Examination of non-involved skin, previously involved skin, and peripheral blood for herpes simplex virus DNA in patients with recurrent herpes-associated erythema multiforme

The association between infection with HSV and the subsequent, development of erythema multiforme is well established, although the role that the virus plays in the pathogenesis of this disorder is not known. HSV DNA has been detected in cutaneous lesions of herpes-associated erythema multiforme (HAEM), and it has been suggested that the tissue damage seen in these lesions is virus-specific. In the current, prospective study, we examined biopsies of lesional, non-involved, and previously involved but healed skin, in addition lo specimens of peripheral blood, from patients with HAEM, for HSV DNA by using the polymerase chain reaction. HSV DNA was detected in lesional skin of 10 of 11 patients compared to 2 of 11 non-involved skin biopsies obtained at the same time. I ISV was present in 4 of 6 blood specimens obtained during the acute episode. Five patients returned 3 months after the acute episode resolved for biopsies of previously involved skin. HSV was detected in 4 of these 5 biopsies. Thus, the presence of HSV DNA in the skin of pal inns with HAEM appears lo be predominantly in areas of clinical involvement; the virus remains in those cutaneous sites for up to 3 months without evidence of clinical disease; and HSV DNA may be detected in the peripheral blood cells during acute HAEM. Based on these findings, we suggest that the virus plays a role in lesion development, that the skin may function as a site of viral persistence, and that hemalogenous spread of viral DNA may bean important factor in the development of HAEM.     

Detection of herpes simplex virus DNA in cutaneous lesions of erythema multiforme

The association between erythema multiforme (EM) and herpes simplex virus (HSV) infection has long been appreciated, although the exact role which HSV may play in the pathogenesis of this herpes-associated EM (HAEM), is unknown. Previous studies have suggested, but not definitively demonstrated, the presence of HSV in lesions of HAEM. The presence of HSV would support the hypothesis that an immune-mediated response directed against HSV-specific antigens in the skin is central to lesion development in HAEM. The purpose of this study was to examine lesions of EM for the presence of HSV DNA by using the polymerase chain reaction (PCR). In addition, in situ hybridization using an HSV-specific RNA probe was performed to further localize the HSV nucleic acids within the skin. DNA was extracted from formalin-fixed, paraffin-embedded specimens of cutaneous lesions of HAEM and also from EM for which no precipitating factor could be documented, otherwise known as idiopathic EM (IPEM). DNA from lesions of bullous pemphigoid served as a negative control. Using PCR to specifically amplify HSV sequences which might be present, and then performing Southern analysis, we demonstrated HSV DNA in 9/13 HAEM and 6/9 IPEM biopsies. No HSV was detected in six lesions of bullous pemphigoid. In situ hybridization of three cutaneous HAEM lesions using an 35S-labeled HSV-specific RNA probe localized the HSV nucleic acids predominantly to the epidermis. Three biopsies of chronic dermatitis, used as negative controls, did not demonstrate this specific hybridization. These findings confirm the presence of HSV in lesions of HAEM and are consistent with the concept of an HSV-specific immune-mediated pathogenesis for this disease. In addition, most cases of IPEM appear to be herpes associated despite the absence of clinically apparent HSV infection.     

Herpes simplex virus (HSV)-associated erythema multiforme (HAEM): a viral disease with an autoimmune component

Erythema multiforme (EM) is a clinical conundrum the name of which reflects the broad morphological spectrum of the lesions. Molecular and immunologic evidence that herpes simplex virus (HSV) causes a subset of EM lesions [herpes-associated EM (HAEM)] is reviewed, and new data are presented which suggest that autoreactive T-cells triggered by virus infection play an important role in HAEM pathogenesis. Disease development begins with viral DNA fragmentation and the transport of the DNA fragments to distant skin sites by peripheral blood mononuclear cells (PBMCs). HSV genes within DNA fragments deposited on the skin [notably DNA polymerase (Pol)] are expressed, leading to recruitment of HSV-specific CD4+ Th1 cells that respond to viral antigens with production of interferon-gamma (IFN-gamma). This step initiates an inflammatory cascade that includes expression of IFN-gamma induced genes, increased sequestration of circulating leukocytes, monocytes and natural killer (NK) cells, and the recruitment of autoreactive T-cells generated by molecular mimicry or the release of cellular antigens from lysed cells. The PBMCs that pick up the HSV DNA [viz. macrophages or CD34+ Langerhans cells (LC) precursors], their ability to process it, the viral proteins expressed in the skin and the presence of epitopes shared with cellular proteins may determine whether a specific HSV episode is followed by HAEM development. Drug-associated EM (DIEM) is a mechanistically distinct EM subset that involves expression of tumor necrosis factor alpha (TNF-alpha) in lesional skin. It is our thesis that the polymerase chain reaction (PCR) assay for HSV DNA detection in lesional skin and staining with antibodies to IFN-gamma and TNF-alpha, are important criteria for the diagnosis of skin eruptions and improved patient management.     

Recurrent herpes simplex virus infections and erythema multiforme: a report of three patients

The widespread preoccupation of the media and patients with herpes prompts this report of two patients who developed erythema multiforme, an allergic response to the herpes simplex virus. While this complication is not new, physicians and nurses working in clinics for sexually transmitted diseases should be aware of this allergic response to infection with herpes simplex virus.     

Identification of herpes simplex virus DNA in lesions of erythema multiforme by the polymerase chain reaction.

An association between erythema multiforme and herpes simplex virus infection has been supported by clinical studies and by the detection by immunofluorescence of herpes viral antigen in sera and skin biopsy specimens of patients with erythema multiforme. In rare cases, the virus has also been isolated in cultures of skin biopsy specimens of erythema multiforme. To investigate further the association between erythema multiforme and herpes simplex virus, we used the polymerase chain reaction for herpes simplex virus to examine skin lesions from patients with erythema multiforme. In this study herpes simplex virus DNA was detected in 11 of 31 biopsy specimens of erythema multiforme; six additional cases showed equivocal amplification results, which is suggestive of low amounts of viral DNA. Seven skin and mucosal biopsy specimens with the histologic changes of herpes virus infection served as positive controls: all were positive for herpes simplex virus DNA. Viral DNA was not detected in control biopsy specimens from skin excised for unrelated conditions. These studies support the association of herpes simplex virus in the pathogenesis of some cases of erythema multiforme. The polymerase chain reaction provides a quick and effective method of detecting herpes simplex virus in lesions of herpes-associated erythema multiforme. Furthermore, the polymerase chain reaction may delineate those cases of erythema multiforme that are etiologically related to herpes virus infection and therefore might be treated with acyclovir to prevent recurrence.     

HLA typing of patients with herpes simplex recidivans and postherpetic erythema exsudativum multiforme

62 patients suffering from recurrent herpes simplex, with ten out of them showing postherpetic erythema multiforme, have been investigated with special reference to histocompatibility standardisation. In comparison with a control group (n = 170), our patients revealed a high and statistically significant level of HLA-B5 (p less than 0,01). Postherpetic erythema multiforme seems to be associated with HLA-A9.     

Detection of herpes simplex virus DNA in the peripheral blood during acute recurrent herpes labialis.

BACKGROUND: Although herpes simplex virus (HSV) has been detected in the peripheral blood of immunocompromised patients and in neonates with disseminated disease, the extent to which this virus may be present in the blood during a localized infection in otherwise healthy adults is unknown. OBJECTIVE: The purpose of this study was to determine whether HSV may be detected in the peripheral blood during acute recurrent herpes labialis. METHODS: Peripheral blood mononuclear cells (PBMCs) were obtained from otherwise healthy adults with recurrent herpes labialis, both during an acute episode and several weeks after the lesions had healed. The PBMCs were examined for the presence of HSV with the polymerase chain reaction (PCR) and viral culture. RESULTS: By PCR, HSV DNA was detected in 7 of 34 specimens from an acute episode but in none of 24 specimens in the convalescent stage (p less than 0.004). PBMCs from seven donors, who were seronegative for HSV, were also negative for HSV by PCR. Viral cultures of 22 PBMC specimens were negative (including four specimens that were positive by PCR). CONCLUSION: The presence of HSV DNA in the blood is a transient phenomenon limited to the period of active infection in a minority of patients with herpes labialis, although it may be important in the development of disseminated disease as well as in the pathogenesis of herpes-associated cutaneous processes such as erythema multiforme.     

复发性生殖器疱疹患者外周血CD4+CD25+Foxp3+调节性T细胞的检测

目的 研究外周血CD4⁺CD25⁺调节性T细胞(Treg细胞)表型和功能改变所致的细胞免疫抑制效应在复发性生殖器疱疹(RGH)发病机制中的作用.方法 采用免疫荧光标染技术,经流式细胞仪检测34例RGH患者和33例正常人外周血CD4⁺CD25⁺T细胞转录因子Foxp3及细胞毒T细胞相关抗原4(CTLA-4)、糖皮质激素诱导的肿瘤坏死因子受体家族相关基因(GITR)、程序性死亡1(PD-1)等抑制性膜分子的表达水平.同时,用免疫磁珠细胞分选技术从外周血中分选出高纯度的CD4⁺CD25⁺T细胞,经体外刺激后分析胞内分泌白介素10(IL-10)和转化生长因子β(TGF-β)阳性细胞数.结果 RGH患者外周血CD4⁺CD25⁺Foxp3⁺Treg细胞数量比正常人对照组显著增加(P<0.001),CD4⁺CD25⁺T细胞表面抑制性膜分子CTLA-4、GITR、PD-1表达明显增强(P分别<0.001、<0.001及<0.01),胞内IL-10和TGF-β阳性细胞增多(P值分别<0.01及<0.001).结论 单纯疱疹病毒感染可诱导CD4⁺CD25⁺Treg细胞活化增殖,高表达负性共刺激分子和分泌抑制性细胞因子,通过多种机制抑制机体的抗病毒免疫应答.     

Association of herpes simplex virus-induced erythema multiforme with the human leukocyte antigen DQw3

We studied the genetic markers in human leukocyte antigen (HLA) region HLA-A, -B, -C, -DR, -DQ, C2, Bf, C4A, and C4B of 31 patients with erythema multiforme (EM) and their families. In contrast to the complement allotypes, which showed no deviation from the distribution in the normal population, two HLA class II antigens occurred in much higher frequency in patients with EM. The frequency of HLA-DQw3 (77.4%) increased with high significance compared with normal Caucasian control individuals (41.2%). An even stronger DQw3 association was found in the patient group with postherpetic EM (88.8%; relative risk, 9.41). Interestingly, all patients suffering from frequently recurrent EM were found to have the DQw3 allele (relative risk 44.2). The previously reported association to HLA-B15 was also seen and may be due to a linkage disequilibrium with the HLA-DR4 allele (66.6% in recurrent EM). Our data provide further evidence that classic recurrent EM is related to herpes simplex virus infection and should be regarded as a distinct entity within the enigmatic EM syndrome. DQw3 may serve as a helpful marker for distinguishing this entity from other diseases with EM-like lesions.     

Complement deposition in the skin of patients with herpes-associated erythema multiforme

Granular staining for C3 by direct immunofluorescence is a frequent finding along the dermoepidermal junction and in papillary blood vessels in the early skin lesions of erythema multiforme. In order to evaluate whether the complement cascade is activated by the classical or alternative pathway, ten biopsies from patients with herpes-associated erythema multiforme, which were positive for granular C3 along the dermoepidermal junction, were stained by an immunofluorescence technic for other complement components. Staining for the components of classical pathway, C1q and C4, were found in none of the ten biopsies. However, in nine of ten biopsies, granular staining for properdin was present along the dermoepidermal junction. These findings suggest complement activation by the alternative complement pathway in herpes-associated erythema multiforme.     

寻常型进展期白癜风患者外周血 CD4+CD25+调节性T细胞检测

目的：探讨寻常型进展期白癜风患者外周血T细胞亚群及CD4^＋CD25^＋调节性T细胞（Treg）的水平变化及其意义。方法：采用流式细胞分析技术对45例寻常型进展期白癜风患者和24例正常人外周血CD4^＋、CD25^＋、Foxp^3＋,CD^4＋CD25^＋,CD4^＋CD25^＋Foxp3^＋T细胞水平。结果：寻常型进展期白癜风患者外周血CD4^＋CD25^＋Treg和CD4^＋CD25^＋Foxp3^＋nTreg细胞数量显著高于正常对照组（P〈0．05）。结论：寻常型进展期白癜风患者外周血Treg水平发生变化,使体内免疫功能处于失衡状态,细胞免疫尤其是Treg可能参与了白癜风的发病。     

Detection of spirochaetal DNA simultaneously in skin biopsies, peripheral blood and urine from patients with erythema migrans

Lyme borreliosis is an emerging zoonosis transmitted by infected hard-bodied ticks. The disease is multisystemic. In the initial stage its typical manifestation is the erythema migrans, a cutaneous lesion that occurs in up to 90% of patients. In order to investigate the presence of the specific agent, Borrelia burgdorferi, in the early stages of the disease, DNA from skin biopsies, urine and peripheral blood of 30 patients with clinically documented erythema migrans and without apparent systemic involvement was analysed by polymerase chain reaction. Borrelia DNA in both blood and skin biopsies was detected in 23 patients, while in 9 patients it was discovered in urine and skin biopsies. These results demonstrate that Borrelia DNA is detectable systemically also in patients with early Lyme borreliosis and strongly suggest a possible dissemination of the causative agents even when only a local infection is assumed.     

扁平苔藓患者外周血CD4+CD25+Treg细胞的检测

目的:检测扁平苔藓患者外周血CD4+CD25+调节性T细胞(regulatory T cells,Treg)的数量及FOX3 mRNA的表达.方法:采用流式细胞术对25例扁平苔藓患者和25例健康对照者外周血CD4+CD25+Treg细胞进行检测,并通过RT-PCR方法检测FOXP3转录因子在外周血单个核细胞的表达.结果:泛发性扁平苔藓和口腔扁平苔藓患者外周血CD4+CD25+ Treg细胞数分别为(2.68±2.21)%和(2.38±2.67)%,均低于健康对照者(7.87±1.77)%;扁平苔藓患者的FOPX3表达水平也低于健康对照(P<0.05).结论:CD4+CD25+ Treg细胞数量和功能下降,可能通过导致扁平苔藓患者免疫功能紊乱参与了扁平苔藓的发病.     

Immune responses to herpes simplex virus in patients with facial herpes simplex and those with eczema herpeticum

The immune response to herpes simplex virus (HSV) was studied in 59 patients with primary and recrudescent facial HSV infections. The patients included nine with atopic eczema, seven of whom had eczema herpeticum (EH). All patients had antibodies to HSV (measured by ELISA) and all but three had HSV-specific cell mediated immunity (CMI) (measured by in vitro lymphoproliferation). Thirteen control subjects were negative for both tests. All three patients with absent CMI to HSV had suffered from severe EH and had depressed CMI to HSV for several months following an attack. In two of these EH patients, a positive CMI response was produced by in vitro removal of CD8 + ve T lymphocytes from peripheral blood mononuclear cells using a panning technique. Thus the absence of CMI to HSV in these patients was due to suppressor cell function rather than a lack of specifically responsive cells. The other four EH patients with normal CMI to HSV had suffered less severe EH, but no association between the absence of CMI to HSV and serum IgE level or activity of the eczema was apparent in the atopic patients. No specific anti-HSV IgE antibody was detectable.     

梅毒患者外周血CD4+CD25bright调节性T细胞的检测

目的 探讨CD4^＋CD25^bright调节性T细胞在梅毒患者免疫学变化中的作用。方法 利用流式细胞仪和单克隆荧光抗体检测梅毒患者外周血中CD4^＋CD25^bright调节性T细胞在CD3^＋CD4^＋辅助性T细胞（Th）中的比例。结果 血清固定组梅毒患者外周血中调节性T细胞比例较正常人对照组显著增高（P〈0．01），而一期、二期及潜伏梅毒患者调节性T细胞比例与正常人对照组差异无统计学意义（P〉0．05）。各组梅毒患者外周血调节性T细胞比例与血清RPR滴度无相关性（P〉0．05）。结论 梅毒患者外周血中CD4^＋CD25^bright调节性T细胞比例增高造成的免疫抑制可能在梅毒血清固定的发生中起着一定的作用。