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。     

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。     

皮肤非朗格汉斯树突细胞组织细胞增生症：一个新病种？

患者男，57岁，全身斑块进行性增多7年，面部、躯干和四肢多发大小不等红色结节和浸润性斑片。组织病理学检查示真皮全层弥漫散在分布的多形性肿瘤细胞，呈低密度增生状态。部分可见细胞突起，使细胞呈多角状。免疫组化CD45、FXIIIa、CD14、MHC-Ⅱ、CD68、lysozyme阳性，并有细胞外间质表达，CD1a、S-100阴性。电镜下细胞突起，核形不规则，胞质及基质中弥漫分布膜包绕低密度囊泡结构，无Birbeck颗粒。本例为一种罕见的皮肤非朗格汉斯组织细胞增生症，临床呈惰性侵袭性经过，可能起源于不成熟的真皮树突细胞。     

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。     

以皮肤单核细胞肉瘤为首发表现的急性单核细胞白血病的特征研究

患儿4个月始发病,以额头、四肢皮肤出现多发皮色结节为特征,一般情况好,未触及肝、脾、淋巴结肿大、睾丸无明显异常、牙龈无异常增生。皮损组织病理：表皮大致正常,真皮及皮下弥漫浸润的异形细胞,体积中等偏大,胞质少,核椭圆形,不规则形,染色质细,个别细胞可见核沟及核仁。免疫组化：S-100蛋白阳性,CD68阳性;CD1阴性,Ki-67阳性率40%;CD21阴性,CD4部分细胞弱阳性。髓过氧化物酶（MPO） 阴性,CD56阳性,CD123阳性,CD163阳性。骨髓片见原、幼单核细胞占0.245。诊断：以皮肤单核细胞肉瘤为首发的急性单核细胞白血病。     

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。     

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。     

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。     

肉芽肿性蕈样肉芽肿

报告1例肉芽肿性蕈样肉芽肿。患者女,49岁。因躯干及四肢红斑、斑块1.5年,间断发热半个月就诊。皮损组织病理:表皮萎缩变薄,真皮内片状密集分布淋巴细胞、组织细胞及多核巨细胞,部分淋巴细胞核异形,真皮内可见肉芽肿病变。免疫组化:淋巴细胞CD3、CD4、CD5阳性,CD8散在阳性,组织细胞CD68阳性,T细胞样受体(TCR)基因克隆重排:TCRγ重排+。诊断:肉芽肿性蕈样肉芽肿。     

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。     

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.     

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.     

Heterogeneity within tissue-specific macrophage and dendritic cell populations during cutaneous inflammation in atopic dermatitis

BACKGROUND: Macrophages and dendritic cells may play a role in chronicity of atopic dermatitis (AD); however, so far only limited data are documented on the distribution of these cells in the skin during cutaneous inflammation. OBJECTIVES: To gain better insight into the presence and distribution of macrophage and dendritic cell (sub)populations in acutely and chronically inflamed skin of AD patients. METHODS: Chronic inflammatory reactions were studied in lesional AD skin biopsies; the atopy patch test was used as a model for the initiation of AD lesions, representing acute inflammation. To determine the number and phenotype of different dermal macrophage and dendritic cell populations immunohistochemistry and digital imaging were used. RESULTS: There was an increase in macrophage numbers in acutely and chronically inflamed AD skin, whereas absolute dendritic cell numbers were unchanged, compared with non-lesional AD skin. Furthermore, phenotypically heterogeneous and overlapping macrophage and dendritic cell populations were present in inflamed AD skin. The classic macrophage marker CD68 and prototypic dendritic cell marker CD1a could bind to the same cell subpopulation in the dermis of inflamed AD skin. Mannose receptors were expressed mainly by macrophages in inflamed AD skin. CONCLUSIONS: In this study we observed changes in macrophage number and phenotype during cutaneous inflammation in AD. Dendritic cell numbers did not change; however, phenotypically dendritic cell and macrophage subpopulations showed increasing overlap during inflammation in AD skin. We show for the first time that within tissue-specific macrophage populations further subpopulations are present, and that monocyte-derived cells may express markers for both dendritic cells and macrophages. Our results point to the existence of a heterogeneous pool of macrophage/dendritic cell-like cells, from which subpopulations of dermal macrophages and dendritic cells arise.     

Cd1d is expressed on dermal dendritic cells and monocyte-derived dendritic cells

CD1 proteins are a family of cell surface molecules that present lipid antigens to T cells. We investigated skin dendritic cells and monocyte-derived dendritic cells for expression of CD1 molecules using a panel of 10 different monoclonal antibodies focusing on the recently described CD1d molecule. By immunohistochemical analysis, CD1d expression in normal human skin was restricted to dendritic appearing cells in the papillary dermis mainly located in a perivascular localization. Langerhans cells did not show detectable CD1d expression in situ. Epidermal/dermal cell suspensions analyzed by flow cytometry demonstrated distinct subpopulations of HLA-DR positive dermal dendritic cells expressing CD1a, CD1b, and CD1c. CD1d was expressed on HLA-DRbright dermal antigen-presenting cells in dermal suspensions (16% +/- 3.6%), as well as on highly enriched dermal dendritic cells migrating out of skin explants (60.5% +/- 8.0%). Migrated mature dermal dendritic cells coexpressed CD83 and CD1d. Western blot analysis on microdissected skin sections revealed the presence of a 50-55 kDa CD1d molecule in dermis, suggesting that CD1d is highly glycosylated in skin. Both immature and mature monocyte-derived dendritic cells cultured in autologous plasma expressed CD1d molecules. In contrast, culture in fetal bovine serum downregulated CD1d expression. In conclusion, antigen-presenting cells in skin express different sets of CD1 molecules including CD1d and might play a role in lipid antigen presentation in various skin diseases. Differential expression of CD1 molecules depending on culture conditions might have an impact on clinical applications of dendritic cells for immunotherapy.     

Langerhans cell hyperplasia and IgE expression in canine atopic dermatitis

Langerhans cells appear to be critical for IgE-mediated allergen capture and presentation in human atopic dermatitis. The present study sought to determine whether epidermal (i.e Langerhans cells) and dermal dendritic cells in the skin of dogs with atopic dermatitis are hyperplastic and expressed surface IgE. Frozen sections of lesional or nonlesional atopic and normal control canine skin were immunostained with CD1a-, CD1c-, and IgE-specific monoclonal antibodies. The enumeration of cells was performed by morphometry in both the epidermis and the dermis. Cell counts were compared with each individual’s total serum IgE levels. Higher numbers of epidermal and dermal dendritic cells were present in atopic dogs than in normal control animals. Epidermal Langerhans cell counts were significantly higher in lesional than in nonlesional atopic specimens. IgE⁺ dendritic cells were observed in lesional atopic epidermis and dermis, and nonlesional atopic dermis, but not in normal control skin specimens. The percentages of IgE⁺ dendritic cells were correlated with each patient’s total serum IgE levels. These results demonstrate dendritic cell hyperplasia and IgE expression in canine atopic dermatitis. Increased epidermal Langerhans cell counts in lesional specimens suggest an epidermal allergen contact in canine atopic dermatitis.     

Expression of Fc receptors for IgG during acute and chronic cutaneous inflammation in atopic dermatitis

Atopic dermatitis is an allergic skin disease characterized by elevated total and antigen-specific serum IgE and IgG4 levels. In acute and chronic cutaneous inflammation, large cellular infiltrates including T cells, dendritic cells and macrophages are found, especially in the dermis. These cells play an important part in the regulation of local inflammatory reactions. Receptors binding IgG (FcgammaR) are involved in dendritic cell and macrophage function. In this study, we examined the in vivo distribution and cellular expression of the three classes of leucocyte FcgammaR in human skin during acute and chronic cutaneous inflammation in atopic dermatitis. Atopy patch test skin was used as a model for acute inflammation in atopic dermatitis, while chronic lesional skin was used to investigate FcgammaR expression in chronically inflamed skin. In atopy patch test sites no increase in the number of CD1a+ dendritic cells and a slight increase in macrophages compared with non-lesional skin was observed. Our results showed increased expression of FcgammaRI (CD64) and FcgammaRIII (CD16) in acutely inflamed skin as well as in chronically inflamed lesional skin, compared with healthy and non-lesional atopic dermatitis skin. FcgammaRI was expressed by RFD1+, RFD7+ and CD68+, but not by CD1a+ dermal dendritic cells. RFD1+ dendritic cells and CD68+ macrophages were the main FcgammaRIII-expressing cells during the acute inflammatory reaction. The significant increase in expression of FcgammaRIII (CD16) and FcgammaRI (CD64) probably results from upregulation of the receptors on resident cells. Insight into the presence of FcgammaR+ cells in human skin during inflammation is important both for our understanding of skin immune reactions and the development of new therapeutic concepts.     

T lymphocytes in lesional skin of patients with dermatitis herpetiformis

Ten patients with dermatitis herpetiformis had biopsies taken from involved skin.Monoclonal antibodies and the avidin-biotin peroxidase staining technique were used to stain for T cells and Langerhans cells in skin sections. A significant increase in the number of CD3-positive T cells was observed in the upper dermis of involved compared with uninvolved skin (P<0.0005). Most of the T cells in involved skin were CD45RO-positive memory cells; CD4-positive T cells exceeded the number of CD8-positive T cells by a ratio of 4:1. In addition, CD1a-positive dendritic cells were observed within the clumps of T cells in involved dermis in nine of the 10 patients, but were absent from the dermis of uninvolved skin. Double immunofluorescent staining demonstrated that approximately 20–40% of the CD3-positive T cells were activated, and expressed the HLA-DR antigen. These findings suggest that activated T cells are involved in the pathogenesis of dermatitis herpetiformis skin lesions.     

Fc epsilon R11/CD23 receptor distribution in patch test reactions to aeroallergens in atopic dermatitis.

There is increasing evidence that exposure to organic allergens may induce or exacerbate lesional skin in patients with atopic dermatitis. In this study, patients with atopic dermatitis were patch tested to 11 common organic allergens and to control chambers containing 0.4% phenol and 50% glycerin in 0.9% saline. In biopsies from positive patch test reactions, patch test control skin, lesional eczematous and non-lesional skin from atopic individuals, and normal skin from non-atopic volunteers, the presence and distribution of macrophages (RFD7+), dendritic cells (RFD1+), and Langerhans cells, and the expression of the low-affinity receptor for IgE (CD23) were investigated. In patch test reactions and lesional skin samples, inflammatory infiltrates of diffusely distributed macrophages (RFD7+), dendritic cells (RFD1+), T lymphocytes (RFTmix+), and Langerhans cells (CD1+) were seen, the latter being present in both the epidermis and the dermis. The numbers of Langerhans cells were reduced in the epidermis and increased in the dermis in patch test reactions and lesional skin compared to their controls. Double staining revealed a change in the distribution of CD23 antigen. In patch test control and non-lesional biopsies many macrophages and only a few Langerhans cells within the dermal infiltrates expressed this antigen. In patch test reaction and lesional skin samples, however, the proportion of CD23+ dermal Langerhans cells had increased compared to macrophages. Furthermore, in these latter samples an increased proportion of dermal CD1+ cells expressed the dendritic cell (RFD1+) marker. These results show that following antigen challenge there are marked similarities between the phenotype of the cellular infiltrate in patch test reaction and lesional skin biopsies, and also demonstrate a changing distribution of CD23 on antigen-presenting cells.