Cellular immune responses.

<p>IL-5, IL-4 and IFN-γ levels in supernatants of restimulated spleen cell cultures of mice, treated with 1 µg (black bars), 5 µg (grey bars) or 25 µg (striped bars) of (A), thioredoxin or (B), arginine kinase, or sham-treated (white bars). Groups were compared to sham-treated controls. *p<0.05, **p<0.01.</p

Immunoblot inhibition experiment.

<p>Extracts from <i>E. kuehniella</i> were separated by polyacrylamide electrophoresis, blotted onto nitrocellulose, and strips were probed with serum samples preincubated with or without 10 µg of recombinant allergen overnight. Lane 1, thioredoxin (Plo i 2) positive serum; lane 2, same serum preincubated with recombinant <i>P. interpunctella</i> thioredoxin; lane 3, arginine kinase (Plo i 1) positive serum; lane 4, same serum preincubated with recombinant Plo i 1; lane 5, nonallergic control serum, lane 6, same serum preincubated with Plo i 2; lane 7, same serum preincubated with Plo i 1; lane 8, buffer control. Molecular weights [kDa] are indicated at the left side, the positions of thioredoxin (TRX) and arginine kinase (AK) at the right side.</p

IgE immunoblots with patients' sera.

<p>Top panels: extracts from whole mediterranean flour moths (<i>E. kuehniella</i>) were separated by polyacrylamide electrophoresis, blotted onto nitrocellulose, and probed with the sera from various groups of patients with indoor allergy or seafood allergy as well as from control individuals. The indoor allergic patient groups were: M, housedust mite allergic patients (n = 69), M+D, patients allergic to mites and animal dander (n = 43), D−M, patients with allergy to animal dander but not to mites (n = 20). Patients SF, seafood allergic patients (n = 22), NC, nonallergic control individuals (n = 2), BC, buffer control without serum, only secondary anti-IgE antibody. Molecular weights [kDa] are indicated at the left side. Bottom panels, the recombinant <i>P. interpunctella</i> 40 kDa arginine kinase (AK) and 12 kDa thioredoxin (TRX) were probed with those patients' sera containing IgE to <i>E. kuehniella</i>, and only the parts of the strips containing the two recombinant allergens are shown.</p

In vitro splenocyte stimulation.

<p>Stimulation index [SI] of thioredoxin or arginine kinase restimulated spleen cell cultures of mice, treated with 1 µg (black bars), 5 µg (grey bars) or 25 µg (striped bars) antigen, compared to sham-treated controls (white bars). **p<0.01.</p

Comparison of thioredoxin sequences.

<p>The deduced amino acid sequence of <i>P. interpunctella</i> thioredoxin Plo i 2 (Pi) was aligned with <i>Bombyx mori</i> thioredoxin (ABM92269) (Bm) with 83% identity. The other arthropod thioredoxin from the shrimp <i>Litopenaeus vannamei</i> (ACA60746) (Lv) was 58% identical, and human thioredoxin (NP_003320) (Hs) shared 47% amino acid identity with Pi. The allergenic fungal thioredoxin (2J23_A) Mala s 13 from <i>Malassezia sympodialis</i> (Ms) and wheat (<i>Triticum aestivum</i>) thioredoxin Tri a 25 (Ta) (CAB96931) were 46% and 41% identical, respectively with Plo i 2. The residues equal to Plo i 2 are marked with dashes. The important thioredoxin consensus sequence WCGPC with the two active site cysteines was found in all the sequences (bold type).</p