AIM: To investigate whether non-canonical autophagy transport receptor cell cycle progression 1(CCPG1) is involved in the corneal antifungal immune response.METHODS: Human corneal epithelial cells(HCECs) and human mye...AIM: To investigate whether non-canonical autophagy transport receptor cell cycle progression 1(CCPG1) is involved in the corneal antifungal immune response.METHODS: Human corneal epithelial cells(HCECs) and human myeloid leukemia mononuclear cells(THP-1) macrophages stimulated by Aspergil us fumigatus(A. fumigatus) were used as cell models. The expression of CCPG1 m RNA was detected by q RT-PCR. Western blot was used to determine the protein expression of CCPG1 and interleukin-1β(IL-1β). The dectin-1 neutralizing antibody was used to detect the association between dectin-1 and CCPG1. Immunofluorescence was used to observe the colocalization of CCPG1 and C-type lectin-like receptor-1(CLEC-1) in THP-1 macrophages.RESULTS: The expression of CCPG1 started to increase at 4 h after infection and increased in a time-dependent manner in HCECs and THP-1 macrophages. With dectin-1 neutralizing antibody pretreatment, the expression of IL-1β was down-regulated. CCPG1 up-regulation in response to A. fumigatus infection was independent of dectin-1. Immunofluorescence showed the colocalization of CCPG1 and CLEC-1 in THP-1 macrophages.CONCLUSION: As a specific autophagy protein of noncanonical autophagy pathway, CCPG1 is involved in corneal infection with A. fumigatus.展开更多
基金Supported by the National Natural Science Foundation of China(No.82171019)the Natural Science Foundation of Shandong Province(No.ZR2021MH368)+1 种基金Traditional Chinese Medicine Research Project of Qingdao(No.2020-zyy055)Shandong Qingdao Outstanding Health Professional Development Fund。
文摘AIM: To investigate whether non-canonical autophagy transport receptor cell cycle progression 1(CCPG1) is involved in the corneal antifungal immune response.METHODS: Human corneal epithelial cells(HCECs) and human myeloid leukemia mononuclear cells(THP-1) macrophages stimulated by Aspergil us fumigatus(A. fumigatus) were used as cell models. The expression of CCPG1 m RNA was detected by q RT-PCR. Western blot was used to determine the protein expression of CCPG1 and interleukin-1β(IL-1β). The dectin-1 neutralizing antibody was used to detect the association between dectin-1 and CCPG1. Immunofluorescence was used to observe the colocalization of CCPG1 and C-type lectin-like receptor-1(CLEC-1) in THP-1 macrophages.RESULTS: The expression of CCPG1 started to increase at 4 h after infection and increased in a time-dependent manner in HCECs and THP-1 macrophages. With dectin-1 neutralizing antibody pretreatment, the expression of IL-1β was down-regulated. CCPG1 up-regulation in response to A. fumigatus infection was independent of dectin-1. Immunofluorescence showed the colocalization of CCPG1 and CLEC-1 in THP-1 macrophages.CONCLUSION: As a specific autophagy protein of noncanonical autophagy pathway, CCPG1 is involved in corneal infection with A. fumigatus.
