Skin inflammation arising from cutaneous regulatory T cell deficiency leads to impaired viral immune responses.

Individuals with atopic dermatitis immunized with the small pox vaccine, vaccinia virus (VV), are susceptible to eczema vaccinatum (EV), a potentially fatal disseminated infection. Dysfunction of Forkhead box P3 (FoxP3)-positive regulatory T cells (Treg) has been implicated in the pathogenesis of atopic dermatitis. To test whether Treg deficiency predisposes to EV, we percutaneously VV infected FoxP3-deficient (FoxP3(KO)) mice, which completely lack FoxP3(+) Treg. These animals generated both fewer VV-specific CD8(+) effector T cells and IFN-gamma-producing CD8(+) T cells than controls, had higher viral loads, and exhibited abnormal Th2-polarized responses to the virus. To focus on the consequences of Treg deficiency confined to the skin, we generated mixed CCR4(KO) FoxP3(KO) bone marrow (CCR4/FoxP3) chimeras in which skin, but not other tissues or central lymphoid organs, lack Treg. Like FoxP3(KO) mice, the chimeras had impaired VV-specific effector T cell responses and higher viral loads. Skin cytokine expression was significantly altered in infected chimeras compared with controls. Levels of the antiviral cytokines, type I and II IFNs and IL-12, were reduced, whereas expression of the proinflammatory cytokines, IL-6, IL-10, TGF-beta, and IL-23, was increased. Importantly, infection of CCR4/FoxP3 chimeras by a noncutaneous route (i.p.) induced immune responses comparable to controls. Our findings implicate allergic skin inflammation resulting from local Treg deficiency in the pathogenesis of EV.

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

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 T(H)1 cytokines in response to VV, decreased VV specific serum IgG(2a), increased VV specific serum IgG(1), and impaired upregulation of IFN-alpha, 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 T(H)1 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.

Intraperitoneal stability of alginate-polyornithine microcapsules in rats: an FTIR and SEM analysis.

Alginate-polycation microcapsule systems have been used over decades as delivery vehicles for cell and protein therapy. These systems have been unpredictable across a range of indications with questions resulting around the inherent stability of the alginate polysaccharide and failure mode of the delivery system. The current study focuses on such a system using 5 different alginates, 2 of which are commercially purified, which are crosslinked by polyornithine. Capsules formed by frequency-generated droplet formation were studied in the peritoneal cavity of Long-Evans rats over the course of 3 months by morphometry, Fourier-transform infrared spectroscopy (FTIR), and scanning electron microscopy of the surface. Individual capsule components were also investigated on FTIR and a relative stability index was generated by titration for comparison to explanted samples over time. Using these techniques, a distinct degradation pattern was noted and is compared between the 5 alginate sources.

Safety and efficacy of radiofrequency wire recanalization of chronic central venous occlusions.

PURPOSE: The purpose of this study is to evaluate the safety and efficacy of using a radiofrequency wire (RF) for central venous occlusion (CVO) recanalization after failure using conventional techniques. MATERIALS AND METHODS: A retrospective analysis of all central venous recanalization procedures using an RF wire from January 2007 to December 2012 was performed. This comprised 13 consecutive procedures in 12 patients. The electronic medical record and radiologic imaging studies were reviewed to obtain information regarding patient demographics, indication for revascularization, duration of vascular occlusion, procedure outcome, and complications. RESULTS: Technical success was achieved in nine of 13 (69%) occluded vessels in nine of 12 (75%) patients. Lengths of successfully crossed lesions (mean 29.8 ± 29.3 mm) were significantly less than those of unsuccessfully crossed lesions (mean 90 ± 73.7 mm), p = 0.039. Of the nine patients with technically successful procedures, three died with patent stents of causes unrelated to the procedure, one had a patent stent at 547 days, two had stent patency of 94 days and 345 days, and three were lost to follow-up. One patient (8.3%) experienced a major complication wherein tracheal perforation by the RF wire contributed to the patient's death. CONCLUSIONS: RF recanalization is a viable option in patients with central vein occlusion refractory to traditional procedures. However, it does not guarantee successful revascularization and is not without the potential for harm.

Skin inflammation in RelB(-/-) mice leads to defective immunity and impaired clearance of vaccinia virus.

BACKGROUND: Atopic dermatitis (AD) is an inflammatory skin disorder occurring in genetically predisposed individuals with a systemic T(H)2 bias. Atopic dermatitis patients exposed to the smallpox vaccine, vaccinia virus (VV), occasionally develop eczema vaccinatum (EV), an overwhelming and potentially lethal systemic infection with VV. OBJECTIVE: To establish a murine model of EV and examine the effects of skin inflammation on VV immunity. METHODS: The skin of RelB(-/-) mice, like that of chronic AD lesions in humans, exhibits thickening, eosinophilic infiltration, hyperkeratosis, and acanthosis. RelB(-/-) and wild-type (WT) control mice were infected with VV via skin scarification. Viral spread, cytokine levels, IgG2a responses and VV-specific T cells were measured. RESULTS: Cutaneously VV-infected RelB(-/-), but not WT mice, exhibited weight loss, markedly impaired systemic clearance of the virus and increased contiguous propagation from the inoculation site. This was associated with a dramatically impaired generation of IFN-gamma-producing CD8(+) vaccinia-specific T cells along with decreased secretion of IFN-gamma by VV-stimulated splenocytes. The T(H)2 cytokines-IL-4, IL-5, IL-13, and IL-10-on the other hand, were overproduced. When infected intraperitoneally, RelB(-/-) mice generated robust T cell responses with good IFN-gamma production. CONCLUSION: Allergic inflammation in RelB(-/-) mice is associated with dysregulated immunity to VV encountered via the skin. We speculate that susceptibility of AD patients to overwhelming vaccinia virus infection is similarly related to ineffective T cell responses. CLINICAL IMPLICATIONS: The susceptibility of patients with AD to EV following cutaneous contact with VV is related to ineffective antiviral immune responses.