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

Abstract

Previous reports have shown that transcutaneous immunization (TCI) with proteins or peptides in combination with adjuvants efficiently induces specific cellular and humoral immune responses. However, depending on the kind of skin pretreatment, induction of cellular immune responses was restricted to generation of either specific cytotoxic T lymphocytes (CTLs) or T-helper (Th) cells. In this study, we induced antigen-specific CTL responses together with the appropriate Th responses by TCI of C57BL/6 mice. We applied ovalbumin protein or an ovalbuminderived fusion peptide containing a CTL and Th epitope together with a combination of cholera toxin (CT) and CpG oligodeoxynucleotide (CpG) onto cold wax-depilated and hydrated bare skin. TCI with the ovalbumin fusion peptide induced more robust CTL and Th responses than that with ovalbumin protein. The fusion peptide in combination with the nontoxic CT derivative CTA1-D2D1 and CpG induced an antigen-specific CTL response, albeit less efficiently than in combination with complete CT. Further, we compared the potency of HPV-16 E7 oncoprotein-derived peptides containing single (CTL) or multiple (CTL 1 Th 1 B cell) epitopes to induce effective CTL responses. Strong E7-specific CTL responses were detected only after TCI with the E7 multiepitope peptide. This peptide was also shown to protect mice against tumor growth after challenge with HPV-16 E7-positive tumor cells. TCI with E7 protein and CT/CpG led to formation of an E7-specific humoral immune response. ' 2005 Wiley-Liss, Inc.Key words: transcutaneous immunization; human papillomavirus type 16 E7 (HPV-16 E7); CTA1-D2D1; cytotoxic T lymphocyte; T helper cell; ovalbumin; tumor protection TCI is a novel strategy in modern vaccinology. By this application, blood-borne transmission of diseases through the reuse of needles, which is a common practice in remote parts of the world, can be circumvented and better compliance of vaccinees obtained. Apart from its benefits toward vaccine safety, TCI targets LCs, a dense population of highly accessible APCs within the skin. 1 An important feature of vaccination against viral infections and virus-based tumors is the induction of CTLs. 2-4 Upon activation by APCs, predominantly DCs, CTLs are able to selectively kill antigen-expressing cells. 5 For clearance of certain viral infections, the aid of Th cells for CTL activation is necessary. 6 The bias to one of the 2 Th subsets (Th1 > Th2) is a prerequisite for specific CTL induction. 7 Generation of specific CTLs against proteinaceous or peptidebased vaccines in combination with adjuvants after TS of the skin is possible. 8-10 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 ...

Human B Cells Secrete Granzyme B When Recognizing Viral Antigens in the Context of the Acute Phase Cytokine IL-21

“…Furthermore, the BCR can recognize epitopes within larger structures such as virus particles, and the spectrum of Ags recognized is not limited to peptide Ags but also includes carbohydrate, phospholipid, and nucleic acid Ags (3,4). Not surprisingly in this context, the presence of MHC-independent epitopes in addition to MHC-restricted epitopes could significantly enhance cellular immune responses in a recently described mouse model of human papillomavirus-induced tumors (25). With the intent to combine the advantages of both TCR and BCR, another group designed genetically engineered T cells with chimeric receptors.…”

Initiation of adaptive immune responses by transcutaneous immunization

“…The bacterial products CT and LT [31], which are able to serve remarkably as antigen and adjuvant, have been used predominantly to evoke humoral and/or cellular immunity. Other studies take advantage of the well-described model antigen Ovalbumin [9][10][11]. However, as discussed above, approaches involving such large molecules require an 'adequate' pre-treatment of the skin, in order to allow the passage through the skin barrier.…”

“…The majority of established TCI protocols use large immunogenic molecules like CT (86 kDa) or LT (84 kDa) as adjuvants and/or antigen [8] in combination with whole protein antigens like bovine serum albumin [8][9][10][11], diphteria toxoid (DT) [12], tetanus toxoid (TT) [12,13] among others. Therefore, they rely on an initial step of skin barrier disruption, generally either by tape-stripping [14,15], degreasing with acetone [9,10] or hydration [8,16].…”

Transcutaneous Immunization withClostridium difficileToxoid A Induces Systemic and Mucosal Immune Responses and Toxin A-Neutralizing Antibodies in Mice

“…Interestingly, TCI induces both systemic and mucosal immune responses (6,22,23,28,41,42,48). TCI has been shown to be safe and effective in animals and humans (9,21,23,42,47,58). In order to assess whether TCI would induce immune responses against C. difficile toxin A, we therefore transcutaneously immunized mice with a toxoid derivative of C. difficile toxin A (CDA), with or without the immunoadjuvant CT, and measured systemic and mucosal anti-CDA immune responses, including induction of toxin Aneutralizing antibodies in immunized mice.…”