Abstract
Detailed analysis of the cells that infiltrate lesional skin cannot be performed in skin biopsy specimens using immunohistochemistry or cell separation techniques because enzyme treatments applied during the isolation step can destroy small amounts of protein and minor cell populations in the biopsy specimen. Here, we describe a method for isolating T cells from drops of whole blood obtained from lesions during skin biopsy in patients with cutaneous T-cell lymphoma. Lesional blood is assumed to contain lesional resident cells, cells from capillary vessels, and blood overflowing from capillary vessels into the lesion area. The lesional blood showed substantial increases in distinct cell populations, chemokines, and the expression of various genes. The proportion of CD8+CD45RO+ T cells in the lesional blood negatively correlated with the modified severity-weighted assessment tool scores. CD4+CD45RO+ T cells in the lesional blood expressed genes associated with the development of cancer and progression of cutaneous T-cell lymphoma. In addition, CD8+CD45RO+ T cells in lesional blood had unique T-cell receptor repertoires in lesions of each stage. Assessment of lesional blood drops might provide new insight into the pathogenesis of mycosis fungoides and facilitate evaluation of the treatment efficacy for mycosis fungoides as well as other skin inflammatory diseases.
Highlights
The tissue environment surrounding skin lesions has an important role in skin disease
Cytometry by time-of-flight (CyTOF) revealed that the lesional blood contained more granulocytes, and fewer monocytes and B cells than the peripheral blood (Fig. 1d,e). These findings indicate that the cell populations in lesional blood might differ from that in peripheral blood
Analysis showed that the same T-cell receptor (TCR) (TRAV1-2–TRAJ33) were increased (Fig. 4i,k,l). These findings indicate that CD8+ T cells with different TCR repertoires are directed toward each skin lesion, which is expected given the heterogeneity of malignant T cells in skin lesions
Summary
The tissue environment surrounding skin lesions has an important role in skin disease. Lesional blood samples could provide valuable information about the surrounding environment, including the levels and types of cytokines and inflammatory cells, without the need for enzyme treatment. Low awareness and diagnostic difficulties of MF, many patients seek dermatologic consultation for the first time after their condition has already progressed to the mycotic or tumor stage. This delay in diagnosis may lead to tumor formation, ulceration, leukemic transformation, visceral invasion, and death within a few months. The development of an effective method of analyzing lesional blood drops may be useful for evaluating other skin inflammatory diseases, such as atopic dermatitis and psoriasis, as well as the efficacy of their treatments
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