Percutaneous peptide immunization via corneum barrier-disrupted murine skin for experimental tumor immunoprophylaxis

Abstract

H-2K b -restricted tumor epitope peptides, including tyrosinase-related protein 2 residues 181-188 (TRP-2) and connexin 37 residues 52-59 (MUT1), were applied to permeability barrier-disrupted C57BL͞6 (B6) mouse skin from which the stratum corneum of the epidermis had been removed by tape-stripping. This procedure primed tumor-specific cytotoxic T lymphocytes (CTLs) in the lymph nodes and spleen, protected mice against subsequent challenge with corresponding tumor cells, and suppressed the growth of established tumors. Preventive and therapeutic effectiveness was correlated with the frequency of tumor-specific CTL precursors. MHC class II Ia b؉ cells separated from tape-stripped skin, compared with those from intact skin, exhibited a strong antigen-presenting capacity for CTL, suggesting that CTL expansion after peptide application is primarily mediated by epidermal Langerhans cells. Thus, percutaneous peptide immunization via barrier-disrupted skin provides a simple and noninvasive means of inducing potent anti-tumor immunity which may be exploited for cancer immunotherapy. One of the principal goals in tumor immunoprophylaxis and therapy is induction of anti-tumor responses by generating sufficient numbers of tumor antigen-specific cytotoxic T lymphocytes (CTLs). In tumor bearers, however, not only effective CTL priming but also recognition and effective lysis of tumor cells by CTLs are often blocked or attenuated by down-regulated expression of major histocompatibility complex (MHC) class I antigens and of costimulatory and adhesion molecules on tumorcell surfaces (1), suggesting one evasion mechanism of tumor cells which affects immune surveillance. Therefore, establishment of efficient CTL priming strategies is an important issue for cancer immunotherapy.Dendritic cells are potent antigen-presenting cells distributed widely at epithelial surfaces which interface with the external environment to facilitate efficient antigen trapping (2, 3). Because of high levels of expression of MHC class I antigen and costimulatory and adhesion molecules, tumor-specific CTLs are efficiently induced with tumor antigen-or peptide-loaded dendritic cells (4-7). Studies in animal models have shown the potential of dendritic cell-based tumor immunotherapy to elicit protective anti-tumor immune responses, and the validity of such an approach has also been confirmed in humans (8-10). While understanding of the mechanism of propagation of dendritic cells under cytokine influence has progressed at a rapid pace, the intricate manipulations required for in vitro preparation of large numbers of dendritic cells in a form appropriate for immunotherapy seem to be a major drawback in current immunotherapeutic strategies.We have reported that disruption of the skin barrier results in not only enhanced permeability but also alterations in the immunoregulatory function of the skin in such a way that epidermal Langerhans cells (LCs) function as vigorous antigen presenters for T-helper cells (11). Various studies have shown that LCs target tumo...

Layer-by-Layer-Assembled Multilayer Films for Transcutaneous Drug and Vaccine Delivery

“…46,[48][49][50] An additional advantage is that physical barrier disruption itself has been shown to trigger activation of keratinocytes and LCs in skin, providing some level of 'built-in' adjuvant response. [44][45][46][49][50][51][52] We thus tested a model where the dorsal ears of anesthetized mice were tape-stripped followed by application of as-assembled, airdried (Poly-1/ova) 40 films on either glass or PDMS substrates, fixed in place by surgical tape against the exposed ear skin, mimicking a common strategy used in skin vaccination studies 45,46 (Figure 1a). Tape-stripping reproducibly removed significant portions of the SC but did not disrupt the viable epidermis, as assessed by histology ( Figure S3).…”

Langerhans cells cross-present antigen derived from skin

“…Epicutaneous or transmucosal immunization with protein and peptides has been reported to be efficient in inducing IFN-␥ production and cytotoxicity in T cells (38), especially when combined with inflammatory stimuli like tape stripping, cholera toxin, and Toll-like receptor ligands, such as oligonucleotides and imiquimod (16)(17)(18)(19). Seo et al (16) showed that LCs from tape-stripped skin are activated and express higher levels of costimulatory molecules and MHC-class I. When these LCs were peptide pulsed, they were able to induce cyotoxicity in CD8 ϩ T cells, indicating that they might be involved in this process (16).…”

“…Seo et al (16) showed that LCs from tape-stripped skin are activated and express higher levels of costimulatory molecules and MHC-class I. When these LCs were peptide pulsed, they were able to induce cyotoxicity in CD8 ϩ T cells, indicating that they might be involved in this process (16). However, this was not formally proven, and therefore, the precise role of LCs still remains unclear.…”

“…In vitro generated human LCs derived from CD34 ϩ precursors are able to cross-present exogenous antigen to CD8 ϩ T cells (13,14), and human Langerhans-like cells are more potent than dermal-like and monocyte-derived DCs (15). Two recent reports showed that epicutaneous immunization with an MHC-class I-restricted peptide or with ovalbumin protein onto tape-stripped skin induces cytotoxic T cell activity in the draining lymph node which can be further increased by coadministration of cholera toxin (16,17). Others reported that these responses can also be boosted with Toll-like receptor ligands, such as oligonucleotides and imiquimod (18,19).…”

Transcutaneous immunization in mice: Induction of T-helper and cytotoxic T lymphocyte responses and protection against human papillomavirus-induced tumors

“…[8][9][10] TS leads to removal of the outermost horny layer of the skin, resulting in enhanced permeability and maturation and activation of LCs via inflammatory cytokines secreted from keratinocytes. 11,12 The barrier function of the SC can also be bypassed by hydration of the skin, which leads to swelling of the keratinocytes and pooling of fluid into intercellular spaces. This kind of pretreatment of the skin leads to development of humoral 13,14 and Th responses after TCI with proteins or peptides.…”