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。     

成人正常皮肤CD68阳性单核-巨噬细胞分布的研究

目的 观察正常真皮内CD68阳性单核-巨噬细胞的分布、形态和密度。方法 正常成人8例,每例均取面部、躯干、四肢近端、四肢远端、手掌和足跖6个部位皮肤进行表皮铺片和纵行与水平连续切片。CD68单克隆抗体染色,观察单核-巨噬细胞的分布。结果 真皮浅层CD68阳性细胞可表现为树枝状及非树枝状,呈网状分布。真皮浅层CD68阳性细胞密度为:四肢远端(562±230)个/mm²,腹部(517±162)个/mm²,面部(509±235)个/mm²,手掌(507±192)个/mm²,四肢近端(472±138)个/mm²,足跖(361±78)个/mm²。真皮深层CD68阳性细胞密度低,多为树枝状,散在于胶原纤维间。结论 CD68阳性单核-巨噬细胞在真皮浅层形成较致密网状分布。提示单核-巨噬细胞在真皮内有明确的方向性,其防御的方向是穿透表皮进入真皮的入侵物。     

系统性红斑狼疮患者CD1a、CD68、HLA-DR等的研究

目的 研究系统性红斑狼疮(SLE)患者外观正常及病变皮肤中朗格汉斯细胞(LC)一些重要表面标志的变化。方法 应用CD1a、CD68和HLA-DR等单克隆抗体和ABC免疫组化技术对9例SLE患者外观正常和皮损部位的组织进行了免疫表型检测。结果 ①SLE皮损中LC的数量减少,且其形态与表面标志亦有变化;②SLE病损处的角质形成细胞(KC)强弱不等地表达HLA-DR抗原,个别病例的外观正常皮肤KC也可局灶性表达HLA-DR抗原;③SLE外观正常皮肤或皮损的表皮中均未见细胞间粘附分子1和CD4阳性LC,仅在真皮的浸润细胞中见到较多的阳性细胞;④发现在SLE外观正常皮肤和皮损表皮内出现两类CD68阳性的树枝状细胞;在SLE皮损的浸润细胞中CD68阳性树枝状细胞大量增加;⑤细纤维状CD68阳性物质呈网状围绕基底部的KC,这些细纤维状阳性物质有些与表皮树枝状细胞相连,有些则没有明显的关系。结论 SLE外观正常和病变皮肤中LC一些重要表面标志的变化有所不同。外观正常皮肤和皮损表皮内出现两类树枝状细胞,一类可能为LC,而另一类则来源不清;在SLE皮损的浸润细胞中这些CD68阳性树枝状细胞大量增加,表皮内存在CD68阳性纤维状染色,其意义尚需进一步研究。     

人表皮内CD1a和CD68阳性细胞的检测

目的观察人表皮铺片内及切片中CD68阳性细胞的密度,并与朗格汉斯细胞(LC)进行比较。方法正常人8例,取其面部、躯干、四肢近端、四肢远端、手掌和足跖6个部位皮肤,进行铺片和切片。CDla和CD68单克隆抗体进行ABC过氧化酶技术和荧光双标记染色。结果 CDla阳性细胞多位于表皮中部,呈线性排列。CD68阳性细胞一部分为树突状,主要位于表皮的中、下部。在手掌和足跖表皮铺片的LC最少,密度为184～252个／mm~2,CD68阳性细胞密度为232～310个／mm~2。面、躯干及四肢表皮内的CDla阳性细胞密度较多,分别为530～809个／mm~2,这些部位CD68阳性细胞的密度为524～637个／mm~2。CDla和CD68双标记阳性的树突细胞多位于表皮的中层。各个部位的双标记阳性细胞线性密度均小于CDla和CD68阳性细胞线性密度的一半以下(28.6％～49.8％)。面部、躯干和四肢皮肤双标记阳性细胞占CDla阳性细胞的百分比和双标记阳性细胞占CD68阳性细胞百分比差异无统计学意义。结论表皮内存在CDla~ CD68~ ,CDla~ CD68~-和CDla~-CD68~ 3个表面标志不同的细胞群。在表皮内存在较多的CD68阳性而LC标志阴性的细胞。     

CD1a、CD68抗原在湿疹患者皮损中的表达

目的:观察CD1a、CD68抗原在湿疹患者皮损中的表达,探讨其临床意义。方法:采用免疫组织化学方法检测30例湿疹患者皮损表皮、真皮中CD1a、CD68抗原的表达,并与10例正常人皮肤进行比较。结果:正常皮肤中CD1a抗原主要表达于表皮棘层、基底膜朗格汉斯细胞膜上,而湿疹皮损中CD1a抗原主要散在分布于表皮棘层及真皮乳头中。CD68主要为单核-巨噬细胞膜染色,表达在表皮下部及真皮。湿疹皮损表皮及真皮中CD1a阳性、CD68阳性的细胞线性密度均明显高于正常皮肤(t值分别为2.86、4.43,P值均0.01)。结论:CD1a、CD68抗原在湿疹患者皮损处表达较高,提示朗格汉斯抗原提呈细胞增多及单核-巨噬细胞浸润可能与湿疹发病有关。     

银屑病皮损区肥大细胞分布和血管内皮细胞CD34表达的研究

目的探讨肥大细胞(MC)和血管内皮细胞在银屑病发病中的作用.方法采用组织化学和免疫组化的方法,观察寻常性银屑病皮损处MC和CD34标记血管内皮细胞的分布情况.结果经甲苯胺蓝特殊染色发现,银屑病患者真皮区的MC密度在进行期(33.07±14.63)个/mm2,高于静止期(21.80±4.86)个/mm2,静止期又高于正常对照组(15.85±6.93)个/mm2,它们之间的差异均有显著性;免疫组化观察发现,银屑病真皮区CD34标记的微血管密度在进行期(2931±4.04)个/mm2,明显高于静止期(2231±2.07)个/mm2,而静止期又明显高于正常对照组(18.81±2.59)个/mm2.结论寻常性银屑病皮损处真皮内肥大细胞和血管内皮细胞密度明显增加,且与病情变化有关.     

巨噬细胞是正常成人真皮中主要的细胞成分

目的 研究正常人真皮内的巨噬细胞和树枝状细胞占所有有核细胞的比率。方法 正常人8例,每例均取面部、躯干、四肢近端、四肢远端、手掌和足跖6个部位皮肤,进行铺片和纵行与水平连续切片。CD68,CD36单克隆抗体和FXIIIa多克隆抗体染色。观察这些巨噬细胞胞占真皮中所有有核细胞的比例。结果 CD68阳性巨噬细胞占真皮浅层所有有核细胞总数的67.5±17.1%, FXIIIa阳性真皮树枝状细胞为35.2±14.5%,CD36阳性细胞为27.0±11.4%。结论 巨噬细胞不仅是正常成人真皮中的主要的免疫细胞,而且是真皮内皮肤免疫防御功能的主要细胞。在真皮内还有一定数量的抗原提呈细胞。     

局部温热对接触性超敏反应小鼠皮肤朗格汉斯细胞的影响

目的 探讨局部温热对小鼠接触性超敏反应激发部位朗格汉斯细胞（LC）数目和形态的影响。方法 8周龄雌性SPF级Balb/c小鼠60只,随机分为致敏前3 d加热组、致敏同时加热组和致敏2 d后加热组,每组20只。分别于致敏前3 d、致敏同时和致敏2 d后在背部致敏部位施以37 ℃、39 ℃、41 ℃、43 ℃的局部温热（各5只）20 min。同时设立5只致敏-不加热组作为对照。于小鼠右耳背侧激发后2 d,分离右耳背侧表皮,采用免疫组化方法检测表皮LC的数目和形态变化。结果 随所施温度的升高,LC数目在致敏前加热组逐渐减少,37 ℃、39 ℃、41 ℃、43 ℃依次为（321.83 ± 41.81）、（251.12 ± 16.29）、（191.41 ± 28.7）、（128.33 ± 77.61）个/mm2,组间差异有统计学意义（P < 0.05）;在致敏同时加热组逐渐增多,但差异无统计学意义（P > 0.05）;在致敏后加热组也逐渐增多,依次为（320.83 ± 113.6）、（398.33 ± 31.91）、（437.83 ± 29.78）、（477.25 ± 86.79）个/mm2,差异有统计学意义（P < 0.01）。无论致敏前3 d、致敏同时和致敏2 d后施以不同温热条件,LC的树突随所施温度升高呈增多、增长的趋势;但在43 ℃条件下,其趋势略有下降。结论 致敏部位的局部温热对激发部位LC形态和密度有影响,可能与接触性超敏反应的严重程度有关。     

真皮内单核-巨噬细胞形态分析

单核-巨噬细胞是真皮内重要的免疫细胞。CD68是这些单核-巨噬细胞的重要标志。我们发现在正常人的真皮浅层存在呈网状分布的CD68阳性细胞,这些细胞在形态和染色强度上存在一定的差异^[1]。为进一步研究这些细胞的差异,我们采用图像分析方法,对正常真皮内的单核-巨噬细胞的形态及染色强度进行分析,探讨真皮内这类细胞的形态特点。     

斑秃皮损内朗格汉斯细胞及CD8+ T细胞的数量及分布分析

【摘要】 目的 探讨斑秃患者脱发皮损中朗格汉斯细胞在斑秃病理进程中的分布以及与T细胞的关系。 方法 对29例斑秃患者（活动期16例,非活动期13例）头皮脱发皮损进行CD1a免疫组化染色,对其中17例斑秃患者行CD4、CD8免疫组化染色。荧光半定量PCR测定局部皮损浅层和深层CD1a和粒细胞巨噬细胞刺激因子（GM-CSF）的mRNA表达水平。 结果 斑秃患者表皮和真皮各处包括真皮浅层血管周围、毛囊周围,真皮深层血管周围、毛囊周围CD1a阳性LC数量均较健康对照显著增加（Z = 4.354,2.884,4.640,3.217, 3.496,均P ＜ 0.01）,活动期皮损表皮层、深层血管、深层毛囊CD1a阳性LC数量较非活动期皮损高（Z = 2.457, 2.130,1.954,P ≤ 0.05）。斑秃患者CD1a 、GM-CSF mRNA相对表达量在皮损真皮浅层虽与健康对照无差异,但在深层均高于健康对照（Z = 2.702,2.941,均P ＜ 0.01）。斑秃患者浅层血管周围LC与深层毛囊周围CD8+ T细胞数量呈正相关（r = 0.618,P ＜ 0.05）,活动组浅层血管周围LC与深层毛囊周围CD8+ T细胞数量分布呈正相关关系（r = 0.795,P = 0.01）,非活动组浅层血管周LC与深层毛囊周围CD8+ T细胞数量分布则无相关关系。 结论 斑秃患者皮损LC数量增加,且在活动期皮损升高更明显。活动期斑秃皮损中浅层血管周围LC与深层毛囊周围CD8+ T细胞数量呈正相关,推测LC在斑秃疾病进展中发挥作用。     

Distinctive dendritic cell subsets expressing factor XIIIa, CD1a, CD1b and CD1c in mycosis fungoides and psoriasis

The papillary dermis of psoriasis and mycosis fungoides (MF) lesions is characterized by prominent collections of cells with dendritic morphology. Immunophenotypically distinct populations of cutaneous dendritic cells have been identified as CD1a+, FXIIIa-Langerhans cells (LC) and CD1a-, FXIIIa+ dermal dendritic cells (DDC). In this study, antibodies against the human GDI cluster of antigens (i.e. CD1a, CD1b and CD1c) and the DDC) marker (FXIIIa) were used to further characterize the subsets of dendritic cells in normal skin as compared to neonatal foreskin, psoriasis and MF by both immunoperoxidase and double immunofluorescence techniques. Normal skin and foreskin epidermis and dermis contained few CD1b+ or CD1c+ cells along with normal numbers of CD1a+ LC and FXIIIa+ DDC. Both MF and psoriasis were characterized by CD1a+ cells in the epidermis and dermis. FXIIIa+ cells were greatly expanded in the upper dermis of MF lesions and to a lesser degree in psoriasis as has been previously described by our group. MF contained significantly increased epidermal and dermal CD1b+ (15.7/5 high power fields [HPF] and 59.7/5 HPF respectively) and CD1c+ dendritic cells (33.8/5 HPF and 95.9/5 HPF respectively), while in psoriasis these cells were not statistically different from normal skin. Double immunofluorescence studies revealed that some (<25%) FXIIIa+ cells co-expressed CD1b and CD1c in MF>psoriasis> foreskin, while FXIIIa+ DDC never co-expressed CD1a. Thus, in contrast to normal skin in which epidermal or dermal dendritic cells rarely express CD1b and CD1c antigens, these members of the CD1 family are upregulated on both LC and DDC in benign and malignant inflammatory states. Upregulation of CD1b and CD1c on MF epidermal and dermal dendritic cells, as compared to psoriasis, foreskin and normal skin, may be useful in the immunophenotypic recognition of MF, as well as in helping to understand its immunobiology.     

"Dermal dendritic cells" comprise two distinct populations: CD1+ dendritic cells and CD209+ macrophages

A key cell type of the resident skin immune system is the dendritic cell (DC), which in normal skin is located in two distinct microanatomical compartments: Langerhans cells (LCs), mainly in the epidermis, and dermal DCs (DDCs), in the dermis. Here, the lineage of DDCs was investigated using monoclonal antibodies and immunohistology. We provide evidence that "DDC" comprise at least two major phenotypic populations of dendritic-appearing cells, immature DC expressing CD1, CD11c and CD208; and macrophages expressing CD209, CD206, CD163, and CD68. These data suggest that dermal dendritic-appearing macrophages comprise a novel part of the innate immune response in the resident skin immune system.     

CD1a+, CD3+, CD4+, CD8+, CD68+ and cutaneous lymphocyte-associated antigen-positive cells in Bowen's disease

BACKGROUND: Bowen's disease (BD) is a squamous cell carcinoma in situ that rarely invades into the underlying dermis. However, little is known about its immunohistology. Objectives To evaluate the relationship between the cytological properties of the tumour cells in BD and the host immune response. METHODS: We examined the expression of p53, proliferating cell nuclear antigen (PCNA) and Ki67 antigen, and the number of mitotic cells, together with the number of intratumoral and dermal infiltrating CD1a+, CD3+, CD4+, CD8+, CD68+ and cutaneous lymphocyte-associated antigen (CLA)+ cells in 18 cases of genital BD. RESULTS: When compared with normal genital skin (n = 10), there was a significantly higher number of mitotic cells as well as higher expression of p53+, PCNA+ and Ki67+ cells in BD. There was significant mutual correlation between CD3+, CD4+ and CD68+ cells in the tumoral epidermis. The number of CD1a+ Langerhans cells significantly decreased in BD epidermis; however, dermal CD1a+ cells were increased. Interestingly, numbers of dermal CD1a+ cells significantly correlated with those of intratumoral CD3+, CD4+ and CD68+ cells. In situ hybridization for human papillomavirus (HPV) demonstrated that HPV-infected BD had significantly less infiltration of intratumoral CD3+ cells and CLA+ cells. CONCLUSIONS: The present data suggest that dermal CD1a+ cells may participate in the immune surveillance and that HPV infection may interfere with the intratumoral infiltration of CLA+ cells in BD.     

Thrombomodulin expression on dermal cells in normal and psoriatic skin

Abstract The various subsets of dermal cells with a dendritic appearance can be identified by phenotypic differences in cell markers. We report on the morphology and tissue distribution of dermal cells detected with a monoclonal antibody against thrombomodulin in histological sections of normal arm and scalp skin and psoriatic skin. Double staining with antibodies to factor XIIIa, CD34 and CD68 was also employed in scalp biopsies to elucidate the relationship between thrombomodulin⁺ dermal cells and dermal dendrocytes and macrophages described by others. Thrombomodulin⁺ dermal cells in normal arm skin had little cytoplasm with fine branched dendrites and tended to be localized just beneath the epidermis. In scalp skin these cells had longer, more numerous dendrites and were distributed in the papillae and perivascular adventitial dermis primarily in the upper and central reticular dermis. In psoriatic skin, thrombomodulin⁺ dermal cells had an increased cytoplasmic volume with stout, less branched dendrites and appeared in the papillae and among inflammatory cells. Dermal cells detectable by thrombomodulin expression were factor XIIIa^–, CD34^– and CD68^–, and seemed to represent a distinct subset of dermal cells which may function in tissue repair. However, thrombomodulin⁺ dermal cells and factor XIIIa⁺ dendrocytes were frequently seen close together and could act cooperatively to regulate extravascular thrombin homeostasis in both normal and pathological dermal environments. Received: 26 May 1997     

Immunophenotypic analysis of normal human dendritic cells isolated from epidermis and dermis

Background The skin immune system comprises two types of dendritic cells, i.e. CD1a-positive Langerhans cells in the epidermis and CD36-positive dendritic macrophages in the dermis. Dendritic cells can migrate from skin explants into a culture medium.
Methods We have examined the morphology and immunophenotype of the dendritic cells migrating from epidermal and dermal sheets in vitro. The epidermis and dermis of keratomes of normal human skin were separated with dispase and cultured for 72 h. At this time, the non-adherent cells in the medium were removed, enriched on a metrizamide or Lymphoprep gradient, counted, prepared by cytospin, and labeled for CD1a, CD36, and HLADr.
Results Cells migrating from the epidermis and dermis show many thin projections or a few veils from the cell surface. Approximately four times more cells migrate from epidermal than dermal sheets from the same keratome.
Conclusions Using methods to separate the epidermis from the dermis, both CD1a-positive Langerhans cells and CD36-positive dendritic macrophages can be obtained from both tissues, although in different numbers.     

Expression of CD1a and CD86 on scleroderma Langerhans cells

Scleroderma is a chronic autoimmune connective tissue disorder of unknown etiology that affects the microvasculature and loose connective tissue. Langerhans cells play an important role in the immune system of the skin. By immunohistochemistry we investigated the phenotypical characteristics of epidermal and dermal Langerhans cells and their spatial relationship with infiltrating lymphocytes in systemic scleroderma (SSc) and localized scleroderma. Skin samples were obtained from patients by 6 mm punch biopsy. Samples were stained with antibodies against CD1a and CD86. The number of cells stained with both antibodies in the dermal and epidermal infiltration was calculated. In contrast to normal skin, both types of scleroderma skin showed a marked increase in CD1a+ dermal Langerhans cells, whereas the number of CD1a+ cells in localized scleroderma was much higher than that in SSc (p < 0.05) either in the dermis or in the epidermis. The expression of CD86 was increased in the dermis of localized scleroderma compared with that in SSc or normal skin (p < 0.05). This study revealed that Langerhans cells may play an important role in the pathogenesis of scleroderma, especially in localized scleroderma. CD86 is predominantly expressed on dermal Langerhans cells in the lesional skin of localized scleroderma. Therefore, it might play an important role in the pathogenesis of localized scleroderma.     

The development of manifest psoriatic lesions is linked with the invasion of CD8+ T cells and CD11c+ macrophages into the epidermis

Koebner response was studied in 35 psoriatic patients. Two punch biopsies per patient were taken from non-lesional psoriatic skin before, and 6 h, 2 days, 7 days, 14 days and 21 days after, tape stripping. Alterations in the numbers of CD1+ Langerhans cells, CD4+ and CD8+ T cells and CD11c+ macrophages were mapped morphometrically. Results were compared with lesional and non-lesional psoriatic skin, and control skin. Nine of 35 patients were Koebner-positive. No statistically significant differences were noted between non-lesional psoriatic and control skin. CD4+ T cells increased in number 2 days after trauma in both the epidermis and the dermis, whereas epidermal CD8+ T cells and CD11c+ macrophages increased only in the Koebner-positive lesional skin after 7 days. The changes in lesions induced by tapestripping resembled those seen in lesional psoriatic skin (mature plaques). The number of CD1+ cells increased in mature psoriatic lesions only. It seems possible that trauma per se stimulates the accumulation of CD4+ T cells at the site of injury, but the development of manifest psoriatic lesions correlates with invasion of CD8+ T cells and CD11c+ macrophages into the epidermis.     

A model system using tape stripping for characterization of Langerhans cell-precursors in vivo

Little is known about the immigration of bone marrow-derived progenitors of Langerhans cells (LC) into the epidermis. We developed an in vivo system based on the tape stripping method that allowed us to study the immigration of LC into the epidermis after intradermal injection of bone marrow-derived dendritic cells (DC). Tape stripping induced a mechanical disruption of the epidermal barrier that led to skin inflammation and subsequent emigration of LC and dermal DC from the skin. Emigrating LC and dermal DC were observed in lymphatic vessels, and the numbers of LC and dermal DC in the draining lymph node increased. Up to 500 times more injected precursors migrated into tape-stripped epidermis as compared with unstripped epidermis. Newly immigrated cells were slender with one or two dendrites and acquired a more dendritic morphology after 2-4 days. They were both MHC II-positive and negative and they did not express Langerin/CD207, nor macrophage-mannose receptor/CD206 and Fc-epsilon receptor I. In contrast, all cells that had entered the epidermis expressed CD11c and CCR6, suggesting that they were LC. We conclude that this experimental system may serve as a valuable tool for the further characterization of LC-precursors and the conditions necessary for LC-immigration into the epidermis.