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112 Agreement of Total vs. Component Milk-Specific IgE 114 Early predictors of clinician diagnosed eczema at age 4: The
Cincinnati Childhood Allergy and Air Pollution (CCAAP) Birth
J. Bogdanovic1, J. M. Skripak2, E. C. Matsui1, A. W. Burks3, R. A.
Wood1, R. G. Hamilton1; 1Johns Hopkins University School of Medicine,
T. G. Epstein, D. I. Bernstein, L. Levin, P. H. Ryan, J. E. Lockey, G. K. K.
Baltimore, MD, 2Mount Sinai School of Medicine, New York, NY, 3Duke
Hershey, M. Villareal, G. K. LeMasters; University of Cincinnati, Cincin-
University Medical Center, Durnham, NC.
RATIONALE: To determine the agreement between ImmunoCAP cow’s
RATIONALE: Early immunologic and environmental predictors of ec-
milk-specific IgE measurements and IgE antibody responses to individual
zema are not well characterized. This study examines predictive factors
METHODS: Sera from 30 children (6-16 years) with cow’s milk allergy,
METHODS: 636 infants of atopic parents enrolled in a study regarding
20 of which received milk oral immunotherapy (MOIT), were analyzed for
traffic pollution and allergy underwent annual clinical evaluations and
IgE anti-cow’s milk-(F2), alpha-lactalbumin-(ALA), beta-lactoglobulin-
skin prick tests (SPT) to 17 allergens, including milk and egg. Home
(BLG) and casein-(CAS) by Phadia ImmunoCAP. IgE anti-cow’s milk
dust samples were evaluated for endotoxin. Exposure to traffic related pol-
levels alone were compared to the sum of individual IgE anti-alpha-lactal-
lution was estimated. Eczema at age 4 (n 5 115) was defined by erythema
bumin;beta-lactoglobulin;casein in kUa/L. Bias/inaccuracy was assessed
and papulation/infiltration on physical exam; atopic dermatitis (n 5 84)
by deviation from an ideal ratio of 1.0. All IgE antibody levels >50 kUa/L
was defined as eczema and at least 1 positive SPT at age 4. Multivariate lo-
were re-analyzed at dilutions to insure accuracy.
gistic regression analysis was performed.
RESULTS: Assuming the sum of IgE anti-ALA-BLG-CAS should accu-
RESULTS: After adjusting for race, gender, income, pets, endotoxin, traf-
rately reflect, or possibly underestimate the total milk specific IgE, IgE
fic exposure and breast-feeding, the most significant predictors for eczema
anti-milk/summed IgE anti-ALA-BLG-CAS ratios for pre MOIT sera
at age 4 were parental eczema (adjusted[a]OR: 1.9 [1.1-3.3]), SPT positiv-
ranged from 2.17-1.74 (n 5 2; over-estimate possibly due to missing IgE
ity to egg at age 1 (aOR 2.9[1.6-5.4]), SPT positivity to cat/dog at 2 years
anti-BSA/lactoferrin), 1.04-0.90 (n 5 3; equivalence) and 0.77-0.42
(aOR 2.1 [1.1-4.3]), and SPT positivity to mold at 2 years (aOR 2.2 [1.1-
(n 5 25, underestimate). In MOIT-treated subjects, IgE anti-milk/IgE
4.4]). Dog ownership at age 1 trended towards a protective effect from
anti-ALA-BLG-CAS ratios remained generally constant within each sub-
atopic dermatitis at age 4 (aOR 0.6 [0.3-1.0]). Traffic exposure was not pre-
ject over a year of MOIT despite fluctuations in absolute IgE anti-milk
dictive for eczema or atopic dermatitis.
levels and marked increases in IgG anti-milk.
CONCLUSIONS: SPT positivity to egg at 1 year, and to animals and mold
CONCLUSIONS: Patient-dependent bias can occur in which IgE anti-
at 2 years are independent predictors of eczema at 4 years. Dog ownership
milk measurements alone may underestimate the actual level of IgE
in the first year of life may be protective of atopic dermatitis.
antibody reactive with milk components by up to 2.4 fold. While the con-cordance between the levels of milk-specific IgE antibody and the summed
115 Immunomodulatory Effect of Rapamycin on Innate Immune
individual IgE anti-milk components is good for a minority of milk allergic
patients, there is a majority where underestimation, possibly due to limited
M. D. Howell, B. E. Kim, D. Y. Leung; National Jewish Health, Denver,
molar excess of components on the milk ImmunoCAP, may be substantial
RATIONALE: Atopic dermatitis (AD) is complicated by recurrent skininfections. Previous studies have shown that treatment with topical calci-
113 Ligation of GITRL Induces Chemokine Production by neurininhibitorsandcorticosteroidsreduceStaphylococcusaureuscoloni-
zation on AD skin. Therefore, it was of interest to determine whether these
A. M. Byrne, D. Y. Leung, E. Goleva; National Jewish Health, Denver,
agents could induce the innate immune response which plays an important
role in controlling the number of skin microbes.
RATIONALE: We have recently demonstrated surface expression of
METHODS: The HaCaT keratinocyte cell line and normal human kerat-
GITRL on keratinocytes. GITRL is a co-stimulatory molecule, originally
inocytes (NHK) were exposed to triamcinolone or pimecrolimus for 24
recognized on professional APCs. Ligation of GITRL on antigen-present-
hours. Additionally, rapamycin was used as a negative control for pimecro-
ing cells (APCs) by its receptor, GITR, expressed on T cells, leads to bidi-
limus, as it binds to the same intracellular receptor-FKB12 - but targets the
rectional signal transduction. This study investigated signalling and
mTOR pathway, as opposed to the calcineurin pathway. LL-37 and HBD-3
chemokine production by keratinocytes after GITRL ligation in resting
expression was evaluated using real-time RT-PCR.
RESULTS: Exposing keratinocytes to pimecrolimus or triamcinolone did
METHODS: 4 mm skin punch biopsies from male Balb/C mice and PAM
not significantly induce gene expression of either LL-37 or HBD-3.
212 murine keratinocytes were incubated with GITR fusion protein (GITR:
Unexpectedly however, rapamycin significantly induced HBD-3 (p <
Fc FP) or Control fusion protein (Control: Fc FP) (10 mg/ml) for 24 h.
0.05) and LL-37 (p < 0.01) at concentrations of 1 nM or greater.
Biopsies and cell supernatants were analyzed by real-time PCR, immuno-
Treatment with 1 mM of rapamycin increased HBD-3 expression by 93.0
histochemical staining and ELISA for chemokine expression. NFkB nu-
6 22.5% and LL-37 expression by 104.3 6 12.4%. Similarly, rapamycin
clear translocation was assessed by immunofluorescence. Preincubation
significantly (p < 0.05) induced HBD-3 and/or LL-37 expression in 4
of PAM 212 cells with NFkB activation inhibitor was preformed for 1 or
out of 5 NHK lots tested. Treating keratinocytes with a NF-kB inhibitor
3 h at concentrations of 10 nM, 100 nM and 1 mM.
prior to stimulation with 1 mM rapamycin reduced the expression of
RESULTS: Stimulation of murine skin biopsies with GITR: Fc FP resulted
HBD-3 and LL-37 by approximately 40% and 90%, respectively.
in mRNA induction for chemoattractant: cutaneous T cell attracting che-
CONCLUSIONS: Rapamycin induces innate immune response genes in
mokine (CTACK), thymus and activation-regulated chemokine (TARC),
keratinocytes and this effect can be (partially) antagonized by a NF-kB in-
monocyte chemoattractant protein-1 (MCP-1) and murine beta defensin
hibitor. Further studies are needed to elucidate the link between the mTOR
3 (MBD3) (p < 0.05 as compared to Control: Fc FP treatment).
pathway and NF-kB signaling in keratinocytes.
Immunofluorescent studies on mouse biopsies treated with GITR: Fc FPconfirmed corresponding TARC and MCP-1 protein production by kerati-nocytes. PAM 212 cells released TARC into supernatant after GITRL liga-tion. Chemokine induction was shown to be NF-kB mediated.
CONCLUSIONS: We conclude that GITRL expressing keratinocyteshave the potential to influence the cellular milieu in inflamed skin throughchemokine production after ligation by GITR expressing cells.
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Molecular Ecology (2008) 17 , 1238–1247 Recombination and genetic differentiation among natural populations of the ectomycorrhizal mushroom Tricholoma matsutake from southwestern China J I A N P I N G X U ,*†‡ TA O S H A ,† YA N - C H U N L I ,* Z H I - W E I Z H A O † and Z H U L . YA N G ** Key Laboratory of Biodiversity and Biogeography, Kunming Institute of Botany, Chinese