摘要
目的:观察Ⅳ型与Ⅴ型狼疮性肾炎(LN)患者外周血淋巴细胞亚群的变化和肾组织淋巴细胞亚群和单核-巨噬细胞的浸润情况,并进一步分析与临床和病理的关系。方法:52例(男性11例,女性41例,年龄18～55岁)初治活动性系统性红斑狼疮,根据2003年ISN/RPS病理分型标准经肾活检确诊为Ⅳ型(n=32)和Ⅴ型(n=20),流式细胞仪测定外周血淋巴细胞亚群(CD3+、CD4+、CD8+、CD20+细胞)和CD4+CD25+Foxp3+调节性T细胞(Treg细胞)的比例与计数,计算CD4+/CD8+比值。20例年龄、性别匹配的健康志愿者作为对照。同时检测30例Ⅳ型和11例Ⅴ型LN肾组织CD4、CD8、CD20和CD68免疫组化染色,取10例移植肾供肾活检标本作为正常肾组织对照。从中再选取20例(Ⅳ型和Ⅴ型各10例)行CD4和Foxp3双标免疫组化染色。比较LN患者外周血淋巴细胞亚群和Treg细胞与正常对照的差别,分析Ⅳ型和Ⅴ型之间外周血和淋巴细胞亚群和Treg细胞的变化特点。进一步分析Ⅳ型和Ⅴ型LN肾组织淋巴细胞亚群,特别是Treg以及单核-巨噬细胞浸润的差异和临床病理联系。结果:活动性Ⅳ型、Ⅴ型LN患者与正常人比较外周血CD4+细胞比例、计数和CD4+/CD8+细胞比值及Treg细胞显著降低,CD8+细胞及CD20+细胞明显升高(P<0.01)。Ⅳ型与Ⅴ型相比:(1)外周血CD4+细胞比例[(25.9±6.86)%vs(31.0±7.59)%,P<0.05]、计数[(288±173)个/μlvs(420±165)个/μl,P<0.01]和CD4+/CD8+比例降低更明显(0.74±0.31vs1.06±0.57,P<0.05);(2)外周血CD20+细胞比例无明显差异,但Ⅴ型CD20+细胞计数升高较Ⅳ型明显[(185±136)个/μlvs(268±179)个/μl,P<0.05];(3)Treg细胞比例[(0.82±0.4)%vs(1.31±0.7)%,P<0.05]和计数[(8.19±4.26)个/μlvs(17.5±10.0)个/μl,P<0.01]Ⅳ型降低更明显。CD4+细胞比例和计数与AI呈负相关(r分别为-0.281和-0.380,P<0.05),CD4+/CD8+细胞比值与SLEDAI呈负相关(r=-0.307,P<0.05)。Treg细胞比例和计数与SLEDAI呈显著负相关,相关系数分别为-0.411(P<0.01)和-0.480(P<0.01),与肾组织AI呈负相关(r为-0.325和-0.473,P<0.01)。肾组织免疫组化见Ⅳ型肾间质浸润的CD4+、CD8+、CD20+和CD68+细胞均高于Ⅴ型病例,其中肾小管浸润的CD20+细胞Ⅳ型明显高于Ⅴ型(119.3±89.7vs36.0±36.0,P<0.01),肾小球内浸润的CD68+细胞显著高于Ⅴ型(12.4±8.8vs1.7±2.31,P<0.01)。正常肾组织不表达Foxp3+Treg细胞,绝大多数Foxp3表达于CD4+细胞,LN中CD4+Foxp3+Treg细胞主要位于肾间质,Ⅳ型LNCD4+Foxp3+Treg细胞明显高于Ⅴ型(27.6±18.0vs2.8±5.0,P<0.01)。结论:Ⅳ型LN患者外周血CD4+细胞数量、CD4+/CD8+细胞比例和Treg细胞数量较Ⅴ型明显降低,Ⅳ型LN肾组织T、B淋巴细胞,特别是调节性T细胞和巨噬细胞浸润较Ⅴ型明显升高,提示不同病理类型LN的免疫发病机制不同。
Objective：To observe the variation of lymphocyte subsets in peripheral blood and renal tissue of patients with class Ⅳ and class Ⅴ lupus 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 class Ⅳ LN（n =32） or Class Ⅴ LN （n = 20） （ISN/RPS2003 classificatioin criteria） , were enrolled in the study. Another twenty age-and sex-matcbed 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 class Ⅳ and class Ⅴ LN 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. Th^＋ number of CD4 ^＋ , CD8 ^＋ , CD20 ^＋ , Foxp3^＋ and CD68 ^＋ cells was detected in renal tissues by immunohistochemistry. Results： Comparing with HC, the proportion and absolute cell numbers of CD4 ^＋ cells, the ratio of CD4 ^＋/CD8 ^＋ cells and the level of Treg cells in peripheral blood were decreased significantly （P 〈0. 01 ）, meanwhile the proportion of CD8 ^＋ cells and CD20 ^＋ cells raised apparently in active LN patients （P 〈0. 01 ）. Comparing with class Ⅴ, the proportion （25.9% ±6. 86% vs 31.0% ±7. 59% ,P 〈0. 05） and absolute cell numbers [ （ 288 ± 173 ） cells/μl vs （ 420 ± 165 ） cells/μl, 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 class Ⅳ. No significant difference in the ratio of CD20^＋ cells could be found between class Ⅳ and class Ⅴ, but the absolute cell numbers elevated more significantly in class Ⅴ[ （185 ± 136） cells/μl vs （268± 179） cells/μl, P 〈 0. 05 ]. The patients with class Ⅳ LN also had statistically lower levels of Treg cells than did patients with class Ⅴ when 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 [ （98. 19 ±4. 26） cells/μl vs （ 17.5 ±10. 0） cells/μl, P 〈0. 051. 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）. Renal tissues immunohistochemistry showed there was significant increase of infiltrating CD4 ^＋ , CD8 ^＋ and CD68 ^＋ cells in LN. However, infiltrating CD4 ＋ , CD8 ^＋ , CD20^＋ and CD68 ^＋ cells in renal interstitial increased more apparently in class Ⅳ than in class Ⅴ. Especially infiltrating CD68^＋ cells in glomeruli （12.2 ±8.7 vs 1.7±2.13,P〈0.01） and CD20^＋ cells in renal interstitium （119.3 ±89.7 vs 36. 0±36. 0,P 〈0. 01 ） increased more significantly in class Ⅳ. 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 class Ⅳ than in class Ⅴ （ 27.6 ± 18.0 vs 2. 8± 5.0, P 〈 0. 01 ）. Conclusion： The level of CD4 ^＋ cells, CD4 ^＋/CD8 ^＋ ratio and Treg cells decreased more significantly in patients with class Ⅳ LN. The infiltrating lymphocyte subsets, especially Treg cells, and monocytes in renal tissues also increased significantly in class Ⅳ. These results suggested that there be the difference of the immunology pathogenesis in class Ⅳ and class Ⅴ LN.
出处
《肾脏病与透析肾移植杂志》
CAS
CSCD
2008年第2期101-112,共12页
Chinese Journal of Nephrology,Dialysis & Transplantation
