Allergy and autoimmune disease: a registry-based study.

BACKGROUND: Allergy and autoimmunity are two potential outcomes of a dysregulated immune system, but the relationship between them is unclear. It has been hypothesized that they could be inversely associated because of different T helper cell reactivity patterns. However, both positive and negative associations have been reported. Therefore, our aim was to perform a large epidemiological study with a defined allergic disease cohort. METHODS: During the years 1990-2002, 68 770 subjects were tested for total serum IgE (Total-IgE) and 72 228 were tested with Phadiatop for diagnosing allergic disease at Karolinska University Hospital, Stockholm, Sweden. This cohort was then linked with the Swedish Inpatient Registry 1968-2004 for a follow-up with regard to recorded discharges for 28 autoimmune diseases. We then used Cox regression and logistic regression to estimate the risk of autoimmune diseases in general in the allergy-tested subjects. RESULTS: Subjects with positive Phadiatop test were at a statistically decreased risk of subsequent autoimmune disease in comparison with subjects with negative test; hazard ratio (HR): 0.80 [95% (Confidence interval) CI: 0.68-0.94). Prior autoimmune disease was associated with a decreased risk of positive Phadiatop test [odds ratio: 0.83 (95% CI: 0.72-0.96)] in comparison with negative test. Subjects with highly elevated Total-IgE were at a statistically increased risk of a subsequent autoimmune disease in comparison with subjects with normal levels [HR: 1.36 (95% CI: 1.09-1.70)], but no association was found between prior autoimmune disease and different Total-IgE levels. CONCLUSION: The study supports the hypothesis that allergy, defined as positive Phadiatop test, could be inversely related to autoimmune disease but this association is weak.

Elevated expression and genetic association links the SOCS3 gene to atopic dermatitis.

In a systematic analysis of global gene-expression patterns, we found that SOCS3 messenger RNA was significantly more highly expressed in skin from patients with atopic dermatitis than in skin from healthy controls, and immunohistochemical analysis confirmed a similar elevation of SOCS3 protein. Furthermore, we found a genetic association between atopic dermatitis and a haplotype in the SOCS3 gene in two independent groups of patients (P<.02 and P<.03). These results strongly suggest that SOCS3, located in a chromosomal region previously linked to the disease (17q25), is a susceptibility gene for atopic dermatitis.

Allergy and cancer.

BACKGROUND: Conflicting results have provided support for two distinct and contradictory hypotheses: (i) allergy has a protective effect against cancer by enhanced immune surveillance, and (ii) allergy is associated with an increased risk of cancer by chronic immune stimulation. We therefore aimed us to perform a large epidemiological study with a defined allergic disease cohort. METHODS: During the years 1988-2000, 70 136 patients tested for total serum immunoglobulin E (IgE) and 57 815 tested with Phadiatop for diagnosing allergic disease at Karolinska University Hospital, Stockholm, Sweden, were linked with the Swedish Cancer Registry for a virtually complete follow up with regard to cancer. FINDINGS: The total number of observed cancers was normal in the total serum IgE-cohort; standardized incidence ratio (SIR) = 0.98 (95% CI: 0.92-1.04) and in the Phadiatop-cohort: SIR = 0.99 (0.92-1.06) independent of the level of IgE and positive or negative Phadiatop. Specific analysis was done for cancer of the lung, cervix, pancreas, lymphoma, and nonmelanoma skin cancer. None of these forms of cancer had increased risks. INTERPRETATION: The study does not support the hypothesis that allergy has a protective effect against cancer, nor does it support an increased risk.

Atopy patch test with house dust mite allergen--an IgE-mediated reaction?

BACKGROUND: The eczema reaction in the atopy patch test (APT) is proposed to be immunoglobulin (Ig)E mediated, but can take place also in individuals lacking allergen-specific IgE in serum. The purpose of this study was to evaluate the importance of allergen-specific serum IgE for the APT reaction. METHODS: Ten patients with reproducible positive APT to extract of Dermatophagoides pteronyssinus, five patients with (group A) and five patients without (group B) detectable serum-IgE to D. pteronyssinus, were tested with extract of D. pteronyssinus on normal skin for 6, 24, 48 and 72 h. Skin biopsies were taken and analysed for cell infiltrates, eosinophils (EG2), IgE, FcepsilonRI, CD1a, CD4, CD8 and metalloproteinase 9 (MMP9). RESULTS: The number of IgE+, CD4+, EG2+ and MMP9+ cells increased with time in group A. FcepsilonRI+ cells and CD8+ cells increased with time in both groups. A correlation was found between the levels of D. pteronyssinus-specific serum-IgE and the score of dermal cell infiltrates at 72 h. The three patients with the highest values of allergen-specific IgE also had the highest expression of EG2+ cells and the highest APT scores. CONCLUSIONS: Our study strengthens the hypothesis that the IgE molecule has a key role, at least as an amplifier, in the APT reaction.

Atopy patch test reactions to Malassezia allergens differentiate subgroups of atopic dermatitis patients.

BACKGROUND: The yeast Malassezia is considered to be one of the factors that can contribute to atopic dermatitis (AD). OBJECTIVES: To investigate the reactivity to Malassezia allergens, measured as specific serum IgE, positive skin prick test and positive atopy patch test (APT), in adult patients with AD. METHODS: In total, 132 adult patients with AD, 14 with seborrhoeic dermatitis (SD) and 33 healthy controls were investigated for their reactions to M. sympodialis extract and three recombinant Malassezia allergens (rMal s 1, rMal s 5 and rMal s 6). RESULTS: Sixty-seven per cent of the AD patients, but only one of the SD patients and none of the healthy controls, showed a positive reaction to at least one of the Malassezia allergens (extract and/or recombinant allergens) in at least one of the tests. The levels of M. sympodialis-specific IgE in serum correlated with the total serum IgE levels. Elevated serum levels of M. sympodialis-specific IgE were found in 55% and positive APT reactions in 41% of the AD patients with head and neck dermatitis. A relatively high proportion of patients without head and neck dermatitis and patients with low total serum IgE levels had a positive APT for M. sympodialis, despite lower proportions of individuals with M. sympodialis-specific IgE among these groups of patients. CONCLUSIONS: These results support that Malassezia can play a role in eliciting and maintaining eczema in patients with AD. The addition of an APT to the test battery used in this study reveals a previously overlooked impact of Malassezia hypersensitivity in certain subgroups of AD patients.

Positive atopy patch test reactions to Pityrosporum orbiculare in atopic dermatitis patients.

BACKGROUND: Pityrosporum orbiculare, although a part of our normal cutaneous microflora, can cause skin infections and induce specific immunoglobulin (Ig) E antibodies in atopic dermatitis patients. P. orbiculare is therefore considered to be one of the trigger factors for atopic dermatitis. OBJECTIVE: To investigate if P. orbiculare can induce an eczematous reaction in atopic dermatitis patients, seborrhoeic dermatitis patients and healthy controls. METHODS: Fifteen atopic dermatitis patients, eight seborrhoeic dermatitis patients and eight healthy controls were patch tested with extract of P. orbiculare on non-lesional, tape-stripped skin of the back. NaCl was used as a negative control. The patch tests were evaluated after 24, 48 and 72 h. Skin biopsies were taken from P. orbiculare patch test sites at 24 h and 72 h, from NaCl patch test sites at 72 h, from non-lesional skin and, in the atopic dermatitis patients, also from lesional skin. The skin biopsies were investigated with immunohistochemical techniques. P. orbiculare-specific IgE in serum was analysed with RAST. RESULTS: Specific IgE to P. orbiculare was found in serum from 13/15 atopic dermatitis patients and in eight of them a positive patch test reaction to P. orbiculare was observed, with a maximal reaction at 48 h. Significantly higher serum levels of P. orbiculare-specific IgE were detected in patch test-positive compared with patch test-negative atopic dermatitis patients (P < 0. 01). The seborrhoeic dermatitis patients and healthy controls were RAST and patch test-negative for P. orbiculare. In the patch test-positive atopic dermatitis patients an infiltration of CD4+ T cells and eosinophils was observed at the P. orbiculare patch test sites together with an upregulation of ICAM-1 and HLA-DR expression. CONCLUSIONS: P. orbiculare can induce an eczematous reaction in sensitized atopic dermatitis patients and may be an important trigger factor in these patients. The P. orbiculare patch test can be of diagnostic value in this subgroup of atopic dermatitis patients.

Direct Ni2+ antigen formation on cultured human dendritic cells.

The possible direct antigen formation of Ni2+ on antigen-presenting cells (APCs) was studied with cultured human dendritic cells (DCs) obtained from 10 subjects contact allergic to Ni2+ and six non-allergic control individuals. All contact allergic subjects showed a significantly increased peripheral blood mononuclear cell (PBMC) response in vitro to Ni2+. DCs were expanded from the plastic-adherent cell fraction of PBMCs by culturing with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) for 7 days to obtain immature DCs, and with the addition of monocyte-conditioned medium for another 4 days, for DC maturation. The DCs were pulsed for 20 min with Ni2+ (50 micrometers) in protein-free Hank's balanced salt solution (HBSS) and added to freshly prepared autologous responder PBMCs. With five allergic subjects, immature DCs pulsed with Ni2+ demonstrated a significant capacity to activate Ni2+-reactive lymphocytes. With the remaining five patients and the six controls no difference in lymphocyte proliferation was observed between Ni2+-pulsed and non-pulsed immature DCs. In contrast, with mature Ni2+-pulsed DCs from both 'positive responder' (n=4) and 'non-responder' (n=4) patients, there was a significantly stimulated PBMC proliferation, whereas with the controls (n=4) still no activation was observed. Our results indicate that direct formation of the antigenic determinant of Ni2+ on APCs is possible and that Ni2+ uptake and processing mechanisms may not play a major role. Differences in the ease of activation of Ni2+-reactive lymphocytes are discussed in terms of a possible heterogeneity in the availability of Ni2+-reactive groups presented on endogenous peptides bound in the antigen binding groove of human leucocyte antigen (HLA) class-II molecules.

Peripheral blood T-cell receptor beta-chain V-repertoire in atopic dermatitis patients after in vitro exposure to Pityrosporum orbiculare extract.

The yeast Pityrosporum orbiculare belongs to the normal cutaneous flora but is also considered to be one of the factors that may contribute to atopic dermatitis (AD). In the present study we investigated the possibility that P. orbiculare can act with superantigen activity in AD. P. orbiculare-reactive T-cell lines (TCLs) were obtained after stimulation of peripheral blood mononuclear cells (PBMC) with P. orbiculare extract. T-cell receptor beta-chain V-segment (TCRBV) usage was investigated using monoclonal antibodies and flow cytometry. We could not find any difference in TCRBV usage between AD patients (n = 10) and healthy controls (n = 5), either in fresh PBMC or in P. orbiculare-reactive TCLs. Compared with their original PBMCs the P. orbiculare-reactive TCLs showed a decreased usage of several TCRBVs, although increased usage of certain TCRBVs could be seen in some of the individuals. Further analysis of the CDR3-length polymorphism exhibited a shift in CDR3-length distribution, indicating oligoclonal expansion of T cells specific to different antigens in the P. orbiculare extract. In conclusion we have not found any evidence for superantigen activity in P. orbiculare extract, but our data support the importance of classical major histocompatibility complex (MHC)-restricted allergens in P. orbiculare.

Pityrosporum orbiculare-reactive T-cell lines in atopic dermatitis patients and healthy individuals.

The yeast Pityrosporum orbiculare is one of the factors that may contribute to atopic dermatitis (AD). In the present study we compared the T-cell response to P. orbiculare in 12 AD patients with specific immunoglobulin (Ig)E antibodies (Ab) in serum against P. orbiculare with that of six non-atopic healthy controls. Freshly isolated peripheral blood mononuclear cells (PBMC) were cultured for 3 days in the presence of P. orbiculare extract. The proliferative response as measured by [3H]-thymidine incorporation was significantly higher in the AD patients than in the healthy controls (P < 0.05). Furthermore, significantly higher levels of interleukin (IL)-5 (P < 0.05), as analyzed by ELISA, were produced by PBMC from the AD patients compared to the healthy controls. Pityrosporum orbiculare-reactive T-cell lines (TCL) established by P. orbiculare stimulation of PBMC for 11 days produced significantly higher levels of IL-4 and IL-5 after stimulation with anti-CD3 Ab and showed a higher IL-4/interferon (IFN)-gamma ratio (P < 0.05) in the AD patients compared to the healthy controls. The higher proliferative PBMC response to P. orbiculare and the Th2-like cytokine production by P. orbiculare-stimulated TCL from AD patients indicate that P. orbiculare may play a role in maintaining skin inflammation in AD.

Culture of human epidermal Langerhans cells in a skin equivalent.

Langerhans cells (LCs) have been cultured in a skin equivalent (SE). Seventy-two SEs were produced by inserting skin biopsies from nine subjects into dermal equivalents consisting of fibroblasts in a collagen matrix. The SEs were cultured in a serum-free medium containing 2-mercaptoethanol with or without 5 ng/mL granulocyte-monocyte colony-stimulating factor (GM-CSF). The SEs were cultured for 12 or 15 days. In the latter case, 0, 1 or 10 microg/mL cyclosporin A (CyA) was added for the last 3 days. The SEs were then snap frozen for immunohistochemistry. The migration of LCs was evaluated by measuring the distances from the inserted skin biopsy in the SEs to the HLA-DR + and CD1a+ dendritic cells localized at the longest distance from the biopsy in the epidermal outgrowth on both sides of the biopsy. The density of these cells was estimated in 15-day-old SEs by counting them on both sides of the inserted skin biopsy and dividing the number of positive cells by the migrated distances. All epidermal outgrowths (range 0.6-3.7 mm) were well differentiated and displayed HLA-DR+, CD1a+ and Lag+ dendritic cells. Only occasionally were CD83+ cells observed. In the 15-day-old SEs cultured with GM-CSF, a few CD86+ cells were seen in the epidermal outgrowths and occasionally CD80+ cells. The median (n = 4) density of CD1a+ and HLA-DR+ cells in the epidermal outgrowths at day 15 was 5.2 and 9.1 cells/mm, respectively. GM-CSF did not influence migration in 12-day-old SEs, but there was a tendency to increased migration of HLA-DR+ dendritic cells in 15-day-old SEs. CyA did not affect migration or density. We conclude that LCs can be cultured with an in vivo-like density in a SE. They express the phenotype of immature antigen-presenting cells efficient in capturing and processing antigen. This model may be suitable for studies of the initial phase of contact allergic reactions.

Detection of Pityrosporum orbiculare reactive T cells from skin and blood in atopic dermatitis and characterization of their cytokine profiles.

BACKGROUND: Atopic dermatitis (AD) is associated with increased levels of serum IgE, and T-helper (Th) cells are thought to a play role in the pathogenesis. Individuals with AD often develop IgE antibodies against the yeast Pityrosporum orbiculare, a member of the normal cutaneous flora. OBJECTIVE: The role of P. orbiculare in atopic dermatitis was investigated by examining the T-cell reactivity for P. orbiculare. METHODS: Freshly isolated peripheral blood mononuclear cells (PBMC) were isolated from 10 AD patients with serum IgE antibodies against P. orbiculare, and from six healthy controls. The proliferative response after P. orbiculare stimulation, measured by [3H]thymidine incorporation, was examined in the PBMC and in T-cell clones (TCC) obtained from skin and blood of one patient. The cytokine profile of the TCC was determined by enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA) and reverse transcriptase-polymerase chain reaction (RT-PCR) following challenge with either P. orbiculare extract or anti-CD3 antibodies and phytohaemagglutinin. RESULTS: The PBMC response to P. orbiculare was significantly higher in the AD patients than in the control group (P < 0.05). Twenty-nine out of 36 tested TCC derived from one responding patient were reactive for P. orbiculare. The clones were CD2+ and CD4+, except for one CD8+ blood clone. A majority of the TCC derived from lesional skin showed a Th2- or Th2/Th0-like cytokine profile. A co-expression of interleukin-5 (IL-5) mRNA and IL-13 mRNA was detected in five out of six P. orbiculare-reactive clones analysed for their cytokine gene expression with RT-PCR. CONCLUSION: Our data suggest that P. orbiculare can induce a T-cell response in AD patients. The Th2-like profile of P. orbiculare-reactive TCC derived from lesional skin indicates that P. orbiculare may play a role in maintaining IgE-mediated skin inflammation in AD.

Itch and inflammation induced by intradermally injected interleukin-2 in atopic dermatitis patients and healthy subjects.

To explore the pruritogenic and inflammatory effects of cytokines, a single dose of 20 micrograms recombinant human interleukin-2 was injected intradermally into eight patients with atopic dermatitis and eight healthy controls. The study was double-blind and randomized with glucose as a negative control. The effects were evaluated by recording local itch and erythema over 72 h and by examining skin biopsies taken at 24 h and 72 h. In patients and controls, interleukin-2 provoked a low-intensity local itch with maximal intensity between 6 h and 48 h and erythema with maximal extension between 12 h and 72 h. In the atopic dermatitis patients, these reactions tended to appear earlier and were less pronounced than in the healthy controls. Interleukin-2 induced dermal mononuclear cell infiltrates consisting mainly of CD3+ cells. A majority of the T cells were CD4+. The number of dermal CD25+, HLA-DR+ and ICAM-1+ cells was also increased at the interleukin-2 induced spongiosis and exocytosis as well as HLA-DR+ and ICAM-1+ keratinocytes. The microscopic findings tended to be more prominent at 72 h than at 24 h in both groups, but with a somewhat slower onset in the atopic dermatitis patients. In conclusion, a single intradermal injection of interleukin-2 induced local itch, erythema, dermal T-cell infiltrates, spongiosis, exocytosis and activation of keratinocytes both in atopic dermatitis patients and in healthy controls.