delivered through the IM and SC routes elicited robust IFN-γ response in CD4+ T cells (2.655%±0.225%, P=0.0179; 2.205%±0.165%, respectively (percentage of positive cells expressed as mean ± standard error of mean), as a recall-memory response against the immunized antigen, TT (Figure 5A). The IV route did not elicit significant IFN-γ secretion in CD4+ cells. IL-4 production in CD4+ cells was significant in all three routes of immunization (Figure 5B). CD8+ cells did not produce significant IFN-γ or IL-4 compared to the positive control alum TT through any of the immunization routes, but cytokine secretion was significant compared to the negative control (Figure 6). TT-loaded large PCL NPs did not elicit significant cytokine responses in either CD4+ or CD8+ T cells. ELISA for detecting IFN-γ and IL-4 levels in the supernatant of the cultured splenocytes revealed that IFN-γ and IL-4 were produced in significant amounts in cases where immunization with small PCL NPs was done through all the three routes (Figure 7). Thus, the cytokine milieu in the spleen is overall a Th1/Th2 type promoting both CMI and humoral responses if immunized with TT-loaded small PCL NPs.


Fabrication of nanoadjuvant with poly-ε-caprolactone (PCL) for developing a single-shot vaccine providing prolonged immunity.

Prashant CK, Bhat M, Srivastava SK, Saxena A, Kumar M, Singh A, Samim M, Ahmad FJ, Dinda AK - Int J Nanomedicine (2014)

(A and B) Interferon (IFN)-γ and interleukin (IL)-4 levels in supernatant of cultured splenocytes. Significant secretion of IL-4 (A) and IFN-γ (B) is observed from total splenocytes from mice immunized with small poly-ε-caprolactone nanoparticles (PCL NPs) through all three routes.Abbreviations: IM, intramuscular; SC, subcutaneous; IV, intravenous; TT, tetanus toxoid; neg cont, negative control.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3928464&req=5

f7-ijn-9-937: (A and B) Interferon (IFN)-γ and interleukin (IL)-4 levels in supernatant of cultured splenocytes. Significant secretion of IL-4 (A) and IFN-γ (B) is observed from total splenocytes from mice immunized with small poly-ε-caprolactone nanoparticles (PCL NPs) through all three routes.Abbreviations: IM, intramuscular; SC, subcutaneous; IV, intravenous; TT, tetanus toxoid; neg cont, negative control.
Mentions: The flow-cytometry data of isolated splenocytes indicated that antigen-loaded small NPs delivered through the IM and SC routes elicited robust IFN-γ response in CD4+ T cells (2.655%±0.225%, P=0.0179; 2.205%±0.165%, respectively (percentage of positive cells expressed as mean ± standard error of mean), as a recall-memory response against the immunized antigen, TT (Figure 5A). The IV route did not elicit significant IFN-γ secretion in CD4+ cells. IL-4 production in CD4+ cells was significant in all three routes of immunization (Figure 5B). CD8+ cells did not produce significant IFN-γ or IL-4 compared to the positive control alum TT through any of the immunization routes, but cytokine secretion was significant compared to the negative control (Figure 6). TT-loaded large PCL NPs did not elicit significant cytokine responses in either CD4+ or CD8+ T cells. ELISA for detecting IFN-γ and IL-4 levels in the supernatant of the cultured splenocytes revealed that IFN-γ and IL-4 were produced in significant amounts in cases where immunization with small PCL NPs was done through all the three routes (Figure 7). Thus, the cytokine milieu in the spleen is overall a Th1/Th2 type promoting both CMI and humoral responses if immunized with TT-loaded small PCL NPs.

Bottom Line: They did not elicit efficient CD8(+) T-cell responses.This biodegradable nanoadjuvant system may help to develop single-shot immunization for prolonged immunity without booster doses.The capability of enhanced CMI response may have high translational potential for immunization against intracellular infection.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, All India Institute of Medical Sciences, Jamia Hamdard, New Delhi, India.

ABSTRACT

Purpose: The aim of the study was to load a model antigen, tetanus toxoid (TT), in poly-ε-caprolactone nanoparticles (PCL NPs) of two size ranges, ie, mean 61.2 nm (small) and 467.6 nm (large), and study its effect on macrophage polarization as well as antigen presentation in human monocyte-derived macrophages in vitro, along with humoral and cell-mediated immune (CMI) response generated in Swiss albino mice following immunization with the TT-loaded NPs.

Materials and methods: PCL NPs were synthesized by solvent evaporation. The antigen-loaded PCL NPs were characterized for size, zeta potential, and protein-release kinetics. Swiss albino mice were immunized with the antigen-loaded PCL NPs. Flow cytometry was used to quantify interferon-γ- and interleukin-4-secreting cluster of differentiation (CD)4(+) and CD8(+) T cells in the spleen, and enzyme-linked immunosorbent assay was used to quantify anti-TT antibody levels in the serum of immunized mice.

Results: Small PCL NPs generated an M1/M2 type polarization of human blood monocyte-derived macrophages and T helper (Th)1/Th2 polarization of autologous CD4(+) T cells. Efficient CD8(+) T-cell responses were also elicited. Large PCL NPs failed to cause any type of macrophage polarization. They did not elicit efficient CD8(+) T-cell responses.

Conclusion: TT-loaded small PCL NPs were able to generate persistent and strong CMI and humoral responses against TT 2 months after single injection in mice without booster dose. This biodegradable nanoadjuvant system may help to develop single-shot immunization for prolonged immunity without booster doses. The capability of enhanced CMI response may have high translational potential for immunization against intracellular infection.

Show MeSH
Related in: MedlinePlus