Stable incorporation of GM-CSF into dissolvable microneedle patch improves skin vaccination against influenza

Abstract

The widely used influenza subunit vaccine would benefit from increased protection rates in vulnerable populations. Skin immunization by microneedle (MN) patch can increase vaccine immunogenicity, as well as increase vaccination coverage due to simplified administration. To further increase immunogenicity, we used granulocyte-macrophage colony stimulating factor (GM-CSF), an immunomodulatory cytokine already approved for skin cancer therapy and cancer support treatment. GM-CSF has been shown to be upregulated in skin following MN insertion. The GM-CSF-adjuvanted vaccine induced robust and long-lived antibody responses cross-reactive to homosubtypic and heterosubtypic influenza viruses. Addition of GM-CSF resulted in increased memory B cell persistence relative to groups given influenza vaccine alone and led to rapid lung viral clearance following lethal infection with homologous virus in the mouse model. Here we demonstrate that successful incorporation of the thermolabile cytokine GM-CSF into MN resulted in improved vaccine-induced protective immunity holding promise as a novel approach to improved influenza vaccination. To our knowledge, this is the first successful incorporation of a cytokine adjuvant into dissolvable MNs, thus advancing and diversifying the rapidly developing field of MN vaccination technology.

Heterosubtypic influenza protection elicited by double-layered polypeptide nanoparticles in mice

“…Dissolvable MN patch-based skin vaccination has enhanced the immunogenicity of influenza vaccines (56)(57)(58)(59). MN patch vaccination with inactivated influenza virus resulted in more efficient lung virus clearance and enhanced cellular recall responses than IM vaccination after influenza challenge (57).…”

Recent advances of microneedles for biomedical applications: drug delivery and beyond

“… Microneedle type Material Vaccine (type) Key finding Ref. Dissolving microneedle Hydroxyethyl starch 70000 Hepatitis B surface antigen (protein) The antigenicity of the Hepatitis B adjuvant maintained for 6 months at 50 °C with only a 10% loss 101 Dissolving microneedle Hyaluronic acid Live attenuated BCG (bacteria) Vaccination using microneedle array mixed with Bacille Calmette–Guerin (BCG) powder did not provoked severe inflammation and bruise as compared to intradermal injection 102 Dissolving microneedle Sucrose Rabies vaccination (nucleic acid) A rabies DNA vaccine integrated with dissolving microneedle was ten-fold lower vaccine dose than full-dose intramuscular vaccination 103 Dissolving microneedle Poly-vinyl alcohol Influenza (virus) Microneedle patch was capable of improving vaccine immunogenicity and increasing vaccination coverage 104 Hollow microneedle Polyimide IgG and IgG1 (protein) Nanoparticulate vaccines were discovered for protein antigen to control immune response using hollow microneedle 99 Solid microneedle and hollow microneedle Silicone Ovalbumin (nucleic acid) Hollow microneedle patches delivered ovalbumin efficiently for in vivo immunization in contrast to solid microneedle patches 105 Coated microneedle Ceramic Diphtheria and tetanus toxoid (peptide) Ceramic nanoporous microneedle arrays have a better capability of transporting diphtheria toxoid and tetanus toxoid 100 …”

“…testified that ceramic nanoporous microneedle arrays have a better capability of transporting diphtheria toxoid and tetanus toxoid. Obviously, microneedles transcend other methods depending on their overwhelming advantages including painless, portability, individual use and rapid delivery of vaccine (Table 3)99, 100, 101, 102, 103, 104, 105.…”

Cutaneous vaccination ameliorates Zika virus-induced neuro-ocular pathology via reduction of anti-ganglioside antibodies

“…MNPs were fabricated and underwent quality control testing as previously described. 41 Briefly, MNPs containing the vaccine were dissolved in PBS, ran on a 10% SDS-PAGE gel and immune-stained with anti-ZIKV E-protein antibody (EMD Millipore, MAB10216) in order to visualize protein bands and to determine successful incorporation of the inactivated virus. MNPs were fabricated in a two-step process with polydimethylsiloxane (PDMS) molds as previously described.…”

“…Spleens were also collected at 7 and 14 DPV (−53 and −46 DPI) to evaluate the presence of recently activated T-cell populations, as well as for differentiated B-cell subsets. 41 Samples were acquired on an LSR II flow cytometer (BD Biosciences) and data were analyzed with FlowJo (FlowJo LLC, BD, v9.9) (Gating strategies available in SFigure 2A, B, and C).…”