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Background: The development of recombinant house dust mite (HDM) allergens opened the way for the in-depth characterization of these molecules but also provided new opportunities to refine the diagnostic procedures of HDM allergy as well as the allergen-specific immunotherapy through tailor-made treatments., Objective: In the present study, the HDM allergen Der p 21 was expressed in Pichia pastoris under a secreted form. The physico-chemical as well as the allergenic characterizations of recombinant Der p 21 (rDer p 21) were performed., Methods: Purified rDer p 21, secreted from recombinant P. pastoris was characterized by CD and MS analysis and the frequency of IgE reactivity was determined by ELISA using 96 sera of HDM-allergic patients from Bangkok. The direct airway epithelial cell activation by rDer p 21 was also evaluated., Results: rDer p 21 was highly expressed under a secreted form in P. pastoris. The physico-chemical characterization of purified rDer p 21 showed that the allergen displayed appropriate α-helix secondary structure content although a two amino acids truncation at the N-terminus of the protein was evidenced by MS. The prevalence of IgE reactivity to rDer p 21 reached 25% in the cohort of the HDM-allergic patients. rDer p 21 could trigger IL-8 production in airway epithelial cells through TLR2-dependent signaling., Conclusion: Properly folded rDer p 21 produced in P. pastoris is appropriate for HDM allergy diagnosis as well for future recombinant allergen-based specific immunotherapy., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Broad-complex (Br-c) is the early ecdysone responsive gene encoding a family of zinc-finger transcription factors. In this study, the full-length cDNA of the Br-c gene of the giant tiger shrimp (Penaeus monodon) was identified. PmBr-c was 1897 bp in length containing an ORF of 1329 bp deducing to a polypeptide of 442 amino acids. PmBr-c was more abundantly expressed in ovaries than testes of P. monodon broodstock. The expression levels of PmBr-c mRNA in ovaries of juveniles was significantly greater than that in stages II (vitellogenic), IV (mature) and V (post-spawning) ovaries of intact broodstock. The expression level of PmBr-c was significantly increased in stage III (nearly mature) ovaries of intact wild broodstock and in stage IV ovaries of eyestalk-ablated broodstock (P<0.05). In domesticated broodstock, ovarian PmBr-c was expressed lower in 18-month-old shrimp compared to 6-month-old shrimp (P<0.05). In situ hybridization revealed that PmBr-c mRNA was localized in ooplasm of previtellogenic oocytes in various ovarian stages of P. monodon broodstock. Serotonin (5-HT, 50 μg/g body mass; 18-month-old shrimp) and progesterone (0.1 μg/g body mass; 14-month-old shrimp) injection significantly promoted the expression level of PmBr-c in ovaries of domesticated broodstock at 24 and 48-72 h post injection (hpi, P<0.05). The expression levels of PmBr-c in ovaries of juvenile P. monodon was significantly increased following 20-hydroxyecdysone treatment (1 μg/g body mass; 4-month-old shrimp) for 168 hpi (P<0.05). Taken together, PmBr-c seems to play an important role during ovarian development of P. monodon., (Copyright © 2013 Elsevier Inc. All rights reserved.)