小鼠角质形成细胞 MHC-Ⅰ类限制性内源性抗原递呈系统的构建

目的：建立一转染小鼠角质形成细胞系,使其表达MHC－Ⅰ类分子限制的卵白蛋白（ovalbumin,OVA）抗原。方法：将含有融合OVA基因（在角蛋白5的启动下）和小鼠H－2K^b的重组质粒转染于小鼠PAM212细胞（H－2K^d）。采用RT－RCP和流式细胞分析技术检测相应基因的转录及翻译水平的表达。结果：本实验建立了2个表达小鼠H－2K^b分子（命名为PAM－Kb）的转染细胞系,4个可表达OVA mRNA（命名为PAM－OVA）的转染细胞系和2个可同时表达小鼠H－2K^b分子和OVAmRNA的转染细胞系（命名为PAM－OK）。结论：所建立的MHC－Ⅰ限制的内源性抗原系统为进一步理解多肽／MHC－Ⅰ复合体与具有相应特异T细胞受体的CD8^ T细胞之间的相互作用奠定了基础。     

Vitiligo is associated with a significant increase in HLA-DR6 and a decrease in DQw2 antigens in Northern Italian patients.

Background Pathogenesis of vitiligo is still unclear; although the mechanism of melanocyte destruction is not know, reports of abnormalities in immunoregulation and immune reactivity in these patients support the autoimmune theory of the disease. A wide variety of autoimmune disorders have been associated with HLA antigens, and at the present time there are conflicting results about the association between vitiligo and HLA phenotypes. Objective and methods To determine if some increases in certain HLA antigens in the Northern Italian population suffering from vitiligo exist, we studied 33 patients, and we compared the results obtained with those observed in 443 control subjects of the same geographical area. Results HLA-DR and DQ typing showed a statistically significant increase of DR6 antigen and a statistically significant decrease of DQw2 type. The comparison of the phenotype frequencies of the loci A,B and C observed in the patients and in the control group showed no statistically significant differences after the p correction. Conclusion This study showed that genes in or closely linked to the HLA-DR and DO regions of the major histocompatibility complex could influence susceptibility to vitiligo in Northern Italian patients.     

角质形成细胞抗原提呈功能的研究进展

角质形成细胞不仅形成机体的物理屏障,同时具有重要的免疫功能.角质形成细胞在一些疾病状态下或者直接受到干扰素γ等细胞因子的刺激后,可以表达抗原提呈相关分子,如主要组织相容性复合体Ⅱ、细胞间黏附分子1等,同时主要组织相容性复合体Ⅰ也会发生改变.有研究表明,活化后的角质形成细胞可以加工并提呈内源性或外源性抗原,进一步活化抗原特异性T淋巴细胞.角质形成细胞的这一功能对阐明相关皮肤病的发病机制及作出针对性的靶向治疗具有重要意义.但同经典的抗原提呈细胞相比,角质形成细胞发挥抗原提呈功能可能还存在着一定的局限性.     

TAP1-EGFP和TAP2-EGFP融合蛋白在恶性黑素瘤细胞系中的表达

目的 探讨pTAP1-EGFP和(或)pTAP2-EGFP转入恶性黑素瘤细胞株后TAP表达的变化,观察TAP在A375中表达后的业细胞定位.方法 在恶性黑素瘤细胞株A375中转入pTAP1-EGFP和(或)pTAP2-EGFP,G418筛选稳定转染细胞株,检测转染后TAP1和TAP2表达水平的变化.将pDsRed2-ER和pTAP1-EGFP或pTAP2-EGFP共转染入A375细胞内,激光共聚焦显微镜下观察TAP1-EGFP和TAP2-EGFP融合蛋白的亚细胞定位.流式细胞仪检测转染前后细胞表面HIA-Ⅰ的表达.结果 将pTAP1-EGFP和(或)pTAP2-EGFP转染A375细胞株后筛选出稳定克隆.转染pTAP1-EGFP和(或)pTAP2-EGFP后能明显增加A375细胞TAP1和TAP2在蛋白水平的表达,并能增加细胞表面HLA-Ⅰ的表达.共转染pDsRed2-ER和pTAP1-EGFP或pTAP2-EGFP后,在激光共聚焦显微镜下观察,发现TAP1-EGFP和TAP2-EGFP融合蛋白的绿色荧光能够与pDsRed2-ER的红色荧光重叠.结论 构建的pTAP1-EGFP和(或)pTAP2-EGFP质粒在A375细胞系中表达成为TAP1-EGFP和TAP2-EGFP融合蛋白后,能准确定位在内质网上,为研究TAP诱导的后续免疫效应提供基础.     

Human leukocyte antigen class II alleles and natural history of HPV 2/27/57-induced common warts

Epidemiological studies have demonstrated an association between HLA-DQB1*03 alleles and the risk of cervical cancer induced by human papillomavirus (HPV). As persistence of HPV infection is required for developing cervical cancer, we wanted to elucidate the role of HLA-class II allele polymorphisms in the persistence of common warts induced by HPV 2, HPV 27 or HPV 57. Therefore, we determined the distribution of HLA-DQA1, -DQB1, and -DRB1 alleles in 71 patients presenting with HPV 2/27/57-induced common warts which had persisted for at least 18 months as well as in 92 individuals who had never suffered from common warts or whose warts had healed in less than 18 months. Among patients with long-lasting warts, the carriership frequencies and allele frequencies of DQA1*0301, DQB1*0301, DRB1*07 and DRB1*09 were higher, and the allele frequencies of DQA1*0501, DQB1*0603, DRB1*01 and DRB1*03 were lower. Statistically significant differences (Bonferroni adjusted Fishers exact test) were found for carriership frequency of DQA1*0301 (46.5 vs 21.7%, P=0.013) and for carriership frequency (18.3 vs 1.1%, P=0.0015) and allele frequency (12 vs 0.5%, P=0.000013) of DQB1*0301. A greater proportion of patients with long-lasting warts than of subjects without persistent warts were homozygous at the DQA1 (14.1 vs 6.5%) and DQB1 (16.9 vs 8.6%) gene loci. These results suggest that the natural history of cutaneous HPV 2/27/57-induced common warts may be modulated by allele polymorphisms at the HLA-DQA1 and HLA-DQB1 gene loci.     

The major histocompatibility complex and the Fugu aspect of immunity

The major histocompatibility complex (Mhc) is a set of genes that codes for plasma-membrane associated proteins, which are primarily expressed on the cells and tissues of the immune system. The proteins serve as cell markers during the initiation phase of immune response. T lymphocytes recognize Mhc molecules with their receptors when they recognize a foreign antigen on the surface of an antigen presenting cell. The functional Mhc genes are highly polymorphic. One consequence of this polymorphism is the existence of nonresponders to foreign antigens. Certain combinations of self Mhc molecules and nonself antigens fail to stimulate an immune response. A possible explanation for this nonresponsiveness is the need to make an individual unresponsive to (tolerant of) its own proteins. Tolerance leads to the inactivation of certain T cell clones; thus, if the same clones also happen to recognize a particular foreign antigen, the individual becomes unresponsive to this antigen as well. It then contains a blind spot in its repertoire.     

Promoter region polymorphism in the human TNF-α gene is not associated with lichen sclerosus

Abstract The pathogenesis of lichen sclerosus remains unknown. However, it has been frequently associated clinically with autoimmunity. The MHC haplo-type A1, B8, DR3 is associated with many autoimmune conditions and has also been associated with the uncommon allele of the tumour necrosis factor (TNF-α) promoter polymorphism. This allele is also associated with higher production of TNF in vivo and in vitro, and thus it has been speculated that it is the TNF-α gene which underlies the genetic association of many diseases with the autoimmune haplotype. There have been many reports of HLA associations with lichen scleroses, but these have not been concordant. We therefore decided to analyse the TNF-oc polymorphism in patients with lichen scleroses to determine if TNF-α was likely to play a role in susceptibility or severity of lichen scleroses. No association between alleles of the TNF-α polymorphism and lichen scleroses was found.     

Is the human skin a pheromone‐producing organ?

It is controversial whether or not humans convey specific compounds within their body odours which can potentially affect the physiology and behaviour of others. Such compounds are called pheromones and have been discovered in many other species, including mammals. It has been suggested that humans might have a special organ within their nose that can transmit such chemosensory information. However, the evidence for this organ is highly questionable. In any case, the main olfactory system is a highly diverse system, capable of transmitting pheromonal information. So far, no single substance has been found that acts as a chemical messenger for erotic attraction. On the other hand, studies investigating the pheromonal properties of natural complex body odour have proven that it does deliver information about the sender and that it has an effect on the physiology and likely behaviour of other humans. Its significance for human mating preferences probably lies not in driving them to choose the right mate but rather in warning them not to choose the wrong one.     

Polymorphisms in the SEEK1 and SPR1 genes on 6p21.3 associate with psoriasis in the Swedish population

Psoriasis is a chronic skin disease that results in red and scaly lesions. Several psoriasis susceptibility loci have been identified across the genome, of which PSORS1 on 6p21.3 is predominant. There is an ongoing debate regarding whether the HLA-C allele, Cw*0602, can be considered the major predisposing factor in this region. Investigation of other genes in the PSORS1 region with regard to psoriasis may provide alternate candidates to HLA-C. We have characterized two overlapping genes, SEEK1 and SPR1. SEEK1 encodes two putative protein isoforms: the first being one of 152 amino acids from the full-length splice-isoform (exon 1-6), and the second being one of 100 amino acids from an alternate splice-isoform (exon 1 and 6). SPR1 encodes a highly conserved protein of 134 amino acids, and in addition to characterization of human SPR1 we report the cloning of its orthologs in mouse and pig. Both SEEK1 and SPR1 are expressed in normal and psoriasis skin. In a case-control study, five of the nine single nucleotide polymorphisms (SNPs) found in SEEK1 were associated with psoriasis, while one of the four SNPs found in SPR1 showed association. Testing the Cw*0602 confounding status revealed that two of the SEEK1 SNPs showed Cw*0602-independent association, while the SPR1 SNP showed Cw*0602-dependent association. The second exon of SEEK1, containing the two Cw*0602-independent SNPs, showed the highest concentration of the psoriasis-associating SNPs, but did not appear to be translated.     

Essential concept of transplant immunology for clinical practice

Our understanding of transplant immunology has advanced from gross allograft rejection to cellular response and to current molecular level. More sensitive assays have been developed to characterize patient sensitization and to detect pre-existing donor-specific antibodies (DSA) in pre-transplant crossmatch. After a transplant, pre-existing or de novo DSA are increasingly monitored to guide clinical management. Therefore, it is important for clinicians to understand the basic concepts and key components of transplant immunology as well as be familiarized with the modern immunological techniques used in kidney transplantation.     

Dermatitis herpetiformis in a Japanese patient with anaplastic large cell lymphoma

We report a 73-year-old Japanese man with dermatitis herpetiformis which developed after diagnosis of anaplastic large cell lymphoma. The patient suffered fever, sweating, shivering, and multiple enlarged cervical lymph nodes. The diagnosis of anaplastic large cell lymphoma was confirmed by the histologic features of a biopsied cervical lymph node. The patient underwent combination chemotherapy. However, one month after the initial therapy, pruritic erythematous skin lesions with peripheral vesicles appeared on his buttocks. A skin biopsy showed subepidermal blister formation associated with polymorphonuclear and mononuclear cell infiltrates. Direct immunofluorescence examination of the area adjacent to the lesion showed granular deposits of IgA at the dermoepidermal junction. While it is well-known that dermatitis herpetiformis can develop into lymphoma, there have been only a few reports of its appearance after a diagnosis of lymphoma. This case suggests that dermatitis herpetiformis may be induced by anaplastic large cell lymphoma.     

Hereditary angioneurotic edema and HLA types in two Danish families

Summary HLA types were determined in 19 patients and 9 healthy members of 2 Danish families with hereditary angioneurotic edema. The study revealed no connections between hereditary angioneurotic edema and the HLA system.     

The connection between in vitro water uptake and in vivo skin moisturization

Background/purpose: Humectancy or hygroscopy is the water absorption tendency of a substance from the surroundings. Our interest, from the clinical point of view, consists of correlating this tendency in vitro and its effect in vivo for the development of drugs and formulations for the treatment of dry skin syndrome or diseases accompanied by dry skin. Method: In vitro, water absorption was measured using the comparative isopiestic method. This method is based on bringing the vapor of the water to isothermal equilibrium between a reference system and the material to be studied. The in vivo model on guinea‐pigs for the dry skin syndrome tested the therapeutic ability of mono‐, di‐ and tri‐glycerols to provide moisture to dry skin leading to healing. The moisture content in the stratum corneum was measured with a Corneometer CM 825^® PC that measures merely the presence of high dielectric material (humectant or water), whereas the Mexameter MX 16^® measures a pathological parameter – the erythema. Results: Adding hydroxyl groups to a consecutive set of polyhydroxyalkanes increases the humectancy of the polyols in vitro. This elevation was found to be linear at low relative humidities (Relative humidity=31.9% and 37°C). In vivo, moisture was returned to normal within a week in all three groups. However, only glycerol managed to abolish the erythema within 7 days. Conclusion: A rise in water absorption ability in vitro, at a rate of about 0.25 mol water per hydroxyl group was revealed in a consecutive set of glycerols (mono‐, di‐ and tri‐glycerols). One would expect that the better humectant is a material, i.e. in which the higher its physical ability to hold water in vitro the more effective it will be in recovering skin dryness. We have found, however, that glycerol, which has the lowest humectant activity in vitro, from the set of glycerols, di‐ and tri‐glycerol, has been proven to be the best for eliminating the signs of skin dryness. Accordingly, we propose to distinguish between the in vitro humectancy (i.e. the water uptake of a material), and its in vivo moisturizing effect, i.e. its ability to cure skin dryness and erythema. This finding supports our conclusion that the connection between in vitro humectancy and in vivo moisturization is not a simple correlation.     

The International Foundation for Dermatology: an exemplar of the increasingly diverse activities of the International League of Dermatological Societies

The International Foundation of Dermatology (IFD) was established by the International League of Dermatology Societies to promote the care of skin disease in the developing world. Starting from an initial base of the Regional Dermatology Training Centre in Tanzania it has successfully trained a cadre of clinical officers and dermatology residents from different African countries. It has now broadened this approach to an assessment of the effectiveness of focused training in Mali. The IFD is also completing a global assessment of dermatological needs in developing countries with a view to establishing guidelines and programmes for the control of common skin diseases. An ongoing strategy has been to work with other agencies to help ease the burden of other endemic tropical diseases that affect the skin; preventing the development of elephantiasis in filarial lymphoedema has been one such project implemented through a programme of skin hygiene.     

过敏原特异性T细胞检测方法的研究进展

追踪T细胞数量和功能的变化在临床诊断和评估过敏原免疫治疗疗效等方面具有重要的参考价值。本文综述了羧基荧光素琥珀碱亚胺酯连续稀释法、酶联免疫斑点检测法、胞内细胞因子染色法和微阵列免疫传感器等过敏原特异性T细胞检测方法和临床应用的研究进展,为选择和发展合适的检测方法并应用于临床提供参考。     

The human hair follicle immune system: cellular composition and immune privilege

The immunology of the hair follicle, its relationship with the 'skin immune system' and its role in hair diseases remain biologically intriguing and clinically important. In this study, we analysed the immunoreactivity patterns of 15 immunodermatological markers to determine the cellular composition and immune privilege of the human hair follicle immune system in anagen VI (growth phase). The most prominent cells located in or around the hair follicle were Langerhans cells, CD4+ or CD8+ T cells, macrophages and mast cells, whereas B cells, natural killer cells and gammadelta T cells were found very rarely. Langerhans cells (CD1a+, major histocompatibility complex, MHC class II+), and T cells (CD4+ or CD8+) were predominantly distributed in the distal hair follicle epithelium, whereas macrophages (CD68+, MHC class II+) and mast cells (Giemsa+) were located in the perifollicular connective tissue sheath. Transmission electron microscopy confirmed low numbers of immune cells in the proximal hair follicle epithelium, and very few macrophages and Langerhans cells were seen in the dermal papilla. Melanophages were observed in the connective tissue sheath and dermal papilla. MHC class I (HLA-A, -B, -C) and beta2-microglobulin immunoreactivity was found on most skin cells, but was substantially reduced on isthmus keratinocytes and virtually absent in the proximal hair follicle epithelium. Apart from the absence of Fas ligand immunoreactivity, the sharply reduced numbers of T cells and Langerhans cells, and the virtual absence of MHC class I expression all suggest that the anagen proximal hair follicle constitutes an area of immune privilege within the hair follicle immune system, whose collapse may be crucial for the pathogenesis of alopecia areata.