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Adjuvant for transcutaneous immunization

a technology of adjuvant and immunization, applied in the field of adjuvant, can solve the problem of not being expected to achieve immunization

Inactive Publication Date: 2006-11-30
UNITED STATES OF AMERICA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0013] An object of the invention is to provide a system for transcutaneous immunization that induces an immune response (e.g., humoral and / or cellular effectors) in an animal or human. The system provides simple application to intact skin of an organism of a formulation comprised of antigen and adjuvant to induce a specific immune response against the antigen. In particular, the adjuvant may activate antigen presenting cells of the immune system (e.g., Langerhans cells in the epidermis, dermal dendritic cells, dendritic cells, macrophages, B lymphocytes) and / or induce the antigen presenting cells to phagocytose the antigen. The antigen presenting cells then present the antigen to T and B cells. In the instance of Langerhans cells, the antigen presenting cells then may migrate from the skin to the lymph nodes and present antigen to lymphocytes (e.g., B and / or T cells), thereby inducing an antigen-specific immune response.
[0023] Inclusion of an adjuvant may allow potentiation or modulation of the immune response. Moreover, selection of a suitable antigen or adjuvant may allow preferential induction of a humoral or cellular immune response, specific antibody isotypes (e.g., IgM, IgD, IgA1, IgA2, IgE, IgG1, IgG2, IgG3, and / or IgG4), and / or specific T-cell subsets (e.g., CTL, Th1, Th2 and / or TDTH).

Problems solved by technology

Therefore, the reference would teach away from using a molecule like cholera toxin (which is 85,000 daltons) because such molecules would not be expected to penetrate the skin and, therefore, would not be expected to achieve immunization.

Method used

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  • Adjuvant for transcutaneous immunization
  • Adjuvant for transcutaneous immunization

Examples

Experimental program
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Effect test

example 1

[0115] BALB / c mice at 6 to 8 weeks of age were immunized transcutaneously as described above for “Immunization Procedure”, in groups of five mice. The mice were immunized using 100 μl of immunization solution which was prepared as follows: liposomes prepared as described above for “Liposome Preparation” were mixed with saline to form the liposomes. The pre-formed liposomes were then diluted in either saline (liposome alone group) or with CT in saline to yield an immunizing solution containing liposomes at 10-150 mM phospholipid with 100 μg of CT per 100 μl of immunizing solution. CT was mixed in saline to make an immunizing solution containing 100 μg of CT per 100 μg of solution for the group receiving CT alone. Solutions were vortexed for 10 seconds prior to immunization.

[0116] The mice were immunized transcutaneously at 0 and 3 weeks. Antibody levels were determined using ELISA as described above for “ELISA IgG(H+L)” 3 weeks after the boosting immunization, and compared against p...

example 2

[0117] BALB / c mice at 6 to 8 weeks of age were immunized transcutaneously as described above for “Immunization Procedure”, in groups of five mice. The mice were immunized at 0 and 3 weeks using 100 μl of immunization solution prepared as follows: BSA was mixed in saline to make an immunizing solution containing 200 μg of BSA per 100 μl of saline for the group receiving BSA alone; BSA and CT were mixed in saline to make an immunizing solution containing 200 μg of BSA and 100 μg of CT per 100 μl of saline for the group receiving BSA and CT. Where liposomes were used, the liposomes were prepared as described above for “Liposome Preparation”, and were first mixed with saline to form the liposomes. They were then diluted in BSA or BSA and CT in saline to yield an immunizing solution containing liposomes at 50 mM phospholipid with 200 μg of BSA per 100 μl of immunizing solution, or 200 μg BSA+100 μg CT per 100 μl of immunizing solution. Solutions were vortexed for 10 seconds prior to immu...

example 3

[0119] BALB / c mice at 6 to 8 weeks of age were immunized transcutaneously as described above for “Immunization Procedure”, in groups of five mice. The mice were immunized at 0 and 3 weeks using 100 μl of immunization solution prepared as follows: LT was mixed in saline to make an immunizing solution containing 100 μg of LT per 100 μl of saline for the group receiving LT alone. Where liposomes were used the liposomes prepared as described above for “Liposome Preparation”, and were first mixed with saline to form the liposomes. The pre-formed liposomes were then diluted in LT in saline to yield an immunizing solution containing liposomes at 50 mM phospholipid with 100 μg of LT per 100 μl of immunizing solution. Solutions were vortexed for 10 seconds prior to immunization.

[0120] The anti-LT antibodies were determined using ELISA as described above for “ELISA IgG(H+L)” 3 weeks after the second immunization. The results are shown in Table 3. LT was clearly immunogenic both with and with...

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Abstract

A transcutaneous immunization system delivers antigen to immune cells without perforation of the skin, and induces an immune response in an animal or human. The system uses an adjuvant, preferably an ADP-ribosylating exotoxin, to induce an antigen-specific immune response (e.g., humoral and / or cellular effectors) after transcutaneous application of a formulation containing antigen and adjuvant to intact skin of the animal or human. The efficiency of immunization may be enhanced by adding hydrating agents (e.g., liposomes), penetration enhancers, or occlusive dressings to the transcutaneous delivery system. This system may allow activation of Langerhans cells in the skin, migration of the Langerhans cells to lymph nodes, and antigen presentation.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This is a continuation in-part of U.S. application Ser. No. 08 / 749,164 filed Nov. 14, 1996.GOVERNMENT RIGHTS [0002] The U.S. government may retain certain rights in this invention.BACKGROUND [0003] The invention relates to transcutaneous immunization, and adjuvants useful therein, to induce an antigen-specific immune response. [0004] Transcutaneous immunization requires both passage of an antigen through the outer barriers of the skin, which are normally impervious to such passage, and an immune response to the antigen. We showed in U.S. application Ser. No. 08 / 749,164 that using cholera toxin as an antigen elicits a strong antibody response that is highly reproducible; the antigen could be applied in a saline solution to the skin, with or without liposomes. In the present application, we show transcutaneous immunization using adjuvants such as, for example, bacterial exotoxins, their subunits, and related toxins. [0005] There is a repo...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): A61K39/395A61K39/00A61K9/127A01N37/18A61F13/00A61K9/00A61K9/06A61K9/08A61K9/107A61K9/52A61K9/56A61K9/70A61K35/14A61K35/39A61K39/015A61K39/05A61K39/08A61K39/10A61K39/102A61K39/104A61K39/106A61K39/39A61P37/00A61P37/04
CPCA61K9/06A61K9/08A61K9/107A61K9/127A61K9/7023A61K39/39B82Y5/00A61K2039/54A61K2039/55511A61K2039/55527A61K2039/55544A61K2039/55555A61K2039/55566A61K2039/53A61P37/00A61P37/04A61K39/00
Inventor GLENN, GREGORY M.ALVING, CARL R.
Owner UNITED STATES OF AMERICA
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