Allograft inflammatory factor-1 （ AIF-1 ） is a cytoplasmic calcium-binding protein involved in inflammatory response-related diseases in mammals. Previously an identified AIF-1 gene was simply reported in yellow gro...Allograft inflammatory factor-1 （ AIF-1 ） is a cytoplasmic calcium-binding protein involved in inflammatory response-related diseases in mammals. Previously an identified AIF-1 gene was simply reported in yellow grouper. The characterization of AIF-1 gene and its expression at the gene and protein level are further described. Yellow grouper AIF-1 is composed of 147 amino acids, and 64% ~ 84% identical to other homologues. Basal level AIF-1 mRNA expression was noted in spleen, anterior kidney and kidney, using reverse-transcription polymerase chain reaction （RT-PCR）. After stimulation of LPS, the AIF-1 mRNA expression was up-regulated in tissues examined： spleen, anterior kidney, kidney, heart and liver, but not in muscle. The recombinant AIF-1 protein was expressed in Escherichia coli, and then purified for the development of antiserum. Western blotting analysis revealed a band with a molecular mass of about 17 ku.展开更多
Initial ischemia/reperfusion injury (IRI) may have an impact on recipient immune responses after transplantation. Allograft inflammatory factor-1 (AIF-1) has been implicated in the regulation of inflammation associate...Initial ischemia/reperfusion injury (IRI) may have an impact on recipient immune responses after transplantation. Allograft inflammatory factor-1 (AIF-1) has been implicated in the regulation of inflammation associated with organ rejection. We hypothesized that it is either passively released from injured tissues during organ procurement, or actively secreted by allograft infiltrating cells contributing to allograft dysfunction. We investigated the impact of IRI in an in vitro study of human heart tissue during the process of transplantation. The mRNA expression levels for both isoforms of the AIF-1, I2 and I3 were significantly increased after 30 minutes reperfusion (AIF-1 I2: p 0.01 vs. AIF-1 I3: p 0.005). Expression levels for IL-18 and the TLRs were increased after 30 minutes of reperfusion. Only IL-18 and TLR-2 were statistically significant (IL-18: p 0.0001 vs. TLR-2: p 0.01). The mRNA expression levels for AIF-1 I2 and IL-18 were decreased from the original levels of ischemia after 60 and 90 minutes reperfusion. The TLR-2 and -4 were presented with minimal levels of reduction after 60 minutes. However, mRNA expression levels for all were decreased to the original levels of ischemia after 90 minutes, except for AIF-1 I3, but the difference was not statistically significant. AIF-1 and IL-18 were specifically detected in myocytes and interstitial tissues by immunohistochemistry (IHC) stain after IRI. TLR-4 was non-specific, and TLR2 was minimally expressed. The study discusses the evidence supporting that the AIF-1 may have therapeutic potential for strategies in the control of innate immune responses early on, after transplantation.展开更多
基金The National Natural Science Foundation of China under contract No. 30671619
文摘Allograft inflammatory factor-1 （ AIF-1 ） is a cytoplasmic calcium-binding protein involved in inflammatory response-related diseases in mammals. Previously an identified AIF-1 gene was simply reported in yellow grouper. The characterization of AIF-1 gene and its expression at the gene and protein level are further described. Yellow grouper AIF-1 is composed of 147 amino acids, and 64% ~ 84% identical to other homologues. Basal level AIF-1 mRNA expression was noted in spleen, anterior kidney and kidney, using reverse-transcription polymerase chain reaction （RT-PCR）. After stimulation of LPS, the AIF-1 mRNA expression was up-regulated in tissues examined： spleen, anterior kidney, kidney, heart and liver, but not in muscle. The recombinant AIF-1 protein was expressed in Escherichia coli, and then purified for the development of antiserum. Western blotting analysis revealed a band with a molecular mass of about 17 ku.
文摘Initial ischemia/reperfusion injury (IRI) may have an impact on recipient immune responses after transplantation. Allograft inflammatory factor-1 (AIF-1) has been implicated in the regulation of inflammation associated with organ rejection. We hypothesized that it is either passively released from injured tissues during organ procurement, or actively secreted by allograft infiltrating cells contributing to allograft dysfunction. We investigated the impact of IRI in an in vitro study of human heart tissue during the process of transplantation. The mRNA expression levels for both isoforms of the AIF-1, I2 and I3 were significantly increased after 30 minutes reperfusion (AIF-1 I2: p 0.01 vs. AIF-1 I3: p 0.005). Expression levels for IL-18 and the TLRs were increased after 30 minutes of reperfusion. Only IL-18 and TLR-2 were statistically significant (IL-18: p 0.0001 vs. TLR-2: p 0.01). The mRNA expression levels for AIF-1 I2 and IL-18 were decreased from the original levels of ischemia after 60 and 90 minutes reperfusion. The TLR-2 and -4 were presented with minimal levels of reduction after 60 minutes. However, mRNA expression levels for all were decreased to the original levels of ischemia after 90 minutes, except for AIF-1 I3, but the difference was not statistically significant. AIF-1 and IL-18 were specifically detected in myocytes and interstitial tissues by immunohistochemistry (IHC) stain after IRI. TLR-4 was non-specific, and TLR2 was minimally expressed. The study discusses the evidence supporting that the AIF-1 may have therapeutic potential for strategies in the control of innate immune responses early on, after transplantation.
