Prevalence and Viral Loads of Cutaneous Human Polyomaviruses in the Skin of Patients With Chronic Inflammatory Skin Diseases.

BACKGROUND: Human skin microorganisms have been associated with various skin diseases. However, most studies have focused on bacterial communities, and little is known about normally resident skin viruses such as the Polyomaviridae and their association with cutaneous disorders. METHODS: We investigated the infection levels of Merkel cell polyomavirus (MCPyV), human polyomavirus 6 (HPyV6), and human polyomavirus 7 (HPyV7), using triplet skin swabs collected from lesional and nonlesional skins of 86 Japanese patients with inflammatory skin diseases and mycosis fungoides, and from 149 healthy control individuals. RESULTS: This age-matched case-control study provides the first analyses of the loads of polyomaviruses in association with various skin diseases. The viral loads were significantly higher for HPyV6/HPyV7 and lower for MCPyV in patients with psoriasis. The viral load variation was observed not only at lesion sites, but also at clinically unaffected skin sites in most of the patients. The viral strains tested were all of the Asian/Japanese genotype. CONCLUSIONS: Our findings suggest a covariation in the infection levels of cutaneous polyomaviruses in certain inflammatory skin conditions. Worldwide prospective longitudinal studies are warranted to understand the influence of such alterations on the pathogenesis of inflammatory skin disorders.

[Detection of HTLV-1 DNA in biopsies of Chilean patients with cutaneous T-cell lymphoma]. / Búsqueda del ADN del virus HTLV-1 en biopsias de pacientes con linfoma cutáneo de células T.

BACKGROUND: Human T-lymphotropic virus-1 (HTLV-1) infection has been associated with the pathogenesis of cutaneous T cell lymphomas (CTCL). AIM: To search for HTLV-1 DNA in skin biopsies of patients with CTCL. MATERIAL AND METHODS: A retrospective study was conducted using 25 biopsies of patients with CTCL. DNA was extracted from lymphoid tissue by microdissection. A nested PCR was conducted to detect HTLV-1 genome using primers for the tax region. As negative controls, four cases of superficial perivascular dermatitis were chosen. As positive controls, five cases of T-cell leukemia/lymphoma (ATCL) were studied. RESULTS: A positive reaction was found in 3 of 25 cases. These biopsies corresponded to a case of Mycosis Fungoides, a case of CD30 (-) T-cell lymphoma and a case of lymphomatoid papulosis. Search was negative in the four cases of superficial perivascular dermatitis and positive in four cases of adult T-cell leukemia/lymphoma (ATCL). CONCLUSIONS: HTLV-1 DNA search in tissues is a useful tool recommended to study T-cell lymphomas. HTLV-1 infection only occurs in sporadic cases but may contribute to tumor aggressiveness and prognosis.

Merkel cell polyomavirus: its putative involvement in a particular subset of cutaneous lymphoma with possibly unfavorable outcome.

BACKGROUND: A major etiologic hypothesis in primary cutaneous T-cell lymphomas is a defective lymphocyte apoptosis after antigenic activation, which could be induced by various infectious agents. OBJECTIVE: To investigate the presence of the possibly lymphotropic and folliculotropic Merkel cell polyomavirus (MCPyV) DNA and proteins in folliculotropic mycosis fungoides (fMF). STUDY DESIGN: Fresh-frozen and fixed skin biopsies were collected in lesional and non-lesional skin from 24 fMF patients, in lesional skin from 22 patients with various T-cell mediated skin benign infiltrates (TSBI) and in normal-appearing skin from 22 healthy individuals (HI). Detection and quantification of MCPyV DNA were carried out using real-time PCR; MCPyV genome integration status was presumed through a previously described differential real-time PCR (MCPyV ΔC-TAg) targeting a constantly conserved sequence versus an integration-induced deleted sequence. The MCPyV proteins expression was assessed by immunohistochemistry using antibodies targeting a tumoral antigen or a capsid protein. RESULTS: Although MCPyV DNA was similarly detected in lesional versus non-lesional fMF samples (50% each), in 36.4% HI and 40.9% TSBI; viral load was significantly higher in fMF lesional samples versus HI and TSBI. The integration of the viral genome appeared unlikely. The MCPyV proteins expression was exclusively observed inside skin appendages in 18.2% of the fMF lesional skin samples. CONCLUSION: MCPyV genome detection rate was similar in all skin samples, but MCPyV viral load was significantly higher in fMF lesions versus TSBI and HI, although the viral genome was probably not integrated. Episomal MCPyV DNA may be expressed in skin appendages in fMF.

Búsqueda del ADN del virus HTLV-1 en biopsias de pacientes con linfoma cutáneo de células T / Detection of HTLV-1 DNA in biopsies of Chilean patients with cutaneous T-cell lymphoma

Background: Human T-lymphotropic virus-1 (HTLV-1) infection has been associated with the pathogenesis of cutaneous T cell lymphomas (CTCL). Aim: To search for HTLV-1 DNA in skin biopsies of patients with CTCL. Material and Methods: A retrospective study was conducted using 25 biopsies of patients with CTCL. DNA was extracted from lymphoid tissue by microdissection. A nested PCR was conducted to detect HTLV-1 genome using primers for the tax region. As negative controls, four cases of superficial perivascular dermatitis were chosen. As positive controls, five cases of T-cell leukemia/lymphoma (ATCL) were studied. Results: A positive reaction was found in 3 of 25 cases. These biopsies corresponded to a case of Mycosis Fungoides, a case of CD30 (-) T-cell lymphoma and a case of lymphomatoid papulosis. Search was negative in the four cases of superficial perivascular dermatitis and positive in four cases of adult T-cell leukemia/lymphoma (ATCL). Conclusions: HTLV-1 DNA search in tissues is a useful tool recommended to study T-cell lymphomas. HTLV-1 infection only occurs in sporadic cases but may contribute to tumor aggressiveness and prognosis.

EBV+ cutaneous B-cell lymphoproliferation of the leg in an elderly patient with mycosis fungoides and methotrexate treatment.

A 77-year-old man with a 5-year history of mycosis fungoides (MF) who had received several lines of therapy, including intravenous courses of Methotrexate (MTX) for the past 2 years, went on to develop several ulcerated cutaneous nodules on the left leg. Biopsy revealed diffuse sheets of EBV-positive large B cells (CD20+ CD30 ± IgM Lambda), with an angiocentric distribution and a monoclonal IGH gene rearrangement. Although the pathological features were diagnostic for an EBV-positive diffuse large B-cell lymphoma (DLBCL), several possibilities could be considered for assignment to a specific entity: EBV-positive DLBCL of the elderly, methotrexate-induced lymphoproliferative disorder (LPD), lymphomatoid granulomatosis, or the more recently described EBV-positive mucocutaneous ulcer. The development of EBV+ lymphoproliferations has been reported in two other patients with MF under MTX, and occurred as skin lesions of the leg in one of these and in the current case, which may question the relatedness to primary cutaneous DLCBL, leg-type.

Human herpesvirus 8 is not detectable in lesions of large plaque parapsoriasis, and in early-stage sporadic, familial, and juvenile cases of mycosis fungoides.

BACKGROUND: Human herpesvirus (HHV) 8, an essential etiologic agent of Kaposi sarcoma, is also associated with several lymphoproliferative disorders. The involvement of HHV 8 in mycosis fungoides (MF) and large plaque parapsoriasis (LPP) is controversial, with contradictory reports from various countries worldwide. OBJECTIVE: We sought to investigate the presence of the HHV 8 genome in skin lesions of LPP and early-stage sporadic, familial, and juvenile MF in patients in Israel. METHODS: Archival paraffin-embedded and frozen samples from skin biopsies of untreated patients with LPP and early-stage MF performed in 1990 through 2006 were randomly collected from the department of dermatology of a tertiary medical center in central Israel. DNA was extracted, and a TaqMan-based real-time polymerase chain reaction assay specific for the K6 gene region was used to detect the HHV 8 genome. RESULTS: A total of 46 biopsies were sampled from 11 patients with LPP and 35 with early-stage MF (17 adults with sporadic MF, 10 children, and 8 patients with familial MF). In all, 44 samples were negative for HHV 8 DNA; two samples from adults with sporadic MF were positive. LIMITATIONS: The presence of HHV 8 antibodies or virus sequences was not assessed in peripheral blood. CONCLUSION: The results of this study, conducted in a region relatively endemic for HHV 8, support most earlier studies showing a lack of association of HHV 8 infection with LPP and sporadic adult-type MF. To our knowledge, the lack of association of HHV 8 infection with juvenile and familial MF has not been previously reported.

Evidence of multiple infectious agents in mycosis fungoides lesions.

The etiology of mycosis fungoides (MF) remains to be determined. Several studies have proposed a viral etiology with controversial results. In this case-control study we investigated the presence of Epstein-Barr virus (EBV) and the debated presence of Human T-cell lymphotrophic virus I (HTLV-I) sequences, by polymerase chain reaction on nucleic acid extracts from formalin-fixed paraffin-embedded skin biopsies. Moreover, by a multivariate approach we analyzed in the same case-control study also the contribution of two previously examined pathogens: Hepatitis C virus (HCV) and Borrelia burgdorferi (Bb). Significant differences in the frequency of infectious agents in cases and controls were detected for Bb, HTLV-I and EBV. In MF patients we found the concurrent presence of two or three of these pathogen sequences in 21 out of 83 cases, but only in 1 out of 83 healthy controls. Our results suggest that the persistence of multiple infectious agents may cause a long-term antigenic stimulation contributing to the malignant transformation of T lymphocytes, especially when associated with HTLV-I like sequences. However, these infectious agents do not seem to have effects on disease progression.

Cytomegalovirus: its potential role in the development of cutaneous T-cell lymphoma.

To investigate the potential role of CMV in cutaneous T-cell lymphoma (CTCL), we studied cytomegalovirus (CMV) seroprevalence in parapsoriasis (PP), mycosis fungoides (MF) and Sézary syndrome (SS) compared with healthy control patients. In cases where CMV seropositivity was observed, CMV PCR analyses were performed on skin biopsies. CMV seroprevalence was 37.1% in the control group, 50.68% in the PP + MF + SS group (P = 0.08), 56.2% in the MF + SS group (P = 0.07), 40% in the PP group (P = 0.9), 66.67% in the MF group (P = 0.009), 42.86% in the SS group (P = 0.9). CMV PCR in initial skin biopsies were all negative. However, PCR CMV was positive in two SS skin biopsies realized at an advanced stage. Our results show that latent CMV infection may play a role in the susceptibility of MF in predisposed subjects by inducing T-cell proliferation and resistance to apoptosis. Concerning SS, an immunosuppressive state may be responsible for CMV reactivation that in turn may interfere with evolution of the disease.

Evaluation of the role of human herpes virus 6 and 8 in parapsoriasis.

INTRODUCTION: The aetiology of mycosis fungoides and parapsoriasis (which may be considered as an early stage of mycosis fungoides) remains debated. Previous recent studies have suspected the involvement of viral agents and particularly human herpes viruses (HHV).The aim of the present study was to screen for the presence of HHV-6 and HHV-8 genome in parapsoriasis samples. METHOD: Fifty paraffin-embedded samples from skin biopsies of parapsoriasis were retrospectively collected from archival files in our Dermatology department. Total DNA was extracted from samples using the phenol-chloroform method and the presence of viral genomes was screened using real-time PCR. RESULTS: Forty nine out of the fifty tissue samples of parapsoriasis were interpretable, they were all found negative for HHV-6 and HHV-8. DISCUSSION: This study does not confirm the suspected role of HHV-6 or -8 in parapsoriasis. HHV-8 has been the most studied virus in parapsoriasis and more widely in cutaneous lymphoproliferative diseases and our results are in agreement with most of the studies which found none or few HHV-8 in more advanced stages of cutaneous lymphoproliferative diseases. Concerning HHV-6, our study is the first one investigating the presence of this virus in lesional tissue samples of patients with parapsoriasis. In conclusion, parapsoriasis does not seem to be associated with either HHV-6 or HHV-8.

High association of human herpesvirus 8 in large-plaque parapsoriasis and mycosis fungoides.

OBJECTIVE: To investigate the presence of human herpesvirus 8 (HHV-8) in lesional skin of German patients with large-plaque parapsoriasis (LPP) or mycosis fungoides (MF). The pathogenetic relevance of HHV-8 in cutaneous T-cell lymphoma is controversial. Recently, a highly significant association of HHV-8 in LPP was found, which suggests a role in the pathogenesis of this disease. DESIGN: Retrospective study of the presence of HHV-8 in German patients with lymphoproliferative diseases. SETTING: Dermatologic clinic at a university hospital of the Ruhr University Bochum, Bochum, Germany. PATIENTS: Fifty-three patients treated for lymphoproliferative skin diseases were included in the study, including 14 patients with LPP, 31 with different stages of MF, and 8 with lymphomatoid papulosis (LyP). Twenty-three patients with Kaposi sarcoma (KS) made up the positive control group, and 10 patients with atopic dermatitis served as negative controls. MAIN OUTCOME MEASURES: The presence of HHV-8 was analyzed from paraffin-embedded lesional tissue samples using a nested polymerase chain reaction for the open reading frame (ORF) 26 and with immunohistochemical staining for the latency-associated nuclear antigen (LANA) encoded by ORF 73. RESULTS: A high association of HHV-8 infection in both lymphoproliferative skin diseases was observed: 87% of LPP and 70% of MF tissue samples tested positive for HHV-8 DNA from ORF 26. However, HHV-8 was not detectable in LPP and MF by using the immunohistochemical marker LANA. CONCLUSIONS: A virus unambiguously associated with KS, HHV-8 was frequently detected at low amounts in LPP and MF specimens. However, based on the methods of HHV-8 detection used in this study, no conclusion can be drawn on the etiologic and pathogenetic role of HHV-8 in these diseases.

Human T-cell lymphotropic virus type 1 provirus and phylogenetic analysis in patients with mycosis fungoides and their family relatives.

BACKGROUND: Mycosis fungoides (MF) is a cutaneous T-cell lymphoma of unknown aetiology. A pathogenic role of human T-cell lymphotropic virus type 1 (HTLV-1) has been suggested but remains controversial. To determine whether MF is linked to HTLV-1. METHODS: Blood samples were collected from 60 patients, 15 family relatives of patients with MF (MFRs), 20 healthy controls and 10 patients with HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP). The presence of HTLV-1 antibodies in serum was tested by the Western blot rp21e-enhanced test. DNA was extracted from the blood with the Qiagen blood kit. We used 500 ng of DNA either in conventional HTLV-1-specific polymerase chain reaction (PCR) or in real-time PCR using primers sk43 and sk44 together with a tax-specific fluorescent probe. RESULTS: In Western blot, antibodies against three to four HTLV-1 antigens were detected in 52% of patients with MF. All of the patients with HAM/TSP were positive, while only 7% of the MFRs and none of the 20 healthy controls reacted with HTLV-1 antigens in Western blot. One of 60 patients with MF and one of 15 MFRs were positive in HTLV-1 PCR. These two PCR-positive samples which were quantified in real-time PCR showed that fewer than five in 10(6) cells were HTLV-1 infected. We succeeded in amplifying and sequencing the 5' end of the provirus from the blood of the PCR-positive MFR by seminested PCR. A positive result was also obtained in this test. Phylogenetic tree analyses revealed a high homology of this sequence with other HTLV-1 sequences from the Middle East. The above PCR-positive MFR was the brother of a PCR-negative patient with MF. CONCLUSIONS: These findings demonstrate that HTLV-1 is probably not the aetiological agent of MF. However, it may play a role in immunosuppression and in the spreading of the disease.

Conjugal transformed mycosis fungoides: the unknown role of viral infection and environmental exposures in the development of cutaneous T-cell lymphoma.

The etiology of mycosis fungoides (MF) is uncertain, although infectious agents and other environmental exposures have been implicated. We describe what appears to be the first case in which both a husband and his wife were diagnosed with large-cell transformation of MF. After 10 years of having stage I MF, the wife developed tumors that showed sheets of large transformed cells with dysplastic nuclei on skin biopsies, leading to a diagnosis of transformed MF. Her husband was diagnosed 14 months later with transformed MF following a biopsy of his right arm and leg after a 15-year history of presumed psoriasis. The fact that this rare occurrence happened in a couple who had been married for more than 25 years points to a common environmental exposure. Future studies should aim to clarify the potential role of infectious agents, such as human T-lymphotropic virus I and II, cytomegalovirus, Epstein-Barr virus, and other environmental exposures, in the development of MF.