Introduction Mycosis fungoides (MF), the most common form of primary cutaneous T-cell lymphomas (CTCL), originates from a clonal expansion of epidermotropic CD4+ memory T cells and is characterized by an indolent disease course and a stepwise evolution with sequential appearance of patches, plaques, and tumors. The immunesuppression related both to the functional impairment of neoplastic T cells and to chemotherapeutic treatments, determines the development of severe infectious complications which represent the most frequent cause of death. Among those, bacterial sepsis is the most common one and is the cause of about 50% of CTCL deaths. Polymorphonucleate granulocytes (PMNs) constitute an essential part of the innate immune system: their activity against pathogens is mediated by the interactions between Toll-like receptors (TLRs) and PAMPs. The aim of this study was to investigate PMN functional activity and phenotype in MF patients in different disease’s stages. Materials and methods PBMC samples were obtained from 20 MF patients (10 stage I-IIa and 10 in stage IIb-IV) and 20 healthy donors coming from Turin blood bank. All control subjects were age- and sex-matched. The diagnosis of MF was made according to standard clinical and immunopathological findings. MF patients were classified according to the new tumor, node, metastasis (TNM) (stage I to IV). PMN functional activity was evaluated by phagocytosis and intracellular killing (Survival Index) tests towards Klebsiella pneumonia. Quantification of the oxidative burst activity was done by flow cytometry using a commercial kit (Phagoburst) that evaluate the percentage of phagocytic cells producing reactive oxidants and their enzymatic activity. Flow-cytometry was applied to analyze PMN phenotype, the following mouse anti-human antibodies were used: TLR2/CD282 (clone TL2.1); TLR4/CD284 (clone HTA 125); TLR5/CD285 (clone 85B152.5); TLR8/CD288 (clone 44C143); TLR9/CD289 (clone eB72-1665). Results Results obtained showed a functional impairment of PMNs: phagocytosis and intracellular killing values were lesser than those of healthy subjects in the advanced stages of MF, whereas samples form initial stages were superimposable to normal donors. Discussion and conclusions This study has been planned to complete our previous investigations on the innate immune system, that demonstrated PMN’s functional impairment in other malignant and not malignant skin conditions such as Sezary’s Syndrome and atopic dermatitis. The results we obtained with MF patients show that the defects of PMNs’ functional activity are only present in the advanced stages of MF and could therefore explain the increased frequency of infections related to disease progression.
Evaluation of PMNs functional and phenotypical alterations in mycosis fungoides patients.
MERLINO, Chiara;NOVELLI, Mauro;BANCHE, Giuliana;ALLIZOND, VALERIA;MANDRAS, Narcisa;SCALAS, Daniela;ROANA, Janira;FIERRO, Maria Teresa;TULLIO, Viviana Cristina;FAVA, PAOLO;QUAGLINO, Pietro;CUFFINI, Annamaria
2014-01-01
Abstract
Introduction Mycosis fungoides (MF), the most common form of primary cutaneous T-cell lymphomas (CTCL), originates from a clonal expansion of epidermotropic CD4+ memory T cells and is characterized by an indolent disease course and a stepwise evolution with sequential appearance of patches, plaques, and tumors. The immunesuppression related both to the functional impairment of neoplastic T cells and to chemotherapeutic treatments, determines the development of severe infectious complications which represent the most frequent cause of death. Among those, bacterial sepsis is the most common one and is the cause of about 50% of CTCL deaths. Polymorphonucleate granulocytes (PMNs) constitute an essential part of the innate immune system: their activity against pathogens is mediated by the interactions between Toll-like receptors (TLRs) and PAMPs. The aim of this study was to investigate PMN functional activity and phenotype in MF patients in different disease’s stages. Materials and methods PBMC samples were obtained from 20 MF patients (10 stage I-IIa and 10 in stage IIb-IV) and 20 healthy donors coming from Turin blood bank. All control subjects were age- and sex-matched. The diagnosis of MF was made according to standard clinical and immunopathological findings. MF patients were classified according to the new tumor, node, metastasis (TNM) (stage I to IV). PMN functional activity was evaluated by phagocytosis and intracellular killing (Survival Index) tests towards Klebsiella pneumonia. Quantification of the oxidative burst activity was done by flow cytometry using a commercial kit (Phagoburst) that evaluate the percentage of phagocytic cells producing reactive oxidants and their enzymatic activity. Flow-cytometry was applied to analyze PMN phenotype, the following mouse anti-human antibodies were used: TLR2/CD282 (clone TL2.1); TLR4/CD284 (clone HTA 125); TLR5/CD285 (clone 85B152.5); TLR8/CD288 (clone 44C143); TLR9/CD289 (clone eB72-1665). Results Results obtained showed a functional impairment of PMNs: phagocytosis and intracellular killing values were lesser than those of healthy subjects in the advanced stages of MF, whereas samples form initial stages were superimposable to normal donors. Discussion and conclusions This study has been planned to complete our previous investigations on the innate immune system, that demonstrated PMN’s functional impairment in other malignant and not malignant skin conditions such as Sezary’s Syndrome and atopic dermatitis. The results we obtained with MF patients show that the defects of PMNs’ functional activity are only present in the advanced stages of MF and could therefore explain the increased frequency of infections related to disease progression.
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