模拟日光照射后皮肤CD1a、CD68阳性细胞变化的研究

目的 观察正常人皮肤经日光模拟器照射（solar-simulated ultraviolet radiation,ssUVR)后,朗格汉斯细胞（Langerhans cells, LC）和CD68阳性的巨噬细胞的变化。方法14名健康汉族女性志愿者于背部非曝光部位接受ssUVR照射。选择2个正方形部位,一处为正常对照,另一处为每日一次ssUVR照射。第4天照射后的72小时,进行活检取材。对所有标本进行CD1a和CD68免疫组化染色。结果 未照射部位正常表皮内的LC密度为258±61个/mm2,ssUVR照射部位的LC密度明显降低为96±53个/mm2,LC的形态不完整,树突变短而不明显。真皮浅层CD68阳性的巨噬细胞,未照射部位密度为290±22个/mm2,ssUVR照射部位的密度升高为399±65个/mm2。经过照射后真皮这些巨噬细胞数目明显增多位置上移,形态上树突变长并且大多数互相连接紧密。结论ssUVR照射可使LC数目减少,形态破损。真皮内的巨噬细胞则增高,这似有助于弥补紫外线照射对局部免疫的抑制作用。     

热处理、UVB辐射及两者联合作用对人表皮黑素细胞合成热休克蛋白72的影响

目的 探讨热处理、中波紫外线（UVB）辐射及两者联合作用下,体外培养的人表皮黑素细胞热休克蛋白72（heat shock protein 72,HSP72）的变化。 方法 热处理（42 ℃,1 h/d,连续3 d）、UVB辐射（50 mJ/cm2,连续3 d）及两者联合（首先42 ℃,加热1 h,然后 50 mJ/cm2 UVB照射,连续3 d）处理正常人表皮（包皮）黑素细胞2 h、6 h后收集细胞,用实时荧光定量PCR技术和 Western 印迹技术分别检测不同组别间HSP72 mRNA及蛋白表达水平的差异。结果 热处理组（6.584 ± 0.871）及联合作用组（7.269 ± 0.454）与对照组（0.975 ± 0.089）相比,mRNA 表达水平明显增加（P < 0.001）;UVB辐射组（0.832 ± 0.084）与对照组相比,mRNA表达水平差异无统计学意义（P > 0.05）。Western 印迹结果显示,热处理组（2.022 ± 0.058）及联合作用组（2.080 ± 0.045）HSP72蛋白表达水平明显高于对照组（0.532 ± 0.033,P < 0.001）;UVB组（0.546 ± 0.021）与对照组相比,表达水平差异无统计学意义（P > 0.05）。析因设计方差分析结果显示,热处理与UVB辐射对HSP72 mRNA和蛋白表达均无交互作用（F = 2.106,1.399,P < 0.05）。结论 热处理引起HSP72表达显著增多,可能与热处理引起黑素细胞功能增强及UVB辐射后的损伤保护作用有关。     

热处理对UVB辐射后体外培养人黑素细胞活性的影响

目的 探讨热处理与窄谱中波紫外线（NB-UVB）联合作用对正常人黑素细胞活性的影响。 方法 分别以20、30、50、70、90、120、180 mJ/cm2 NB-UVB辐射体外培养的正常人黑素细胞,采用四甲基偶氮唑蓝比色法（MTT）检测细胞增殖活性并选择最佳照射剂量;以42 ℃,1 h为热处理干预剂量,将黑素细胞分为4组：正常对照组、UVB组、加热组、UVB + 加热组,连续干预3 d,以左旋多巴为底物测定酪氨酸酶活性,NaOH法测定黑素含量,流式细胞仪检测细胞周期变化。结果 NB-UVB照射呈剂量依赖性减少黑素细胞存活率,选择50 mJ/cm2作为最佳照射剂量;黑素细胞经不同干预后,UVB组、UVB+加热组的酪氨酸酶活性分别为0.244 ± 0.018、0.310 ± 0.015,较对照组（0.235 ± 0.018）分别增长3.8%和31.9%（P < 0.05）,两组黑素含量分别为0.201 ± 0.016、0.286 ± 0.019,较对照组（0.171 ± 0.016）分别增长17.5%和67.3%（P < 0.05）;UVB组和UVB+加热组处于G1期的黑素细胞较对照组分别减少23.94%和33.51%（P < 0.05）,S期细胞分别增加15.35%（P < 0.05）和17.76%（P > 0.05）,G2期分别增加11.93%（P < 0.05）和16.08%（P > 0.05）。 结论 热处理与NB-UVB可以协同增加黑素细胞的酪氨酸酶活性,促进黑素合成及细胞的增殖分化。     

强功率UVA1对兔耳增生性瘢痕模型影响的机制研究

目的 探讨不同剂量UVA1对全层皮肤缺损诱导的兔耳增生性瘢痕模型的影响的可能机制。方法 24只新西兰白兔双耳腹面手术切除2 cm × 5 cm全层皮肤至筋膜建立兔耳增生性瘢痕模型后,随机分成4组,每组6只,将每只兔左耳分别于伤后即刻（U0）、1个月（U1）、2个月（U2）、3个月（U3）开始强功率UVA1照射,右耳为非照射组;各照射组分为两个剂量组［60 J/cm2（M）,110 J/cm2（H）］,分别在照射前后进行MMP-1、TIMP-1、TGF-β1、PCNA和α-SMA的免疫组织化学染色和透射电镜检查。结果 与照射前比较,中高剂量组照射后瘢痕处MMP-1表达：U1组分别为10.43 ± 1.61、11.16 ± 1.57;U2组分别为8.63 ± 2.61、7.33 ± 1.58;U3组分别为5.74 ± 1.43、3.11 ± 0.27;均显著增加（P < 0.05）。TGF-β1表达：U1组分别为12.51 ± 4.13、12.02 ± 5.02;U2组分别为18.74 ± 6.42、19.69 ± 4.52;U3组分别为20.51 ± 1.78、29.45 ± 6.55。PCNA表达：U1组分别为2.67 ± 0.44、2.04 ± 0.65;U2组分别为4.50 ± 0.97、5.82 ± 0.68;U3组分别为7.45 ± 1.47、8.16 ± 1.07;均显著降低（P < 0.05）。只有高剂量组显著降低TIMP-1表达,U1组为12.74 ± 4.58,U2组为15.17 ± 3.26,U3组为20.72 ± 3.31（P < 0.05）。只有U1H、U1M、U2H组α-SMA表达（1.33 ± 0.34、2.04 ± 0.20、3.60 ± 1.75）显著降低（P < 0.05）。U0组：与对照组比较,高剂量组MMP-1表达（2.25 ± 0.38）显著降低（P < 0.05）,而TGF-β1表达（23.90 ± 2.92）显著增加（P < 0.05）。中、高剂量组PCNA（7.42 ± 0.65、7.59 ± 0.31）、TIMP-1（29.82 ± 1.94、33.51 ± 1.19）及α-SMA表达（6.31 ± 0.61、2.97 ± 0.56）均显著增加（P < 0.05）。透射电镜结果显示：强功率UVA1照射后胶原纤维直径变细;成纤维细胞胞质变少,细胞器欠发达,多数为静止的纤维细胞。结论 UVA1对瘢痕的作用可能与其抑制TGF-β1、TIMP-1、PCNA和α-SMA的表达,同时促进MMP-1的表达,从而促进基质蛋白降解及抑制成纤维细胞和肌成纤维细胞的增值活性有关。如果UVA1过早干预,则会出现相反的结果。     

CD1a、CD68在继发性瘢痕疙瘩中的表达及意义

目的 探讨继发性瘢痕疙瘩皮损中表皮朗格汉斯细胞（LC）和真皮CD68阳性组织细胞的分布和密度。方法 取30例继发性瘢痕疙瘩患者的皮损、14例正常人皮肤组织切片进行CD1a和CD68免疫组化染色。以测微尺标定目镜方格计数方格内阳性细胞数,计算出单位面积内细胞的密度。组间比较采用SPSS软件进行 Student t检验。结果 在继发性瘢痕疙瘩表皮内CD1a阳性LC密度为（61 ± 49）个/mm2,正常表皮为（258 ± 61）个/mm2,两组比较,t = 9.88,P ＜ 0.01;继发性瘢痕疙瘩真皮CD1a阳性细胞密度为（40 ± 65）个/mm2。继发性瘢痕疙瘩表皮中无CD68阳性细胞,真皮内CD68阳性组织细胞密度为（287 ± 73）/mm2,正常皮肤为（290 ± 22）个/mm2,两组比较,t = 0.02,P ＞ 0.05。继发性瘢痕疙瘩真皮浅层CD68阳性组织细胞占真皮中所有细胞的62% ± 12%,而正常皮肤为70% ± 14%,两组比较,t = 2.66,P ＜ 0.05。 结论 继发性瘢痕疙瘩表皮中LC减少,无CD68阳性的细胞。真皮中LC增多;真皮浅层CD68阳性组织细胞占真皮中所有细胞的比例下降。     

小鼠免疫性接触性荨麻疹动物模型的构建

目的 建立小鼠免疫性接触性荨麻疹动物模型。 方法 将60只BALB/c小鼠随机分5组：抗二硝基苯酚IgE单克隆抗体（抗DNP IgE） + 二硝基氟苯（DNFB）组和抗DNP IgE + 偏苯三酸酐（TMA）组小鼠尾静脉注射抗DNP IgE,24 h后分别在小鼠耳部涂DNFB和TMA;DNFB组、TMA组、生理氯化钠液（NS）组小鼠尾静脉注射NS,24 h后分别在小鼠双耳涂DNFB、TMA、NS,观察小鼠在14 d内的风团、瘙痒情况、肥大细胞脱颗粒数、脾脏指数和皮肤病理。 结果 抗DNP IgE + DNFB组全部（12/12）和抗DNP IgE + TMA组部分小鼠（8/12）产生风团;抗DNP IgE + DNFB组及抗DNP IgE + TMA组小鼠的搔抓次数［30 min时分别为（31.58 ± 3.58）次/h,（22.17 ± 3.81）次/h］、肥大细胞脱颗粒率［（70.21 ± 26.01）%,（54.25 ± 39.57）%］及脾脏指数（7.54 ± 1.56,7.87 ± 1.18）与NS组［（2.00 ± 0.85）次/h,（14.45 ± 6.79）%,5.37 ± 1.16］相比均显著增加（F值分别为437.86、14.41、4.29,均P < 0.01）;DNFB组、TMA组及NS组均无风团产生。 结论 通过抗DNP IgE诱导及DNFB激发后可快速、较稳定地建立小鼠免疫性接触性荨麻疹动物模型。     

温热对HPV感染皮肤中朗格汉斯细胞的影响

目的 探讨不同温热条件处理后的正常皮肤/尖锐湿疣皮损组织中朗格汉斯细胞（LC）的形态、数量及迁移。方法 利用37 ℃、42 ℃、45 ℃不同的温热条件处理置于培养液中的离体尖锐湿疣皮损（16例）和正常皮肤组织（15例），采用免疫组化和流式细胞分析方法测定LC形态、数量和迁移的变化。结果 表皮中LC数量随着温度的增高而减少，正常皮肤组织表皮内LC数量分别为：782.40 ± 114.88（37 ℃），649.44 ± 119.40（42 ℃），510.88 ± 118.64（45 ℃）；疣体组织表皮内LC数量分别为：646.04 ± 135.67 （37 ℃）、489.38 ± 118.19 （42 ℃）、387.93 ± 110.15 （45 ℃ ）；LC树突变短、减少或消失。温热促进LC迁移至培养液中。不同温度下，游走的LC表达CD1a的数量不同，正常皮肤组织游走至培养液中LC表达CD1a的比率分别是0.19% ± 0.18%（37 ℃），0.89% ± 0.19%（42 ℃），1.59% ± 0.28%（45 ℃）；尖锐湿疣组织分别为0.62% ± 0.31%（37 ℃），2.31% ± 0.54%（42 ℃），6.33% ± 0.98%（45 ℃）。结论 温热能够促进LC的迁移，籍此有可能增强LC在免疫应答中的抗原提呈作用。     

高脂饮食对小鼠皮肤创面修复影响的实验研究

目的 观察长期高脂饮食对小鼠皮肤创面愈合的影响并初步探讨其相关机制。 方法 10周龄C57BL/6J野生型小鼠16只,随机分为2组,每组8只,分别采用高脂及普通膳食喂养8周后,在小鼠背部建立全层皮肤缺损模型。每日对创面愈合状况进行观察,记录创面愈合率及上皮化水平。于术后第14天处死所有小鼠并切取创面组织进行组织学检测,比较两组创面新生表皮厚度、创面床内胶原沉积率、新生血管数目、细胞增殖及炎症细胞浸润水平。喂养前、喂养8周后及手术后第14天检测体重。术后第14天禁食12 h后检测外周血总胆固醇（TC）、三酰甘油（TG）水平。采用t检验进行统计学分析。 结果 高脂膳食组在高脂喂养8周后及手术后第14天平均体重［分别为（27.3 ± 0.7） g和（28.8 ± 0.7） g］显著高于普通膳食组［分别为（21.2 ± 0.6） g和（23.1 ± 1.1） g］,两组比较,t值分别为21.98和25.22,均P < 0.001。术后第14天,高脂膳食组TG及TC水平［分别为（1.35 ± 0.32） mmol/L和（4.21 ± 0.41） mmol/L］远高于普通膳食组［分别为（0.99 ± 0.28） mmol/L和（2.71 ± 0.31） mmol/L, 两组比较,t值分别为2.24和6.49,均P < 0.05］;高脂膳食组创面愈合时间明显慢于普通膳食组［分别为（13.5 ± 0.5） d和（12.6 ± 1.1） d,t = 1.99,P < 0.05］,创面新生表皮薄于普通膳食组［分别为（47.8 ± 13.8） μm 和（95.7 ± 13.7） μm,t = 5.68,P < 0.001］,CD31阳性血管数低于普通膳食组［分别为（8 ± 1）个和（13 ± 3）个,t = 4.1,P < 0.001］,ki-67阳性细胞数目低于普通膳食组［分别为（21 ± 4）个和（49 ± 10）个,t = 3.33,P < 0.001］,但巨噬细胞及肥大细胞浸润水平明显高于普通膳食组（均P < 0.05）,各组创面床内胶原沉积率差异无统计学意义（P > 0.05）。 结论 长期高脂饮食会影响小鼠创面的愈合,延缓皮肤创面修复。     

氨基酮戊酸光动力疗法对鲜红斑痣动物模型鸡冠的影响

目的 探讨外用氨基酮戊酸光动力疗法（ALA-PDT）对鲜红斑痣（PWS）动物模型鸡冠的作用和治疗鲜红斑痣的可行性。 方法 80只莱亨鸡随机分为10组：空白对照组、单纯ALA组、单纯激光75 J/cm2组、100 J/cm2组、150 J/cm2、200 J/cm2组,ALA-PDT 75 J/cm2组、100 J/cm2组、150 J/cm2组、200 J/cm2组。单纯激光组仅给予630 nm红光照射,ALA-PDT组外用ALA后给予红光照射。干预14 d、28 d分别取材,观察鸡冠形态学、组织学变化,计算毛细血管减少率和血管内皮细胞凋亡情况。 结果 外用ALA-PDT 75 J/cm2、100 J/cm2、150 J/cm2和200 J/cm2组鸡冠实验区域颜色变淡,光镜下真皮毛细血管数目减少、管径变小,部分血管内皮细胞凋亡;毛细血管减少率分别为33.53% ± 4.89%、52.02% ± 2.77%、67.48% ± 5.58%、88.96% ± 2.47%;凋亡指数分别为：63.44 ± 1.09、88.50 ± 6.11、94.32 ± 3.67、113.76 ± 10.57,与其余各组分别比较,均P < 0.01;血管内皮细胞凋亡深度分别为：201.19 μm ± 0.33 μm、266.15 μm± 1.02 μm、546.09 μm ± 2.45 μm、766.37 μm ± 1.08 μm,各组间两两比较,均P < 0.01。 结论 外用ALA-PDT对鸡冠毛细血管有明显损伤,损伤程度和深度与红光能量密度相关;诱导血管内皮细胞凋亡可能是其治疗机制之一。     

梅毒螺旋体不同接种途径对家兔致病性研究初探

目的 探讨不同接种途径建立的梅毒家兔模型的发病及免疫应答情况。 方法 20只家兔随机分为睾丸接种组及背部皮内接种组,每组10只。睾丸接种组家兔睾丸注射2 × 107梅毒螺旋体Nichols株。背部皮内接种组家兔背部剃毛后选取6个点进行皮内注射,6个点共注射2 × 107梅毒螺旋体Nichols株。观察两组家兔的临床症状及血清学反应情况。 结果 睾丸接种组接种后第8天,睾丸开始肿大变硬,13 ~ 16 d肿大最为明显,之后逐渐缩小,至接种后28 d所有家兔睾丸回缩,触诊未见肿大变硬;发病2个月内未发现其他部位出现皮损。背部皮内接种组接种后7 d,接种部位皮肤出现红肿,15 ~ 45 d皮损红肿最为明显,且出现溃疡,有3只家兔共12个接种点的皮肤发生溃疡,暗视野检查阳性,触诊呈软骨样硬度。睾丸接种组接种后（14.50 ± 1.08） d时临床症状最为明显,皮内接种组（29.00 ± 10.30） d时临床症状最为明显,两组比较,t = 5.02,P ＜ 0.05。两组家兔快速血浆反应素环状卡片试验（RPR）最高滴度分布经秩和检验,差异有统计学意义（P ＜ 0.05）,睾丸接种组滴度高于背部皮内接种组。 结论 梅毒螺旋体接种家兔不同部位,引起家兔不同的临床症状及免疫反应。睾丸接种组RPR阳性时间早,且滴度高。背部皮内接种组的临床症状类似于人类一期梅毒硬下疳表现。     

Suppression of contact sensitivity by local hyperthermia treatment due to reduced Langerhans cell population in mice

The effects of local hyperthermia treatment on contact sensitivity (CS) and on the number of Langerhans cells (LCs) were studied in mice. CS was significantly suppressed when mice were sensitized in the hyperthermia treated skin I, 2 or 4 days after treatment (43 degrees C for 45 min). This suppressive effect was not observed 7 or 14 days after the treatment. CS was also suppressed when mice were sensitized in non-treated skin I day after the treatment. The density of LCs detected as ATPase-positive cells also decreased significantly 1, 2, 4 and 7 days after the treatment. There appeared to be a positive correlation between the number of LCs and the extent of CS when mice were sensitized at hyperthermia treated skin. It was observed that this suppressive effect on CS was dose- and temperature-dependent. It could be transferred by spleen cells from the hyperthermia treated and DNFB-sensitized donors, and was antigen specific when spleen cells were transferred before sensitization of the recipient mice. This indicated it was, in part, associated with the induction of suppressor cells. These findings suggest that local hyperthermia treatment reduces the number of LCs with subsequent suppression of the induction phase of delayed-type hypersensitivity by the generation of antigen-specific suppressor cells.     

温热对HPV感染皮肤朗格汉斯细胞磷酸肌醇3-激酶活性的影响

目的 探讨不同局部温热条件对HPV感染皮损游走朗格汉斯细胞(LC)磷酸肌醇3激酶(P13-K)活性的影响.方法 利用37℃,42℃,45℃的温热处理培养液中的离体正常皮肤组织和尖锐湿疣皮损,采用荧光实时定量PCR法检测游走至培养液巾纯化的CD1a~+ LC P13-K p85α、p110α mRNA的表达水平.结果 正常人皮肤组织和尖锐湿疣组织游走纯化的CD1a~+ LC中P13-K p85a、p110α mRNA的表达量均随局部温度升高而逐渐减少.正常皮肤组织游走纯化的CD1a~+LC 中P13-K p85a mRNA的相对表达量37℃为1.00±0.00,42℃为0.78±0.13,45℃为0.54±0.17;P13-K p110α mRNA在不同温度的相对表达量分别为1.00±0.00,0.80±0.11,0.61±0.12;不同温度间比较,差异均有统计学意义(P＜0.01).而尖锐湿疣组织游走纯化的CD1a~+ LC中P13-K p85a mRNA的相对表达量37℃为1.00±0.00,42℃为0.20±0.11,45℃为0.1±0.08;P13-K p110α mRNA的相对表达量分别为1.00±0.00,0.49±0.21,0.09±0.03;不同温度间比较,差异均有统计学意义(P＜0.01).温热对尖锐湿疣组织游走LC中P13-K活性影响显著高于正常皮肤组织.结论 局部温热可能通过抑制LC P13-K的活性,提高机体免疫应答反应能力,消除HPV感染.     

热处理对体外培养正常人黑素细胞活性的影响

目的 探讨热处理对体外培养的正常人黑素细胞的增殖活性、黑素合成及其酪氨酸酶活性的影响。方法 无菌操作获取正常人包皮环切术后的包皮,按照皮肤表皮细胞培养法获取黑素细胞,利用碱性成纤维细胞生长因子（bFGF）为主要有丝分裂原建立正常人表皮黑素细胞培养系,采用Masson-Fontana染色法对培养细胞进行鉴定,确定细胞种系,噻唑蓝（MTT）比色法测定不同温度条件（39、41、42、43、45 ℃）处理对黑素细胞活力的影响并选择最佳的温度条件,以左旋多巴为底物测定酪氨酸酶活性,比色法测定黑素含量,并将实验组与对照组结果进行比较。结果 体外培养的正常人黑素细胞在5个不同的温度条件作用后,以42 ℃作用后的细胞增殖活性最佳。酪氨酸酶活性和黑素含量测定结果显示,42 ℃每天1 h连续干预3 d后,体外培养的正常人黑素细胞的酪氨酸酶活性增长率为36.4%,黑素合成增长率为78%,均明显高于对照组（P值均 < 0.01）。结论 热处理可以增强体外培养的人黑素细胞的增殖活性,促进其黑素合成,并且可提高其酪氨酸酶的活性。     

温热对HPV-6/11型早期基因E6和E7表达的影响

目的 探讨不同温热条件对HPV感染皮损中E6/E7基因表达的影响.方法 采用PCR法确定6例尖锐湿疣标本HPV型别.利用37℃,42℃,45℃不同的温热条件处理置于培养液中的离体尖锐湿疣皮损,采用实时定量PCR法检测HPV-6/11型早期基因E6/E7 mRNA的表达水平.结果 6例标本中.各有1例分别感染HPV-6、HPV-11,4例同时感染HPV-6和HPV-11.E6、E7 mRNA的表达水平均随温度升高而逐渐减少.HPV-6 E6 mRNA的相对表达量分别为37℃(1.00±0.00),42℃(0.61±0.17).45℃(0.27±0.15);HPV-6 E7 mRNA的相对表达量分别为37℃(1.00±0.00),42℃(0.56±0.21),45℃(0.16±0.11);HPV-11 E6 mRNA的相对表达量分别为37℃(1.00±0.00),42℃(0.60±0.22).45℃(0.16±0.08);HPV-11 E7 mRNA相对表达量分别为37℃(1.00±0.00),42℃(0.55±0.15).45℃(0.24±0.06).组间比较差异有统计学意义(P<0.01.结论 温热对HPV-6/11型E6和E7基因表达的抑制作用可能是感染消退的机制之一.     

Induction of cutaneous delayed hypersensitivity reactions in mice sensitized with intragastrically administered hapten: activation of Langerhans cells in the sensitization and elicitation phases

BACKGROUND: As seen in atopic dermatitis, allergic diseases often produce lesions both in the gastrointestinal tract and the skin, suggesting the involvement of an immunological relationship between the two organs in the pathogenesis. OBJECTIVES: To study the role of gastric and epidermal Langerhans cells (LCs) in the sensitization and elicitation phases, respectively, of cutaneous delayed-type hypersensitivity (DTH) reactions to intragastrically administered hapten. METHODS: BALB/c mice, which were subjected to intragastric administration of trinitrochlorobenzene 5 days previously, received an elicitative challenge of the same hapten to the ear skin. Sections of the ear were immunostained for CD4 and CD8. Epidermal sheets of the ear and epithelial sheets of the forestomach were immunostained for I-A and observed under a confocal laser scanning microscope. RESULTS: Cutaneous DTH reactions were induced in mice, as demonstrated by an increase in ear thickness and a prominent infiltration of CD4+ and CD8+ lymphocytes at 24-36 h after the elicitative challenge. In the elicitation phase, epidermal LCs showed a significant increase in size, indicating in vivo activation, at 24 h. In the sensitization phase, gastric LCs increased in size at 2 h, became round at 6 h, and decreased in number at 24 h, possibly representing the sequential events of LC activation and migration from the epithelium. CONCLUSIONS: The present study demonstrated that gastric LCs and epidermal LCs were activated in vivo in the sensitization and elicitation phases, respectively, of cutaneous DTH reactions in orally sensitized mice.     

光动力疗法对尖锐湿疣皮损朗格汉斯细胞的影响

目的观察光动力疗法(PDT)对尖锐湿疣(CA)患者皮损中朗格汉斯细胞(LC)的影响,进一步探寻PDT治疗CA的免疫机制。方法应用免疫组化标记方法,对15例PDT治疗前后的CA皮损及5例正常包皮组织对照进行染色,在高倍光镜下观察LC的形态、分布及数量等的变化。结果 15例CA皮损较正常对照组织[(18.8±4.0)个/高倍视野]LC数量明显降低(P0.01),表皮内LC分布不规则,大部分LC结构不完整,细胞突减少、缩短甚至消失,缺少LC的典型特征,点状或条索状。个别皮损表皮内LC缺失,而其真皮内见到不典型的LC。PDT治疗后即刻组LC数量[(9.2±2.0)个/高倍视野]较治疗前组[(6.2±1.8)个/高倍视野]明显升高(P0.05),治疗后7 d组LC数量[(6.4±1.1)个/高倍视野]回落接近治疗前水平(P0.05),差异无统计学意义。结论 CA患者皮损局部细胞免疫功能低下,作为抗原提呈细胞的LC,其数量和形态发生改变。PDT能诱导CA皮损局部LC数量及形态发生明显改变,具有调节局部免疫的作用。     

Hyperthermic effects on peripheral lymphocytes isolated from a chronic lymphocytic lymphoma/leukemia patient in vitro

Effects of hyperthermia in vitro on the atypical lymphocytes isolated from peripheral blood of a chronic lymphocytic lymphoma/leukemia patient were investigated. Incubation of the isolated lymphocytes at 43^°C for 5 hr killed 15% of the cells, while more than 95% of the lymphocytes isolated from normal individuals and heated under the same conditions survived. Cultivation of the patient's lymphocytes at ^37°C for 4 days after heat treatment at 43^°C for 5 hr resulted in a viability of less than 50%. Proliferative responses of the patient's lymphocytes to lectins showed 70% more inhibition than did the control when heated at 43^°C for 1 hr and were abolished completely after 3 hr of hyperthermia. Although, in normal individuals, complete recovery of (³H)-thymidine uptake of the lymphocytes heated at 43^°C for 3 hr was observed after 4 days of cultivation at 37^°C, this recovery did not occur in the patient's cells. Morphological changes in the atypical lymphocytes heated at 43^°C for 3 hr occurred in the cytoplasmic organelles; in particular, enlargement of mitochondria, increased numbers of cytoplasmic granules and vesicles, probably derived from lysosomes, and hyperproduction of myelin sheath-like structures around the nuclear membrane were seen.     

Subpopulation of murine epidermal Langerhans cells identified by lectin-binding sites

Summary Lectin-binding profiles of epidermal Langerhans cells (LCs) were investigated in three strains of mice using immunofluorescence procedures. Three lectin-binding profiles were observed in each strain of mice. Most epidermal LCs reacted with concanavalin A (Con A) and Ricinus communis agglutinin 1 (RCA-1), whereas none reacted with Dolichos biflorus agglutinin (DBA). Peanut agglutinin (PNA) and wheat germ agglutinin (WGA) reacted with some of the epidermal LCs. These binding profiles were similar from site to site of the body in all strains of mice. We also investigated the lectin-binding profiles of epidermal Ia positive (Ia⁺) cells migrating into the grafted skin up to 165 days after transplantation. BALB/c (H-2^d) murine skin was grafted onto the back of (C3H/He×BALB/c)F1 (H-2^k×H-2^d) mice. The percentages of migrating I-A⁺ epidermal cells reactive with PNA and WGA were different from those of the normal epidermis soon after grafting and reached a normal level at 43 days after grafting. Our results demonstrated that there is a heterogeneous population of epidermal LCs defined by lectin-binding profiles.     

Pimecrolimus does not affect Langerhans cells in murine epidermis

BACKGROUND: Langerhans cells (LCs) function as specialized antigen-presenting cells in the epidermis, and therefore play a critical role in cutaneous immunological reactions. Topical treatment with corticosteroids is associated with a decrease in epidermal LC number and antigen-presenting capacity in laboratory animals and humans. OBJECTIVES: To examine whether pimecrolimus, a nonsteroidal inflammatory cytokine inhibitor recently introduced for the topical treatment of atopic dermatitis, differs from corticosteroids in effects on LCs. METHODS: Groups of BALB/c mice were treated twice daily on one to five consecutive days on the inner surface of the right ear with 10 micro L of ethanolic solutions of the test compounds at their clinically used concentrations (1% pimecrolimus, 0.1% betamethasone-17-valerate, 1% hydrocortisone and 0.05% clobetasol propionate) or with the vehicle (controls) alone. At selected time points after the treatment epidermal sheets were prepared and examined histomorphometrically for LCs immunolabelled with antibodies to major histocompatibility complex (MHC) class II and DEC 205, and adenosine diphosphatase staining. RESULTS: No changes in number or morphology of LCs were observed in epidermal sheets of mice treated for 5 days with pimecrolimus. In contrast, an almost complete depletion of LCs was observed in skin samples treated with hydrocortisone, betamethasone or clobetasol. Even a single-day treatment schedule with hydrocortisone, betamethasone or clobetasol caused a significant reduction in MHC class II+ LCs, by 31%, 62% and 87%, respectively. CONCLUSIONS: It is therefore unlikely that topically applied pimecrolimus affects epidermal LCs, in contrast to corticosteroids.