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Transdermal influenza immunization with vaccine-coated microneedle arrays.

Koutsonanos DG, del Pilar Martin M, Zarnitsyn VG, Sullivan SP, Compans RW, Prausnitz MR, Skountzou I - PLoS ONE (2009)

Bottom Line: Substantial antibody titers with hemagglutination inhibition activity were detected in sera collected two and four weeks after a single vaccine dose.Microneedle vaccination induced a broad spectrum of immune responses including CD4+ and CD8+ responses in the spleen and draining lymph node, a high frequency of antigen-secreting cells in the lung and induction of virus-specific memory B-cells.In view of the convenience of delivery and the potential for self-administration, vaccine-coated metal microneedles may provide a novel and highly effective immunization method.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology & Immunology and Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia, United States of America.

ABSTRACT

Background: Influenza is a contagious disease caused by a pathogenic virus, with outbreaks all over the world and thousands of hospitalizations and deaths every year. Due to virus antigenic drift and short-lived immune responses, annual vaccination is required. However, vaccine coverage is incomplete, and improvement in immunization is needed. The objective of this study is to investigate a novel method for transdermal delivery using metal microneedle arrays (MN) coated with inactivated influenza virus to determine whether this route is a simpler and safer approach than the conventional immunization, capable to induce robust immune responses and confer protection against lethal virus challenge.

Methodology/principal findings: Inactivated A/Aichi/2/68 (H3N2) influenza virus was coated on metal microneedle arrays and applied to mice as a vaccine in the caudal dorsal skin area. Substantial antibody titers with hemagglutination inhibition activity were detected in sera collected two and four weeks after a single vaccine dose. Challenge studies in mice with 5 x LD(50) of mouse adapted Aichi virus demonstrated complete protection. Microneedle vaccination induced a broad spectrum of immune responses including CD4+ and CD8+ responses in the spleen and draining lymph node, a high frequency of antigen-secreting cells in the lung and induction of virus-specific memory B-cells. In addition, the use of MN showed a dose-sparing effect and a strong Th2 bias when compared to an intramuscular (IM) reference immunization.

Conclusions/significance: The present results show that delivery of inactivated influenza virus through the skin using metal microneedle arrays induced strong humoral and cellular immune responses capable of conferring protection against virus challenge as efficiently as intramuscular immunization, which is the standard vaccination route. In view of the convenience of delivery and the potential for self-administration, vaccine-coated metal microneedles may provide a novel and highly effective immunization method.

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Related in: MedlinePlus

IL-4 and IFN-γ secretion by T cells in response to inactivated influenza antigen.The frequency of cellular immune responses is shown in immunized and challenged mice. Spleens and lymph nodes from all groups of immunized mice were individually processed and cells were cultured in the presence of NP Class I or NP Class II peptides for stimulation. (A) Splenocytes and lymphocytes were stimulated with NP Class I peptide and analyzed with ELISA for IL-4 production at 72 hours. Splenocytes and lymphocytes cultured in the presence of NP Class I (B) or NP Class II stimulants (C) were analyzed for IFN-γ production at 72 hours after stimulation. (N) Naïve control, (Inf.) unimmunized infected mice (average±s.e). ND: not detected. ap<0.05 when MN 10 µg compared to the IM 10 µg group. bp<0.05 when IM 10 µg compared to the MN 10 µg group.
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pone-0004773-g006: IL-4 and IFN-γ secretion by T cells in response to inactivated influenza antigen.The frequency of cellular immune responses is shown in immunized and challenged mice. Spleens and lymph nodes from all groups of immunized mice were individually processed and cells were cultured in the presence of NP Class I or NP Class II peptides for stimulation. (A) Splenocytes and lymphocytes were stimulated with NP Class I peptide and analyzed with ELISA for IL-4 production at 72 hours. Splenocytes and lymphocytes cultured in the presence of NP Class I (B) or NP Class II stimulants (C) were analyzed for IFN-γ production at 72 hours after stimulation. (N) Naïve control, (Inf.) unimmunized infected mice (average±s.e). ND: not detected. ap<0.05 when MN 10 µg compared to the IM 10 µg group. bp<0.05 when IM 10 µg compared to the MN 10 µg group.

Mentions: To assess cellular immune responses, we re-stimulated the splenocytes and lymph node cultures (LN) isolated from challenged mice on day 4 with NP Class I and II restricted peptides, and quantified their ability to produce cytokines (IL-2, IL-4, IFN-γ) by ELISA (Figure 6). The production of IL-2 did not show significant differences in splenocyte or LN cultures re-stimulated with Aichi virus or NP Class I and II peptides (data not shown).


Transdermal influenza immunization with vaccine-coated microneedle arrays.

Koutsonanos DG, del Pilar Martin M, Zarnitsyn VG, Sullivan SP, Compans RW, Prausnitz MR, Skountzou I - PLoS ONE (2009)

IL-4 and IFN-γ secretion by T cells in response to inactivated influenza antigen.The frequency of cellular immune responses is shown in immunized and challenged mice. Spleens and lymph nodes from all groups of immunized mice were individually processed and cells were cultured in the presence of NP Class I or NP Class II peptides for stimulation. (A) Splenocytes and lymphocytes were stimulated with NP Class I peptide and analyzed with ELISA for IL-4 production at 72 hours. Splenocytes and lymphocytes cultured in the presence of NP Class I (B) or NP Class II stimulants (C) were analyzed for IFN-γ production at 72 hours after stimulation. (N) Naïve control, (Inf.) unimmunized infected mice (average±s.e). ND: not detected. ap<0.05 when MN 10 µg compared to the IM 10 µg group. bp<0.05 when IM 10 µg compared to the MN 10 µg group.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2651574&req=5

pone-0004773-g006: IL-4 and IFN-γ secretion by T cells in response to inactivated influenza antigen.The frequency of cellular immune responses is shown in immunized and challenged mice. Spleens and lymph nodes from all groups of immunized mice were individually processed and cells were cultured in the presence of NP Class I or NP Class II peptides for stimulation. (A) Splenocytes and lymphocytes were stimulated with NP Class I peptide and analyzed with ELISA for IL-4 production at 72 hours. Splenocytes and lymphocytes cultured in the presence of NP Class I (B) or NP Class II stimulants (C) were analyzed for IFN-γ production at 72 hours after stimulation. (N) Naïve control, (Inf.) unimmunized infected mice (average±s.e). ND: not detected. ap<0.05 when MN 10 µg compared to the IM 10 µg group. bp<0.05 when IM 10 µg compared to the MN 10 µg group.
Mentions: To assess cellular immune responses, we re-stimulated the splenocytes and lymph node cultures (LN) isolated from challenged mice on day 4 with NP Class I and II restricted peptides, and quantified their ability to produce cytokines (IL-2, IL-4, IFN-γ) by ELISA (Figure 6). The production of IL-2 did not show significant differences in splenocyte or LN cultures re-stimulated with Aichi virus or NP Class I and II peptides (data not shown).

Bottom Line: Substantial antibody titers with hemagglutination inhibition activity were detected in sera collected two and four weeks after a single vaccine dose.Microneedle vaccination induced a broad spectrum of immune responses including CD4+ and CD8+ responses in the spleen and draining lymph node, a high frequency of antigen-secreting cells in the lung and induction of virus-specific memory B-cells.In view of the convenience of delivery and the potential for self-administration, vaccine-coated metal microneedles may provide a novel and highly effective immunization method.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology & Immunology and Emory Vaccine Center, Emory University School of Medicine, Atlanta, Georgia, United States of America.

ABSTRACT

Background: Influenza is a contagious disease caused by a pathogenic virus, with outbreaks all over the world and thousands of hospitalizations and deaths every year. Due to virus antigenic drift and short-lived immune responses, annual vaccination is required. However, vaccine coverage is incomplete, and improvement in immunization is needed. The objective of this study is to investigate a novel method for transdermal delivery using metal microneedle arrays (MN) coated with inactivated influenza virus to determine whether this route is a simpler and safer approach than the conventional immunization, capable to induce robust immune responses and confer protection against lethal virus challenge.

Methodology/principal findings: Inactivated A/Aichi/2/68 (H3N2) influenza virus was coated on metal microneedle arrays and applied to mice as a vaccine in the caudal dorsal skin area. Substantial antibody titers with hemagglutination inhibition activity were detected in sera collected two and four weeks after a single vaccine dose. Challenge studies in mice with 5 x LD(50) of mouse adapted Aichi virus demonstrated complete protection. Microneedle vaccination induced a broad spectrum of immune responses including CD4+ and CD8+ responses in the spleen and draining lymph node, a high frequency of antigen-secreting cells in the lung and induction of virus-specific memory B-cells. In addition, the use of MN showed a dose-sparing effect and a strong Th2 bias when compared to an intramuscular (IM) reference immunization.

Conclusions/significance: The present results show that delivery of inactivated influenza virus through the skin using metal microneedle arrays induced strong humoral and cellular immune responses capable of conferring protection against virus challenge as efficiently as intramuscular immunization, which is the standard vaccination route. In view of the convenience of delivery and the potential for self-administration, vaccine-coated metal microneedles may provide a novel and highly effective immunization method.

Show MeSH
Related in: MedlinePlus