RESUMO
Interleukin-12 receptor ß2 (IL-12Rß2) is a signaling subunit of heterodimeric receptors for IL-12 and IL-35. It plays important regulatory functions in the development of Th1 cells and in the expression of inflammatory cytokines in mammals and other higher vertebrates. However, little is known about IL-12Rß2 in teleost fish. In this work, we have cloned and characterized IL-12Rß2 from grass carp (Ctenopharyngodon idella). The full-length cDNA of grass carp IL-12Rß2 is 2875 bp, which encodes a mature protein with 741 amino acids. This mature protein contains three fibronectin type III domains, a transmembrane helix, and CXW and WSXWS-like motifs that are characteristic of the type I cytokine receptor family. Phylogenetic analysis revealed that cyprinid fish IL-12Rß2 formed a single branch, clearly separated from those of other vertebrates. We expressed and purified a recombinant grass carp IL-12Rß2 protein containing major antigenic regions, which was used to raise a polyclonal antibody. The specificity of the antibody was assessed by Western blotting analysis of whole cell lysates from Escherichia coli cells expressing the recombinant IL-12Rß2, grass carp intestinal intraepithelial lymphocytes, and cultured C. idella kidney cells. To explore the potential regulatory role of IL-12Rß2 in inflammation, we generated an intestinal inflammation model by anal intubation of fish with Aeromonas hydrophila. Immunohistochemical staining of the inflamed intestines revealed that IL-12Rß2 expression is consistent with inflammatory cell recruitment during intestinal inflammation. Real-time quantitative PCR revealed that IL-12Rß2 is widely expressed in normal tissues and is up-regulated in most tissues after infecting with A. hydrophila. We found that IL-12Rß2, IL-12p35, and interferon-γ were expressed in similar patterns in the intestines during inflammation. Taken together, our results suggest that IL-12Rß2 is involved in the regulation of intestinal inflammation.
Assuntos
Imunidade Adaptativa/genética , Carpas/genética , Carpas/imunologia , Doenças dos Peixes/imunologia , Regulação da Expressão Gênica/imunologia , Receptores de Interleucina-12/genética , Receptores de Interleucina-12/imunologia , Aeromonas hydrophila/fisiologia , Sequência de Aminoácidos , Animais , Sequência de Bases , Proteínas de Peixes/química , Proteínas de Peixes/genética , Proteínas de Peixes/imunologia , Perfilação da Expressão Gênica/veterinária , Infecções por Bactérias Gram-Negativas/imunologia , Receptores de Interleucina-12/química , Alinhamento de Sequência/veterináriaRESUMO
Objective To simulate leg length discrepancy by unilateral increase in lower limb of normal person, analyze gait features in the case of leg length discrepancy and its effect on walking gait, so as to provide theoretical proofs for chronical musculoskeletal diseases in lower limb amputees due to leg length discrepency. Methods Leg length discrepancy was simulated by subjects wearing shoes to increase the unilateral height of one leg. The time-space parameters, ground reaction forces and joint angles of the subjects during normal walking gait and leg length discrepancy gait were obtained via the 3D motion capture system and the reaction force platform to make comparative analysis. Results Significant differences were found between leg length discrepancy gait and normal gait in terms of step length, stride time and single supporting period. In the case of leg length discrepancy gait, the ground reaction force of both feet significantly increased at heel-strike phase compared with normal gait, and obvious changes were observed in angles of hip, knee and ankle joints. Conclusions Leg length discrepancy is an important cause leading to gait abnormalities, and maybe a cause of leg joint diseases for trans-tibial amputees wearing prosthesis.
