Zhengqi Lu

Interleukin-17-secreting T cells in neuromyelitis optica and multiple sclerosis during relapse

Growing evidence suggests that interleukin (IL)-17 and IL-17-secreting CD4(+)T (Th17) cells are involved in the pathogenic mechanisms of multiple sclerosis (MS). IL-17-secreting CD8(+)T cells were recently identified as a novel subset of CD8(+)T cells. We aimed to analyze the role of Th17 and IL-17 secreting CD8(+)T cells in the pathogenesis of neuromyelitis optica (NMO) as well as MS. Fourteen patients with NMO, 20 with MS and 16 control participants (CTL) were enrolled between November 2008 and December 2009. The proportion of Th17 cells and IL-17 secreting CD8(+)T cells were counted using flow cytometry, and serum levels of IL-6, IL-17, IL-21, IL-23, and transforming growth factor-beta (TGF-β) were measured by enzyme-linked immunosorbent assay. Patients with NMO had a larger proportion of Th17 cells than patients with MS (3.72% versus [vs.] 2.58%, p=0.02) and CTL (3.72% vs. 1.36%, p<0.001). The proportion of Th17 cells in patients with MS was also markedly higher than in the CTL (2.58% vs. 1.36%, p<0.001). IL-17-secreting CD8(+)T cell counts in NMO patients were markedly higher than in MS patients (1.61% vs. 1.09%, p=0.036) and CTLs (1.61% vs. 0.58%, p<0.001). The proportion of IL-17-secreting CD8(+)T cells in MS patients was also higher than in CTLs (1.09% vs. 0.58%, p=0.002). Serum IL-17 and IL-23 levels were increased in patients with NMO and MS, while serum IL-21 concentration was higher only in NMO patients compared to CTL. We concluded that Th17 cells were highly activated in patients with NMO. IL-17-secreting CD8(+)T cells were increased in patients with NMO and MS during relapse and have an important role in the pathological mechanism of NMO and MS.

Overexpression of CNTF in Mesenchymal Stem Cells reduces demyelination and induces clinical recovery in experimental autoimmune encephalomyelitis mice

Human Mesenchymal Stem Cells (MSCs) were previously reported to ameliorate neuronal functional deficits in the MOG35-55-induced experimental autoimmune encephalomyelitis (EAE) mice by inducing T cell anergy. Human Ciliary neurotrophic factor (CNTF) recently was found to promote myelogenesis and reduce inflammation in CNTF-deficient EAE mice. We ectopically overexpressed CNTF in human MSCs to investigate its potential role in promoting remyelination and improving functional recovery in EAE mice. MSCs transfected by Ad-CNTF-IRES-EGFP (MSC-CNTF) were injected intravenously into EAE mice 10 days after the immunization. Neurological functional tests were scored daily by grading clinical signs (score 0-6). Immunofluorescence microscopy was used to detect MSC-CNTF in spinal cord. Expression of NG2, CNTF, and cleaved caspase-3 was measured by immunohistochemistry. CNTF expression was also analyzed by Western blot. Myelin was detected by Solochrome Cyanin staining. Our results found that CNTF concentration in MSC-CNTF cells was 20-fold higher than that in either MSC or Ad-EGFP-transfected MSCs (MSC-EGFP) in vitro. Mice receiving MSC-CNTF cells showed remarkable neuronal functional recovery: the cumulative clinical scores were significantly decreased, and the disease onset was statistically delayed. Mice receiving MSC-CNTF cells showed reduced TNF-alpha, IFN-gamma and increased the level of cytokine IL-10 in peripheral blood and a large number of MSC-CNTF cells were detected in the spleen, but were not detected in other organs such as lung, liver and kidney. In the lesions of these mice, 1) the number of cleaved caspase3-positive cells was significantly reduced; 2) MSC-CNTF- and NG2-positive cells were significantly increased; and 3) the expression of CNTF was dramatically increased. In addition, demyelination was significantly reduced in MSC-CNTF mice. These data indicated that MSC-CNTF may improve functional recovery in EAE mice, possibly by exerting their immunoregulatory activity, inhibiting inflammation, homing MSC-CNTF cells to the lesions, elevating CNTF expression, reducing demyelination, and stimulating oligodendrogenesis.

Soluble egg antigen from Schistosoma japonicum modulates the progression of chronic progressive experimental autoimmune encephalomyelitis via Th2-shift response

Soluble egg antigen (SEA) is strongly antigenic and inherently induces Th2-biased immune responses. In this study, we tested whether SEA from Schistosoma japonicum is able to prevent experimental autoimmune encephalomyelitis (EAE) induced by MOG(35-55) peptide, an established animal model of multiple sclerosis (MS). Intraperitoneal administration with SEA before EAE induction and in the preclinical phase after EAE induction successfully ameliorated the severity and progression of EAE on mice compared with phosphate buffered saline (PBS) controls, while no protective effect was shown when SEA immunization began after disease onset. This effect was associated with reduced interferon gamma (IFN-gamma) production and/or increased interleukin 4 (IL-4) production in spleen and central nervous system (CNS) even at the chronic stage. Similarly, we observed reduced inflammation and demyelination in spinal cords of SEA pretreated EAE mice compared with controls. Our data indicate that immunization with SEA from S. japonicum induces a preestablished Th2-biased microenvironment that provides preventive immune-modulating effects on EAE progression. This study may have important implications for its promising therapeutic use in MS and other autoimmune diseases.

Increased memory Th17 cells in patients with neuromyelitis optica and multiple sclerosis

OBJECT To investigate the clinical relevance of memory Th17 cells in patients with neuromyelitis optica (NMO) or multiple sclerosis (MS). PATIENTS AND METHODS The proportion of peripheral memory Th17 cells was determined by flow cytometry. Sera IL-17A and IL-23 levels were detected by ELISA kits. RESULTS Memory Th17 proportion and IL-17A level were much higher in patients with NMO or MS and were related to clinical features. After high-dose intravenous methylprednisolone (IVMP) therapy, memory Th17 proportion and IL-17A and IL-23 levels were decreased. CONCLUSIONS Memory Th17 is related to the development and relapse of NMO and MS, and IVMP can inhibit memory Th17.

Comparative Brain Stem Lesions on MRI of Acute Disseminated Encephalomyelitis, Neuromyelitis Optica, and Multiple Sclerosis

Background Brain stem lesions are common in patients with acute disseminated encephalomyelitis (ADEM), neuromyelitis optica (NMO), and multiple sclerosis (MS). Objectives To investigate comparative brain stem lesions on magnetic resonance imaging (MRI) among adult patients with ADEM, NMO, and MS. Methods Sixty-five adult patients with ADEM (n = 17), NMO (n = 23), and MS (n = 25) who had brain stem lesions on MRI were enrolled. Morphological features of brain stem lesions among these diseases were assessed. Results Patients with ADEM had a higher frequency of midbrain lesions than did patients with NMO (94.1% vs. 17.4%, P<0.001) and MS (94.1% vs. 40.0%, P<0.001); patients with NMO had a lower frequency of pons lesions than did patients with MS (34.8% vs. 84.0%, P<0.001) and ADEM (34.8% vs. 70.6%, P = 0.025); and patients with NMO had a higher frequency of medulla oblongata lesions than did patients with ADEM (91.3% vs. 35.3%, P<0.001) and MS (91.3% vs. 36.0%, P<0.001). On the axial section of the brain stem, the majority (82.4%) of patients with ADEM showed lesions on the ventral part; the brain stem lesions in patients with NMO were typically located in the dorsal part (91.3%); and lesions in patients with MS were found in both the ventral (44.0%) and dorsal (56.0%) parts. The lesions in patients with ADEM (100%) and NMO (91.3%) had poorly defined margins, while lesions of patients with MS (76.0%) had well defined margins. Brain stem lesions in patients with ADEM were usually bilateral and symmetrical (82.4%), while lesions in patients with NMO (87.0%) and MS (92.0%) were asymmetrical or unilateral. Conclusions Brain stem lesions showed various morphological features among adult patients with ADEM, NMO, and MS. The different lesion locations may be helpful in distinguishing these diseases.

HLA-DPB1 0501 is associated with susceptibility to anti-aquaporin-4 antibodies positive neuromyelitis optica in southern Han Chinese.

OBJECTIVES To analyze the role of HLA-DRB1 and -DPB1 alleles in the pathogenesis of neuromyelitis optica (NMO) and multiple sclerosis in Southern Han Chinese. METHODS Thirty serum anti-aquaporin 4 antibodies (AQP4-Ab)-positive NMO patients, 53 conventional multiple sclerosis (C-MS) patients, and 93 controls (CTLs) were enrolled. The HLA-DRB1 and -DPB1 alleles of the subjects were determined by sequencing-based typing (SBT). RESULTS The frequency of the DRB1 0901 was lower in NMO patients than in CTLs (P(uncorr)=0.022, OR: 0.194, 95% CI: 0.043-0.876), and DRB1 1602 was higher in NMO patients than in C-MS (P(uncorr)=0.038, OR: 3.491, 95% CI: 1.024-11.896) and CTLs (P(uncorr)=0.051, OR: 2.711, 95% CI: 0.971-7.556). The frequency of DPB1 0501 was significant higher in NMO patients than in C-MS (P(uncorr)=0.018, OR: 4.629, 95% CI: 1.235-17.350) and CTLs (P(uncorr)=0.001, P(corr)=0.022, OR: 7.096, 95% CI: 2.011-25.044). CONCLUSIONS DPB1 0501 correlates with risk of AQP4-Ab positive NMO in Southern Han Chinese.

IL-22 secreting CD4 + T cells in the patients with neuromyelitis optica and multiple sclerosis

Interleukin (IL)-22 secreting CD4(+) T (Th22) cells and IL-22 are involved in the pathogenesis of autoimmune disease, but their role in neuromyelitis optica (NMO) and multiple sclerosis (MS) is unclear. We measured the proportion of Th22, Th17, CD4(+)IL-22(+)IL-17A(+) T cells and serum IL-22 in NMO and MS patients. The proportion of Th22 cells, Th17 cells and serum IL-22 were increased in patients with NMO and MS. Our findings suggest that increased Th22 cells may play an important role in the pathogenesis of NMO and MS.

Helicobacter pylori infection in Neuromyelitis Optica and Multiple Sclerosis.

Objective: To determine the Helicobacter pylori infection status in patients with multiple sclerosis (MS) and neuromyelitis optica (NMO) spectrum. Methods:H. pylori infection was certified by indirect immunofluorescence assay. Aquaporin-4 (AQP4) antibody was detected by cell-based assay. H. pylori seroprevalence was measured in 118 patients with NMO (n = 52), high-risk NMO (hrNMO, longitudinally extensive transverse myelitis, n = 17 and recurrent optic neuritis, n = 7), MS (n = 42) and healthy controls (n = 27). Logistic regression analysis was used to determine associations between H. pylori infection and NMO and MS. Results:H. pylori antibodies were present in 119 serum samples (82.1%, 119/145), with antibody positivity in 90.4% (47/52) of the patients with NMO, 95.8% (23/24) of the patients with hrNMO, 73.8% (31/42) of the patients with MS and 59.3% (16/27) of the controls. NMO spectrum patients had greater positivity for H. pylori than MS patients (p < 0.05) and controls (p < 0.05). The frequency of H. pylori seropositivity did not significantly differ between MS patients and controls (p = 0.726). H. pylori seropositivity was significantly higher in AQP4 antibody-positive patients (54/58, 93.1%; p = 0.038) than in AQP4 antibody-negative patients (48/60, 80%). Logistic regression analysis showed that H. pylori seropositivity was significantly associated with hrNMO [odds ratio (OR) = 9.311, p = 0.005] or hrNMO + NMO (OR = 6.350, p = 0.028). Conclusion:H. pylori infection was present in most Chinese patients with NMO and hrNMO, and may be a risk factor for the NMO spectrum.

Effect of high-dose methylprednisolone treatment on Th17 cells in patients with multiple sclerosis in relapse.

Objectives –  Growing evidences have suggested that Th17 cells are involved in the pathogenic mechanisms of multiple sclerosis (MS). Treatment with high‐dose intravenous methylprednisolone (IVMP) has beneficial effects on functional recovery in patients with MS during relapse. The present study was designed to analyze the influences of IVMP on Th17 cells in patients with MS after a 5‐day high‐dose IVMP treatment.

Aquaporin 4 Antibodies in the Cerebrospinal Fluid Are Helpful in Diagnosing Chinese Patients with Neuromyelitis Optica

Objective: It was the aim of this study to compare the diagnostic efficiency of anti-aquaporin 4 (AQP4) antibody detection between serum and cerebrospinal fluid (CSF) samples in Chinese patients with central nervous system demyelinating diseases. Methods: Anti-AQP4 antibodies were detected by a cell-based assay. We calculated the sensitivity, specificity and coherence in 118 patients with neuromyelitis optica (NMO, n = 39), multiple sclerosis (n = 34), longitudinally extensive transverse myelitis (LETM, n = 22), optic neuritis (ON, n = 6), opticospinal multiple sclerosis (n = 8) and acute partial transverse myelitis (n = 9). Results: Forty-four serum samples (33.8%) were positive for anti-AQP4 antibodies. Anti-AQP4 antibody seropositivity was 76.9, 59.1 and 16.7% in patients with NMO, LETM and ON, respectively. Sixty-five CSF samples (50%) were positive for anti-AQP4 antibodies. Anti-AQP4 antibody positivity was 87.1, 81.8, 83.3, 62.5 and 11.8% in patients with NMO, LETM, ON, opticospinal multiple sclerosis and multiple sclerosis, respectively. The ĸ value of the coherence test was 0.585 (p < 0.0001) between the two types of samples. The antibody positivity rate was significantly different between the two body fluids (p = 0.0008, McNemar test). The sensitivity and specificity were 74.3 and 100% in serum, 85.7 and 88.2% in CSF, and 94.3 and 88.2% for serum and CSF combined, respectively. Conclusion: The sensitivity of anti-AQP4 antibodies in the CSF was higher than that in the serum, and their combined use is helpful in diagnosing Chinese patients with NMO.