The Study On The Change Of Lymphocyte Subsets And Clinical Significance In Lupus Nephritis Of Different Pathology Types

Part OneSignificance of lymphocyte subsets' changes in peripheral blood of patients with classⅣand classⅤlupus nephritisObjective:To observe the significance of lymphocyte subsets' changes in peripheral blood of patients with classⅣand classⅤlupus nephritis(LN).Methodology:Fifty-two patients,41 female and 11 male with age from 18 to 55 years old,diagnosed systemic lupus erythematosus(SLE) and classⅣLN(n=32) or ClassⅤLN(n=20) (ISN/RPS2003 classificatioin criteria),were enrolled in the study.Another twenty age-and sex-matched healthy volunteers served as healthy controls(HC).The proportions of peripheral blood lymphocyte subsets(CD4~+,CD8~+,CD20~+ cells) and CD4~+CD25~+Foxp3~+ regulatory T cells(Treg cells) were determined in classⅣand classⅤLN patients as compared to HC by flow cytometry.Correlation analysis of the percentages and absolute cell numbers of Treg cells with clinical disease activity and pathology active index(AI) were performed.Resluts:1.Comparing with HC,the proportion and absolute cell numbers of CD4~+ cells,the ratio of CD4~+/CD8~+ T cells and the level of Treg cells in peripheral blood of active LN patients were decreased significantly(P＜0.01),meanwhile the proportion of CD8~+ cells and CD20~+ cells raised apparently in active LN patients(P＜0.01).2.Comparing with classⅤ,(1) the proportion(25.9%±6.86%vs 31.0%±7.59%, P＜0.05) and absolute cell numbers(288±173cells/ul vs 420±165cells/ul,P＜0.01) of CD4~+ cells and CD4~+/CD8~+ ratio(0.74±0.31 vs 1.06±0.57,P＜0.05) decreased more significantly in classⅣ.(2) No significant difference in the ratio of CD20~+ cells could be found between classⅣand classⅤ,but the absolute cell numbers elevated more significantly in class Ⅴ(185+136cells/ul vs 268±179cells/ul,P＜0.05).(3) Patients with classⅣLN also had statistically lower levels of Treg cells than did patients with classⅤwhen expressed as either percentages of peripheral blood lymphocyte cells(0.82±0.40%vs 1.25±0.70%, P＜0.05) or absolute cell numbers(8.19±4.26cells/ul vs 17.5±10.0 cells/ul,P＜0.05).3.Pearson correlation analysis showed the proportion and the number of CD4~+ cells were correlated with AI(r=-0.281,P＜0.05 and r=-0.380,P＜0.01 respectively).With correlation analysis the proportion and the absolute cell numbers of Treg cells were correlated with SLE-DAI(r=-0.376,P＜0.01 and r=-0.504,P＜0.01 respectively ) and AI(r=-0.278 and r=-0.489,P＜0.01 respectively).Conclusion:The level of CD4+ celIs,CD4+/CD8+ ratio and Treg cells decrease more significantly in classⅣLN patients than did in classⅤLN patients.These results suggest the difference of the immunology pathogenesis of classⅣand classⅤLN.Part twoAlterations of infiltrating lymphocyte subsets and CD68~+ cells in renal tissue of patients with ClassⅣand ClassⅤlupus nephritisObjective:To observe the status of lymphocyte subsets and CD68+ cells infiltrating in renal tissues of patients with classⅣand classⅤlupus nephritis(LN).And to analysis the difference and correlation with clinico-pathologic characteristics.Methodology:Forty-one patients(female 31 and male 10 with age from 18 to 55 years old) suffered from active systemic lupus erythematosus(SLE) and renal biopsy proved classⅣLN(n=30) or classⅤLN(n=11)(ISN/RPS2003 classificatioin criteria) were enrolled in the study.All cases were selected from the patients enrolled in the study of part one. CD4~+,CD8~+,CD20~+ and CD68~+ cells were detected in renal tissues by immunohistochemisty.Meanwhile,twenty patients' renal biopsies(including LN-Ⅳand LN-V) with double immunohistochemical marker for the Treg transcription factor Foxp3,combined with a second marker for CD4 or CD8.Comparing the differences of infiltrating cells in classⅣand classⅤLN,the correlation with clinicopathology was evaluated as well.Resluts:1.Comparing with HC,the numbers of CD4~+,CD8~+,CD20~+ and CD68~+ cells in renal tissues of active LN patients were increased significantly(P＜0.01),but the level of lymphocyte subsets in peripheral blood and in diseased renal tissue was not parallel.2.Meanwhile,infiltrating CD4~+,CD8~+,CD20~+ and CD68~+ cells in renal interstitial increased more apparently in classⅣthan in classⅤ.Especially infiltrating CD68~+ cells in glomerular(12.2±8.7 vs 1.7±2.13,P＜0.01) and CD20+ cells in renal interstitial(119.3±89.7 vs 36.0±36.0,P＜0.01) increased more significantly in classⅣ.3.Infiltrating CD20~+ cells in interstitial and the degree of CD68~+ cells in glomerular were correlated with SLE-DAI(r=0.507 and 0.393,respectively).The level of infiltrating CD20~+ cells in interstitial was correlated with AdsDNA.4.Infiltrating CD20~+ cells in interstitial and the degree of CD68~+ cells in glomerular were correlated with AI(r=0.657 and 0.444,respectively).The degree of CD68~+ cells infiltrating in glomerular was correlated with proliferation of intrinsic cells in glomerular(r=0.624,P＜0.001).5.No Foxp3~+ Treg cells were found in normal renal tissue.Most Foxp3~+ cells were CD4~+.Foxp3~+ Treg cells were located in renal interstitium.Foxp3~+ Treg cells increased more significantly in classⅣthan in classⅤ(27.6±18.0 vs 2.8±5.0,P＜0.01).Conclusion:1.Infiltrating CD4~+,CD8~+,CD20~+ and CD68~+ cells in renal tissue of LN increased significantly.2.The number of infiltrating lymphocyte subsets,especially Treg cells,and macrophages in renal tissue of classⅣLN was higher than that did in classⅤLN.3.The study suggested the difference of lymphocyte subsets and macrophages infiltrating in renal tissue might participate the pathogenesis in classⅣand classⅤLN.Part three A study on clinico-pathology correlation in different types of T lymophocyte subsets' abnormality of lupus nephritis with diffuse proliferative lesionsObjective:To analyze the clinico-pathology characteristics and prognosis in different changes of CD4~+/CD8~+ T cells ratio of lupus nephritis(LN) with diffuse proliferative lesions.Methodology:Seventy female patients(age from 16 to 54 years old) suffered from active LN and renal biopsy proved classⅣ,including IVG,IVS and classⅤ+Ⅳ(ISN/RPS2003 classificatioin criteria),were enrolled in the study.Proportions of peripheral blood lymphocyte subsets(CD4~+,CD8~+,CD20~+ cells) were determined by flow cytometry. According to CD4~+/CD8~+ ratio(RT) these patients were divided to low RT group(RT≤0.5,n=45)and high RT group(RT≥1.5,n=25).CD4~+,CD8~+,CD20~+ and CD68~+ cells were detected in renal tissues by immunohistochemisty.The clinico-pathology characteristics and prognosis were compared between the two groups.Resluts:1.In low RT group,the level of CD4~+ T cells decreased significantly,CD8~+ T cells increased apparently.However,in high RT group,the level of CD4~+ T cells raised slightly, CD8~+ T cells decreased apparently and CD20~+ B cells increased significantly.2.Comparing to high RT group,in low RT group onset age of SLE was younger(26.5±10.4y vs 32.7±9.4y,P＜0.05),SLEDAI(17.2±6.4 vs 14.8±4.4,P=0.09) and the prevalence of fever(51.1%vs 24.0%),malar rash(64.4%vs 32%),skin vasculitis(22.2% vs 0%) and gorss hematuria(26.7%vs 8.0%) were higher;and the level of serum IL-10 was increased significantly(15.5±11.8 vs 7.0±6.1,P＜0.01).3.Renal biopsy showed in low RT group the percentage of class IVS was higher,meanwhile,the proportion of patiens with renal vascular lesion(20%vs 4%,P＜0.05) and AI(9.16±4.34 vs 7.17±3.10,P＜0.05) were increased more signifcantly than in high RT group.4.There were no significance differences of infiltrating CD4~+,CD8~+,CD20~+ and CD68~+ cells in renal tissues of the two groups. 5.Follow-up study showed there were no difference in complete or partial remission rate at the 6th and 12th months in different RT groups.To patients with class IVG LN,in both low RT group and high RT group MMF induction therapy was more effective than CTX induction therapy,especially for the patients in high RT group.Accompany with disease remission,the abnormal RT was ameliorated.Conclusion:The imbalance of CD4~+,CD8~+ T lymphocytes induce the different characteristics of renal lesions in LN-Ⅳ.ClassⅣwith CD8 predominance manifests inflammation and vasculitis,and the increase of CD8~+ T cells is associated with vascular lesion.ClassⅣwith CD4 predominance frequently accompany with membranous lesion,and the level of CD20~+ cells increase significantly at same time,that is suggested Th cells and B cells participate the development of membranous lesions.Our study suggests distinct abnormal RT probably participate the onset and development of LN through different pathophysiology,and these might be associated with the selection of immunotherapy protocols.