RESUMO
BACKGROUND: House dust mites are an important source of indoor allergens. More than 30 allergens of Dermatophagoides farinae (D. farinae) have been identified. Yet there may be many other allergens in mites remain to be characterized. METHODS: α-Tubulin (also named Der f 33) was cloned, expressed and purified. Reaction to specific-IgE, skin prick test and a mouse asthma model were employed to determine the allergenicity of Der f 33. RESULTS: The recombinant Der f 33 reacted to the serum of patients with mite allergy. The positive rate of skin prick test (SPT) was 23.5%. In an asthma mouse model, Der f 33 induced the airway allergy-like responses. Moreover, serum specific IgE and IgG1, interleukin-4 (IL-4) from bronchoalveolar lavage fluid (BALF) and spleen cell culture supernatant were markedly increased. In addition, Der f 33 upregulated the CD80 and TNF-α levels in dendritic cells (DCs). CONCLUSIONS: Der f 33 is a novel allergen of D. farinae. It modulates the functions of DCs and induces airway allergy.
Assuntos
Alérgenos/imunologia , Antígenos de Dermatophagoides/imunologia , Proteínas de Artrópodes/imunologia , Asma/imunologia , Dermatophagoides farinae/imunologia , Tubulina (Proteína)/imunologia , Alérgenos/genética , Alérgenos/toxicidade , Animais , Antígenos de Dermatophagoides/genética , Antígenos de Dermatophagoides/toxicidade , Proteínas de Artrópodes/toxicidade , Asma/induzido quimicamente , Dermatophagoides farinae/genética , Feminino , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Tubulina (Proteína)/genética , Tubulina (Proteína)/toxicidadeRESUMO
Undimerized beta-tubulin is toxic in the yeast S. cerevisiae. It can arise if levels of beta-tubulin and alpha-tubulin are unbalanced or if the tubulin heterodimer dissociates. We are using the toxicity of beta-tubulin to understand early steps in microtubule morphogenesis. We find that deletion of PLP1 suppresses toxic beta-tubulin formed by disparate levels of alpha- and beta-tubulin. That suppression occurs either when alpha-tubulin is modestly underexpressed relative to beta-tubulin or when beta-tubulin is inducibly and strongly overexpressed. Plp1p does not affect tubulin expression. Instead, a significant proportion of the undimerized beta-tubulin in plp1Delta cells is less toxic than that in wild-type cells. It is also less able to combine with alpha-tubulin to form a heterodimer. As a result, plp1Delta cells have lower levels of heterodimer. Importantly, plp1Delta cells that also lack Pac10, a component of the GimC/PFD complex, are even less affected by free beta-tubulin. Our results suggest that Plp1p defines a novel early step in beta-tubulin folding.