Transcutaneous immunization without heterologous adjuvant

Abstract

Transcutaneous immunization can deliver antigen to the immune system through the stratum corneum without physical or chemical penetration to the dermis layer of the skin. This delivery system induces an antigen-specific immune response without the use of a heterologous adjuvant. Although perforation of intact skin is not required, superficial penetration or micropenetration of the skin can act as an enhancer; similarly, hydration may enhance the immune response. This system can induce antigen-specific immune effectors after epicutaneous application of a formulation containing one or more antigens. The formulation may initiate processes such as antigen uptake, processing, and presentation; Langerhans cell activation, migration from the skin to other immune organs, and differentiation to mature dendritic cells; contacting antigen with lymphocytes bearing cognate antigen receptors on the cell surface and their stimulation; and combinations thereof. Systemic and / or regional immunity may be induced. Immune responses that provide prophylactic and / or therapeutic treatments are preferred. Antigenic activities in the formulation may be found in the same molecule, two or more different molecules dissociated from each other, or multiple molecules in a complex formed by covalent or non-covalent bonds. For antigens which are proteinaceous, they may be provided in the formulation as a polynucleotide for transcutaneous genetic immunization. Besides simple application of a dry or liquid formulation to the skin, patches and other medical devices may be used to deliver antigen for immunization.

Description

DESCRIPTION OF RELATED APPLICATIONS [0001] This application is a continuation in-part of U.S. appln. Ser. No. 08 / 896,085 (filed Jul. 17, 1997 and pending); U.S. application Ser. No. 09 / 157,395 (filed Sep. 21, 1998 and pending) which is a divisional of U.S. application Ser. No. 08 / 749,164 (filed Nov. 14, 1996, now U.S. Pat. No. 5,910,306); U.S. application Ser. No. 09 / 257,188 (filed Feb. 25, 1999 and pending); U.S. application Ser. No. 09 / 309,881 (filed May 11, 1999 and pending); and U.S. application Ser. No. 09 / 311,720 (filed May 14, 1999 and pending) which is a continuation in-part of PCT / US97 / 21324 (filed Nov. 14, 1997 designating the U.S., now abandoned). [0002] This application also claims priority benefit from provisional U.S. Appln. No. 60 / 090,169 (filed Jun. 22, 1998) and U.S. Appln. No. 60 / 128,370 (filed Apr. 8, 1999).[0003] All patent applications cited herein, as well as patents issued therefrom, are incorporated by reference in their entirety. BACKGROUND OF THE INVENTION ...

AI Technical Summary

Benefits of technology

[0009] The stratum corneum, a layer of dead skin cells and lipids, has traditionally been viewed as a barrier to the hostile world, excluding organisms and noxious substances from the viable cells below the stratum corneum (Bos, 1997a). The secondary protection provided by skin antigen presenting cells such as Langerhans cells has only recently been recognized (Celluzzi and Falo, 1997). Moreover, the ability to immunize through the skin using the crucial concept of a skin-active adjuvant has only been recently described (Glenn et al., 1998a). Scientific recognition of this important advance in vaccination was prompt. “It's a very surprising result, and it's lovely,” said vaccine expert Barry Bloom of the Howard Hughes Medical Institute and the Albert Einstein College of Medicine in New York, the strategy sounds “very easy, very safe, and certainly inexpensive” (CNN News, Feb. 26, 1998).

[0035] In contrast to the expectations of the art, our delivery system provided by transcutaneous immunization is capable of achieving efficient delivery of at least antigen and / or polynucleotide encoding antigen through the skin to the immune system.

Problems solved by technology

These homologous proteins cause intestinal fluid secretion and massive diarrhea (Spangler, 1992), and are viewed as dangerous toxins.

Thus, one could have reasonably expected that CT would be extremely reactogenic when placed on the skin or inserted through the stratum corneum, and would cause similar redness and swelling.

Craig (1965) cautioned, “The absence of skin lesions in clinical cholera certainly does not preclude the possibility that the noxa responsible for gut damage could also have a deleterious effect upon the skin provided it is applied to skin in sufficient concentration.” The extreme reactogenicity of cholera toxin in the skin was used as a test for its toxicity and such prior art evidenced an expectation that cholera toxin would be reactogenic if applied to the skin, producing an undesirable reaction.

This lack of reactogenicity when cholera toxin was placed on the skin for transcutaneous immunization was surprising and contradicted conclusions one would have drawn from the prior art.

A liquid formulation of CT placed on the skin acted as a non-toxic, non-reactogenic adjuvant, in contrast to the expectations of Craig, while injection of CT into the skin results in swelling and redness.

Our findings, however, unexpectedly showed that such formulations are devoid of reactogenicity.

Besides the physical restriction of limiting passage through the skin of low molecular weight, passage of polypeptides was believed to be limited by chemical restrictions.

(U.S. Pat. No. 5,679,647) stated that “it is believed that the bioavailability of peptides following transdermal or mucosal transmission is limited by the relatively high concentration of proteases in these tissues.

Yet unfortunately, reliable means of delivering peptides .

Images