Near infrared photoimmunotherapy targeting the cutaneous lymphocyte antigen for mycosis fungoides.

Background: Mycosis fungoides (MF) is a low-grade T-cell lymphoma with primary cutaneous involvement accounting for more than half of all primary cutaneous lymphomas. The treatment of MF is very challenging due to the limited therapies available. Near-infrared photoimmunotherapy (NIR-PIT) is a newly developed and highly selective cancer treatment that employs a monoclonal antibody conjugated to a photo-absorber dye, the hydrophilic phthalocyanine IRdye 700DX® (IR700), and near infrared light. In this study, we investigated the effect of NIR-PIT on MF targeting the cell-surface antigen cutaneous lymphocyte antigen (CLA)Matherial and methods: MF derived My-La CD4+ cells were incubated with the anti-CLA antibody conjugated to IR700 and then irradiated with a 690 nm near-infrared light. Cell death was evaluated by propidium iodide staining and flow cytometry 24 hours after irradiation.Results: Treatment with anti-CLA or light irradiation exhibited very modest pro-death effects, whereas treatment with the anti-CLA antibody conjugated to IR700 and then irradiation with a 690 nm near-infrared light induced a substantial increase in death in the MF cell line.Conclusions: NIR-PIT targeting CLA to treat MF showed marked antitumour effects. As such, CLA-targeted NIR-PIT could be a promising treatment for MF and, possibly, other cutaneous diseases characterized by CLA+ skin infiltrating T-cells.

Salivary Samples for the Diagnosis of Pemphigus vulgaris Using the BIOCHIP Approach: a Pilot Study.

Pemphigus vulgaris (PV) is a rare autoimmune intraepithelial blistering skin disease characterized by the presence of circulating autoantibodies against desmoglein 3 (DSG3) and desmoglein 1 (DSG1), resulting in loss of the normal epithelial cell-to-cell adhesion, through a process called acantholysis. In recent years, a BIOCHIP-based indirect immunofluorescence technique for the determination of anti-DSG3 and anti-DSG1 autoantibodies has been described. Even though, the use of saliva anti-DSG3 and anti-DSG1 ELISA for the diagnosis of PV has been already reported, there are no studies concerning the utilization of saliva by the BIOCHIP approach. In the present pilot study, ELISA and BIOCHIP were performed, using salivary and serum samples from the same patients to investigate if the detection of anti-desmoglein autoantibodies in salivary samples by BIOCHIP could be used as a test for the diagnosis of PV. There was a strong correlation between ELISA and BIOCHIP results both for anti-DSG3 and anti-DSG1 serum autoantibodies. Autoantibodies to DSG3 were detected in 8 out of 8 salivary samples by ELISA and in 6 out of 8 salivary samples by the BIOCHIP approach. Autoantibodies to DSG1 were negative in all salivary samples using both ELISA and BIOCHIP. There were no positive results in the negative control group. In conclusion, the results of this pilot study indicate lack of correlation between serum and salivary results using both ELISA and BIOCHIP, indicating that saliva may not be the ideal substrate for the laboratory diagnosis of PV using these approaches.

TLR7 Gln11Leu single nucleotide polymorphism and response to treatment with imiquimod in patients with basal cell carcinoma: a pilot study.

AIM: The Toll-like receptor 7 (TLR7) agonist, imiquimod, offers a topical and noninvasive therapeutic method for the clinical treatment of superficial basal cell carcinoma (BCC). In this study we explored the relationship between the functional X-linked TLR7 rs179008/Gln11Leu polymorphism and the response to imiquimod in patients with BCC. PATIENTS & METHODS: Thirty-four BCC patients treated with imiquimod were included in the study. SNP genotyping of the TLR7 promoter polymorphism was performed by TaqMan allelic discrimination assay. RESULTS: In the group of female nonresponders to imiquimod a higher frequency of the altered genotype compared with responders was observed. Similarly, in the group of male nonresponders to imiquimod both patients with the mutated genotype were nonresponders. CONCLUSION: The results of this study show that patients carrying at least one T allele of the TLR7 promoter polymorphism are associated with an increased probability to be resistant to imiquimod therapy.

Effects of TNF-alpha inhibitors on the number of epidermal Langerhans cells in uninvolved skin of psoriatic patients: a pilot study.

Only limited data are available on the effects of TNF-alpha inhibitors on dendritic cells. However, TNF-alpha plays a central role in the biology of dendritic cells, both with regard to their maturity process and mobilization to secondary lymphoid organs. In particular, the effects of TNF-alpha inhibitors on Langerhans cells in healthy skin have never been investigated. In this pilot study, we aimed to assess the change of the density of Langerhans cells within the normal, not photo-exposed, skin of 17 psoriatic patients, before and after 16 weeks of treatment with TNF-alpha inhibitors. Most of the patients (88%) showed an increase or a similar density of Langerhans cells after 16 weeks of therapy with TNF-alpha inhibitors compared with baseline values. Only 2 patients (12%) showed a reduction of these cells following therapy with TNF-alpha inhibitors.

Biochip technology for the serological diagnosis of bullous pemphigoid.

Bullous pemphigoid is an autoimmune blistering skin disease characterized by the presence of circulating autoantibodies which recognize specific proteins of the epidermis and dermoepidermal junction. Diagnosis is based on clinical criteria and laboratory investigations, notably histology, direct and indirect immunofluorescence, and ELISA. This study describes a new immunofluorescence assay for parallel determination of anti-BP180 and anti-BP230 based on recombinant antigenic substrates. The aim of the study was to detect BP180 and BP230 autoantibodies by BIOCHIP technology using both a specially designed recombinant BP180-NC16A protein and cells expressing the BP230-gc antigen fragment. 18 patients with bullous pemphigoid were included in the study. Autoantibodies to BP180 were detected by the BIOCHIP technique in 83.33% of patients with clinical, serological, and immunohistological confirmed bullous pemphigoid while autoantibodies against BP230-gC were detected only in 39% of patients. The detection of anti-BP180-NC16A and anti-BP230-gC by a new biochip-based immunoassay is a suitable alternative to indirect immunofluorescence and ELISA. This method has the advantage of easily discriminating the different autoantibody specificities. The BIOCHIP method is faster, cheaper, and easy to use when compared with the ELISA approach. For this reason, the new method could be used as an initial screening test to identify patients with bullous pemphigoid, and doubtful results could then be confirmed by ELISA.

CD7 expression in reactive and malignant human skin T-lymphocytes.

CD7, a molecule normally expressed on 90% of CD4+ T cells, is often deficient on the malignant T cells of cutaneous T cell lymphoma. Therefore deletion of CD7 is considered a specific marker for the diagnosis of cutaneous T cell lymphomas. Because an expansion of CD4+CD7- cells may also be observed in benign lymphocyte-mediated dermatoses, we present our experience concerning CD7 expression in both cutaneous T cell lymphomas and a broad variety of T cell-mediated inflammatory dermatoses using an immunohistochemical approach on frozen sections from 45 patients. No, or at most scarce, expression of CD7 was detectable in the inflammatory skin conditions investigated as compared to CD3-positive cells. In cutaneous T cell lymphomas, a striking reduction of CD7 reactive cells was found in either reactive or malignant T cell components. Our findings indicate that CD7-negative T cells are more common within both benign and neoplastic T cell infiltrates than was previously demonstrated and suggest that, using a conventional immunoenzymatic technical approach on fresh-frozen sections, CD7 deletion is an unreliable criterion for the immunohistological diagnosis of T cell-mediated skin infiltrates.