OBJECTIVE: IgA(1) aberrant O-glycosylation is one of the main pathogeneses of IgA nephropathy (IgAN), and the core I beta3-Gal-T-specific molecular chaperone (Cosmc) mRNA expression of IgAN patients was significantly decreased. This study tried to clarify whether the down-regulation was a result of genetic disorders or external suppressions. METHOD: Sixty-five IgAN patients, 23 non-IgAN glomerulonephritis patients and 21 normal controls were recruited. Genomic DNA was extracted and the Cosmc gene was PCR amplified and directly sequenced. Peripheral B lymphocytes of IgAN patients and normal controls were isolated, and cultured with RPMI-1640 alone or with lipopolysaccharide (LPS) for 72 h. The Cosmc mRNA expression levels at baseline, after RPMI culture or RPMI + LPS treatment were measured by real-time RT-PCR. RESULTS: (1) The whole coding frame region of the Cosmc gene was successfully amplified and directly sequenced. Four single nucleotide polymorphisms were detected in two IgAN patients. Two were missense mutations and the others were silent mutations. However, they are different from each other, and unrelated to expression levels; (2) the baseline Cosmc mRNA expression in IgAN patients was significantly lower than normal controls (Ct(COSMC/GAPDH) 1.29 +/- 0.08 versus 1.20 +/- 0.01, 31% of normal controls); (3) the Cosmc mRNA expression level of IgAN patients was remarkably increased after the RPMI culture (1.22 +/- 0.12 versus 1.29 +/- 0.08, 219% of the baseline level), while not in normal controls and (4) treatment with LPS (culture with RPMI + LPS) could strongly inhibit the expression of Cosmc mRNA (1.25 +/- 0.01 versus 1.22 +/- 0.12, 61% of the RPMI treatment group). CONCLUSION: No common Cosmc gene mutation was detected. Significantly increased Cosmc expression was observed in plasma-free culture, while LPS could significantly inhibit it, which suggested that it might not be genetic disorders but external suppression that causes the low Cosmc mRNA expression in IgAN.
OBJECTIVE: IgA(1) aberrant O-glycosylation is one of the main pathogeneses of IgA nephropathy (IgAN), and the core I beta3-Gal-T-specific molecular chaperone (Cosmc) mRNA expression of IgANpatients was significantly decreased. This study tried to clarify whether the down-regulation was a result of genetic disorders or external suppressions. METHOD: Sixty-five IgANpatients, 23 non-IgANglomerulonephritispatients and 21 normal controls were recruited. Genomic DNA was extracted and the Cosmc gene was PCR amplified and directly sequenced. Peripheral B lymphocytes of IgANpatients and normal controls were isolated, and cultured with RPMI-1640 alone or with lipopolysaccharide (LPS) for 72 h. The Cosmc mRNA expression levels at baseline, after RPMI culture or RPMI + LPS treatment were measured by real-time RT-PCR. RESULTS: (1) The whole coding frame region of the Cosmc gene was successfully amplified and directly sequenced. Four single nucleotide polymorphisms were detected in two IgANpatients. Two were missense mutations and the others were silent mutations. However, they are different from each other, and unrelated to expression levels; (2) the baseline Cosmc mRNA expression in IgANpatients was significantly lower than normal controls (Ct(COSMC/GAPDH) 1.29 +/- 0.08 versus 1.20 +/- 0.01, 31% of normal controls); (3) the Cosmc mRNA expression level of IgANpatients was remarkably increased after the RPMI culture (1.22 +/- 0.12 versus 1.29 +/- 0.08, 219% of the baseline level), while not in normal controls and (4) treatment with LPS (culture with RPMI + LPS) could strongly inhibit the expression of Cosmc mRNA (1.25 +/- 0.01 versus 1.22 +/- 0.12, 61% of the RPMI treatment group). CONCLUSION: No common Cosmc gene mutation was detected. Significantly increased Cosmc expression was observed in plasma-free culture, while LPS could significantly inhibit it, which suggested that it might not be genetic disorders but external suppression that causes the low Cosmc mRNA expression in IgAN.
Authors: Tongzhong Ju; Yingchun Wang; Rajindra P Aryal; Sylvain D Lehoux; Xiaokun Ding; Matthew R Kudelka; Christopher Cutler; Junwei Zeng; Jianmei Wang; Xiaodong Sun; Jamie Heimburg-Molinaro; David F Smith; Richard D Cummings Journal: Proteomics Clin Appl Date: 2013-09-09 Impact factor: 3.494
Authors: R Coppo; R Camilla; A Amore; L Peruzzi; V Daprà; E Loiacono; S Vatrano; C Rollino; V Sepe; T Rampino; A Dal Canton Journal: Clin Exp Immunol Date: 2009-11-05 Impact factor: 4.330