Dissertation
Dissertation > Medicine, health > Internal Medicine > Systemic disease > Autoimmune diseases > Autoimmune diseases, connective tissue disease > Lupus erythematosus > Systemic lupus erythematosus

The Effect of Oligodeoxynucleotides on the Expression of Toll-like9in Peripheral Blood B Cells and the Secretion of Type Ⅰ Interferon in the Patients with Systemic Lupus Erythematosus

Author FangJiao
Tutor LiZhiJun
School
Course Internal Medicine
Keywords lupus erythematosus systemic Oligodeoxynucleotides Toll like receptor9
CLC R593.241
Type Master's thesis
Year 2011
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Objective: To investigate the immunity regulation effect of immunostimulatoryCpG-containing oligodeoxynucleotides (CpG–ODN) and immunosuppressive ODNon the expression of Toll like receptor9(TLR9) on peripheral blood B lymphocytesand the secretion of type I interferon (IFN) in vitro. We also wish this study couldcontribute to the new strategy for systemic lupus erythematosus (SLE) by exploring thepossible mechanisms of the effect.Methods: Isolated and collected peripheral blood mononuclear cell (PBMC) fromSLE and health control.60cases with SLE (study group) and20cases healthy donorswere enrolled. The PBMC were incubated with RPMI-1640, CpG-ODN andCpG-ODN+immunosuppressive-ODN for48hours in vitro. Then we detected theexpression of TLR9on peripheral blood CD19+B lymphocytes by flow cytometry. Atthe same time the cell-free supernatants were collected and assayed for IFN-α byenzyme-linked immunosorbent assay (ELISA). Compared the differences of the threegroups. According to the SLE Disease Aetivity Index (SLEDAI), SLE patients weredivided into two groups: the slightly active group (S group, n=30, SLEDAI score<10)and the disease moderately/highly active group (MH group, n=30, SLEDAI score≥10). The differences of intracellular TLR9expression and IFN-α level in three groupswere further compared on the basis of the SLEDAI.Results:1.①In the SLE patients, compared with CpG-ODN group, the RPMI-1640group of the proportion of B cells expressing TLR9was lower (mean±SD:11.71±9.62vs19.51±11.23, P<0.001), immunosuppressive-ODN group was also lower(15.04±7.91vs19.51±11.23, P=0.011), although immunosuppressive-ODN groupwas higher than the RPMI-1640group, there was no significant difference betweentwo groups(P=0.057); the mean fluorescence intensity (MFI) was no significant difference among three groups (P>0.05);②In the slightly active patients, comparedwith CpG-ODN group, the RPMI-1640group of the proportion of B cells expressingTLR9was lower (9.19±5.24vs15.92±7.82, P<0.001), immunosuppressive-ODNgroup was also lower (12.54±6.18vs15.92±7.82, P=0.049), althoughimmunosuppressive-ODN group was higher than the RPMI-1640group, and therewas also no significant difference between the two groups(P=0.064); the MFI was nosignificant difference among three groups (P>0.05); In the moderately/highly activepatients, the consequences were similar to the slightly active patients;③In the health people,the CpG-ODN group of the proportion of B cells expressing TLR9was higher thanthe RPMI-1640group (11.59±4.83vs5.43±3.33, P<0.001), also higher than theimmunosuppressive-ODN group (11.59±4.83vs7.77±4.49, P=0.006); to the MFI,both the CpG-ODN group and immunosuppressive-ODN group were higher than theRPMI-1640group, and there was different.(P=0.026, P=0.019).2.The secretion ofIFN-α in the supernatants:①In the SLE patients, the IFN-α level of with CpG-ODNgroup was higher than RPMI-1640group (6.98±0.39vs4.53±0.32, P<0.001), andimmunosuppressive-ODN group was lower than CpG-ODN group(4.63±0.34vs6.98±0.39, P<0.001), but the between the immunosuppressive-ODN group andRPMI-1640group was no statistics difference(P=0.102);②In the slightly activepatients, compared with CpG-ODN group, the RPMI-1640group of the IFN-α waslower (4.37±0.25vs6.82±0.24, P<0.001), and the immunosuppressive-ODN groupwas still lower (4.48±0.27vs6.82±0.24, P<0.001). In the moderately/highly activepatients, the consequences were similar to the slightly active patients.③In the health people,the CpG-ODN group was higher than the other groups and the differences weresignificant.(4.06±0.19vs1.44±0.15, P<0.001;4.06±0.19vs1.55±0.19, P<0.001),but there was no difference between the RPMI-1640group and immunosuppressive-ODNgroup(P=0.060).3.The expression of TLR9on peripheral blood B lymphocytes waspositive correlated with the secretion of IFN-α in PBMC.(Pearson r=0.332, P <0.001).Conclusions: In vitro, our data indicate immunosuppressive-ODN can significantinhibit the expression of TLR9on peripheral blood B lymphocytes and the IFN-αrelease form the supernants induced by CpG-ODN. Suggesting that the mechanism may be related to TLR9on peripheral blood mononuclear cell. Modulating theCpG-ODN–TLR9pathway may offer new opportunities for the understanding andtreatment of systemic lupus erythematosus.

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