| Objective: Multiple sclerosis (MS) is a chronic autoimmuneinflammatory demyelinating disease of central nervous system (CNS). Atpresent, the pathogenesis of MS is still not clear. The domestic and abroadscholars tended to study the immunological mechanism of T cell. Theyconsidered that T cell-mediated immune response was the main factor oflesion initiation in acute MS. But there were a few scholars believed that theimmune response of B lymphocytes also played an important role in thepathogenesis of multiple sclerosis. Researches showed that there were specificautoimmune antibodies generate in CSF from clonally expanded B cells inpatient with MS. Glyceraldehyde-3-phosphate dehydrogenas (GAPDH)-reactive autoantibody was more significant in CSF. This study investigated:1.The expression level of GAPDH-reactive autoantibody in CSF was detectedin patient with MS;2.The difference of the expression level ofGAPDH-reactive autoantibody in CSF was compared between patients withMS and patients with OND；3.The relationship between the expression levelof GAPDH-reactive autoantibody in CSF and sex, age as well as clinical scorewas investigated in patients with MS.Methods: According to the McDonald diagnostic standard in2010,23patients with MS were selected as patient group, meanwhile EDSS score wasrecorded.30patients with other neurological diseases were selected as controlgroup, which included17patients with other inflammatory neurologicaldiseases (OIND) and13patients with other noninflammatory neurologicaldiseases (ONIND). All the selected patients in hospital accepted the lumberpuncture in the first two days after the docter obtained the consent from thepatients and their families.(The patients didn’t get the treatment withcorticosteroids or IVIG).3ml CSF was taken from every patient (The CSF with obvious bleeding was abandoned). The CSF was divided into threegroups and marked as well as cryopreserved in fridge with the temperature of-80℃for the following test. All the samples were collected and thecorresponding reagents were bought. The samples were examined by ELISAand got the value of OD450nm, which represented the mean values obtainedfrom three independent determinations. Statistical analysis was performed bySPSS13.0and compared significant difference of the OD value between MSgroup and other groups. The mean+2SD of ONIND group was defined as thecut-off level. The positive rate of the elevated levels of GAPDH-reactiveautoantibody in CSF was calculated and compared in each group. If there wassignificant difference, the relationship between the expression level ofGAPDH-reactive autoantibody in CSF and sex, age as well as clinical scorewould be analyzed in patients with MS.Results:1The result of ELISAThe result of ELISA showed that: Under the wavelength of450nm, theOD values was0.7246to0.8473, the mean±SD was0.7865±0.0376in MSgroup; the OD values was0.6953to0.8375, the mean±SD was0.7522±0.0354in ONID group; the OD values was0.7042to0.8265, the mean±SDwas0.7578±0.0395in NIND group. The OD value was in normal distributionin each group, The method of independent-sample student t test was used, anddefined α=0.05. The result showed there was significant difference in theexpression of the GAPDH-reactive antibody between MS group andOIND/ONIND group. The p value was0.006/0.038.The mean+2SD of ONIND group was defined as the cut-off level, andthe OD value> the cut-off level was considered to be positive. It showed thatthe patients number of elevated level of GAPDH-reactive antibody in CSFwere2cases in MS group,8.7%;0case in OIND group,0%;0case inONIND group,0%.2The relationship between the expression level of GAPDH-reactiveautoantibody in CSF and clinical indices in patients with MS All data accorded with normal distribution, the independent-samplestudent t test showed there was no significant difference of the expressionlevel of GAPDH-reactive autoantibody in CSF between males and females(F=2.184，P=0.103) in patients with MS. The Spearman correlation analysisshowed that the expression level of GAPDH-reactive autoantibody in CSF hadno correlation with age（r=-0.197，P=0.367）or the clinical score of EDSS(r=0.327，P=0.127) in patients with MS.Conclusions: The expression level of GAPDH-reactive autoantibody inCSF was elevated in patients with MS, and had significant differencecompared with the patients with OIND or ONIND. It means GAPDH may bea candidate Ag for an autoimmune response to neurons and axons in MS. Thiswill provide a new direction for Chinese scholars to study the immunologicalmechanism of MS in the future. Detection of the GAPDH-reactive antibody inCSF may be an auxiliary means for the diagnosis of MS in clinic. Theexpression level of GAPDH-reactive autoantibody in CSF had no significantdifference between males and females and also had no correlation with age orthe clinical score of EDSS in patients with MS. |
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