The Journal of Allergy and Clinical Immunology
Volume 120, Issue 6 , Pages 1382-1388, December 2007

Impaired immune response to vaccinia virus inoculated at the site of cutaneous allergic inflammation

  • Jordan E. Scott, MD

      Affiliations

    • Department of Pediatrics, Division of Allergy and Immunology, Children's Hospital, Harvard Medical School, Boston, Mass
    • These authors contributed equally to this work.
    • ,
  • Abdallah ElKhal, PhD

      Affiliations

    • Department of Pediatrics, Division of Allergy and Immunology, Children's Hospital, Harvard Medical School, Boston, Mass
    • These authors contributed equally to this work.
    • ,
  • Eva-Jasmin Freyschmidt, PhD

      Affiliations

    • Department of Pediatrics, Division of Allergy and Immunology, Children's Hospital, Harvard Medical School, Boston, Mass
    • ,
  • Daniel H. MacArthur, BS

      Affiliations

    • Department of Pediatrics, Division of Allergy and Immunology, Children's Hospital, Harvard Medical School, Boston, Mass
    • ,
  • Douglas McDonald, MD, PhD

      Affiliations

    • Department of Pediatrics, Division of Allergy and Immunology, Children's Hospital, Harvard Medical School, Boston, Mass
    • ,
  • Michael D. Howell, PhD

      Affiliations

    • National Jewish Medical and Research Center, Denver, Colo
    • ,
  • Donald Y.M. Leung, MD, PhD

      Affiliations

    • National Jewish Medical and Research Center, Denver, Colo
    • ,
  • Amale Laouar, PhD

      Affiliations

    • Center for Blood Research, Institute for Biomedical Research, Harvard Medical School, Boston, Mass
    • ,
  • N. Manjunath, PhD

      Affiliations

    • Center for Blood Research, Institute for Biomedical Research, Harvard Medical School, Boston, Mass
    • ,
  • Teresa Bianchi, PhD

      Affiliations

    • Brigham and Women's Hospital and Department of Pediatrics and Dermatology, Harvard Medical School, Boston, Mass
    • ,
  • Marianne Boes, PhD

      Affiliations

    • Brigham and Women's Hospital and Department of Pediatrics and Dermatology, Harvard Medical School, Boston, Mass
    • ,
  • Hans C. Oettgen, MD, PhD

      Affiliations

    • Department of Pediatrics, Division of Allergy and Immunology, Children's Hospital, Harvard Medical School, Boston, Mass
    • ,
  • Raif S. Geha, MD

      Affiliations

    • Department of Pediatrics, Division of Allergy and Immunology, Children's Hospital, Harvard Medical School, Boston, Mass
    • Corresponding Author InformationReprint requests: Raif S. Geha, MD, Division of Immunology, Karp 10th floor, One Blackfan Street, Boston, MA 02115.

Received 9 April 2007; received in revised form 30 July 2007; accepted 1 August 2007. published online 24 September 2007.

Article Outline

Background

Patients with atopic dermatitis (AD) exposed to the vaccinia virus (VV) smallpox vaccine have an increased risk of developing eczema vaccinatum.

Objective

To investigate the effects of local allergic skin inflammation on vaccinia immunity.

Methods

BALB/c mice were epicutaneously sensitized with ovalbumin (OVA) to induce allergic skin inflammation or with saline control, then inoculated with an attenuated VV strain by skin scarification or intraperitoneally. After 8 days, serum IgG anti-VV and cytokine secretion by splenocytes were measured.

Results

Mice inoculated with VV at sites of epicutaneous sensitization with OVA, but not control mice inoculated at saline exposed sites, developed satellite pox lesions and had impaired secretion of TH1 cytokines in response to VV, decreased VV specific serum IgG2a, increased VV specific serum IgG1, and impaired upregulation of IFN-α, but not the cathelicidin-related antimicrobial peptide, at the infection site. The VV immune response of OVA-sensitized mice inoculated with VV at distant skin sites or intraperitoneally was normal.

Conclusion

Local immune dysregulation at sites of allergic skin inflammation underlies the impaired TH1 immune response to VV introduced at these sites and the increased susceptibility to develop satellite pox lesions, a characteristic of eczema vaccinatum in patients with AD.

Clinical implications

In a mouse model of AD, inoculation of VV at inflamed skin sites is associated with increased numbers of satellite pox lesions and an abnormal immune response to the virus. This may contribute to the susceptibility of patients with AD to virus dissemination after smallpox vaccination.

Key words: Eczema vaccinatum, allergy, vaccinia virus, smallpox vaccination, viral response, TH1/TH2 cells

Abbreviations used: AD, Atopic dermatitis, CRAMP, Cathelicidin-related antimicrobial peptide, DC, Dendritic cell, DMEM, Dulbecco modified Eagle medium, EV, Eczema vaccinatum, LN, Lymph node, OVA, Ovalbumin, pDC, Plasmacytoid dendritic cell, PFU, Plaque-forming unit, VV, Vaccinia virus

 

Smallpox, which is caused by infection with variola virus, has a fatality rate as high as 30%.1 Prophylactic smallpox vaccination with vaccinia virus (VV) is the only known protection. Smallpox was eradicated in 1979 by mass immunization with VV. The only known stocks of variola virus are stored near Novosibirsk, Russia, and at the Centers for Disease Control and Prevention in Atlanta. Concern has recently increased about the potential misuse of variola virus as a bioterrorism weapon and has led to consideration of resuming mass immunization against smallpox. Although effective, VV immunization is associated with significant complications.2 One of these complications is eczema vaccinatum (EV), which is caused by localized and/or systemic viral dissemination that can be lethal. Patients with AD are predisposed to develop EV, which occurs in roughly 10 to 40 per million vaccinees with an approximately equal number of secondary cases in contacts of vaccinees.2, 3 Although EV is usually confined to the skin and self-limited, it can progress to systemic disease. Mortality may be as high as 6% in cases involving vaccinees, and as high as 4% in secondary cases.4, 5 Although smallpox vaccination is contraindicated in patients with atopic dermatitis (AD), they may develop EV after contact with a recently immunized individual, as has been documented lately in a child whose father had received the vaccine.6 The immunologic basis of the enhanced susceptibility of patients with AD to EV is not well understood.

Atopic dermatitis is a genetically determined pruritic chronic relapsing inflammatory skin disease that currently affects as many as 20% of children in the United States.7 In acute lesions, CD4+ cells are predominantly of the TH2 type and express IL-4, IL-5, and IL-13, whereas in chronic lesions, IFN-γ–producing TH1 cells may dominate.7 AD is associated with a systemic allergic response evidenced by elevated serum IgE levels, peripheral blood eosinophilia, and a propensity of T cells to secrete TH2 cytokines in response to anti-CD3 stimulation.7

Both the TH1 cytokine IFN-γ and the IgG2a antibody response it drives have been shown to be important for viral clearance and containment in mice.8, 9, 10 In contrast, the TH2 cytokines IL-4 and IL-10 inhibit viral clearance and increase susceptibility for VV dissemination.11, 12 We hypothesized that the susceptibility of patients with AD to EV may result from an altered TH immune response to VV. This could be caused by local immune dysregulation in the inflamed skin where the virus is introduced. Alternatively, it may result from a globally altered systemic immune response.

To address these questions, we took advantage of a murine model of allergic skin inflammation we have established that mimics many aspects of the allergic skin inflammation and the systemic TH2 response observed in patients with AD.13, 14, 15 We compared the clinical and immune response of epicutaneously sensitized mice to VV inoculation at sites of allergic skin inflammation, distant skin sites, and intraperitoneally. We used an attenuated VV strain because efforts are currently aimed at examining the safety and efficacy of attenuated VV strains (eg, Modified Vaccinia Ankara) for the immunization of immunodeficient subjects and patients with AD.16 Our results show that inoculation of an attenuated VV strain at sites of allergic skin inflammation, but not at distant skin sites or intraperitoneally, was associated with a impairment of the TH1 immune response to the virus and the development of satellite pox lesions, a characteristic feature of EV.

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Methods 

Virus source and expansion 

The VV Western Reserve Thymidine kinase mutant strain (VTK-79L) was obtained from American Type Culture Collection (Manassas, Va; VR-2056). CV1 cells (American Type Culture Collection; CCL-70) grown to 80% confluence in Dulbecco modified Eagle medium (DMEM) supplemented with 10% FCS for propagation of the virus were rinsed and overlaid with DMEM/2.5% FCS containing VV at a ratio of 1 plaque-forming unit (PFU) per cell. Then a 10-fold volume of DMEM/2.5% FCS was added, and infected cells were incubated at 37°C in 5% CO2 for 2 days. VV was harvested after disruption of the cells by 3 freeze-thaw cycles, and its titer was assessed by a standard plaque-forming assay using CV1 cells.17

EC sensitization and vaccinia infection protocols 

Female BALB/c mice 4 to 6 weeks old were EC sensitized as described previously.13 After five 1-week exposures to the patches separated by 2-week intervals, mice were inoculated with VV at the site of EC sensitization or at a site on the upper back removed as far as possible from the sensitization patch (Fig 1, A). Ten microliters of PBS containing 1 × 107 PFU VV was placed on the skin, followed by administration of 20 superficial scratches using a 27½-gauge needle, and the inoculation site was covered with a gauze patch and Tegaderm (Westnet Inc, Canton, Mass). Mice were weighed daily and examined for lethargy and the development of satellite lesions and were killed 8 days after infection or at days 1, 2, 4, and 8 to determine viral loads in skin. In other experiments, EC sensitized mice were inoculated with VV intraperitoneally using 5 × 106 PFU in 200 μL DMEM, and their immune response was examined 8 days later. All experiments were reviewed and authorized by the Children's Hospital Institutional Animal Care and Use Committee.

  • View full-size image.
  • Fig 1. 

    A, EC sensitization and VV infection protocol. B and C, Skin infiltration with CD4+ T cells and eosinophils and expression of TH2 and TH1 cytokines at the time of VV inoculation. D, Representative satellite skin lesion. E, Satellite lesions in mice inoculated with VV at OVA versus saline-sensitized sites. ∗P < .05. HPF, High-power field; Eos, eosinophils; SAL, saline; ns, not significant.

Microscopic analysis of skin 

Skin sections of 4 μm were stained with hematoxylin and eosin. CD4 staining of frozen skin sections was performed as previously described.13 Positive cells were counted blinded in 10 to 15 high-power fields.

Quantitative PCR analysis of VV genome copies 

Tissue samples were immediately frozen and stored at –80°C. Quantification of VV genomes was performed as previously described.18

In vitro cytokine synthesis of splenocytes 

One million splenocytes were incubated with 1 × 106 A-20 (H2d) B cells derived from a mouse B-cell lymphoma line that had been infected with VV for 24 hours or left uninfected, then irradiated. As positive control, cells were incubated in plates coated with 0.2 μg/mL anti-CD3ɛ chain (clone 145-2C11) mAb (BD Bioscience Pharmingen, San Diego, Calif). Supernatants were collected after 24 hours (for IFN-γ and IL-2 detection) and 72 hours (for IL-4, IL-5, and IL-13 detection), and cytokine concentration was determined by ELISA (BD Bioscience Pharmingen).

Vaccinia specific IgG2a and IgG1 antibody ELISA 

Flat-bottom 96-well microtiter plates (Immulon 2HB, Thermo Scientific, Milford, Mass) were coated overnight with 1 × 106 PFU/well of purified inactivated VV (Advanced Biotechnologies, Columbia, Md). After blocking with 1% BSA/PBS for 2 hours, sera were added, diluted 1:10 for IgG2a and 1:1000 for IgG1 in duplicate, and incubated overnight at 4°C. Biotin-conjugated goat antibodies against mouse IgG2a and IgG1 (BD Bioscience Pharmingen) were used for detection at 2 μg/mL. Washing steps between incubations were performed with 0.05% Tween/PBS. ODs were measured at 405 nm.

Quantitative PCR for cytokines, cathelicidin-related antimicrobial peptide, IFN-α, and IFN-γ 

RNA was prepared from skin homogenates, and quantitative PCR was performed as previously described19 by using primers and an ABI Prism 7300 sequence detector from Applied Biosystems, Foster City, Calif. Relative expression levels were calculated by the relative standard curve method as outlined in the manufacturer's technical bulletin (Applied Biosystems). Quantities of all targets in test samples were normalized to the corresponding glyceraldehyde 3-phosphate dehydrogenase (GAPDH) levels in the skin and expressed as Target Gene normalized to GAPDH.

Statistical analyses 

Differences in values between experimental groups were examined for significance with GraphPad Prism software using the 2-tailed Student t test.

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Results 

Clinical manifestations of BALB/c mice infected with vaccinia by skin scarification at the site of EC sensitization 

Fig 1, B and C, shows that skin sites EC sensitized with ovalbumin (OVA) for five 1-week cycles exhibit allergic skin inflammation similar to that previously documented in skin sites EC sensitized with OVA for three 1-week cycles.13, 14 This allergic inflammation is characterized by infiltration with eosinophils and CD4+ cells (Fig 1, B) and increased expression of the TH2 cytokines IL-4 and IL-13, with no increase in IFN-γ expression (Fig 1, C).

After VV inoculation, mice in both OVA and saline-sensitized groups maintained baseline weights (data not shown). However, only OVA-sensitized mice developed lethargy and exhibited satellite skin lesions (Fig 1, D and E). VV genomes were detected at the inoculation site on days 1, 2, 4, and 8, with no significant differences between OVA and saline-sensitized groups (data not shown). We were unable to detect VV genomes in satellite skin lesions, suggesting that the virus was cleared from these sites by day 8. Viral loads in lung and kidney at day 8 were not significantly different between the 2 groups (data not shown).

Cytokine expression by splenocytes of vaccinia infected BALB/c mice 

There is evidence that IFN-γ promotes whereas TH2 cytokines impair the clearance of VV.11, 12, 20, 21 Splenocytes from mice inoculated with VV at saline-exposed sites mounted a robust IFN-γ and IL-2 response to VV stimulation. In contrast, splenocytes from mice inoculated with VV at OVA-sensitized sites had significantly diminished production of IFN-γ and IL-2 in response to VV (Fig 2, A) but comparable production of IL-4 and IL-13 (Fig 2, B). Production of IFN-γ and IL-2 in response to anti-CD3 was comparable between OVA and saline-sensitized mice infected with VV (data not shown), ruling out a generalized impairment of TH1 cytokine production in OVA-sensitized mice.

Decreased vaccinia specific IgG2a antibody response in mice inoculated with VV in OVA-sensitized skin sites 

Immunization with live VV induces protective IgG antibody.9 The vaccinia-specific serum IgG2a antibody level was significantly decreased in mice inoculated at OVA-sensitized sites relative to mice inoculated at saline-exposed sites (P = .02), whereas the vaccinia-specific serum IgG1 antibody level was modestly increased with borderline significance (P = .047; Fig 3). Because IFN-γ is important for IgG2a responses and inhibits IgG1 responses,22 these findings support the observation that the IFN-γ response to VV is impaired in mice inoculated with VV in OVA-sensitized skin.

Vaccinia virus inoculation at a skin site distant from the site of EC sensitization with OVA or intraperitoneally elicits a virtually normal immune response to VV in EC sensitized mice. The impaired TH1 response of mice to VV infection at the site of EC sensitization with OVA may be a result of a global predisposition of these mice to mount a TH2 response or/and to a local effect of allergic skin inflammation on the response to VV introduced in EC sensitized skin. To distinguish between these possibilities, we examined the response to VV inoculated at a distant skin site. OVA-sensitized mice inoculated at a distant skin site exhibited no lethargy and did not develop satellite skin lesions, and their splenocytes exhibited only a modest, albeit significant, 2-fold decrease in IFN-γ production in response to VV (Fig 4, A). Secretion of the TH2 cytokines IL-4 and IL-13 as well as IgG2a and IgG1 responses were comparable in the 2 groups (Fig 4, B, and data not shown).

To confirm that EC sensitization has little effect on the response to VV introduced at sites other than the sensitization site, we examined the immune response to VV inoculation in the peritoneal cavity. Splenocytes from mice EC sensitized with OVA and intraperitoneally inoculated with VV produced similar amounts of IFN-γ and IL-2 in response to VV stimulation as splenocytes from similarly inoculated mice EC sensitized with saline (Fig 5, A). Furthermore, the VV specific IgG2a and IgG1 Ab responses were similar in these 2 groups (Fig 5, B).

Expression of IFN-α, but not cathelicidin-related antimicrobial peptide (CRAMP) or IFN-γ mRNA, is impaired after VV infection. Members of the cathelicidin family of antimicrobial peptides exhibit antiviral activity against VV,23 are downregulated by the TH2 cytokines IL-4 and IL-13 and in skin lesions of atopic dermatitis,24, 25 and augment TH1 responses.26 Decreased cathelicidin expression at sites of OVA sensitization could contribute to the decreased TH1 response after VV inoculation. Fig 6, A, shows that there is significantly less CRAMP mRNA expression in OVA-sensitized compared with saline-exposed skin sites before VV inoculation. VV infection, however, significantly upregulated CRAMP expression in both sites to a comparable degree with no significant difference between the 2 groups. These results were confirmed at the protein level by immunoperoxidase staining (data not shown).

Type I IFNs are important for innate immunity to viral infection27 and may promote TH1 responses.28 Fig 6, B, shows that there was no significant difference in IFN-α mRNA expression between OVA versus saline-exposed skin sites before VV inoculation. VV infection of saline-sensitized skin, but not of OVA-sensitized skin, resulted in significant upregulation of IFN-α expression. These results suggest that allergic skin inflammation may interfere with local induction of IFN-α expression by VV. Plasmacytoid dendritic cells (pDCs) are a major source of type I IFN production and have been reported in some studies, but not others, to be decreased in AD skin lesions.29, 30 Ears of EC sensitized mice were challenged with the sensitizing agent (OVA or saline) and evaluated 48 hours later for infiltration by dendritic cells (DCs). The total number of cells recovered from skin were comparable between the 2 groups. The percentage of ClassIIloCD19-B220+CD11b−CD11clow pDCs was significantly increased in the dermis, but not epidermis, of OVA-challenged skin compared with saline-challenged skin. The percentages of ClassII+CD19−B220−CD11b+CD11chigh myeloid DCs in dermis and epidermis were significantly higher in OVA-challenged skin (see this article's Fig E1 in the Online Repository at www.jacionline.org). We also investigated the possibility that increased TH2 cytokine expression in OVA-sensitized sites inhibits IFN-α production. To test this hypothesis, we examined the effect of IL-4 on IFN-α production by human blood mononuclear cells stimulated with the Toll-like receptor (TLR) 3 ligand PolyI:C and TLR9 ligand CpG. In 2 experiments, IL-4 caused a modest (∼25%) inhibition of TLR-triggered IFN-α production (see this article's Fig E2 in the Online Repository at www.jacionline.org). We also examined IFN-γ expression in the skin after VV inoculation. IFN-γ expression was comparable in VV-infected OVA-sensitized sites and saline-sensitized sites (Fig 6, C).

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Discussion 

Our results demonstrate that inoculation of VV at the site of cutaneous allergic inflammation results in an impaired immune response to the virus characterized by decreased production of TH1 cytokines, decreased IgG2a antibody response, and development of satellite pox lesions.

Mice inoculated with VV at sites of skin allergic inflammation induced by EC sensitization with OVA were impaired in their capacity to secrete the TH1 cytokines IL-2 and IFN-γ in response to VV (Fig 2, A). IFN-γ inhibits VV replication in mouse cells31 and promotes the development of VV-specific cytotoxic T cells and of VV-specific IgG2a.9 Consistent with previous observations that IFN-γ promotes the IgG2a response and downregulates the IgG1 response,22 mice inoculated with VV at sites of allergic skin inflammation had decreased IgG2a and increased IgG1 antibody levels against VV. Surprisingly, these mice expressed IFN-γ in their VV-infected skin sites at levels comparable to those found in VV-infected saline-sensitized sites of controls (Fig 6, C). This finding suggests that local factors in OVA-sensitized skin promote the expression of IFN-γ. A potential candidate is increased numbers of myeloid DCs, which are a potent source of IL-12.32

The impaired TH1 response of mice inoculated with VV at sites of allergic skin inflammation was associated with the appearance of satellite pox lesions (Fig 1, B and C). However, these mice exhibited neither weight loss nor increased viral loads in skin or internal organs examined. This may be a result of the attenuated nature of the virus strain used and of intact innate immunity, which is known to play a key role in viral clearance. Mice EC sensitized with OVA mounted a normal TH1 response to intraperitoneal inoculation with VV and, more importantly, a virtually normal TH1 response to VV inoculation at distant skin sites (Fig 4, Fig 5). We have recently observed that mice deficient in the nuclear factor-κB family member RelB, which exhibit skin lesions that share features with chronic AD,33 have impaired IFN-γ production in response to VV inoculated cutaneously, but not intraperitoneally.18 However, RelB deficiency is associated with dysregulated function of multiple cell types, which could have contributed to this impairment. Our current results demonstrate that in otherwise normal EC sensitized mice, local allergic skin inflammation, rather than a global dysregulation of the systemic immune response, is responsible for the impaired TH1 response to VV.

Several factors may have caused the impaired TH1 response to VV introduction at sites of allergic inflammation. One possibility is that skin inflammation caused rapid local elimination of the virus and/or interfered with carrying the virus to regional draining lymph nodes (LNs). This is unlikely because mice inoculated with VV at sites of OVA versus saline sensitization had comparable viral loads in skin and draining LNs and mounted comparable TH2 responses to the virus (Fig 2, B). An alternative and more likely possibility is that DCs in LNs that drain sites of allergic sensitization skew the response of naive T cells away from TH1. In support of this notion, DCs isolated from LNs that drain skin sensitized with Aspergillus fumigatus skew the TH response of naive T-cell receptor-OVA transgenic T cells away from TH1 and toward TH2 (Oyoshi, unpublished data, December 2006). Finally, we found decreased local expression of CRAMP at the time of VV inoculation. Therefore, failure to upregulate local expression of IFN-α after infection, in part a result of increased local expression of IL-4 and IL-13, may have contributed to the decreased TH1 response to VV inoculation at OVA-sensitized sites because both CRAMP and IFN-α promote TH1 responses.26, 28 Although it is possible that host cell (CV1)–derived contaminants may contribute to the inflammatory reaction in response to VV inoculation, this would not account for the differences between mice inoculated at OVA-sensitized sites versus saline-sensitized sites.

Our model of skin allergic skin inflammation shares many features with AD. These include skin inflammation characterized by infiltration of CD4+ T cells and eosinophils and expression of mRNA for TH2 cytokines and a systemic TH2 response to antigen characterized by elevated antigen specific and total IgE. Given these shared features, our findings suggest that the increased susceptibility of patients with AD to EV may be related to local immune dysregulation in the skin rather than their systemic TH2-dominated immune response. They also suggest that to achieve an optimal immune response after immunization of patients with AD with attenuated VV strains, care should be taken to avoid inoculation of the virus in inflamed skin sites.

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Appendix. Supplementary data 

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References 

  1. Esposito JJ, Fenner F. Poxviruses. In:  Knipe DM,  Howley PM editor. Fields virology. Philadelphia: Lippincott Williams & Wilkins; 2001;p. 2885–2922
  2. Lane JM, Ruben FL, Abrutyn E, Millar JD. Deaths attributable to smallpox vaccination, 1959 to 1966, and 1968. JAMA. 1970;212:441–444
  3. Neff JM, Lane JM, Fulginiti VA, Henderson DA. Contact vaccinia: transmission of vaccinia from smallpox vaccination. JAMA. 2002;288:1901–1905
  4. Lane JM, Millar JD. Risks of smallpox vaccination complications in the United States. Am J Epidemiol. 1971;93:238–240
  5. Shirasawa K, Akai K, Kawaguchi Y, Maeda S, Nagahara S, Toyoda H, et al. Widespread eczema vaccinatum acquired by contacts: a report of an autopsy case. Acta Pathol Jpn. 1979;29:435–455
  6. Marris E. Dramatic rescue relieves rare case of smallpox infection. Nat Med. 2007;13:517
  7. Leung DY, Bieber T. Atopic dermatitis. Lancet. 2003;361:151–160
  8. Karupiah G, Blanden RV, Ramshaw IA. Interferon gamma is involved in the recovery of athymic nude mice from recombinant vaccinia virus/interleukin 2 infection. J Exp Med. 1990;172:1495–1503
  9. Xu R, Johnson AJ, Liggitt D, Bevan MJ. Cellular and humoral immunity against vaccinia virus infection of mice. J Immunol. 2004;172:6265–6271
  10. Kohonen-Corish MR, King NJ, Woodhams CE, Ramshaw IA. Immunodeficient mice recover from infection with vaccinia virus expressing interferon-gamma. Eur J Immunol. 1990;20:157–161
  11. Sharma DP, Ramsay AJ, Maguire DJ, Rolph MS, Ramshaw IA. Interleukin-4 mediates down regulation of antiviral cytokine expression and cytotoxic T-lymphocyte responses and exacerbates vaccinia virus infection in vivo. J Virol. 1996;70:7103–7107
  12. van Den Broek M, Bachmann MF, Kohler G, Barner M, Escher R, Zinkernagel R, et al. IL-4 and IL-10 antagonize IL-12-mediated protection against acute vaccinia virus infection with a limited role of IFN-gamma and nitric oxide synthetase 2. J Immunol. 2000;164:371–378
  13. Spergel JM, Mizoguchi E, Brewer JP, Martin TR, Bhan AK, Geha RS. Epicutaneous sensitization with protein antigen induces localized allergic dermatitis and hyperresponsiveness to methacholine after single exposure to aerosolized antigen in mice. J Clin Invest. 1998;101:1614–1622
  14. Spergel JM, Mizoguchi E, Oettgen H, Bhan AK, Geha RS. Roles of TH1 and TH2 cytokines in a murine model of allergic dermatitis. J Clin Invest. 1999;103:1103–1111
  15. Laouini D, Elkhal A, Yalcindag A, Kawamoto S, Oettgen H, Geha RS. COX-2 inhibition enhances the TH2 immune response to epicutaneous sensitization. J Allergy Clin Immunol. 2005;116:390–396
  16. Phelps AL, Gates AJ, Hillier M, Eastaugh L, Ulaeto DO. Comparative efficacy of modified vaccinia Ankara (MVA) as a potential replacement smallpox vaccine. Vaccine. 2007;25:34–42
  17. In:  Earl PL,  Cooper N,  Wyatt LS,  Moss B,  Carroll MW editor. Preparation of cell cultures and vaccinia virus stocks. Mississauga, Ontario, Canada: John Wiley & Sons, Inc; 1998;
  18. Freyschmidt EJ, Mathias CB, Macarthur DH, Laouar A, Narasimhaswamy M, Weih F, et al. Skin inflammation in RelB(-/-) mice leads to defective immunity and impaired clearance of vaccinia virus. J Allergy Clin Immunol. 2007;119:671–679
  19. Elkhal A, Pichavant M, He R, Scott J, Meyer E, Goya S, et al. CD1d restricted natural killer T cells are not required for allergic skin inflammation. J Allergy Clin Immunol. 2006;118:1363–1368
  20. Ennis FA, Cruz J, Demkowicz WE, Rothman AL, McClain DJ. Primary induction of human CD8+ cytotoxic T lymphocytes and interferon-gamma-producing T cells after smallpox vaccination. J Infect Dis. 2002;185:1657–1659
  21. Huang S, Hendriks W, Althage A, Hemmi S, Bluethmann H, Kamijo R, et al. Immune response in mice that lack the interferon-gamma receptor. Science. 1993;259:1742–1745
  22. Snapper CM, Paul WE. Interferon-gamma and B cell stimulatory factor-1 reciprocally regulate Ig isotype production. Science. 1987;236:944–947
  23. Howell MD, Jones JF, Kisich KO, Streib JE, Gallo RL, Leung DY. Selective killing of vaccinia virus by LL-37: implications for eczema vaccinatum. J Immunol. 2004;172:1763–1767
  24. Ong PY, Ohtake T, Brandt C, Strickland I, Boguniewicz M, Ganz T, et al. Endogenous antimicrobial peptides and skin infections in atopic dermatitis. N Engl J Med. 2002;347:1151–1160
  25. Howell MD, Gallo RL, Boguniewicz M, Jones JF, Wong C, Streib JE, et al. Cytokine milieu of atopic dermatitis skin subverts the innate immune response to vaccinia virus. Immunity. 2006;24:341–348
  26. Davidson DJ, Currie AJ, Reid GS, Bowdish DM, MacDonald KL, Ma RC, et al. The cationic antimicrobial peptide LL-37 modulates dendritic cell differentiation and dendritic cell-induced T cell polarization. J Immunol. 2004;172:1146–1156
  27. van den Broek MF, Muller U, Huang S, Zinkernagel RM, Aguet M. Immune defence in mice lacking type I and/or type II interferon receptors. Immunol Rev. 1995;148:5–18
  28. Manetti R, Annunziato F, Tomasevic L, Gianno V, Parronchi P, Romagnani S, et al. Polyinosinic acid: polycytidylic acid promotes T helper type 1-specific immune responses by stimulating macrophage production of interferon-alpha and interleukin-12. Eur J Immunol. 1995;25:2656–2660
  29. Wollenberg A, Wagner M, Gunther S, Towarowski A, Tuma E, Moderer M, et al. Plasmacytoid dendritic cells: a new cutaneous dendritic cell subset with distinct role in inflammatory skin diseases. J Invest Dermatol. 2002;119:1096–1102
  30. Guttman-Yassky E, Lowes MA, Fuentes-Duculan J, Whynot J, Novitskaya I, Cardinale I, et al. Major differences in inflammatory dendritic cells and their products distinguish atopic dermatitis from psoriasis. J Allergy Clin Immunol. 2007;119:1210–1217
  31. Karupiah G, Xie QW, Buller RM, Nathan C, Duarte C, MacMicking JD. Inhibition of viral replication by interferon-gamma-induced nitric oxide synthase. Science. 1993;261:1445–1448
  32. Hogg KG, Kumkate S, Anderson S, Mountford AP. Interleukin-12 p40 secretion by cutaneous CD11c+ and F4/80+ cells is a major feature of the innate immune response in mice that develop Th1-mediated protective immunity to Schistosoma mansoni. Infect Immun. 2003;71:3563–3571
  33. Barton D, HogenEsch H, Weih F. Mice lacking the transcription factor RelB develop T cell-dependent skin lesions similar to human atopic dermatitis. Eur J Immunol. 2000;30:2323–2332

 Supported by federal funds from the National Institute of Allergy and Infectious Diseases, National Institutes of Health, Department of Health and Human Services, under contract number HHSN266200400030C (Atopic Dermatitis Vaccinia Network).

 Disclosure of potential conflict of interest: J. Scott has consulting arrangements with Alicare Medical Management, owns Pharmaceutical holder trust stock, and is on the speakers' bureau for Verus Pharmaceuticals. The rest of the authors have declared that they have no conflict of interest.

PII: S0091-6749(07)01457-1

doi:10.1016/j.jaci.2007.08.004

The Journal of Allergy and Clinical Immunology
Volume 120, Issue 6 , Pages 1382-1388, December 2007

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