Human CD4+CD25+ Regulatory T Cells Suppress Anti-Porcine Xenogeneic Responses

Due to the shortage of human organs, xenotransplantation is being explored as an alternative to allotransplantation, but immune rejection remains a major hurdle to its implementation. We tested the ability of human CD4+CD25+ T cells (Treg cells) to suppress CD4+ T cell-mediated anti-porcine xenoresponses usingin vitroassays. Human Treg cells were hyporesponsive to porcine cell stimulation and suppressed the proliferative response of CD4+CD25- T cells in a dose-dependent manner, and comparison of the allo- and xenoresponses indicated that more Treg cells might be required to suppress the xenogeneic response than the allogeneic response. Stimulation of CD4+CD25- T cells with porcine cells resulted in secretion of IFN-gamma, TNF-alpha, IL-10, IL-6 and IL-2, and Treg cells suppressed the secretion of these cytokines, as well as the CD4+CD25- T-cell cytolytic response against porcine cells. These results suggest a potential role for Treg cells in promoting xenograft survival.     

Expression of the T-cell markers CD2 and CD28 in healthy and atopic children during the first 18 months of life

Atopy may be associated with a reduced T-cell function early in life, particularly regarding maturation of Th1 responses. The T-cell surface molecules CD2 and CD28 are involved in important T-cell activation pathways. Stimulation via the CD2 receptor increases the responsiveness to interleukin (IL)-12, which is a potent inducer of Th1 responses, whereas CD28 stimulation is critical for Th2 differentiation. Our aim was to prospectively study the expression of the cell-surface markers CD2 and CD28 on T-cells in relation to development of atopic disease. Children (n = 172) were followed from birth to 18 months and the cumulative history of atopic disease was recorded. Blood samples were obtained at birth and at 18 months, and in a subgroup of 78 infants also at 3, 6 and 12 months. Flow cytometry was used to analyze the T-cell markers CD2 and CD28, the latter also within the subsets of T-helper (CD4+) and T-cytotoxic (CD8+) cells. At 18 months, 31 children had and 118 did not have atopic symptoms. At this age, skin prick test (SPT) positive children with atopic symptoms with or without an atopic family history (AFH) showed a lower expression of CD2 mode fluorescence intensity (FI) as well as a lower proportion of CD2+ cells, as compared with non-sensitized children with neither atopic symptoms nor AFH. This was accompanied by a higher expression of CD28 FI on CD2+CD8+CD28+ cells. No significant differences were seen at time points before 18 months, although the proportion of CD2+ tended to be low also earlier in life. In conclusion, the observed reduced expression of CD2 in atopic infants may support previous findings that atopy is associated with a reduced CD2 function. The high CD28 FI in SPT positive children with atopic symptoms may possibly be a consequence of a TH2-skewed immune system.     

应用羟基荧光素二醋酸琥珀酰亚胺脂探讨新型免疫抑制剂J2作用机制

目的探讨基于CIM立体构型设计的新型免疫抑制剂（J2）在小鼠体内的免疫抑制作用以及其作用机制。方法将羧基荧光素二醋酸琥珀酰亚胺酯（CFSE）标记的C57BL／6（H-2b）小鼠的淋巴细胞4×10^6～6×10^6经尾静脉注入经印钴照射（900拉德）后的DBA／2（H-2d）小鼠体内构建模型。将DBA／2小鼠随机分为5组，每组5只，模型建立后即刻腹腔注射J2，每天1次。对照组注射生理盐水、CsA组注射环孢素A（10mg／kg体重）；实验组J2A组（1mg／kg体重）、J2B组（4mg／kg体重）、J2C组（8mg／kg体重）。给药3d后分组处死DBA／2小鼠取脾制备淋巴细胞，用CD3、CD4、CD8单抗分别标记受检T细胞，FACS流式细胞仪检测CD4^＋T细胞和CD8^＋T细胞在小鼠体内分裂增殖的情况，并检测CD4^＋T细胞的凋亡情况。结果CFSE标记C57BL/6小鼠的脾淋巴细胞着染率大于99％。对照组分裂的CD4^＋T细胞（75．34±1．58）％、分裂的CD8^＋T细胞（83．48±1．25）％明显多于CsA组和实验组（P〈0．01）。J2B组和J2C组分裂的细胞数分别与CsA组比较差异无统计学意义（P〉0．05），但J2A组多于CsA组（P〈0．05）。对照组CD4^＋T细胞早期凋亡的比例（41．1±3．4）％明显高于其他各组（P〈0．01）。J2A组（35．6±4．1）％、J2B组（24．0±3．7）％和J2C组（13．6±2．3）％CD4^＋T细胞早期凋亡的比例显著高于CsA组（7．4±1．9）％（P〈0．05）。结论J2可能抑制T细胞亚群CD4^＋T细胞的活化，从而进一步抑制CD8^＋T细胞的分裂增殖而具有免疫抑制作用。     

CD44v6、P27蛋白在乳腺癌组织中的表达及意义

目的 探讨乳腺癌中CD44v6、p27蛋白表达的意义。方法 应用免疫组化S-P法检测65例乳腺癌中CD44v6、p27蛋白表达。结果 65例乳腺癌中CD44v6、p27蛋白表达总阳性率分别为58.4％、38.4％。CD44v6蛋白阳性表达率与乳腺癌组织学类型、腋淋巴结转移呈正相关(P<0.05,P<0.01),与预后呈负相关(P<0.01),与组织学分级无明显相关性(P>0.05);p27蛋白阳性表达与乳腺癌组织学类型、组织学分级及腋淋巴结转移呈负相关(P<0.05),与预后呈正相关(P<0.05)。结论 CD44v6、p27蛋白表达与乳腺癌浸润、转移及预后显著相关,检测乳腺癌中CD44v6、p27蛋白表达水平,对判断乳腺癌恶性程度、复发转移潜能,评估患者预后及术后选择合理治疗方案有一定参考价值。     

The immunohistology of CD40 in human oral epithelium in health and disease

BACKGROUND: CD40 has a role in the regulation of immune responses, cell proliferation and migration, and apoptosis. Little is known of its distribution in oral mucosal pathology. METHODS: Oral keratinocyte lines were tested for CD40 protein by Western blotting. Immunohistochemistry was used to stain paraffin sections of oral mucosa in health and in inflammatory, reactive, dysplastic and malignant disease. RESULTS: Western blotting confirmed the presence of CD40 in oral keratinocytes. CD40 was generally expressed by keratinocytes in the basal layer, with variable parabasal expression. Langerhans cells also stained positively. Expression was lost in nine of 33 (27%) epithelial dysplasias, seven of which were severe. Eighty-one percent of well, 69% of moderately and 50% of poorly differentiated oral squamous cell carcinomas (OSCC) expressed CD40. Overall, 45 of 65 (69%) OSCC were positive. The pattern of expression was unrelated to tumour differentiation. CONCLUSION: CD40 expression by basal and parabasal oral keratinocytes is physiological. Expression is lost in approximately one-third of oral epithelial dysplasias and OSCC. The significance of such loss remains unknown, but may be related to immunological or other abnormalities of keratinocyte homeostasis.     

Effect of electro-acupuncture combined with early rehabilitation on motor function and expressions of CD11b/CD18 and tumor necrosis factor-α in patients with acute cerebral infarction

Objective To observe the effect of electro-acupuncture combined with early rehabilitation on the motor function and expressions of the adhesion molecules CD11b and CD18 in the polymorphonuclear leucocytes (PMN) and monocytes and serum tumor necrosis factor-α (TNF-α) levels in patients with acute cerebral infarction (ACI). Methods A total of 165 ACI patients were randomly divided into control group (group A, n=50), conventional rehabilitation group (group B, n=50) and comprehensive rehabilitation group (group C, n=65). The expressions of CD11b and CD18 in the PMN and monocytes and serum TNF-α levels were determined before and at 1, 2, and 4 weeks after the treatment. Thirty-two healthy subjects were also recruited as the normal control group (group N). The neurological function of the subjects was evaluated by modified Edinburgh-Scandinavia stroke scale (MESSS) and Fugi-Meyer Assessment (FMA), and their activity of daily living (ADL) was assessed using Barthel index (BI). Results The CD11b/CD18 expression in the PMN and MN and serum TNF-α level in groups A, B and C were significantly higher than those in group N before and 1 week after the treatment (P<0.05). CD11b/CD18 expression and serum TNF-α level were significantly lower in groups B and C than in the group A at 1 week after the treatment, and significantly lower in group C than in group B (P<0.05). At 2 weeks of treatment, CD11b/CD18 and TNF-α were significantly lower in groups B and C than in the group A, being the lowest in group C (P<0.05). The scores of mESSS in both groups B and C were lower than that in group A, and the scores were lower in group C than in group B. Group C showed higher FMA scores than group B, both having higher scores than group A. At 4 weeks of treatment, the mESSS scores were significantly lower, hut the FMA and ADL score significantly higher in groups B and C than in the control group (P<0.05), and the differences were more obvious in group C. Groups B and C had greater effective rate than group A (P<0.05), and the rate was the highest in group C (P<0.05). Conclusion Electro-acupuncture combined with early rehabilitation promotes the recovery of motor function in ACI patients probably by regulating the expressions of the adhesion molecules CD11b and CD18 on the PMN and monocytes and the serum levels of TNF-α.     

RNA干扰阻断BT／CD28共刺激通路在小鼠心脏移植中的抗排斥作用

目的探讨RNA干扰（RNAi）技术阻断B7／CD28共刺激通路对小鼠异体心脏移植排斥反应的影响及其机制。方法经体外转录合成针对CD80 mRNA和CD86 mRNA序列特异性小片段干扰RNA（siRNA），转染供者骨髓来源的树突状细胞（DC），半定量逆转录聚合酶链反应、流式细胞仪检测DC转染CD80siRNA、CD86siRNA前后CD80 mRNA和CD86 mRNA的表达水平以及细胞表面CD80及CD86的表达情况。在小鼠异位心脏移植前7d，经静脉给受者输注经siRNA干扰后的DC（干扰DC组），同时设立同种异体对照组、环孢素A（CsA）治疗组（术后皮下注射CsA 5mg／d）、同系移植对照组和未干扰DC组（移植前输注未转染DC），观察各组移植心脏的存活时间，对移植物的排斥反应进行病理分级，并测定移植物组织中白细胞介素2（IL-2）、γ干扰素（IFN-γ）及IL-10的mRNA表达水平。结果siRNA转染DC后，其CD80 mRNA及CD86 mRNA的表达受到明显抑制，CD80、CD86的阳性率分别由84％和67％下降至35％和30％。与同种异体对照组和未干扰DC组比较，干扰DC组移植心脏存活时间明显延长（P〈0．01），组织排斥反应病理分级显著降低（P〈0．01），移植心脏组织中IL-2 mRNA和IFN-γ mRNA的表达水平明显降低（P〈0．01），而IL-10 mRNA的表达水平明显升高（P〈0．01）。结论利用RNAi敲减供者骨髓来源的DC表面B7分子的表达，以阻断BT／CD28共刺激通路，具有抑制小鼠心脏移植排斥反应的作用，其机理可能是通过诱导T淋巴细胞无能并使T辅助细胞分化向Tn2型方向偏移。     

E-钙黏附蛋白、CD44v6在卵巢上皮性肿瘤及卵巢癌转移灶中的表达

目的：在基因表达水平上研究黏附分子(E-cadherin，CD44v6)与卵巢癌转移的关系。方法：用免疫组化S-P法，选取1997～2002年间石蜡包埋的卵巢上皮性肿瘤组织块107块，其中良性上皮性肿瘤30例，交界瘤13例，恶性34例(Ⅰ、Ⅱ期4例，Ⅲ、Ⅳ期30例)，对30例恶性晚期(Ⅲ、Ⅳ期)病例选取对应的大网膜转移组织。结果：30例良性上皮性肿瘤组织中，E-钙黏附蛋白100％强阳性表达，CD44v6100％阴性。13例交界瘤中，E-钙黏附蛋白有11例(84．6％)强阳性，CD44v6有1例弱阳性表达。34例恶性上皮性肿瘤中，E-钙黏附素有24例(70．6％)阳性表达，CD44v6有11例(32．4％)阳性表达。二指标在良性上皮性肿瘤与卵巢癌之间的阳性表达有显著性差异(P=0．001，P=0．000)。二指标在原发灶和大网膜转移灶之间表达无显著差别(P=1．000，P=1．000)。结论：E-cadherin及CD44v6在上皮性卵巢肿瘤中的表达呈相反趋势，E-cadherin及CD44v6与卵巢癌组织学分级及预后无明显相关性。     

Reduced expression of CD69 and adhesion molecules of T lymphocytes in asthmatic children receiving immunotherapy

T lymphocytes play a fundamental role in the initiation and regulation of chronic inflammatory responses in patients with asthma. CD69 is an early marker of T‐cell activation. The levels of intercellular adhesion molecule‐1 (ICAM‐1, CD54) and L ‐selectin have been reported to increase in patients with allergic diseases and asthma. The present study was therefore undertaken to investigate the expression of CD69, CD54, and L ‐selectin by T lymphocytes of children with asthma, before and after immunotherapy. Eighteen children newly diagnosed with asthma, 11 good and nine poor responders to immunotherapy, and 16 normal subjects, were enrolled in this study. The percentages of CD69⁺, CD54⁺, and CD62L⁺ cells in T lymphocytes were measured by using flow cytometry. The levels of CD69, CD54, and CD62L in serum and culture supernatants were determined by using enzyme‐linked immunosorbent assay (ELISA). The expression of CD69 and CD54 on CD3⁺ T lymphocytes was significantly higher in children with asthma than in control patients. All the patient groups expressed (spontaneously and following stimulation with phorbol myristate acetate and ionomycin together with mite‐extract proteins) greater amounts of CD69 and CD54 than did control subjects. With long‐term immunotherapy, the percentages of CD69⁺ and CD54⁺ T lymphocytes were significantly lower in patients with a good response to immunotherapy. Our results also showed significantly lower serum L ‐selectin levels following immunotherapy. In conclusion, successful immunotherapy resulted in decreased expression and production of CD69 and CD54. These results may explain, in part, the clinical efficacy of immunotherapy.     

Monocyte adhesion molecule expression in interstitial inflammation in patients with renal failure.

BACKGROUND: Patients with renal failure have an increased susceptibility to infections. We therefore studied the recruitment of monocytes and their expression of adhesion molecules CD11b and CD62L at the site of interstitial inflammation in patients with renal failure. Furthermore, we studied if the capacity of monocytes to up-regulate CD11b in interstitial inflammation was determined by the interstitial concentration of chemotactic factors. METHODS: Three intensities of interstitial inflammation (0, intermediate and intense) were established in skin blister chambers. Leukocyte count, CD11b/CD62L expression, monocyte chemotactic protein-1 (MCP-1) and blister activity in terms of CD11b mobilization were determined. RESULTS: The CD62L expression on monocytes was lower in the peripheral circulation in patients with renal failure compared with healthy subjects (P<0.005 and P<0.001). At the site of interstitial inflammation patients had a higher expression of CD62L (intermediate, P<0.05; intense, P<0.005). Furthermore, monocytes from patients had an impaired capacity to mobilize CD11b both in the peripheral circulation (P<0.005) and at the intermediate and intense sites of interstitial inflammation (P<0.005 and P<0.001, respectively) compared with cells collected from healthy subjects. We incubated monocytes in blister exudates, in order to explore whether this phenomenon is caused by cellular factors and/or to the interstitial concentration of chemotactic mediators. The expression of CD11b on monocytes from healthy blood donors incubated in blister exudates from either patients or healthy subjects in vitro was similar. The interstitial concentration of MCP-1 at the site of intermediate inflammation was significantly lower in patients with renal failure compared with the corresponding blister exudate collected from healthy subjects (P<0.05), but no differences were observed at the site of intense inflammation. Furthermore, neutralizing the action of MCP-1 in blister exudates with monoclonal antibodies did not have any impact on monocyte CD11b expression following incubation in blister exudates. CONCLUSION: These studies indicate that the impaired capacity of monocytes to mobilize CD11b at the site of inflammation in patients with renal failure is more dependent on constitutive cellular factors than the concentration of CD11b mobilizing factors in the interstitium.