Analysis of different lymphocyte subpopulations in patients with nonspecific ulcerative colitis

A complex of methods based on a rosette forming reaction, was used to study the content of different lymphocytic populations in the peripheral blood of persons with nonspecific ulcerative colitis. Imbalance in the system of immunocompetent cells was shown to develop in this disease: a decrease in the amount of suppressors (T gamma and theophylline sensitive lymphocytes) and a increase in the amount of immature postthymic cells: cells forming rosettes with autologous erythrocytes, and theophylline dependent lymphocytes.     

Synthesis of eicosanoids in the colonic mucosa in patients with ulcerative colitis

The synthesis of cyclooxygenase (CO) and lipoxygenase (LO) metabolites of arachidonic acids, such as prostaglandins (PG) E2, F2 alpha, 6-keto-P1 alpha, thromboxane B2 (TB2), leukotriene B4 (LTB4) and hydroxyeicosatetraenic acids (HETEA) in the biopsy specimens of the colonic mucosa (CM) was studied in vitro in 30 patients aged 17-66 years who had ulcerative colitis (UC) of various severity and extent. The biopsy specimens of CM from 10 patients with the irritable bowel syndrome were used for comparison. A proportional severity of the disease and increased synthesis of CO and LO metabolites in CM was ascertained in a phase of UC. In the early phase of clinical remission (on the average, following 4 weeks of therapy), there was a comparative reduction in the level of eicosanoids with the preserved high production of TB2 and LO derivatives (LTB4 and HETEA). At the same time, in patients with severe UC, a higher synthesis of LTB4 and HETEA and PG was preserved. The predominance of CM production of eicosanoids having aggregative, vasoconstrictor, and proinflammatory effects (less coefficients of 6-keto-PGF1 alpha/TB2 and PG/LTB4 + ETEA), which had been detected in a phase of UC exacerbation, was preserved in a phase of remission development, by forming the metabolic basis for recurrence of the process.     

Phospholipase A2 in serum and colonic mucosa in ulcerative colitis.

Group II phospholipase A2 is involved in the pathogenesis of various inflammatory diseases and in the host defence against bacteria. The enzyme is expressed in the epithelial cells of colonic mucosa in ulcerative colitis. In this study, we measured the concentration of group II phospholipase A2 in serum and colonic mucosa of patients with ulcerative colitis of different severity and of control patients without any inflammatory disease. The activity of ulcerative colitis was assessed by endoscopy. The concentration of group II phospholipase A2 was measured with an immunoassay. The concentrations of group II phospholipase A2 in serum and colonic mucosa were significantly higher in patients with active and inactive ulcerative colitis than in controls. However, the group II phospholipase A2 levels did not separate patients with different disease activity. The concentration of group II phospholipase A2 in colonic mucosa corresponded with the mucosal inflammatory activity (higher in active colonic areas) intra-individually, but not between different patients with ulcerative colitis. Serum group II phospholipase A2 values were above the normal reference range more often than the values of 11 standard laboratory blood tests widely used for the follow-up of inflammatory activity in ulcerative colitis. These results indicate that the concentration of group II phospholipase A2 is increased in serum and colonic mucosa of patients with ulcerative colitis. The clinical value of the measurement of group II phospholipase A2 in the follow-up of ulcerative colitis remains to be clarified.     

Depletion of non specific esterase activity in the colonic mucosa of patients with ulcerative colitis

BACKGROUND: Non specific esterases (NSE) are a group of cellular carboxylesterases, enzyme markers of monocytes/macrophages, whose tissue distribution in the human body and changes in various disease states have not been adequately studied. We investigate the presence and localization of NSE, in the normal and inflamed human colonic mucosa. DESIGN: NSE were studied histochemically and biochemically using alpha-naphthyl acetate as the substrate, in the colonic mucosa from 67 patients with colitis of various aetiologies and 10 normal controls. In addition, esterase activity was studied biochemically in serum from colitic patients and normal controls. RESULTS: Histochemical study of the colonic tissue demonstrated that NSE were localised in the epithelial brush border, the goblet cells of the glands and a macrophage population of the lamina propria in the colonic mucosa of normal controls and patients with non specific colitis. In active ulcerative colitis, esterase depletion and esterase negative macrophages were identified in parallel with goblet cell disappearance. Gradual reappearance of esterase activity was found after successful treatment. Biochemical study of NSE activity showed that serum and colonic tissue esterase levels were greatly (P < 0.001) reduced in active ulcerative colitis compared to the normal controls or non specific colitis patients and they were increased after successful treatment. Despite this increase, the esterase activity in the colonic tissue from ulcerative colitis patients after treatment was significantly reduced compared to the normal controls. Interestingly, the enzyme levels from non-inflamed areas of the bowel of patients with ulcerative colitis were also significantly (P < 0.01) decreased compared to the normal controls. CONCLUSIONS: These data suggest that esterase reduction in ulcerative colitis is not a simple result of the inflammatory process but rather it precedes its development. This enzyme depletion might have an important pathogenetic implication in the inflammatory process.     

单核细胞趋化蛋白-1在溃疡性结肠炎黏膜中的表达

目的探讨单核细胞趋化蛋白-1(MCP-1)在溃疡性结肠炎(UC)发病机制中的作用。方法32例UC黏膜标本取自行结肠镜检查的患者,按病理组织学对炎症进行分级,Ⅲ、Ⅳ级20例,Ⅰ、Ⅱ级12例;对照组为15名健康成人。应用半定量RT-PCR检测对照组和UC组肠黏膜MCP-1 mRNA表达水平,免疫组织化学方法检测MCP-1蛋白的表达。结果与对照组相比,UC肠黏膜MCP-1 mRNA和其蛋白过度表达,MCP-1 mRNA表达与疾病严重程度成正相关(P<0.01)。MCP-1蛋白表达位于UC肠黏膜固有层单核细胞和T淋巴细胞,病理分级Ⅲ、Ⅳ级肠黏膜MCP-1蛋白表达较病理分级Ⅰ、Ⅱ级组明显增加(t=7.31,P<0.01)。结论UC黏膜组织中MCP-1表达水平明显上调,其表达水平与病情轻重和炎症严重程度呈正相关。     

表皮生长因子受体在溃疡性结肠炎结肠黏膜修复中的作用

目的：观察表皮生长因子受体在活动性和非活动性溃疡性结肠炎患者结肠黏膜的表达，以探讨表皮生长因子受体表达的临床意义。方法：于2001-05／2005-05选择南京医科大学第一附属医院消化科住院的溃疡性结肠炎患者32例作为观察对象。根据改良Williams疾病活动指数将之分为活动性溃疡性结肠炎组（n=22）和非活动性溃疡性结肠炎组（n=10）。对照组10例为同期门诊健康体检者或肠易激综合征患者。所有参与者均同意结肠镜检查及结肠黏膜活检。溃疡性结肠炎组取至炎症最明显处，对照组在直乙交界处取黏膜组织。采用免疫组化SP法检测参与者结肠黏膜表皮生长因子受体及增殖细胞核抗原的表达。阳性反应物呈棕黄色颗粒，表皮生长因子受体免疫阳性反应位于胞浆及胞膜，增殖细胞核抗原免疫阳性反应位于胞核。每张切片随机选取5个高倍视野观察，计数阳性反应的上皮细胞。评估标准：①表皮生长因子受体阳性反应细胞数／所有上皮细胞数〈5％时记为0，〈50％为1，〉50％为2，根据染色强度记为不着色0，黄色为1，棕黄色为2；两者相加：0为阴性（-），1为弱阳性（1＋），2-3为阳性（2＋），4为强阳性（3＋）。②增殖细胞核抗原阳性核染色细胞数／所有上皮细胞数〈25％时为Ⅰ级，26％～50％为Ⅱ级，51％～75％为Ⅲ级，〉76％为Ⅳ级，Ⅲ级和Ⅳ级为增殖细胞核抗原过表达。结果：所有参与者均完成各项检查，无脱落者，全部进入结果分析。对照组表皮生长因子受体阳性表达频率为4／10，增殖活性最低，7例增殖细胞核抗原表达为Ⅰ级；活动性溃疡性结肠炎组表皮生长因子受体阳性表达频率为19／22，增殖活性亦增强，增殖细胞核抗原过表达频率为5／22；非活动性溃疡性结肠炎组表皮生长因子受体全部表达为阳性，增殖细胞核抗原过表达频率为9／10。对照组、活动性溃疡性结肠炎组、非活动性溃疡性结肠炎组表皮生长因子受体阳性表达率、增殖细胞核抗原过表达率依次上升，组间比较差异显著（25％，86％，100％，P〈0．01）；（10％，23％，90％，P〈0．01）。结论：非活动性溃疡性结肠炎患者结肠黏膜表皮生长因子受体表达明显增加，细胞增殖活跃，表皮生长因子受体在溃疡性结肠炎结肠黏膜炎症修复过程中起一定作用，适当的表皮生长因子受体表达在损伤修复中起中关键作用，过度不恰当的表达可能在细胞的恶性转化中起作用，促进溃疡性结肠炎相关性结直肠癌的发生。     

表皮生长因子受体在溃疡性结肠炎结肠黏膜修复中的作用

目的:观察表皮生长因子受体在活动性和非活动性溃疡性结肠炎患者结肠黏膜的表达,以探讨表皮生长因子受体表达的临床意义。方法:于2001-05/2005-05选择南京医科大学第一附属医院消化科住院的溃疡性结肠炎患者32例作为观察对象。根据改良Williams疾病活动指数将之分为活动性溃疡性结肠炎组(n=22)和非活动性溃疡性结肠炎组(n=10)。对照组10例为同期门诊健康体检者或肠易激综合征患者。所有参与者均同意结肠镜检查及结肠黏膜活检。溃疡性结肠炎组取至炎症最明显处,对照组在直乙交界处取黏膜组织。采用免疫组化SP法检测参与者结肠黏膜表皮生长因子受体及增殖细胞核抗原的表达。阳性反应物呈棕黄色颗粒,表皮生长因子受体免疫阳性反应位于胞浆及胞膜,增殖细胞核抗原免疫阳性反应位于胞核。每张切片随机选取5个高倍视野观察,计数阳性反应的上皮细胞。评估标准:①表皮生长因子受体阳性反应细胞数/所有上皮细胞数<5%时记为0,<50%为1,>50%为2,根据染色强度记为不着色0,黄色为1,棕黄色为2;两者相加:0为阴性(-),1为弱阳性(1+),2～3为阳性(2+),4为强阳性(3+)。②增殖细胞核抗原阳性核染色细胞数/所有上皮细胞数<25%时为Ⅰ级,26%～50%为Ⅱ级,51%～75%为Ⅲ级,>76%为Ⅳ级,Ⅲ级和Ⅳ级为增殖细胞核抗原过表达。结果:所有参与者均完成各项检查,无脱落者,全部进入结果分析。对照组表皮生长因子受体阳性表达频率为4/10,增殖活性最低,7例增殖细胞核抗原表达为Ⅰ级;活动性溃疡性结肠炎组表皮生长因子受体阳性表达频率为19/22,增殖活性亦增强,增殖细胞核抗原过表达频率为5/22;非活动性溃疡性结肠炎组表皮生长因子受体全部表达为阳性,增殖细胞核抗原过表达频率为9/10。对照组、活动性溃疡性结肠炎组、非活动性溃疡性结肠炎组表皮生长因子受体阳性表达率、增殖细胞核抗原过表达率依次上升,组间比较差异显著(25%,86%,100%,P<0.01);(10%,23%,90%,P<0.01)。结论:非活动性溃疡性结肠炎患者结肠黏膜表皮生长因子受体表达明显增加,细胞增殖活跃,表皮生长因子受体在溃疡性结肠炎结肠黏膜炎症修复过程中起一定作用,适当的表皮生长因子受体表达在损伤修复中起中关键作用,过度不恰当的表达可能在细胞的恶性转化中起作用,促进溃疡性结肠炎相关性结直肠癌的发生。     

Diagnosis of acute ulcerative colitis and colonic Crohn's disease by colonic sonography

By instilling water into the large intestine, sonographic visualization of the whole length of the colon from the rectosigmoid to the cecum can be achieved. Furthermore, using this method, it is possible to evaluate the lumen, the intestinal wall, and the surrounding connective tissue in detail. In our study, severe, active colonic Crohn's disease and ulcerative colitis could be detected by diagnostic sonography of the colon with a sensitivity of 91% and 89%, respectively. Pathological changes were subsequently confirmed by colonoscopy. Differing echo patterns made differentiation of these two diseases possible. Our results thus show that colonic sonography is a diagnostic procedure that promises to greatly facilitate the evaluation and differentiation of inflammatory large bowel diseases.     

溃疡性结肠炎结肠粘膜HLA－DR抗原表达

本文检测了６０例溃疡性结肠炎和３０例正常结肠粘膜上皮细胞ＨＬＡ－ＤＲ抗原表达。结果显示，３０例正常结肠粘膜上皮及腺体不表达ＨＬＡ－ＤＲ抗原；而６０例ＵＣ中有３２例结肠粘膜上皮和腺体不同程度表达该抗原。其中，４２例活动性ＵＣ中２９例表达，１８例非活动性ＵＣ仅３例表达。同时发现ＵＣ结肠粘膜上皮表达ＨＬＡ－ＤＲ抗原还与粘膜炎症程度成正比。结果提示，细胞免疫机制在ＵＣ的发病机理中起重要作用。     

白细胞介素-8、白细胞介素-15、白细胞介素-18在溃疡性结肠炎患者结肠黏膜的表达及意义

目的研究溃疡性结肠炎(UC)患者结肠黏膜中白细胞介素-8(IL-8)、IL-15、IL-18的表达情况以及与UC疾病严重程度的关系。方法采用免疫组化法检测UC患者及正常对照组结肠黏膜中IL-8、IL-15、及IL-18的表达情况。结果中重度组IL-8阳性细胞数(216.8±24.9)多于轻度组(165.7±27.6)(P<0.01),轻度组多于缓解期组(88.5±21.6)(P<0.01),缓解期组又多于正常对照组(52.1±18.7)(P<0.05)。中重度组IL-15阳性细胞数(29.5±4.2)多于轻度组(18.7±2.9)(P<0.05),轻度组多于缓解期组(11.3±2.6)(P<0.05),缓解期组又多于正常对照组(3.9±1.1)(P<0.01)。中重度组IL-18阳性细胞数(115.7±24.6)多于轻度组(68.7±18.6)(P<0.01,轻度组多于缓解期组(47.3士12.6)(P<0.05),缓解期组又多于正常对照组(21.5±7.9)(P<0.05)。结论活动期UC患者结肠黏膜中IL-8、IL-15及IL-18的表达较缓解期、单纯结肠息肉者升高,且与病情严重程度有关。IL-8、IL-15及IL-18在UC的发病过程中起到了重要作用。     

Macrophage migration inhibitory factor in the sera and at the colonic mucosa in patients with ulcerative colitis: clinical implications and pathogenic significance

BACKGROUND: Inflammatory cytokines produced by activated macrophages are implicated in the pathogenesis of ulcerative colitis (UC). With the theory that macrophage migration inhibitory factor (MIF) may have a role in the accumulation of macrophages, we studied MIF in UC. MATERIALS AND METHODS: A total of 27 patients with UC, 14 patients with Crohn's diseases (CD), 11 patients with other forms of colitis and 26 normal controls were enrolled in the study. The levels of MIF in the sera and culture supernatant were measured by an enzyme-linked immunosorbent assay. MIF, macrophages and T cells were localized at the colonic mucosa by immunohistochemistry. RESULTS: The levels of MIF in the sera were significantly higher in UC than in normal controls (P < 0.05), in serum C-reactive protein (CRP) -positive cases with UC than in CRP-negative cases with UC (P < 0.05), and in patients with severe colitis with UC than in mild colitis with UC (P < 0.05). There was a positive relationship between serum MIF levels with the CRP levels and activities of colitis. However, the levels of MIF in patients with CD and other forms of colitis were not significantly different from their levels in normal controls and UC. Infiltrating cells at the colonic mucosa in UC and CD expressed MIF. CONCLUSIONS: These data suggest a role of MIF in the pathogenesis of UC. MIF may be used as a marker of disease activity in UC and control of MIF production may have therapeutic implications.     

Radiological detection of colonic dysplasia (precarcinoma) in chronic ulcerative colitis

Epithelial dysplasia occurring in long-standing ulcerative colitis is a precancerous lesion. Macroscopically it has a nodular or villous appearance or may be indistinguishable from the surrounding mucosa. An investigation into the radiological diagnosis of dysplasia, using in vivo and in vitro doublecontrast examinations with magnification radiographic studies, correlated with the histological analysis, has been made in four patients. Characteristic radiological abnormalities have been identified in the areas of the mucosa associated with histologically proven dysplasia. These appearances include nodularity and irregular areas with sharply angulated edges which may represent enlarged areae colonicae. The demonstration of these changes is an indication for endoscopie examination and biopsy of the suspicious area.     

Hyposplenism and T lymphocyte subpopulations in coeliac disease and after splenectomy

An imbalance in immunoregulatory cells might explain the increase in autoimmune phenomena associated with hyposplenism. Using the monoclonal antibodies OKT3, OKT4 and OKT8 to identify T-lymphocyte subsets, we have studied 19 patients with treated coeliac disease, 9 of whom had hyposplenism, 10 splenectomized subjects, 10 gastrointestinal control patients and 10 normal subjects. There was no significant difference in the proportion of lymphocytes, OKT3+, OKT4+ or OKT8+ cells, nor in the OKT4+/OKT8+ ratio, between any of the patient groups and normal or control subjects. Splenectomized subjects had higher total lymphocyte counts than normals (p less than 0.01). Hyposplenic coeliac patients had higher total lymphocyte counts than other coeliacs (p less than 0.05). Hyposplenic coeliac patients and splenectomized subjects tended to have an increase in the absolute number of OKT8+ cells, which in the latter group was statistically significant (p less than 0.05). We conclude that the increased frequency of autoimmune phenomena associated with hyposplenism due to coeliac disease or after splenectomy is not the result of an altered ratio of OKT4+/OKT8+ cells or a deficiency of OKT8+ cells.     

中性粒细胞/淋巴细胞比值、血小板/淋巴细胞比值对溃疡性结肠炎的诊断价值

目的探讨中性粒细胞/淋巴细胞比值(NLR)和血小板/淋巴细胞比值(PLR)在溃疡性结肠炎(UC)患者外周血中的水平,评估两者对UC的诊断效能。方法收集87例UC患者纳入UC组,收集65例肠易激综合征(IBS)患者纳入对照组。分析患者临床资料,比较两组NLR和PLR水平差异;采用Pearson相关分析NLR、PLR与临床常用指标白细胞计数(WBC)、血小板计数(PLT)、C反应蛋白(CRP)、红细胞沉降率(ESR)的相关性;采用受试者工作特征(ROC)曲线计算NLR和PLR最佳临界值及曲线下面积(AUC),并与常用炎性指标进行比较。结果UC组患者外周血NLR和PLR均明显高于对照组(P<0.05)。相关性分析发现,NLR、PLR均与WBC、CRP和ESR呈正相关(P<0.05)。NLR用于诊断UC的最佳临界值为2.64,灵敏度和特异度分别为81.9%和62.6%,AUC为0.758,PLR用于诊断UC的最佳临界值为163.40,灵敏度和特异度分别为75.0%和60.6%,AUC为0.759,两者均优于WBC(AUC:0.687)和PLT(AUC:0.745),稍逊于ESR(AUC:0.783)和CRP(AUC:0.830)。NLR联合CRP、PLR联合CRP对UC的诊断价值均优于CRP单独检测。结论NLR和PLR在UC患者外周血中的水平升高。NLR、PLR诊断UC的效能优于常用指标WBC和PLT,其与CRP联合应用可提高UC的诊断效能。     

Effect of smoking and transdermal nicotine on colonic nicotinic acetylcholine receptors in ulcerative colitis

BACKGROUND: Ulcerative colitis (UC) is a disease largely of non-smokers, in which nicotine is of therapeutic value. The mode of action is unknown, but may involve nicotinic acetylcholine receptors (nAChRs) in the bowel wall. AIM: To investigate the presence of nAChRs in rectal mucosa, and the effect of smoking and nicotine on their expression. DESIGN: Prospective case-control study. METHODS: In situ hybridization (ISH) and immunocytochemistry (ICC) were used to show alpha3 nAChRs in colonic mucosa. Rectal mucosa was examined from controls (n=55) and patients with inactive UC (n=62), both smokers and non-smokers, by ICC, using two antibodies to show the density and distribution of receptors in the mucosa. Non-smokers with UC (n=43) were given transdermal nicotine or placebo patches for 6 months, and rectal biopsies, taken before and after treatment, were examined by ICC to show nAChRs. RESULTS: In normal colon, ISH and ICC showed alpha3 subunit in a wide variety of cells, including mucosal epithelium. In rectal biopsies, neither smoking nor nicotine influenced the expression of alpha3 immunoreactivity in epithelium, either in controls or UC. However, controls had a significantly greater density of immunodetectable mucosal epithelium alpha3 subunit, compared with UC patients. DISCUSSION: The presence of nAChRs in colonic epithelium may be pertinent to the beneficial effect of nicotine in UC, but since neither smoking nor nicotine treatment is associated with any change in the expression of epithelial alpha3 nAChRs, the effect may be due to functional changes in the receptor. The decreased number of alpha3 nAChRs in UC compared with controls may be related to an increased cell turnover in UC.     

Cancer in chronic ulcerative colitis. Diagnostic role of segmental colonic lavage.

Saline colonic lavage in 74 patients with chronic ulcerative colitis was performed utilizing a commercially available dental irrigating unit through a polyethylene catheter in the biopsy channel of a colonoscope or through a sigmoidoscope via a lavage-aspirating double-lumen probe. Six patients were found with colonic carcinoma. Two diagnoses of malignancy were established by cytologic smears and cell block alone. Two patients had positive mucosal biopsies and cell block. One patient with a hepatic flexure carcinoma and a second patient with a malignancy proximal to the left colon stricture were missed by these techniques. Considering the established proclivity for carcinoma in these patients, it is felt that segmental lavage in areas of stricutre, grossly dostorted mucosa, or endoscopically inaccessible areas represents a valuable adjunct in the diagnosis of carcinoma in chronic ulcerative colitis.     

Monoclonal antibody-defined T lymphocyte subpopulations in monoclonal gammapathy of undetermined significance

Summary We used monoclonal antibodies of the OK series to study T lymphocyte subpopulations in 55 patients with monoclonal gammapathy of undetermined significance (MGUS) and in 40 healthy control subjects, with the aim to investigate if alterations in T lymphocyte subpopulations occur also in MGUS. Mean OKT3⁺ and OKT8⁺ cell counts were higher (p<0.01 and p<0.001, respectively) and the mean OKT4/OKT8 ratio was lower (p<0.02) in MGUS patients than in the control subjects. MGUS with the IgM-type monoclonal immunoglobulin (IgM-MGUS) showed the most evident derangement of T lymphocyte subpopulations, i.e., a significant increase of OKT8⁺ cells and a significant reduction of the OKT4/OKT8 ratio. OKT4⁺ and OKT8⁺ cells were significantly increased in patients with high paraprotein concentration (above 16 g/l). Our data suggest that alterations of T lymphocytes are present in MGUS, and that they are similar to those observed in malignant lymphoproliferative disorders. This work was supported by a grant fromRegione Lombardia, project n№ 341.