NUCLEOLAR ORGANIZER REGIONS IN CUTANEOUS T CELL LYMPHOMAS

The present work studied the application of AgNOR count to differential diagnosis between cutaneous T cell lymphoma (CTCL) and cutaneous pseudolymphoma (CPL). Paraffin sections from 50 mycosis fungoides (22 MFI-Premycotic stage, 24 MF Ⅰ infiltrative stage and 4 MF Ⅲ - tumor stage), 2 nonepidermotropic cutaneous T cell lymphoma (NECTCL) and 9 CPL were investigated. In each case, 200 cells randomly selected were examined using a × 100 oil immersion lens. The mean number, standard deviation and standard error of the mean of AgNOR counts were as follows: MFⅠ 1.17±0.09, SEM = 0.01; MⅡ 1.17±0.01, SEM = 0.01; MF Ⅲ. 3.55±0.87, SEM = 0.43; NECTCL 4.5±0.28, SEM -0.199; CPL 1.17±0.1, SEM ± 0.03. The results revealed a highly significant difference between CTCL (MFⅢ+NECTCL) and CPL (t = 4.75, P<0.001), tumor stage (MF Ⅲ) and pretumor stage (MFI, MF Ⅱ) of mycosis fungoides (t = 4.75, P<0.001). Thus. AgNOR count is valuable in differential diagnosis.

T-CELL RECEPTOR GENE REARRANGEMENT ANALYSIS IN THE PRIMARY CUTANEOUS T-CELL LYMPHOMA

Object: The present paper is to evaluate the significance of T cell receptor (TCR) gene rearrange ments in primary cutaneous T cell lymphomas (PCTCL) as detected by analysis of Southern Blot (SBA) and polymerase chain reaction (PCR). Patients and Methods: Skin specimens and peripheral blood samples were taken from 44 patients with PCTCL, including 30 patients with mycosis fungoides (MF), 2 patients with Sezary's syndrome (SS), and 12 patients with PCTCL other than MF and SS (PNCTCL). 11 patients with a presumptive diagnosis of MF, 23 patients with lymphoproliferative dermatoses including lymphomatoid papulosis (LyP) and 8 patients with benign cutaneous lymphoid infiltrates were simultaneously studied by the amplification of junctional V (variable) J (joining) sequences of the rearranged TCRγ genes by PCR(TCRγPCR) and the analysis of TCRb chain genes by SBA(TCRβSBA) for detection of clonal gene rearrangements (GR). One lymph node specimen of a case with MF IIA was also detected by TCRγ PCR and TCRβSBA. Results: In MF, GR were detected by TCRγPCR and TCRβSBAb in 83.3 85.7% and 66.7% 71.4% of skin specimens of cases IIA IIB and in 57.1% 70.0% and 14.3% 10.0% of those of cases IA IB, respectively. GR were seen in 66.7% 71.4% and 33.3% 43.0.% of blood samples of cases IIA IIB, and 42.9% 40.0% and 0 10.0% of those of cases IA IB, respectively. GR was confirmed by TCRγ PCR and TCRβSBA in one lymph node showing dermato pathic lymphadenopathy of a case with MF IIA. In 11 patients of clinically suspected MF, GR were present in skin specimens of 5 cases (45.4%) and in blood samples of 3 cases ( 27.3% ) by TCRγ PCR. In PNCTCL, GR were found in 9 skin specimens (90.0%) from 10 patients detected by TCRγ PCR and in 6 skin specimens (75.0%) from 8 patients detected by TCRβSBA. GR were also seen in 6 blood samples (72.8%) from 11 patients detected by TCRγ PCR, and in 7 blood samples (70.0%) from 10 patients by TCRβSBA. In SS and LyP, GR were detected by TCRγ PCR and TCRβSBA in each of the two skin specimens of two cases with LyP and in each of the two blood samples of two cases with SS. GR were seen in one skin specimen of one case with SS and one blood sample of one case with LyP detected by TCRγPCR. Conclusions: This study demonstrated that TCRγ PCR is a rapid, more sensitive tool than TCRβSBA, can be used in the analysis of T cell clonality in skin, lymph node and blood samples of patients with PCTCL and indicated that this method forms a useful supplement to other methods for diagnosis of early and suspected MF, confirmation of PNCTCL and determination of extracutaneous involvement of lymph node and blood.

Aberrant expression of CD56 by circulating Sézary syndrome malignant T lymphocytes

Sézary syndrome(SS) is an aggressive variant of cutaneous T cell lymphoma characterized by the presence of malignant T cells in the skin, peripheral blood and lymph nodes. The tumoral population typically displays a CD3+ CD4+ CD45RO+ memory T cell phenotype. We report a case of SS with an aberrant CD56+ immunophenotype. This patient presented with a generalized erythroderma and palpable small axillary lymph nodes.SS(stage IVA) was diagnosed on histological criteria and by the detection of a major T cell clone in skin and blood, an elevated CD4/CD8 T cell ratio and Sézary cells count > 1000/mm3. Beside the Sézary cell marker KIR3DL2, immunostainings revealed that two third of the malignant cells expressed CD56 but no other natural killer(NK) cell marker such as CD16, CD160 or NKp46. This atypical expression was not linked to an activation-dependent process and remained stable during the time course of the disease. No loss of the panT-cell markers CD2, CD3 or CD4 was detected while a complete down-modulation of CD26 was observed. Despite several lines of treatment, no durable amelioration was observed and patient died after 10 mo of follow-up. Because this CD4+ CD56+ SS case is the only one reported so far, the functional significance of CD56 expression remained difficult to assess in terms of aggressiveness and prognosis.

Clinicopathologic,immunophenotypic and ultrastructrual analyses of ATLL patients with cutaneous involvement

Objective To study 4 cases of adult T cell leukemia/lymphoma (ATLL) as sociated with cutaneous lesions for clinicopathology, immunophenotype, human T cell leukemia/lymphoma virus type Ⅰ (HTLV Ⅰ) provirus DNA and their ultrastru cture. At the same time, HTLV Ⅰ provirus DNA of ATLL patients were also compar ed with 18 cases of cutaneous lymphoma (CL), two cases of actinic reticuloid as well as two cases of lymphocytic infiltration.Methods Immunohistochemistry studies were carried out on the infiltrati ng cells using monoclonal antibodies against CD45 RO, CD20, CD68 on paraffin e mbedded sections by ABC method and using monoclonal antibodies against CD3, CD4 and CD8 with indirect immunofluorescence (IIF) on frozen sections. Skin biopsies were examined by electron microscope. Serum and bone marrow cells were tested f or antibodies against HTLV Ⅰ associated antigen by IIF, and HTLV Ⅰ provirus DNA was examined by PCR method.Results The research showed four patients with ATLL manifesting cutaneo u s lesions, were subsequently found with additional systemic symptoms, as extensi vely enlarged superficial lymph node, abnormal increased IL 2 receptor, flower like cells in their peripheral blood and marrow. The HTLV Ⅰ provirus DNA was positi ve in the peripheral blood, bone marrow, cutaneous lesions and lymph node biopsy specimens by using PCR amplification of specific HTLV Ⅰ fragment while 18 cas es of the CL were negative for HTLV Ⅰ. The special ultrastructure of skin lesi ons was also found in ATLL patients.Conclusions The cutaneous involvement in ATLL is a type of cutaneous T cell lymphoma (CTCL) but shows some differential immunological markers for diffe rential diagnosis. The examination of HTLV Ⅰ antibodies or HTLV Ⅰ provirus D NA is necessary for diagnosis of ATLL. The ultrastrustural characteristics in skin lesions of ATLL were of atypic al lymphocytes and mononuclear cells invading the epidermis, and the mononuclear cells are possessing the phagocytic function and phagocytizing the degenerated epidermic cells or lymphocytes.