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Vaccination is an effective way to reduce the morbidity and mortality of infectious diseases. As a needle-free transcutaneous immunization (NF-TCI) vaccination technology, microneedles (MNs), composed of multiple micro-needles orderly attached to a substrate, can overcome the problems of low immune efficiency, poor compliance and waste of resources that exists in the conventional vaccination by injection, thus becoming a research hotspot in biomedicine. The microneedle vaccine can directly break through the stratum corneum barrier of the skin without touching nerves and blood vessels in the dermis, and effectively delivers the vaccine to the immune cells in the skin tissue to initiate the immune response of the body, thus triggering strong humoral and cellular immune processes. Vaccine delivery via the MNs system possesses the advantages of high safety, satisfying immune effect and practical economy, and shows great prospect in the prevention and treatment of infectious diseases and antineoplastic therapy. This article reviews the development background of MNs in transcutaneous immunization, the types of vaccine delivery, the factors affecting the immune effect, the problems to be solved and development direction in the future.
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An essential step for cancer vaccination is to break the immunosuppression and elicit a tumor-specific immunity. A major hurdle against cancer therapeutic vaccination is the insufficient immune stimulation of the cancer vaccines and lack of a safe and efficient adjuvant for human use. We discovered a novel cancer immunostimulant, trichosanthin (TCS), that is a clinically used protein drug in China, and developed a well-adaptable protein-engineering method for making recombinant protein vaccines by fusion of an antigenic peptide, TCS, and a cell-penetrating peptide (CPP), termed an "all-in-one" vaccine, for transcutaneous cancer immunization. The TCS adjuvant effect on antigen presentation was investigated and the antitumor immunity of the vaccines was investigated using the different tumor models. The vaccines were prepared
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Objective To screen the potent permeation enhancers used in transcutaneous immunization with inactivated highly pathogenic avian influenza vaccine. Methods Five different permeation enhancers, ethanol, propylene glycol, dimethyl sulfoxide, ratinoic acid, oleic acid, were used to treat the skin of BALB/c mice before transcutaneous immunization. Sera were collected before the flist transcutaneous immunization and every two weeks post immunization. The titers of influenza virus-specific humoral IgG and IgA were assayed in serum, lung and nasal lavages by ELISA. The titers of hemagglutination inhibition ( HAI), IFN-γand IL-4 produced by splenic lymphocytes were also detected. Except that, clinical symptom of the skin in different time points and skin pathological changes were observed. Results The serum IgG titers, HAI titers and the influenza virus-specific lgA and IgG in lung and nasal lavages in the groups of HA +CT + DMSO, HA + CT + RA and HA + CT + OA were significantly higher than those of HA and HA + CT groups( P ＜0.05). Moreover, the numbers of splenic lymphocytes producing IFN-γ and IL-4 were increased in the above three groups than those in control groups. In addition, no evident clinical symptoms were observed, but stratum corneum of the skin in different groups showed different changes. Conclusion DMSO,RA and OA are potent permeation enhancers in mouse model inoculated with inactivated high pathogenic avian influenza vaccine transcutaneously.
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OBJECTIVE: Previous reports have shown that transcutaneous immunization (TCI) with proteins or peptides in combination with adjuvants efficiently induces specific cellular and humoral immune responses. We compared the immune response after TCI with new construct which was derived from HPV-16 E7opt+K and pK6hf promoter instead of pCMV promoter and various adjuvant. METHODS: First, we made new construct ligated with HPV-16 E7 opt+K to Hair-follicle Specific pK6hf Promoter. Second, we applied pk6hf-E7 opt+K DNA with or without Lipofectamine 2000 and a combination of cholera toxin (CT) and CpG oligodeoxynucleotide (CpG) onto cold wax-depilated and hydrated bare skin of C57 BL/6 mice. To assess the ability of CTL(cytotoxic T-lymphocyte) activity, we performed intracellular cytokine staining with flow cytometric analysis to determine the number of E7-specific IFN-gamma- secreting CD8+ T cells generated in vaccinated mice with the DNA vaccine. RESULTS: Female C57BL/6 mice immunized by TCI methods with 30 microgram of pk6hf-E7 opt+K DNA with Lipofectamine2000 and CT efficiently generated E7-specific CD8(+) T cells compared with the group of pk6hf-E7 opt+K DNA only or DNA with Lipofectamine2000. CONCLUSION: Our results demonstrate that TCI of the linkaged-E7 DNA , E7 opt+K DNA to pk6hf, and Lipofectamine2000 and CT induced an antigen-specific CTL response. This result is of potential relevance for the development of therapeutic HPV-specific DNA vaccines with TCI and pK6hf promoter can be used safely.
Subject(s)
Animals , Female , Humans , Mice , Cholera Toxin , DNA , Human papillomavirus 16 , Immunity, Humoral , Immunization , Peptides , Skin , T-Lymphocytes , Vaccines, DNAABSTRACT
OBJECTIVE: Transcutaneous immunization (TCI) is a novel vaccination based on the application onto bare skin. We compared the immune response after TCI with the model DNA (OVALBUMIN) to HPV E7 and various adjuvant with intramuscular injection. We investigated the efficacy of immunization with new construct driven by K6hf promoter and compared with CMV promoter. METHODS: First, we make new construct ligated with OVA to Hair-follicle Specific pK6hf Promoter and evaluated the expression. Mouse skin was transfected with pCMV-OVA, pK6hf-OVA, pCMV-beta gal and pK6hf-beta gal and expression was determined by RT-PCR and X-Gal staining. OVA protein expression was analyzed by Western blot. Second, we immunized C57/ BL6 mice with pCMV-OVA or pK6hf-OVA DNA and cholera toxin (CT) and/or CpG. CTL was measured by ELISPOT assay of the splenocytes from the mmunized mice with the DNA vaccine. RESULTS: The beta-Galactosidase activity by X-Gal staining was detected in the epithelium of the mice skin after pK6hf-beta gal application. The mRNA and protein expression from pK6hf-OVA were evident following transcutaneous methods. Those were weaker than pCMV-OVA. TCI with pCMV-OVA and LipofectAMINE 2000 trigered an speicific CTL and Th2 response. CpG was the adjuvant for CTL after pCMV-OVA. CT and CpG did increase the CTL after pK6hf-OVA. CONCLUSION: Our data demonstrate that TCI of DNA is possible methods of CTL. CpG and CT were useful in the adjuvant for CTL. The pK6hf-OVA can induce specific CTL. This result is of potential relevance for the development of therapeutic HPV- specific DNA vaccines with TCI and pK6hf promoter can be used safely.