移植物抗宿主病皮肤科表现研究进展

80年代以来,随着同种骨髓移植(BMT)等治疗方法在临床上的应用,对移植物抗宿主病(GVHD),特别是其皮肤表现(GVHDS)的研究有了很大进展,综述如下.GVHD的病因和发生机理GVHD发生于BMT和免疫缺陷者接受组织不相容性供体的免疫活性淋巴细胞(LC)时.一般认为发病与供体LC和宿主HLA有关,其中HLA-D的遗传位点决定移植细胞是否与组织不相容性抗原发生反应.把实验皮肤与同种LC一起培养,能引起     

皮肤移植物及嵌合体内郎格罕细胞的研究

本文报告以抗Ia单克隆抗体检查表皮内LC的方法,观察了皮肤移植物内及嵌合体内不同基因型LC的结果.结果显示,移植物内原有LC的大部分可被宿主LC所迅速更换,而小部分能持续存在而不被更换,从而提示LC可能存在二个亚群,其一经常移动,另一则较长期(>249天)地固定.结果又显示,输注骨髓细胞或脾、淋巴绪细胞后的嵌合体的表皮内,无例外地可检测到供体基因型的LC;从而提出了一种国内外迄今未使用过的简便准确的检测嵌合体的新方法,建议使用这一方法于动物实验及临床观察.     

花斑癣患者表皮郎格罕细胞的观察

作者采用ATP酶染色方法对10例花斑癣患者皮损表皮内郎格罕细胞(LC)进行了观察,并对4例患者与皮损相对应部位的正常皮肤进行了同样检查.与此同时,取4名正常人胸腹部正常皮肤作对照检查.结果表明:花斑癣皮损表皮内LC密度明显减少,与正常对照组相比,P<0.001,有灶性集聚现象,许多LC的树枝突缩短、减少或消失.4例花斑癣患者与皮损相对应部位的正常皮肤表皮内LC的变化与皮损表皮内LC的变化相类似.作者认为,LC在花斑癣的免疫病理中可能起着重要作用.     

反应停治疗骨髓移植后的移植物抗宿主病

作者报道以反应停300mg/d,连用6个月治疗1例严重广泛的硬化期慢性皮肤移植物抗宿主病,使皮肤硬化逐步好转。患者男,28岁,患严重再生障碍性贫血,在第1次试用骨髓移植后引起迟发性移植物排斥。9个月后第2次接受HLA 相匹配的姐妹同种异体骨髓移植,发生急性Ⅱ度皮肤移植物抗     

急性白血病骨髓移植后并发泛发性带状疱疹1例

患者男,52岁.确诊为急性淋巴细胞性白血病-L2型(ALL-L2)伴ph染色体阳性2年,于2007年8月12日行基因造血干细胞移植术,移植后予环孢素抗排斥反应,半年前患者出现口腔破溃、疼痛等皮肤黏膜移植物抗宿主病(GVHD)表现,加用环孢素和泼尼松抗GVHD治疗,口腔疼痛好转后停用泼尼松及环孢素减量,复查骨髓处于完全血液学及分子生物学缓解中.     

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。     

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。     

色素障碍性皮肤病

941539 白癜风患者表皮内郎格罕细胞观察/朱铁君//中国皮肤性病学杂志。-1994,8(2).-74 对13例进展期白癜风病人,用改良的Juhlin-ShelIeyATP酶细胞化学染色法检查其白斑、白斑边缘及对 侧相应正常皮肤中郎格罕细胞(LC)情况。结果,白斑内LC数目较相对应正常皮肤虽有增加,但无统计学意义(P>0.05),白斑边缘皮肤LC数目比对应正常皮肤显著增多(P<0.005)。白斑及白斑边缘的LC除密度变化     

尖锐湿疣中人乳头瘤病毒抗原的检测及郎格罕细胞的变化

业已公认,机体排斥外来抗原的特异性免疫必须有抗原提呈细胞的参与,皮肤内主要的抗原提呈细胞是郎格罕细胞(LC)^[1].为了探讨尖锐湿疣发病机制中人乳头瘤病毒(HPV)与郎格罕细胞(LC)的关系.我们应用抗生物素-生物素-过氧化酶复合物技术(ABC技术)对85例常规病理诊断为尖锐湿疣的石蜡切片进行HPV抗原检测,并对22例尖锐湿疣的表皮LC进行了观察.     

实验性接触性变态反应中郎格罕氏细胞的动态观察

作者应用DNCB诱发琢鼠实验性接触性皮炎定时活检,制备表皮片,用改进的ATP酶细胞化学方法染色,连续观察表皮LC的动态变化.在变态反应性接触性皮炎,抗原激发后12小时内.LC密度显著下降,树枝状突减少:2~5天内,表皮LC几乎完全消失;以后LC开始重建.14天内密度恢复正常.而在原发刺激性接触性皮炎2小时内LC树技状突即减少,6小时LC密度即显著降低.LC恢复亦较迅速,作者认为这种差异主要是因为两种皮炎的发病机制不同.实验所见提示LC在接触性变态反应的早期、初期以及恢复期均起重要作用.此外,本文还就实验动物的选择、诱发试剂的浓度以及ATP酶细胞化学染色的特异性等问题进行了讨论.     

Immunology of skin transplantation

Untreated viable allogeneic skin is highly immunogenic. Epidermal Langerhans migrate after transplantation out of the donor skin into the lymph node of the recipient where they can activate T cells capable to mediate rejection. Allogeneic skin is used as a temporary coverage of burn wounds, often in combination with autologous skin grafts. Several methods to pretreat the allogeneic skin have been used to delay the rejection process. Processing of allogeneic skin in 85% glycerol results in a non-viable skin with a well-preserved structure. Experiments in a full thickness porcine wound model showed that rejection of glycerol treated allogeneic skin grafts was up to six days delayed. Viable, untreated allogeneic skin grafts were rejected predominantly by CD8 positive T cells whereas in the glycerol treated grafts the influx of host cells was lower and the majority of the cells were macrophages. The outgrowth of the autologous skin grafts underneath glycerol treated allogeneic skin was three days earlier completed when compared to grafts in combination with untreated allogeneic skin. Thus, by processing the allogeneic skin into 85% glycerol, the direct route to induce graft rejection is blocked since the Langerhans cells are non-viable. The glycerol-preserved skin grafts are finally rejected via an indirect route mediated by macrophages; this process is less disturbing for the outgrowth of autologous cells.     

大鼠郎格罕氏细胞Ia抗原及ATP酶的研究

本文报告于大鼠LC表面也具有Ia抗原及ATP酶活性;用Ia抗原间接免疫荧光染色可清晰地显示成年大鼠表皮内的LC:ATP酶染色可用于新生大鼠表皮内LC的检查.本文详细叙述了用I成原间接免疫荧光染色、Ia抗厚间接免疫过氧化酶染色及ATP酶染色显示大鼠表皮内LC的步骤与注意事项.     

IL‐1 signalling determines the fate of skin grafts expressing non‐self protein in keratinocytes

Please cite this paper as: IL‐1 signalling determines the fate of skin grafts expressing non‐self protein in keratinocytes. Experimental Dermatology 2010; 19 : 723–729. Abstract: Although IL‐1 is a known inflammatory cytokine during pathogen infection, the role of IL‐1 in skin graft rejection, particularly where foreign antigen is expressed exclusively in keratinocytes, is less understood. Here, we use a syngeneic skin graft system, where antigens are expressed in epithelial cells via either a keratin 14 or keratin 5 promoter, to explore the role of IL‐1 in graft rejection and induction of epithelial antigen‐specific effector CD8⁺ T‐cell function. Keratin 5 ovalbumin (K5mOVA) transgenic skin grafts destined for rejection demonstrated increased expression of IL‐1β and its receptors compared to K14 HPV16 E7 transgenic grafts that do not reject spontaneously. Rejection of OVA grafts lacking the IL‐1 receptor (IL‐1R1) was delayed and associated with decreased numbers of antigen‐specific CD8 T cells. In contrast, K14E7 grafts survived on immunocompetent, syngeneic recipients with decreased graft levels of IL‐1β and IL‐1R1 and 2. However, in the absence of the IL‐1 receptor antagonist, IL‐1Ra, skin grafts were spontaneously rejected and an E7‐specific CD8 T‐cell response was primed. Thus, expression of the HPV16E7 oncoprotein in epithelial cells prevents IL‐1β‐associated skin graft rejection and induction of antigen‐specific CD8 T‐cell responses. Enhancing IL‐1β signalling, via blocking of the IL‐1 receptor antagonist, may represent an alternative strategy for treatment of HPV16E7‐associated cancers.     

Is epidermal cell proliferation in psoriatic skin grafts on nude mice driven by T-cell derived cytokines?

Plasminogen activity and DNA synthesis by epidermal cells have been reported to be doubled in psoriatic skin grafts compared with grafts of normal skin 6 weeks after transplantation to nude mice. In our study human lymphocytes disappeared from such grafts within 48 h whilst some DR-positive human dendritic cells were retained in the grafts for up to 4 weeks. However, the grafts were infiltrated by Thy 1.2+ mouse lymphocytes within 6 days and this infiltration persisted at a moderate level throughout the observation period. It consisted of perivascular aggregates, scattered dermal and papillary T cells, and some mouse T cells were also found in the epidermal compartment. Grafts of psoriatic and non-psoriatic control skin were infiltrated to a similar extent, suggesting a low-grade rejection response against the human xenografts. These findings raise the possibility that psoriatic keratinocytes are responding abnormally to inflammatory cytokines released by mouse lymphocytes reacting against the skin grafts.     

Inhibition of antiskin allograft immunity by infusions with syngeneic photoinactivated effector lymphocytes

Induction of tolerance for skin allotransplantation requires selective suppression of the host response to foreign histocompatibility antigens. This report describes a new approach that employs pretreatment of effector cells with 8-methoxy-psoralen (8-MOP) and ultraviolet A light (UVA) to render the effector cells of graft rejection immunogenic for the syngeneic recipient. Reinfusion of photodamaged cells resulted in an immunosuppressive host response that permitted prolonged retention of histoincompatible skin grafts and specifically inhibited in vitro and in vivo responses that correlate with allograft rejection. Eight days after BALB/c mice received CBA/j skin grafts, their splenocytes served as a source of alloreactive effector cells. The splenocytes were treated with 100 ng/ml 8-MOP and 1 J/cm2 UVA before reinfusion into naive BALB/c recipients. Recipient mice were tested for tolerance to alloantigens in mixed leukocyte culture (MLC), cytotoxicity (CTL), delayed type hypersensitivity assays (DTH), and challenge with a fresh CBA/j graft. Splenocytes from BALB/c recipients of photoinactivated splenocytes containing the effector cells of CBA/j alloantigen rejection proliferated poorly in MLC and generated lower cytotoxic T cell responses to CBA/j alloantigens in comparison with sensitized and naive controls. Splenocytes from these hyporesponsive mice suppressed the MLC and CTL response to alloantigen from sensitized and naive BALB/c mice. In vivo the DTH response was specifically suppressed to the relevant alloantigen in comparison with controls. Moreover, BALB/c mice treated in this fashion retained a CBA/j skin graft for up to 42 d posttransplantation without visual evidence of rejection. These results indicate that the in vivo and in vitro response to alloantigen can be attenuated by pretreating the host with photoinactivated splenocytes containing the effector cells of alloantigen rejection.     

Langerhans cells are not required for efficient skin graft rejection

The mechanism of skin allograft rejection has been thought to require presentation of graft antigen by resident epidermal Langerhans cells (LCs). We have previously engineered mice that have a selective and constitutive absence of epidermal LCs. By using donor skin from these LC-deficient mice, we show that LCs are not required for rejection of major (FVB --> B6) or minor (H-Y, male --> female on B6 background) antigen-mismatched skin grafts. On the FVB background, where H-Y mismatched grafts are normally maintained indefinitely, grafts lacking LCs are efficiently rejected. Thus, LCs in the donor graft are required for long-term skin engraftment, which supports a regulatory role for LCs in skin graft acceptance.     

Comparison of alloimmunogenicity of Langerhans cells and keratinocytes from mouse epidermis

Among lymphoreticular cells, Langerhans cells and splenic dendritic cells stand alone in their capacity, when hapten-derivatized, to induce vigorous immune responses, irrespective of route of inoculation, including intravenous. We have examined the comparative efficiency of relatively purified populations of Langerhans cells and their epidermal companions, keratinocytes, to induce alloimmunity when injected intravenously into adult mice. It was found that as few as 100 BALB/c Langerhans cells injected intravenously into C3H mice are capable of inducing specific sensitization as evidenced by subsequent accelerated rejection of BALB/c skin grafts. By contrast, 10,000 BALB/c keratinocytes failed to immunize similarly injected C3H recipients. These results emphasize the unparalleled capacity of Langerhans cells to induce sensitization, and they point to Langerhans cells, among cells within the epidermal compartment, as dominant in the alloimmunogenic potential of skin grafts.     

Inhibition of skin allograft rejection and acute graft-versus-host disease by cis-urocanic acid.

Ultraviolet B radiation initiates a suppression of the delayed-type hypersensitivity response accompanied by a generation of antigen-specific suppressor cells and an alteration of antigen-presenting function. In previous studies we and other investigators could achieve a prolongation of graft survival by a treatment of the recipient with UVB light or 8-methoxy-psoralen plus UVA light (PUVA). One of the mediators of the systemic immunomodulatory effects of ultraviolet light or PUVA may be urocanic acid, which is isomerized in the skin by ultraviolet light from its cis- to the trans-isomer. In this work we present evidence that cis-urocanic acid, generated in vitro by a treatment with UVB light or PUVA, is able to prolong the survival of allogeneic MHC disparate skin grafts in mice. In contrast, the rejection of second set grafts was not suppressed. Unirradiated (trans-urocanic acid) had no effect on allograft rejection. In a murine model cis-, but not trans-urocanic acid, prevented or delayed an acute lethal graft-versus-host disease. These experiments demonstrate the potent systemic immunomodulatory effects of cis-urocanic acid in vivo.     

Prolonged viability of human skin xenografts in rats by cyclosporine

The immunosuppressant cyclosporine (CSA) has shown usefulness in both animal and human transplantation. The present study investigated the effect of CSA in human to rat skin xenografts. Recipient rats received either a fresh split-thickness (0.020 in.) or full-thickness graft obtained from plastic surgery, or frozen cadaver skin. The graft bed of recipient Lewis rats was prepared by full-thickness excision. Animals were maintained on CSA 25 mg/kg/day X 50 days, followed by 12.5 mg/kg 2 X/week. Control animals received an equivalent volume of vehicle. All animals receiving split-thickness grafts and treated with CSA maintained their grafts significantly longer (up to 255 days) than controls. The 2 CSA-treated full-thickness grafts and the 10 vehicle-treated controls showed clinical and microscopic signs of rejection at a mean of 6.4 days. Histologic examination of successful grafts showed areas of viable epidermis with a negligible inflammatory infiltrate. There was some loss of normal polarity and occasional apoptotic pigmented basal cells. The dermis revealed moderate fibrosis, probably secondary to the surgical procedure. Graft viability was confirmed by autoradiography. Immunohistochemical staining for S-100 protein revealed morphologic alteration of suprabasilar dendritic (Langerhans-indeterminate) cells, as well as their existence in xenografts at 12 weeks posttransplantation. Toxicities reflected by weight loss and blood chemistries were felt to be dose-dependent. This in vivo model may provide a means for testing percutaneous drug penetration and pharmacokinetics in human skin, and for observing the immune component of explanted cutaneous neoplasms and dermatoses.     

Lympho-epidermal interactions in patients receiving human Langerhans cell free cultured epidermal allografts as a skin substitute

Human Langerhans cell free epithelia can be cultured in vitro, and then can be used as epidermal allografts (EAG) without evidence of rejection. We studied the cellular basis of this phenomenon with mixed epidermal cell lymphocyte reactions (MELR). The capacity of donor-derived epidermal cells to stimulate allogeneic control or recipient cells was abolished when stimulatory cells were Langerhans cell-free cultured keratinocytes and respondors were obtained prior to grafting. Donor-type cultured keratinocytes were able to induce a low response by host-derived cells in assays conducted 2, 4 and 6 weeks after grafting, but not thereafter. They were unable to stimulate allogeneic cells unrelated to the recipient. The ability of crude epidermal suspensions with 2–4% Langerhans cells to stimulate host-derived cells did not increase with time after grafting. No secondary type MELR could be evidenced, suggesting that grafting of EAG did not induce an in vivo immunization to class I antigens expressed by cultured epidermal cells. Lastly, host-derived Langerhans cells were not able to restore the allostimulatory ability of Langerhans cell free epidermal cells from EAG donors when tested against host-derived cells. This suggests that host-derived Langerhans cells which colonize the grafts during the first few weeks following grafting cannot act in the presentation of foreign keratinocyte-bound antigens, which may account for the absence of rejection noted.