J Immunol Res. 2015;2015:856340. Epub 2015 Dec 16. doi: 10.1155/2015/856340.
Flow cytometric analysis of T, B and NK cells antigens in patients with Mycosis Fungoides
Serkan YAZİCİ, Emel BÜLBÜL BAŞKAN, Ferah BUDAK*, Barbaros ORAL*, Şaduman Balaban ADIM¥, Güven ÖZKAYAƥ, Kenan AYDOĞAN, Hayriye SARICAOĞLU, Şükran TUNALI,
Uludag University School of Medicine Departments of Dermatology and Venereology, Immunology*, Pathology¥ and Biostatisticsƥ, 16059 Bursa, TURKEY
We retrospectively analyzed the clinicopathological correlation and prognostic value of cell surface antigens expressed by peripheral blood mononuclear cells in patients with Mycosis Fungoides (MF). 121 consecutive MF patients included study. All patients had peripheral blood flowcytometry as part of their first visit. TNMB and histopathological staging of the cases were retrospectively performed in accordance with International Society for Cutaneous Lymphomas/European Organization of Research and Treatment of Cancer (ISCL/EORTC) criteria at the time of flow cytometry sampling. To determine prognostic value of cell surface antigens, cases were dived into two groups as stable and progressive disease. 17 flowcytometric analysis of 17 parapsoriasis (PP) and 11 analysis of 11 benign erythrodermic patients were included as control groups. Fluorescent labeled monoclonal antibodies were used to detect cell surface antigens: T cell (CD3+, CD4+, CD8+, TCRαβ+, TCRγδ+, CD7+, CD4+CD7+, CD4+CD7–,CD71+), B cell (HLA-DR+, CD19+, HLA-DR+CD19+), NKT cell (CD3+CD16+CD56+ , CD3– CD16+CD56+). The mean value of all cell surface antigens was not statistically significant between parapsoriasis and MF groups. Along with an increase in cases of MF stage statistically significant difference was found the mean values of cell surface antigens. Flowcytometric analysis of peripheral blood cell surface antigens in patients with mycosis fungoides may contribute to predict disease stage and progression.
Keywords: CD antigens, Flow cytometry, Mycosis Fungoides, Prognosis
The Cluster of Differentiation ( termed as Cluster of Designation or Classification Determinant) (CD) is a protocol used for the identification and investigation of cell surface molecules for immunophenotyping of cells. Neoplastic T cells frequently have an altered level of expression of various surface T-cell markers compared with normal T cells. Flow cytometric analysis of the cell surface antigens expressed by peripheral blood cells, is widely used in the diagnosis and also management of hematologic malignancies, however, has not been used routinely in the evaluation of MF patients. (Figure-1) Aberrant expression of T cells, and also B and NK cells markers may be expressed in patients with MF. We aimed to evaluate the prognostic value of cell surface antigens expressed by T, B and NK cells, and correlated with the clinicopathological ISCL/EORTC MF diagnostic score, by four-color flow cytometry.
To determine prognostic value of flowcytometry, according to the course of the follow-up process of the patients were dived into two groups as stable and progressive disease as recommended. ROC curve analysis was performed to determine the cut off values between progressive and stable disease groups.
Figure-1. Flow cytometric analysis has proven to be an efficient and sensitive method to detect and enumerate MF/SS cells in the peripheral blood.
- The mean value of all cell surface antigens was not statistically significant between Parapsoriasis and MF groups. This finding may show that, flow cytometry is not enough alone to differentiate mycosis fungoides from benign dematoses and parapsoriasis.
- According to histopathological and clinical stage in cases of MF, statistically significant difference were found between the mean values of all cell surface antigens with out TCRγδ and CD3+CD16+CD56+.
- While increasing stage of disease the number of CD3+,CD4+, TCRαβ+, CD4+CD7+, CD4+CD7-, CD71+, cells and CD4+/CD8+ cell ratio were significantly increased and the number of CD8+ , CD7+ , HLA-DR+, CD19+, HLA-DR+, CD19+, CD3–CD16+, CD56+, cells decreased significantly. These findings, may be useful in identifying an advanced stage cases in patients with MF.
- Although, in cases of plaque or tumoral stage of MF immunophenotypical abnormalities are significant, the significance of flow cytometric analysis in early stage MF is limited. Our findings suggest that T-cell phenotype affects the clinical behaviour of MF.
- When MF patients examined in two groups as stable and progressive; all cell surface antigens except CD4+CD7- cells may play a role in determining the progression. According to ROC analysis results, Flow cytometric results suggestive of disease progression in peripheral blood at any time(Table-1).
- Flow cytometry is less time consuming and prognostic tool for the management of MF
- We described the mean value of all cell surface antigens of our study population may basic for future investigations.
This study showed that flowcytometric analysis of peripheral blood cell surface antigens in patients with MF may contribute for identifying an advanced stage, and predict the disease progression.
Table-1. ROC analysis of the investigated cell surface antigens: According to ROC curve analysis results, flow cytometric results in peripheral blood at any time suggestive of disease progression were as follows; mean CD3+ cell percentage >79; mean CD4+ cell percentage >55.3; mean CD8+ cell percentage ≤10.6; CD4+/CD8+ percentage >2.4; mean TCRαβ+ cell percentage >73.4; TCRγδ+ cell percentage ≤ 2.2; CD7+ cell percentage ≤76.6; CD4+CD7+ cell percentage >48.2; CD4+CD7– cell percentage >10.7; CD71+ cell percentage >3.5; HLA-DR+ cell percentage ≤18.5; CD19+ cell percentage ≤6; HLA-DR+CD19+ cell percentage ≤6.1; CD3+CD16+CD56+ cell percentage ≤1.9; CD3–CD16+CD56+ cell percentage ≤3.9
- Olsen EA, Whittaker S, Kim YH, et al. International Society for Cutaneous Lymphomas; United States Cutaneous Lymphoma Consortium; Cutaneous Lymphoma Task Force of the European Organisation for Research and Treatment of Cancer. Clinical end points and response criteria in mycosis fungoides and Sézary syndrome: a consensus statement of the International Society for Cutaneous Lymphomas, the United States Cutaneous Lymphoma Consortium, and the Cutaneous Lymphoma Task Force of the European Organisation for Research and Treatment of Cancer. J Clin Oncol. 2011 Jun 20;29(18):2598-607.
- Olsen E, Vonderheid E, Pimpinelli N, et al. ISCL/EORTC. Revisions to the staging and classification of mycosis fungoides and Sezary syndrome: a proposal of the International Society for Cutaneous Lymphomas (ISCL) and the cutaneous lymphoma task force of the European Organization of Research and Treatment of Cancer (EORTC). Blood 2007;110:1713-22.
- Meyerson HJ. Flow cytometry for the diagnosis of mycosis fungoides. G Ital Dermatol Venereol. 2008 Feb;143(1):21-41.
Serkan Yazici, MD
Uludag University School of Medicine
Department of Dermatology and Venereology
Phone: +90 224 42950741
Fax: +90 224 4429229