| Both cell-mediated immunity and humoral factors are involved in the pathogenesis of Guillain-Barre syndrome (GBS). A deregulation of cytokines is supposed. OBJECTIVES The mRNA expressions of cytokines were assessed in PBMNC of patients with GBS in acute and recovery stages。To investigate whether the mRNA of IFN-γ, IL-18, IL-4, IL-10, TNF-α, TGF-β1 are expressed in peripheral blood mono-nuclear cell of patients with GBS ,and to discover which of them are predominant cytokines associated with the clinical symptoms and signs, and to find which of them express high in the recovery stage, and whether any differences in the mRNA expression exist depending on the different types of GBS,and to investigate that IL-18,TNF-α and IFN-γ are pro-inflammatory factors which express high during the acute or active stage, and that IL-4, IL-10 and TGF-β1 are anti-inflammatory cytokines which seem to terminate and limit the inflammatory reactions and to promote the healing course of GBS. METHODS The study was carried out during the period of John 2001 to November 2002 on 20 patients (10 male and 10 female)who had diagnosed as Guillain-Barre syndrome. The mean age of patients was 9.3 years, 18 patients came from country-side. Most of the clinical cases were diagnosed in the summer. All patients need each 5ml blood in acute phrase (beneath 7 days between symptom onset and therapy beginning) and recovery phrase(≥2weeks from symptom onset). PBMNC were divided into 1×106, it was reserved in –70℃ icebox. To asses the mRNA expressions of cytokines, total RNA from the 1×106 PBMNC were isolated. The sample was isolated by one step method of Trizol. The last precipitated pellet was resuspended in diethylpyrocarbonate-treated water. Total RNA per sample was reserve transcribed using Maloney marine leukemia virus reserve transcriptase and oligo-dt primer according to the manufacturers instruction, at 37℃ for 1 hour. Amplification with specific primer was performed in a Gene Amp PCR system 9600 (Perking Elmer) for 35 cycles with a 30s/94℃ denaturation, 45s/54℃ annealing, 1min/72℃ extension profile in the case of β-actin; 1.15min/94℃ denaturation, 1.30min/58℃ annealing, 1.30min/72℃ extension profile in the case of IL-4; 1min/94℃ denaturation, 1.30min/55℃ annealing, 1.15min/72℃ extension profile in the case of IFN-γ and TGF-β1; 1min/94℃ denaturation, 1min/58℃ annealing, 1min/72℃ extension profile in the case of TNF-α. 30s/94℃ denaturation, 45s/55℃ annealing, 1min/72℃ extension profile in the case of IL-10; 1min/94℃ denaturation, 1.30min/53℃ annealing ,1.15min/72℃ extension profile in the case of IL-18. Amplified products were electrophoresed on 1.5-2% agarose gel stained with 0.5μg/ml ethidium bromide. Result. Every of cytokines was expressed in different stage. The mRNA of TGF-β1 was only expressed in recovery stage, the expression was not different in types of GBS, TGF-β1 mRNA expression was in 35%. The mRNA of IL-18,TNF-α and IFN-γ were only expressed in acute stage, its mRNA expression was in 30%,70%and 10% in acute stage. The mRNAs of IL-4 and IL-10 were both expressed in two phases, IL-4 mRNA expression was in 35% in acute phase and in 85% in recovery phase. The mRNA of IL-10 was expressed in 20% in acute stage and in 80% in recovery stage. CONCLUSION.1,IL-18 and TNF-αwere pro-inflammatory cytokines in GBS,the mRNA of IL-18Was expressed earlier than the mRNA of IFN-γ. The level of IL-18 mRNA expression paralleled clinical GBS. They were the major roles in peripheral demyelination. 2,We considered the mRNA of IFN-γ was only expressed in PNS with GBS, was negotive in PBMNC with GBS. 3,The time course of TNF-α expression in PBMNC roughtly paralleled that of clinical GBS score. The mRNA expression of TNF-α related to demyelinated axons in PNS with GBS. 4,IL-4,IL-10 and TGF-βwere anti-inflammatory cytokines, associated with disease recovery and promoted the healing course of GBS. They were immune response downregulating cytokines in GBS. 5,The mRNA expr...
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