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Detecting Anti-aquaporin-4 Antiboday in Neuromyelitis Optica Patients’ Serum

Author: ZhaoMangSuo
Tutor: GengTongChao
School: Tsinghua University
Course: Internal Medicine
Keywords: neuromyelitis optica multiple sclerosis NMO-IgG aquaporin-4
CLC: R744.3
Type: Master's thesis
Year: 2009
Downloads: 31
Quote: 0
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Abstract


Neuromyelitis optica (NMO) is an idiopathic inflammatory demyelinating disease in the central nervous system. Based on the epidemiological, clinical, radiological, pathological and laboratory findings, especially serum NMO-IgG which had been determined as anti-aquaporin-4 autoantibodies, recently most abroad researchers have changed the concept that NMO was a subset of multiple sclerosis(MS) and considered it as a new example of autoimmunological channelopathy in the central nervous system. So far, anti-AQP4 antibody in the serum of NMO patients in the mainland of China have not been reported.. In order to confirm if there is anti-AQP4 antibody in NMO patients’serum in Chinese mainland, HEK293 cell line stably expressing human AQP4 was set up. Total RNA was extrated from brain tissue of a patient diagnosed as temporal epliepsy. Human aquaporin-4 cDNA was amplied by RT-PCR and cloned into the plasmind pEGFP-N1. The recombinant plasmid was tansfected into HEK293, the stable cell line expressing AQP4-GFP fusion protein was screened by antibiotics and flow cytometry sorting. The result show that the rate of stable expression is 80~90%. The expression of AQP4 was detected by RT-PCR and immunofluorescence. The location of AQP4-GFP fusion protein was identified at the membrance under the laser confocal microscopy. Detect serum of NMO, MS, other neurological diseases and normal healthy people as negative control in the mainland of China with the cell line as“antigen”in immunofluorescence, it corfirmed the anti-AQP4 antibody in NMO serum in Chinese mainland, the sensitivity was 88%, specificity was 96.1%. Next,in order to set up a new simple assay to detect anti-AQP4 antibody in the serum of NMO patients which is accessible to clinic practice, we explored the ELISA with synthetic polypeptides as antigen for detecting AQP4 antibody in the serum of NMO patients and firstly testfied that some outmembrane polypeptides of AQP4 protein could be used as antigen in ELISA which can be used to detect anti-AQP4 antibody in the serum of NMO patients. In conclusions, the research identitied the existence of anti-AQP4 antibody in the serum of NMO patients in the mainland of China, and set up a new simple assay for detecting anti-AQP4 antibody in the serum of NMO patients, it would be helpful for diagnosis and therapy of NMO in China.

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CLC: > Medicine, health > Neurology and psychiatry > Neurology > Spinal cord disease > Myelitis
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