类风湿性关节炎患者外周血TH17细胞测定

目的 探讨新的效应T细胞TH17在类风湿性关节炎(RA)患者外周血的分布及意义.方法 选取PHA或PMA+Ion作为刺激剂,建立流式细胞术胞内细胞因子染色的方法,应用该方法检测了35例活动期、30例稳定期RA患者及正常人外周血TH17和TH1细胞百分率.结果 PlVIA+Ion联合刺激的效果优于PHA.RA患者及正常人T细胞经PMA+Ion刺激后,CD3+CD8-T细胞IL-17表达水平较相应未刺激组显著增加(P<0.05).而不论是否加刺激剂,活动期RA患者外周血CD3+CD8-IL-17+细胞明显多于稳定期,二者均多于正常对照组(P<0.05).RA患者外周血TH1细胞呈现与T<,H>17细胞类似的分布特点.结论 活动期RA患者外周血存在TH17细胞的分布异常,T1H17细胞可作为评价RA患者细胞免疫功能状态的新指标.     

Notch信号通路相关微小RNA在胶原蛋白诱导的关节炎小鼠免疫细胞中的表达

目的研究胶原蛋白诱发关节炎(CIA)小鼠T细胞中Notch信号通路相关微小RNA(miRNA)的表达情况。方法用牛Ⅱ型胶原蛋白和佐剂免疫DBA/1J小鼠建立CIA小鼠模型,分离外周血单个核细胞(PBMC),采用流式细胞术分选CIA小鼠和正常对照小鼠外周血、脾脏、淋巴结CD4~+T细胞和CD8~+T细胞。实时荧光定量PCR检测PBMC及T细胞中Notch信号通路相关miRNA分子的表达水平。结果与正常对照相比,CIA小鼠PBMC中miR-200b-3p、miR-30c-5p、miR-30b-5p及miR-23b-3p的表达水平均明显降低,miR-200b-3p、miR-30c-5p及miR-30b-5p在CIA小鼠外周血、脾脏、淋巴结CD4~+T细胞和CD8~+T细胞中的表达与正常对照相比未见明显差异,而CIA小鼠外周血、脾脏CD4~+T细胞中miR-23b-3p的表达水平显著低于正常对照小鼠,外周血CD4~+T细胞中的miR-23b-3p来源于脾脏。结论 miR-23b-3p可能参与Notch信号对CIA小鼠T细胞的免疫调控。     

风湿性关节炎患者T细胞表达CCR5和CXCR3的三色流式细胞分析

目的 在单细胞水平上，研究类风湿性关节炎（RA）患者滑液及外周血中T淋巴细胞上CCR5及CXCR3的表达，并结合临床资料分析其在RA发病中的可能作用机制及临床应用。方法 分离15例RA患者的滑液单个核细胞（SFMC）、外周血单个核细胞（PBMC），及15正常人PBMC（对照），以三色荧光素标记物进行流式细胞术分析T细胞上CCR5及CXCR3的表达。结果 与PBMC相比较，RA患者SFMC中T细胞上CCR5及CXCR3的表达显著增高（特别是CCR5）；而CXCR3的表达个体差异较大；与正常人相比较，RA患者初发或活动期未治时，PBMC中CCR5及CXCR3的表达明显增多。CCR5及CXCR3的表达率与患者的ESR及CRP相关。结论 CCR5^ T细胞积聚在RA患者的病变关节内，且与RA的病情密切相关。     

类风湿关节炎患者外周血和滑液中CD4~+CD25~+调节性T细胞的水平变化

了解具有抑制功能的CD4+CD25+调节性T细胞(Treg)在类风湿关节炎(RA)中的水平变化。分离32例RA患者及35例正常对照者外周血和15例RA关节滑液中的单个核细胞,用荧光抗体标记细胞膜表面CD4、CD25分子和细胞内Foxp3转录因子,进行流式细胞分析,同时用RT-PCR方法测定单个核细胞中Foxp3 mRNA水平。实验发现RA外周血中CD4+CD25hT细胞比例(1.90±1.68)与健康人(1.81±1.79)无明显差异,而RA关节滑液中CD4+CD25+和CD4+CD25hT细胞含量却明显增高(14.98±12.52,8.94±9.67,P<0.01)。RA患者外周血单个核细胞中Foxp3+/CD4+T细胞比值(2.35±2.06)较正常人(7.25±3.98)明显降低(P<0.01),RA外周血中Foxp3 mRNA含量较正常人Treg减少,而RA关节液中Foxp3 mRNA含量较RA外周血更为低下(P<0.01)。RA患者存在CD4+CD25+Treg的异常改变,其外周血和关节液中具有抑制作用的Treg含量明显降低提示RA患者Treg数量减少及抑制功能下降可能是RA自身免疫反应亢强不能控制的原因之一。RA关节液中CD4+CD25hT细胞增高考虑与RA炎症反应造成T细胞过度活化有关。     

类风湿性关节炎患者CD4+CD25+Foxp3+调节性T细胞检测及意义

目的:研究CD4 CD25 FoxP3 调节性T细胞在类风湿关节炎患者(RA)外周血中的比例改变,并探讨其在疾病进程中的意义.方法:选取活动期及稳定期RA患者,采用细胞内染色的流式细胞术及定量PCR的方法,分别在蛋白质和mRNA水平检测FoxP3表达,并与正常人进行比较.结果:RA患者CD4和CD25双阳性细胞所占比例与对照组没有明显差异,而活动期患者外周血CD4 CD25high和CD4 CD25 FoxP3 细胞明显低于稳定期和对照组(P＜0.05).FoxP3 mRNA表达水平与蛋白质表达水平变化相一致.结论:类风湿性关节炎活动期时CD4 CD25 FoxP3 调节性T细胞明显减少,这群调节性T细胞可能参与了类风湿性关节炎的病理进程.     

高血压视网膜病变患者外周血单个核细胞Notch1水平及其与血管内皮功能的相关性

目的 探讨高血压视网膜病变(HRP)患者外周血单个核细胞Notch1表达情况及其与血管内皮功能的关系。方法 选择HRP患者42例为研究对象,另以同期健康体检人群30例作为对照组,收集外周血血清、单个核细胞,通过RT-PCR测定Notch1 mRNA水平,免疫印迹测定Notch1蛋白水平,测定血清VEGF、IGF-1、ET-1、NO水平,并进行统计学分析。结果 HRP组Notch1的mRNA(P<0.01)、蛋白水平(P<0.05)均明显低于对照组,Ⅲ期、Ⅳ期患者Notch1的mRNA、蛋白水平下降更为明显(P<0.01,P<0.05);HRP患者VEGF、IGF-1、ET-1明显高于对照组(P<0.05),NO明显低于对照组(P<0.05);Notch1与VEGF、IGF-1、ET-1呈显著负相关(P<0.05),与NO呈显著正相关(P<0.05);Notch1低水平是HRP发病的独立危险因素(OR:0.733,95%CI:0.601～0.835,P<0.001)。结论 HRP患者外周血单个核细胞Notch1水平下降,血管内皮功能紊...     

RA患者外周血CD8＋CD28-和CD4＋CD25＋调节性T细胞的表达及疾病活动性指标相关性分析

目的：检测类风湿性关节炎（RA）患者外周血CD8＋CD28-、CD4＋CD25＋调节性T细胞亚群,探讨其与临床活动性指标的关系。方法：采用流式细胞术检测台州医院RA患者外周血CD8＋CD28-、CD4＋CD25＋ T细胞亚群比例,探讨调节性T细胞与RA活动性、类风湿因子（RF）、免疫球蛋白（Ig）、C反应蛋白（CRP）、补体C3、抗CCP抗体、抗核抗体（ANA）、血小板（PLT）及血沉（ESR）的关系。结果：活动期RA患者外周血CD4＋CD25＋调节性T细胞亚群比例显著低于正常对照组（P〈0.01）,但稳定期RA患者与正常对照组结果差异无统计学意义（P〉0.05）。活动期和稳定期RA患者CD8＋CD28-与正常对照组相比较,结果无统计学意义（P〉0.05）;CD4＋CD25＋与CRP密切相关（r=-0.593,P〈0.05）,CD8＋CD28-与ESR相关系数呈弱相关。CD4＋CD25＋和CD8＋CD28-细胞与RF、IGG、C3、ANA、anti-CCP和PLT未见明显相关性。结论：活动期RA患者外周血CD4＋CD25＋ T细胞亚群比例减少,CD4＋CD25＋ T细胞可能与类风湿性关节炎疾病进展有关。     

淋巴细胞共刺激信号和凋亡信号异常与RA免疫效应亢进的研究

目的：探究T淋巴细胞表面多种细胞信号分子所介导的细胞活化或凋亡信号在RA患者免疫功能紊乱中的作用。方法：采用流式细胞术检测RA患者外周血T细胞亚群及其表面共刺激分子cD154（cD40L）、CD30和凋亡受体CD95（Fas）的表达。结果：RA患者外周血T细胞亚群偏移，CD4^＋T细胞增加，CD8^＋T细胞减少；共刺激分子CD154在CD4^＋和CD8^＋T细胞上的表达均上调，但CD30分子的表达均降低，并以CD4^＋T细胞降低更为明显。同时，凋亡受体CD95分子在T细胞亚群上的表达均明显增加。结论：RA患者T淋巴细胞表面多种信号分子表达异常，共同导致了RA患者免疫功能紊乱。     

类风湿性关节炎患者滑液中CD4~+CD25~-FOXP3~+T细胞比例增加且抑制功能缺陷

目的研究类风湿性关节炎(RA)患者外周血及关节滑液中CD4~+CD25~-FOXP3~+ T细胞[CD25~-调节性T细胞(Treg)]数量及功能分子表达情况及其与疾病活动度之间的关系。方法收集RA患者60例及健康对照者69例,采用流式细胞术对RA患者外周血和滑液(33例)以及健康对照者外周血CD25~- Treg比例及部分功能分子表达情况进行检测,并将相关数据与炎性指标及病情活动度进行相关性分析。采用Mann-Whitney U检验、 Spearman相关分析进行统计学分析。结果 RA外周血中CD25~- Treg占CD4~+细胞比例较健康对照者外周血无明显差异, RA滑液中CD25~- Treg比例较RA及健康对照者外周血均显著增高。将RA患者按疾病活动度及类风湿因子(RF)、抗环瓜氨酸肽(CCP)抗体、抗突变型瓜氨酸波形蛋白(MCV)抗体、抗角蛋白抗体(AKA)分组, RA外周血CD25~- Treg比例在各组间比较无显著差异,并且与血沉(ESR)和C反应蛋白(CRP)水平无明显相关性。功能分子表达上, RA滑液CD25~- Treg中CD39~+细胞比例显著低于健康对照者外周血, CD73~+细胞比例和转化生长因子β1阳性(TGF-β1~+)细胞比例显著低于RA患者及健康对照者外周血;细胞毒性T细胞相关蛋白4(CTLA4)和白细胞介素10(IL-10)表达水平在各组间均无明显差异。结论 RA滑液中CD25~- Treg比例增加,但CD25~- Treg抑制功能分子CD39、 CD73和TGF-β1表达下降,提示其抑制功能可能存在缺陷,进而导致局部炎症未得到有效控制。     

炎症性肠病患者外周血CD4+CD25+Treg细胞及其特异标志物Foxp3表达水平的研究

目的：通过检测炎症性肠病（IBD）不同病期患者以及对照组外周血CD4^＋CD25^＋Treg及其特异标志物Foxp3的表达，来分析与IBD疾病活动性关系，探讨CD4^＋CD25^＋Treg和Foxp3在IBD发病机制中的作用。方法：52例IBD患者和35例正常对照组分别应用流式细胞术和逆转录．聚合酶链反应（RT-PCR）检测外周血中CD4^＋CD25^＋T细胞亚群的百分率测定和外周血单个核细胞Foxp3mRNA的表达水平。结果：IBD患者外周血CD4^＋CD25^＋Treg细胞比例明显低于疾病缓解期患者和正常对照组（P〈0．01）；活动期IBD患者中使用激素和／或免疫抑制剂与未使用激素和／或免疫抑制剂结果差异有统计学意义；IBD患者外周血单个核细胞（PBMC）中Foxp3mRNA表达水平低于缓解期和正常人，差异有显著性（P〈0．05）；缓解期Foxp3mRNA水平与正常人差异无显著性（P〉0．05）；IBD患者外周血CD4^＋CD25^＋Treg细胞表达率及PBMC中的Foxp3mRNA表达水平与疾病活动指数评分呈负相关性。结论：活动期IBD患者外周血CD4^＋CD25^＋Treg细胞及Foxp3mRNA表达下调，而恢复期其表达回升，且二者呈正相关，并与临床活动评分呈负相关，因此认为CD4^＋CD25^＋Treg细胞和Foxp3可能参与疾病的发生发展，与疾病的活动性密切相关。     

类风湿性关节炎患者外周血TH1/TH2细胞的研究

目的探讨CD4+TH1/TH2细胞在类风湿性关节炎(RA)发生发展过程中的作用.方法采用酶联免疫斑点法(ELISPOT)对15例RA患者和30例健康人外周血中T淋巴细胞亚群及CD4+TH1/TH2功能亚型进行检测.结果RA患者外周血中TH1细胞的百分率较正常对照组升高(P＜0.05).结论 TH1细胞介导的细胞免疫可能与RA的发生发展有关.     

Increased miR‐223 expression in T cells from patients with rheumatoid arthritis leads to decreased insulin‐like growth factor‐1‐mediated interleukin‐10 production

We hypothesized that the aberrant expression of microRNAs (miRNAs) in rheumatoid arthritis (RA) T cells was involved in the pathogenesis of RA. The expression profile of 270 human miRNAs in T cells from the first five RA patients and five controls were analysed by real‐time polymerase chain reaction. Twelve miRNAs exhibited potentially aberrant expression in RA T cells compared to normal T cells. After validation with another 22 RA patients and 19 controls, miR‐223 and miR‐34b were over‐expressed in RA T cells. The expression levels of miR‐223 were correlated positively with the titre of rheumatoid factor (RF) in RA patients. Transfection of Jurkat cells with miR‐223 mimic suppressed insulin‐like growth factor‐1 receptor (IGF‐1R) and transfection with miR‐34b mimic suppressed cAMP response element binding protein (CREB) protein expression by Western blotting. The protein expression of IGF‐1R but not CREB was decreased in RA T cells. The addition of recombinant IGF‐1‐stimulated interleukin (IL)‐10 production by activated normal T cells, but not RA T cells. The transfection of miR‐223 mimic impaired IGF‐1‐mediated IL‐10 production in activated normal T cells. The expression levels of SCD5, targeted by miR‐34b, were decreased in RA T cells after microarray analysis. In conclusion, both miR‐223 and miR‐34b were over‐expressed in RA T cells, but only the miR‐223 expression levels were correlated positively with RF titre in RA patients. Functionally, the increased miR‐223 expression could impair the IGF‐1‐mediated IL‐10 production in activated RA T cells in vivo, which might contribute to the imbalance between proinflammatory and anti‐inflammatory cytokines.     

原因不明复发性流产中CD4+ Notch1+T细胞与IL-10的相关性分析

探讨原因不明复发性流产（URSA）中CD4＋Notch1＋T细胞（Notch1＋占CD4＋T细胞的比例）与白介素-10（IL-10）表达水平的相关性。以雌性CBA/J×雄性Balb/c为正常妊娠模型,以雌性CBA/J×雄性DBA/2J为自然流产模型,采用流式细胞术检测6例未孕CBA/J雌性小鼠、6例正常妊娠模型孕14天CBA/J雌性小鼠和6例自然流产模型孕14天CBA/J雌性小鼠脾细胞中CD4＋Notch1＋T细胞,同时运用ELISA法检测其血清中IL-10的表达水平。相比于未孕组,正常妊娠模型中CD4＋Notch1＋T细胞比例减少,而自然流产模型中CD4＋Notch1＋T细胞比例增加,正常妊娠模型与自然流产模型中CD4＋Notch1＋T细胞比例差异有统计学意义（P〈0.05）。CD4＋Notch1＋T细胞比例与IL-10负相关（r=-0.568,P〈0.05）。结论：CD4＋Notch1＋T细胞可能参与原因不明复发性流产发病机制,拮抗Notch1＋表达有可能成为治疗URSA新途径。     

Expression of TLiSA1 on T cells from patients with rheumatoid arthritis and systemic lupus erythematosus

The expression of a new activation antigen, T cell lineage specific activation antigen (TLiSA1) on peripheral blood T cells from 16 rheumatoid arthritis (RA) and 8 systemic lupus erythematosus (SLE) patients and synovial fluid T cells from RA patients was determined in the context of T cell activation. The percentages of TLiSA1 positive T cells from inactive (4.6 +/- 5.2, mean +/- SE) or active RA (19.3 +/- 8.6) or inactive (1.7 +/- 2.1) or active SLE (8.7 +/- 2.7) were significantly increased compared with that of normal controls (0.7 +/- 0.4) (P less than 0.01). All patients with vasculitis showed relatively high positive percentages. The mean fluorocytometric intensity of TLiSA1 positive T cells from RA and SLE patients was significantly higher than that from normals. Percentages of TLiSA1 positive T cells from synovial fluids (21.8 +/- 4.9%) were significantly increased compared with those from peripheral blood of the same patients, indicating the local activation of T cells in patients with RA. An increase in the expression of TLiSA1 with no increase in the expression of the very late activating antigen 1 (VLA-1) was found in peripheral blood from RA, suggesting a difference in the stage of T cell activation in RA. In RA, there was a clinical correlation with levels of TLiSA1 expression on peripheral T cells. After stimulation with PHA, TLiSA1 positive percentages were increased on Day 2 and continued to increase through 5 days of culture. The maximum expression was obtained on Day 5. An increased number of TLiSA1 positive T cells belonged to OKT8. These results suggest that there is the systemic and the local activation of T cells in RA, following antigen stimulation, or a generalized nonspecific activation of immune system that could provide a means to monitor the abnormal immunologic activity in RA.     

Selective recruitment of CXCR3+ and CCR5+ CCR4+ T cells into synovial tissue in patients with rheumatoid arthritis

The inflamed synovial tissue (ST) of rheumatoid arthritis (RA) is characterized by the selective accumulation of interferon gamma-producing Th1-type CD4+ T cells. In this study, we investigated whether the predominance of Th1-type CD4+ cells in the ST lesion is mediated by their selective recruitment through Th1 cell-associated chemokine receptors CXCR3 and CCR5. The lymphocyte aggregates in the ST of RA contained a large number of CD4+ T cells, which mostly expressed both CXCR3 and CCR5, but not CCR4. In contrast, the frequencies of CD4+ and CD8+ T cells expressing CXCR3 and CCR5 in the blood were significantly decreased in RA patients, compared with healthy controls (HC), although there was no difference in the frequencies of CCR4-expressing CD4+ and CD8+ T cells between RA and HC. CXCR3, CCR5, and CCR4 expression in blood CD4 + T cells and CXCR3 expression in CD8+ T cells were increased after interleukin-15 (IL-15) stimulation. Therefore, the distribution of Th1-type CD4+ T cells into the ST from the blood in RA may be associated with the local expression of chemokines, both CXCR3 and CCR5 ligands, and IL-15 may play a role in enhancing these chemokine receptors on CD4+ T cell infiltrates.     

Expression of allograft inflammatory factor-1 in peripheral blood monocytes and synovial membranes in patients with rheumatoid arthritis

ObjectiveAllograft inflammatory factor-1 (AIF-1) is a cytoplasmic protein expressed in various human cells such as monocyte/macrophages and activated T lymphocytes. A recent study showed that AIF-1 is strongly expressed in infiltrating mononuclear cells and synovial fibroblasts in rheumatoid arthritis and that AIF-1 induces the proliferation of cultured synovial cells. In this study we analysed the expression of AIF-1 in peripheral blood monocytes and synovial membranes from patients with rheumatoid arthritis (RA).MethodsWe examined 71 patients with rheumatoid arthritis and 25 control subjects.ResultsUsing flow cytometry we found significantly increased numbers of circulating AIF-1⁺ monocytes in peripheral blood from RA patients compared with controls. Moreover, there were statistically significant positive correlations between AIF-1⁺ monocytes, DAS28 and the Sharp erosion score.Immunofluorescence staining showed strong expression of AIF-1 by infiltrating mononuclear cells – predominantly macrophages in RA synovial tissues – compared with tissues derived from joints affected by osteoarthritis.ConclusionThe results of this study suggest that AIF-1 may be associated with the pathogenesis of RA and may be a novel cytokine involved in the immunological process underlying RA.     

Expression of Activation Markers on CD4+ and CD8+ Cells from Synovial Fluid, Synovial Tissue, and Peripheral Blood of Patients with Inflammatory Arthritides

We studied the expression of the Tac antigen, the transferrin receptor (Tfr-R), HLA class II antigens (DR, DQ, DP), CD30, and Act 1 on purified CD4+ and CD8+ cells isolated from synovial fluid (SF), synovial tissue (ST), and peripheral blood (PB) of patients with rheumatoid arthritis (RA) and with non-RA inflammatory arthritides (not ST). Subfractionated T cells of PB from healthy individuals served as controls. SF CD4+ cells from RA and non-RA arthritides expressed the Tac antigen much more frequently than corresponding CD8+ cells (54 and 58% versus 16 and 17%). In contrast, SF CD8+ cells of both patient groups expressed the HLA class II antigens rather more frequently than the corresponding CD4+ cells (88 and 68% versus 72 and 40%). Tfr-R expression was low on CD4+ and CD8+ SF T cells from both patient groups. SF T cells did not express CD30, and their expression of Act 1 did not differ from that of normal PB T cells. The RA ST findings were similar to those of RA SF. The overall expression of activation markers on PB T cells of patients was slightly higher than on those of normal controls, and the RA group was slightly higher than the non-RA group. The results show that intra-articular T cells in arthritis are activated and that CD4+ and CD8+ subsets differ in their expression of Tac antigen and HLA class II antigens. There were also similar patterns of activation markers on both CD4+ and CD8+ SF cells from RA and non-RA arthritis patients, suggesting that several types of arthritis display a similar immunopathogenesis in the joints.     

Interleukin-15 up-regulates the expression of CD154 on synovial fluid T cells

To investigate the role of the CD40-CD154 interaction in rheumatoid arthritis (RA), we analysed the expression of CD154 on CD3+ and CD4+ T cells in synovial fluid (SF) from patients with RA and in peripheral blood (PB) from patients and normal controls. As interleukin (IL)-15 is a potent activator of synovial T cells we wanted to study whether IL-15 also regulated the expression of CD154 on these T cells. Freshly isolated synovial T cells did not express significant levels of CD154, as evaluated using flow cytometry, whereas the expression of CD86 and human leucocyte antigen (HLA)-DR was significantly elevated on SF T cells when compared with PB T cells from patients or controls. Synovial T cells could up-regulate their CD154 expression following activation with phorbol 12-myristate 13-acetate (PMA) + ionomycin or anti-CD3 + anti-CD28 monoclonal antibodies (mAbs), but the maximal level of expression remained lower than in control T cells. IL-15 significantly increased the expression of CD154 on SF and PB T cells from patients, whereas IL-2 had minimal effects. Furthermore, IL-15 induced extensive proliferation in SF T cells. Our results show that SF T cells up-regulate the expression of CD154 in the presence of IL-15, a cytokine present in the synovium of patients with RA. These results further emphasize the role of IL-15 in the pathogenesis of RA.     

Increased CCR4 expression on circulating CD4(+) T cells in ankylosing spondylitis, rheumatoid arthritis and systemic lupus erythematosus

Previous studies have suggested that CCR4 is particularly important in the selective recruitment of various subsets of leucocytes in rheumatoid arthritis (RA) and systemic lupus erythematosus (SLE). In this study, we examined the percentage of CD4(+)/CCR4(+) T cells within circulating lymphocytes in active ankylosing spondylitis (AS), RA and SLE patients. The clinical significance of CCR4 expression as well as possible associations between the expression and serum levels of tumour necrosis factor (TNF)-alpha, interferon (IFN)-gamma and interleukin (IL)-10 were also examined. Our results showed that the percentage of CD4(+)/CCR4(+) T cells was significantly elevated in AS and RA patients as compared with normal controls. The percentage was also significantly higher in SLE patients who had received no treatment with glucocorticoids or cytotoxic drugs (untreated SLE) than that in controls. In addition, the percentage of CD4(+)/CCR4(+) T cells showed significant positive correlations with the Bath ankylosing spondylitis disease activity index (BASDAI) in AS and with the SLE disease activity index (SLEDAI) in untreated SLE. Of all the cytokines examined, the elevated serum IL-10 level was closely correlated with the percentage of CD4(+)/CCR4(+) T cells in AS, RA and untreated SLE. These results suggest that CCR4 may be crucial in the pathogenesis of AS, RA and SLE. The percentage of CD4(+)/CCR4(+) T cells can serve as a useful marker for the activity of AS and untreated SLE.