FRINK Linked Data Fragments server

Matches in SemOpenAlex for { <https://semopenalex.org/work/W2023862568> ?p ?o ?g. }

• W2023862568 endingPage "193" @default.
• W2023862568 startingPage "188" @default.
• W2023862568 abstract "Cutaneous T cell lymphomas are considered to represent a clonal malignancy of mature T lymphocytes of the T helper memory subtype. A method enabling the direct identification of clonal malignant cells in tissue and, at the same time, identification of the surface molecules they express has not been available, however. We have developed an application of the FICTION technique (simultaneous fluorescence immunophenotyping and interphase cytogenetics) to be used on fresh blood, skin, and lymph node samples. A prerequisite for this method is the characterization of a moleculocytogenetic clone in order to select the proper probes. With this method, we demonstrate that the true malignant cells express CD3, CD4, and CD45RO in the blood, skin, and lymph nodes of two Sezary syndrome patients. The majority of these cells express also CD45RA (albeit of varying intensity) and CDw150. The cytokine expression pattern of the clonal cells in skin and lymph nodes was interleukin-2 and interferon-γ negative and interleukin-4 positive. Interleukin-10 expression varied. The malignant cells did not express granzyme B, thus indicating that they do not have cytotoxic properties. Clonal cells with the same constant phenotype could be found even in lymph nodes with not yet morphologically identifiable malignant cells. This is the first report of the FICTION method applied directly on skin tissue. With this method we demonstrated that the chromosomally clonal cells in these two cases of Sezary syndrome could be intermediate forms between naïve CD45RA+ and CD45RO+ Th2 cells. Cutaneous T cell lymphomas are considered to represent a clonal malignancy of mature T lymphocytes of the T helper memory subtype. A method enabling the direct identification of clonal malignant cells in tissue and, at the same time, identification of the surface molecules they express has not been available, however. We have developed an application of the FICTION technique (simultaneous fluorescence immunophenotyping and interphase cytogenetics) to be used on fresh blood, skin, and lymph node samples. A prerequisite for this method is the characterization of a moleculocytogenetic clone in order to select the proper probes. With this method, we demonstrate that the true malignant cells express CD3, CD4, and CD45RO in the blood, skin, and lymph nodes of two Sezary syndrome patients. The majority of these cells express also CD45RA (albeit of varying intensity) and CDw150. The cytokine expression pattern of the clonal cells in skin and lymph nodes was interleukin-2 and interferon-γ negative and interleukin-4 positive. Interleukin-10 expression varied. The malignant cells did not express granzyme B, thus indicating that they do not have cytotoxic properties. Clonal cells with the same constant phenotype could be found even in lymph nodes with not yet morphologically identifiable malignant cells. This is the first report of the FICTION method applied directly on skin tissue. With this method we demonstrated that the chromosomally clonal cells in these two cases of Sezary syndrome could be intermediate forms between naïve CD45RA+ and CD45RO+ Th2 cells. simultaneous fluorescence immunophenotyping and interphase cytogenetics fluorescence in situ hybridization mycosis fungoides signaling lymphocytic activation molecule Sezary syndrome In order to understand the pathomechanism and biology of primary cutaneous T cell lymphomas (CTCLs), the phenotype and function of the malignant cells in various compartments of the body should be known. The malignant lymphocytes in most CTCLs are considered to represent a malignancy of mature T lymphocytes of the T helper memory subtype (Haynes et al., 1981Haynes B.F. Metzgar R.S. Minna J.D. Bunn P.A. Phenotype characterization of cutaneous T-cell lymphoma. Use of monoclonal antibodies to compare with other malignant T-cells.N Engl J Med. 1981; 28: 1319-1323Crossref Scopus (263) Google Scholar;Ralfkiaer et al., 1991Ralfkiaer E. Thomsen K. Vejlsgaard G. Expression of a cell adhesion protein (VLAβ) in normal and diseased skin.Br J Dermatol. 1991; 124: 527-532Crossref PubMed Scopus (18) Google Scholar). Previous studies of the phenotype of these cells have been based on morphologic identification of the malignant cells or on flow cytometric identification of clonal T cell receptor (TCR) α, β, or γ locus rearrangement 1Urban A, Asadullah K, Friedrich M, Volk H-D, Sterry W, Döcke W-D: CD34RA-expressing malignant T-helper cells in Sezary-syndrome (SS). 29th annual meeting of the European Society for Dermatological Research, Montpellier 22–25 September 1999. J Invest Dermatol 113:474 1999 (abstr.)1Urban A, Asadullah K, Friedrich M, Volk H-D, Sterry W, Döcke W-D: CD34RA-expressing malignant T-helper cells in Sezary-syndrome (SS). 29th annual meeting of the European Society for Dermatological Research, Montpellier 22–25 September 1999. J Invest Dermatol 113:474 1999 (abstr.) (Haynes et al., 1981Haynes B.F. Metzgar R.S. Minna J.D. Bunn P.A. Phenotype characterization of cutaneous T-cell lymphoma. Use of monoclonal antibodies to compare with other malignant T-cells.N Engl J Med. 1981; 28: 1319-1323Crossref Scopus (263) Google Scholar;Longley et al., 1995Longley J. Tyrrell L. Lu S.-Z. et al.Malignant and normal T cells show random use of T-cell receptor α chain variable regions in patients with cutaneous T-cell lymphoma.J Invest Dermatol. 1995; 105: 62-64Crossref PubMed Scopus (9) Google Scholar;Dummer et al., 1996Dummer R. Heald P.W. Nestle F.O. Ludwig E. Laine E. Hemmi S. Burg G. Sezary syndrome T-cell clones display T-helper 2 cytokines and express the accessory factor-1 (interferon-α receptor β-chain).Blood. 1996; 88: 1383-1389PubMed Google Scholar;Bakels et al., 1997Bakels V. van Oostveen J.W. van der Putte S.C.J. Meijer C.J.L.M. Willemze R. Immunophenotyping and gene rearrangement analysis provide additional criteria to differentiate between cutaneous T-cell lymphomas and pseudo-T-cell lymphomas.Am J Pathol. 1997; 150: 1941-1949PubMed Google Scholar;Bagot et al., 1998Bagot M. Echchakir H. Mami-Chouaib F. et al.Isolation of tumor-specific cytotoxic CD4+ and CD4+CD8dim+ T-cell clones infiltrating a cutaneous T-cell lymphoma.Blood. 1998; 91: 4331-4341PubMed Google Scholar). Based on functional in vitro assays of peripheral blood mononuclear cells, Sezary syndrome (SS), the leukemic subtype of CTCL, has been suggested to present the Th2-type T cell profile whereas mycosis fungoides (MF) would represent a Th1-type profile (Vowels et al., 1992Vowels B.R. Cassin M. Vonderheid E.C. Rook A.H. Aberrant cytokine production by Sezary syndrome patients: cytokine secretion pattern resembles murine Th2 cell.J Invest Dermatol. 1992; 99: 90-94Crossref PubMed Scopus (213) Google Scholar;Saed et al., 1994Saed G. Fivenson D.P. Naidu Y. Nickoloff B.J. Mycosis fungoides exhibits a Th1-type cell-mediated cytokine profile whereas Sezary syndrome expresses a Th2-type profile.J Invest Dermatol. 1994; 103: 29-33Abstract Full Text PDF PubMed Google Scholar;Dummer et al., 1996Dummer R. Heald P.W. Nestle F.O. Ludwig E. Laine E. Hemmi S. Burg G. Sezary syndrome T-cell clones display T-helper 2 cytokines and express the accessory factor-1 (interferon-α receptor β-chain).Blood. 1996; 88: 1383-1389PubMed Google Scholar). These methods may not always have identified specifically the true malignant cells, however, as the CTCL lesions also contain clonal T cells reacting against the tumor cells (Bagot et al., 1998Bagot M. Echchakir H. Mami-Chouaib F. et al.Isolation of tumor-specific cytotoxic CD4+ and CD4+CD8dim+ T-cell clones infiltrating a cutaneous T-cell lymphoma.Blood. 1998; 91: 4331-4341PubMed Google Scholar;Thor Straten et al., 1998Thor Straten P. Ralfkiaer E. Hendriks J. Serement T. Vejlsgaard G.L. Zeuthen J. T-cell variable region genes in cutaneous T-cell lymphomas.Br J Dermatol. 1998; 138: 3-12Crossref PubMed Scopus (28) Google Scholar). Also, our recent studies with fluorescent in situ hybridization (FISH) have shown that chromosomal clones can be detected in the blood of CTCL patients even in the absence of morphologically identifiable Sezary cells (Karenko et al., 1997Karenko L. Hyytinen E. Sarna S. Ranki A. Chromosomal abnormalities in cutaneous T-cell lymphoma (CTCL) and its premalignant conditions as detected by G-banding and interphase cytogenetic methods.J Invest Dermatol. 1997; 108: 22-29Crossref PubMed Scopus (57) Google Scholar). To reliably identify the membrane markers and cytokines expressed by the true malignant cells, we have identified these cells directly in the tissues with in situ hybridization using chromosome centromere-specific probes in combination with simultaneous immunohistochemical staining. A prerequisite for this method is the characterization of the chromosomal clone to be able to select the proper probes. We have modified the FICTION technique (simultaneous fluorescence immunophenotyping and interphase cytogenetics) ofWeber-Matthiesen et al., 1992Weber-Matthiesen K. Winkemann M. Müller-Hermelink A. Schlegelberger B. Grote W. Simultaneous fluorescence immunophenotyping and interphase cytogenetics: a contribution to the characterization of tumor cells.J Histochem Cytochem. 1992; 40: 171-175Crossref PubMed Scopus (130) Google Scholar,Weber-Matthiesen et al., 1993Weber-Matthiesen K. Pressi S. Schlegelberger B. Grote W. Combined immunophenotyping and interphase cytogenetics on cryostat sections by the new FICTION method.Leukemia. 1993; 7: 646-649PubMed Google Scholar) by using new, intense colors visible with a standard (8300) filter set, and, in the study of solid tissue, by using touch preparations instead of cryosections, which allow the observation of intact, whole cells. This is the first report of skin studied with such a method and also the first report where the true malignant cells of CTCL are identified directly in a tissue with immunophenotyping and chromosomal markers. The material consisted of tissue samples obtained from two patients who developed SS and showed a dominant clone of malignant cells with supernumerary copies of chromosome 8, in near-diploid or in near-tetraploid cells in blood (Karenko et al., 1999Karenko L. Kähkönen M. Hyytinen E.-R. Lindlöf M. Ranki A. Notable losses at specific regions of chromosomes 10q and 13q in the Sezary syndrome detected by comparative genomic hybridization.J Invest Dermatol. 1999; 112: 392-395https://doi.org/10.1046/j.1523-1747.1999.00519.xAbstract Full Text PDF PubMed Scopus (50) Google Scholar; see also below). The clones contained also other complex chromosomal abnormalities, which were partly different in the two patients but remained remarkably stable during the course of the disease. Case 1 was a 64-y-old male patient, when he became erythrodermic after mild psoriasis, localized to elbows, knees, and the scalp. In a skin biopsy atypical CD4-positive T lymphocytes were seen in the upper dermis but the biopsy was regarded as nondiagnostic of CTCL because no epidermotropism was seen. There were 10% of Sezary cells in the blood. Six months later, enlarged lymph nodes were detected in his left armpit and, in a biopsy of such a lymph node, nonspecific hyperplasia was seen. In some areas the paracortical area (T zone) was expanded but no definite lymphoma was seen (specimen L 1-1 in the FICTION study). The patient was then treated with psoralen and ultraviolet A. After an additional 6 mo, Pautrier's microabscesses were seen in a skin biopsy, and the histologic diagnosis of MF was made. One month later, 25% of Sezary cells were found in his blood, and the diagnosis of SS was made (T4N0B1). His cultivated blood lymphocytes showed 45% of clonally very aberrant mitoses in G-banding. Further studies showed a gain of chromosome 8 in comparative genomic hybridization, and supernumerary copies of chromosome 8 in near-diploid or near-tetraploid cells in FISH (Karenko et al., 1999Karenko L. Kähkönen M. Hyytinen E.-R. Lindlöf M. Ranki A. Notable losses at specific regions of chromosomes 10q and 13q in the Sezary syndrome detected by comparative genomic hybridization.J Invest Dermatol. 1999; 112: 392-395https://doi.org/10.1046/j.1523-1747.1999.00519.xAbstract Full Text PDF PubMed Scopus (50) Google Scholar). In computerized tomography, enlarged lymph nodes were detected in armpits and groin. He was then sequentially treated with etretinate, total skin electron beam irradiation, and cladribine. The disease progressed, and the treatment was discontinued after 6 mo (18 mo after the first biopsy) because of thrombocytopenia. He died 2 y and 10 mo after the diagnosis of CTCL. A blood specimen (B 1-1) taken 1 wk before death showed clonal cells similar to those detected 1 y 10 mo earlier (25 aberrant out of 25 mitoses after phytohemagglutinin stimulation). This blood sample and postmortem skin (S 1-1) and lymph node specimens (L 2-1) were used for the modified FICTION. Case 2 was a 52-y-old male patient who presented with an eczematous dermatitis on his soles, ankles, breast, and back. Four months later, a skin biopsy showed typical histology of MF, and he had 30% of Sezary cells in peripheral blood (T2N0B1). Clinically, he had patch stage skin lesions, and he was treated with psoralen and ultraviolet A. Ten months after the diagnostic skin biopsy, he developed erythrodermia, and the diagnosis of secondary SS was made. A blood sample showed a near-tetraploid aberrated mitosis (1 of 100 mitoses), that later was observed to be clonal. Two months later, his groin lymph nodes were enlarged and showed histologically dermatopathic lymphonoditis with a slight suspicion of malignant (T4N1B1) affision. This biopsy (L 1-2) was studied with the modified FICTION. He was then treated with whole body and local electron beam irradiation (2000 Gy and up to 20 Gy, respectively), cladribine, and, finally, chlorambucil. In blood samples taken 2 y 7 mo after the diagnosis of MF, and 2 mo thereafter (B 1-2), and in a skin tumor biopsy taken at the latter time point (S 1-2), very similar cytogenetic or moleculocytogenetic findings were detected (Karenko et al., 1999Karenko L. Kähkönen M. Hyytinen E.-R. Lindlöf M. Ranki A. Notable losses at specific regions of chromosomes 10q and 13q in the Sezary syndrome detected by comparative genomic hybridization.J Invest Dermatol. 1999; 112: 392-395https://doi.org/10.1046/j.1523-1747.1999.00519.xAbstract Full Text PDF PubMed Scopus (50) Google Scholar). They included gain of chromosome 8 in comparative genomic hybridization (blood and skin tumor) and supernumerary copies of its centromere in FISH both in near-diploid cells and in the last sample also in near-tetraploid cells (blood). The latter blood (B 1-2) and skin samples (S 1-2) were examined with the modified FICTION method. The number of Sezary cells in the latter blood specimen (B 1-2) was 95%, and 95% of phytohemagglutinin-induced mitoses (19 of 20 mitoses) showed the clonal aberrations. He died 2 y 10 mo after the diagnosis of MF. Blood lymphocyte preparations of three healthy, nonsmoking persons were used as controls. Sections of frozen and formalin-fixed paraffin-embedded skin lesion biopsies of one patient with histopathologically confirmed lymphomatoid papulosis, and with a favorable course, were examined for granzyme B positivity (see below). Cytospin preparations of Ficoll-enriched blood mononuclear cells or touch preparates of frozen skin or lymph node biopsies were fixed in ice-cold acetone, and an immunohistologic staining was performed using antibodies to CD3, CD4 (Dako, Glostrup, Denmark), CD45RA (Caltag, Burlingame, CA), CD45RO, CD8 (Dako), granzyme B, interleukin-2 (IL-2), IL-4, IL-10 (Santa Cruz Biotechnology, Santa Cruz, CA), interferon-γ (IFN-γ) (Neomarkers, Fremont, CA), and signaling lymphocytic activation molecule (SLAM, CDw150; A12 antibody kindly provided by Dr. G. Aversa, DNAX Research Institute of Molecular and Cellular Biology, Palo Alto, CA). The primary antibodies were monoclonal mouse antibodies except for the polyclonal rabbit antibodies against CD3 and IL-2, and goat antibodies against granzyme B, IL-4, and IL-10. The secondary antibodies were antirabbit or antimouse goat conjugates of Texas Red X or Alexa 594 R (Molecular Probes, Leiden, The Netherlands), or antirabbit, antigoat, or antimouse donkey conjugates of Rhodamine Red X (Jackson Immuno Research Laboratories, West Grove, PA). When necessary, for detection of weak mouse primary antibodies, Rhodamine Red X conjugate was followed with antirhodamine rabbit (Molecular Probes) and an antirabbit donkey conjugate of Rhodamine Red X (Jackson Immuno Research Laboratories). All antibody layers were preceded by a layer of normal serum from the animal species, in which the secondary antibody was raised. Through every step of the process, each slide was accompanied by a similar control slide with no primary antibody. After immunohistologic staining, the samples were hybridized with a chromosome 8 centromere-specific probe (D8Z2[pJM128]) labeled (Karenko et al., 1997Karenko L. Hyytinen E. Sarna S. Ranki A. Chromosomal abnormalities in cutaneous T-cell lymphoma (CTCL) and its premalignant conditions as detected by G-banding and interphase cytogenetic methods.J Invest Dermatol. 1997; 108: 22-29Crossref PubMed Scopus (57) Google Scholar) with a green fluorescent color (Alexa 488, Molecular Probes) or digoxigenin, or a chromosome 1 centromere-specific probe (1q12[pUC177]) labeled with biotin (both probes were kindly provided by Dr. T. Visakorpi, University of Tampere, Finland). The hybridization was performed as previously described (Karenko et al., 1997Karenko L. Hyytinen E. Sarna S. Ranki A. Chromosomal abnormalities in cutaneous T-cell lymphoma (CTCL) and its premalignant conditions as detected by G-banding and interphase cytogenetic methods.J Invest Dermatol. 1997; 108: 22-29Crossref PubMed Scopus (57) Google Scholar). After posthybridization washes, the biotin-labeled probes were detected with deglycosylated avidin conjugated with a blue color (NeutrAvidin conjugated with Alexa 350 or Cascade Blue; Molecular Probes), followed by biotinylated antiavidin (goat, Vector Laboratories, Burlingame, CA, or mouse, Sigma, St. Louis, MO), and a new layer of the same deglycosylated avidin conjugate. The digoxigenin-labeled probes were detected with sheep antidigoxigenin antibody (Roche Molecular Biochemicals, Mannheim, Germany), followed by donkey antibody conjugated with fluorescein isothiocyanate (FITC) (Jackson Immuno Research Laboratories). Alternative stainings were used to confirm the results. The immunohistology was performed by detecting the primary mouse antigen with biotinylated antimouse raised in horse, followed by avidin conjugated with FITC (Vector Laboratories), and hybridization with a chromosome 8 centromere-specific probe labeled with a red color (Alexa 594, Molecular Probes). Alternatively, the primary mouse antibody was detected with rabbit antimouse antibody (Sigma), followed by antirabbit raised in swine (Dako), antiswine raised in rabbit (Rockland, Gilbertsville, PA), and finally goat antirabbit conjugated with a blue color (Alexa 350, Molecular Probes). The probes used were chromosome 8 centromere-specific probe labeled with Alexa 488 and chromosome-1-specific probe labeled with Texas Red X (colors, Molecular Probes). The formalin-fixed, paraffin-embedded skin biopsies of cases 1 and 2 were examined for CD30 and a skin biopsy of the third patient with lymphomatoid papulosis for granzyme B with the standard immunoperoxidase technique (for CD30, StreptABC Complex/HRP kit, Dako, followed by 3-amino-ethylcarbazole, for granzyme B).2 Preparations were analyzed with an ultraviolet microscope (Olympus BX 50, Tokyo, Japan; equipped with filter set 8300 and tripleband exciter 83103x, Chroma Technology, Brattleboro, VT). Pictures were taken with a cooled CCD camera (Sensi Cam, PCO, Computer Optics, Kelheim, Germany) connected to a computer (Dell, Limerick, Ireland; with software Image pro Plus, Media Cybernetics, Silver Spring, MD). A similar and constant phenotype over time was found in malignant cells with clonal chromosomal aberrations (i.e., supernumerary copies of chromosome 8 in near-diploid or near-tetraploid cells) in the blood, skin, and lymph nodes of two SS patients (Table 1). The malignant cells were CD3 and CD4 positive but CD8 negative (Table 1 and Figure 1a–c). Both CD45RA and CD45RO positivity was observed in the majority of clonally malignant cells in both patients, although in the blood of both patients and in the skin of case 2 the staining with CD45RA was of weaker intensity (Table 1, Figure 1d–i). In the postmortem lymph node of case 1 (L 2-1), both antigens showed intense staining in the clonal cells (Figure 1d,i).Table 1Phenotype of cells with clonal chromosomal aberrations (extra copies of chromosomes 1 and 8aKarenko et al. 1999, and Kähkönen M, personal communication.) in two patients with Sezary syndromeSurface markerTissueSample n: o – Patient n: oMonths after CTCL diagnosisTCR gene rearrangementiCD3CD4CD8CD45ROCD45RACDw150Granzyme BBlood1-134β+bAll clonal cells positive if not otherwise indicated.+-cFew weakly positive cells.75%dPercentage of clonal cells positive for the marker.85%dPercentage of clonal cells positive for the marker.+NDeND, not done.1-233β, α++ND85%dPercentage of clonal cells positive for the marker.dimfWeakly positive cells.NDNDSkin lesionsgBoth samples negative for CD30 antigen.1-134++–+dimfWeakly positive cells.50%dPercentage of clonal cells positive for the marker.–1-233β. α++–+dimfWeakly positive cells.50%dPercentage of clonal cells positive for the marker.< 10%eND, not done.Lymph node,1-1-6NDNDND+++–no lymphoma histology1-212NDNDND+++–Lymph node, post mortem2-134β++–cFew weakly positive cells.90%dPercentage of clonal cells positive for the marker.90%dPercentage of clonal cells positive for the marker.+–a Karenko et al., 1999Karenko L. Kähkönen M. Hyytinen E.-R. Lindlöf M. Ranki A. Notable losses at specific regions of chromosomes 10q and 13q in the Sezary syndrome detected by comparative genomic hybridization.J Invest Dermatol. 1999; 112: 392-395https://doi.org/10.1046/j.1523-1747.1999.00519.xAbstract Full Text PDF PubMed Scopus (50) Google Scholar, and Kähkönen M, personal communication.b All clonal cells positive if not otherwise indicated.c Few weakly positive cells.d Percentage of clonal cells positive for the marker.e ND, not done.f Weakly positive cells.g Both samples negative for CD30 antigen. Open table in a new tab The lymph node biopsies of both patients obtained before histologically verified lymphoma involvement showed chromosomally clonal cells in hybridization (25%-35% of the cells in the touch preparate in L 1-1, and 24%-42%, respectively, in L 1-2). In both cases, the clonal cells expressed both CD45RA and CD45RO markers (Table 1, Figure 1g). The CDw150 (SLAM) antigen, characterizing activated memory T cells (CD45ROhigh) was detected in all tissues of case 1 (Table 1). In the skin of both patients, the expression was observed in about half of the clonal cells (Figure 1j). In contrast to positive control samples (blood lymphocytes of a healthy person, and the patient with lymphomatoid papulosis), granzyme-B-positive cells were observed only occasionally in the skin or lymph node samples of case 1, and none of these cells represented the malignant clone (Figure 1k–m). A few (less than 10%) clonal skin cells of case 2 expressed granzyme B very weakly (Table 1). The majority of clonal cells expressed IL-4 (Figure 2). The staining intensity was variable with all colors used (Rhodamine Red X, Alexa 594 R), showing very bright color intensities in up to 50% of the clonal cells. At most 20% of the clonal cells were IL-4 negative (Table 2). The majority of the clonal cells of case 1 were IL-10 negative (Table 2), but the majority of the cells of case 2 were IL-10 positive (Table 2, Figure 2). The majority of the clonal cells of both patients were IL-2 negative and none expressed IFN-γ (Table 2, Figure 2). Thus, the cytokine expression pattern of the clonal cells in skin and lymph node was IL-2 negative, IL-4 positive, variable for IL-10, and IFN-γ negative.Table 2Chromosomally clonal cells in two patients with Sezary Syndrome expressed IL-4, but the expression of IL-10 was variable between the patientsTissueSample n: oIL-2IL = 4IL-10IFN-gammaSkin lesion1-1+3%a+, positive immunostaining; ++, brightly positive immunostaining.++50%, +50%+5%NDbND, not done.1-2+2%+80%+80%0%Lymph node, post mortem2-1+2%++25%, +75%+8%0%a +, positive immunostaining; ++, brightly positive immunostaining.b ND, not done. Open table in a new tab Phenotypically, Sezary cells have been considered to be CD4+, CD45RA-, CD45RO+, and functionally of Th2 type (Vowels et al., 1992Vowels B.R. Cassin M. Vonderheid E.C. Rook A.H. Aberrant cytokine production by Sezary syndrome patients: cytokine secretion pattern resembles murine Th2 cell.J Invest Dermatol. 1992; 99: 90-94Crossref PubMed Scopus (213) Google Scholar;Saed et al., 1994Saed G. Fivenson D.P. Naidu Y. Nickoloff B.J. Mycosis fungoides exhibits a Th1-type cell-mediated cytokine profile whereas Sezary syndrome expresses a Th2-type profile.J Invest Dermatol. 1994; 103: 29-33Abstract Full Text PDF PubMed Google Scholar;Dummer et al., 1996Dummer R. Heald P.W. Nestle F.O. Ludwig E. Laine E. Hemmi S. Burg G. Sezary syndrome T-cell clones display T-helper 2 cytokines and express the accessory factor-1 (interferon-α receptor β-chain).Blood. 1996; 88: 1383-1389PubMed Google Scholar). The cells of our patients commonly expressed also CD45RA, however, normally observed in naive T cells (Clement et al., 1988Clement L.T. Yamashita N. Martin A.M. The functionally distinct subpopulations of human CD4+ helper/inducer T lymphocytes defined by anti-CD45R antibodies derive sequentially from a differentiation pathway that is regulated by activation-dependent post-thymic differentiation.J Immunol. 1988; 141: 1464-1470PubMed Google Scholar). They also expressed SLAM (CDw150), which is normally expressed on CD45RO+ peripheral blood memory cells and rapidly upregulated on activated T cells (Cocks et al., 1995Cocks B.G. Chang C.-C.J. Carballido J.M. Yssel H. de Vries J.E. Aversa G. A novel receptor involved in T-cell activation.Nature. 1995; 376: 260-263Crossref PubMed Scopus (414) Google Scholar). SLAM directs the immune response towards the Th0-Th1 pathway (Aversa et al., 1997Aversa G. Carballido J. Punnonen J. Chang C.C.J. Cocks B.G. de Vries J.E. SLAM and its role in T-cell activation and Th cell responses.Immunol Cell Biology. 1997; 75: 202-205Crossref PubMed Scopus (92) Google Scholar). Normally, naive CD45RA+ cells express IL-2, and mature CD45RO+ cells express IL-4, IL-5, IL-10 (Th2), or IFN-γ (Th1). A strong coexpression of both CD45 isoforms occurs during transition from CD45RA+ to CD45RO+ (Dbright in flow cytometry), with expression of IL-2 and IFN-γ (LaSalle and Hafler, 1991LaSalle J.M. Hafler D.A. The coexpression of CD45RA and CD45RO isoforms on T-cells during the S/G2/M stages of cell cycle.Cell Immunol. 1991; 138: 197-206Crossref PubMed Scopus (26) Google Scholar;Picker et al., 1993Picker L.J. Treer J.R. Ferguson-Darnell B. Collins P.A. Buck D. Terstappen L.W.M.M. Control of lymphocyte recirculation in man.J Immunol. 1993; 150: 1105-1121PubMed Google Scholar;Hamann et al., 1996Hamann D. Baars P.A. Hooibrink B. van Lier R.A.W. Heterogeneity of the human CD4+ T-cell population: two distinct CD4+ T-cell subsets characterized by coexpression of CD4RA and CD45RO isoforms.Blood. 1996; 88: 3513-3521PubMed Google Scholar). Cells with weaker coexpression of CD45RA/RO (Ddull) express mainly RO+ type cytokines, IL-4, IL-5, IL-10, or IFN-γ, and have been suggested to represent some stage in T cell differentiation or resting primed T cells (Hamann et al., 1996Hamann D. Baars P.A. Hooibrink B. van Lier R.A.W. Heterogeneity of the human CD4+ T-cell population: two distinct CD4+ T-cell subsets characterized by coexpression of CD4RA and CD45RO isoforms.Blood. 1996; 88: 3513-3521PubMed Google Scholar). The differentiation of naive to Th1 or Th2 cells seems to go through a phase where naive cells express small amounts of IL-4 with IL-2 (Kamogawa et al., 1993Kamogawa Y. Minasi L.E. Carding S.R. Bottomly K. Flavell R.A. The relationship of IL-4 and INF gamma producing T-cells studied by lineage ablation of IL-4-producing cells.Cell. 1993; 75: 985-995Abstract Full Text PDF PubMed Scopus (265) Google Scholar;Bullens et al., 1999Bullens D.M.A. Rafiq K. Kasran A. van Gool S.W. Ceuppens J.L. Naive human T-cells can be a source of IL-4 during primary immune responses.Clin Exp Immunol. 1999; 118: 384-391https://doi.org/10.1046/j.1365-2249.1999.01072.xCrossref PubMed Scopus (21) Google Scholar). The chromosomally clonal cells of our patients characteristically expressed IL-4, typical of RO+ type 2 Th cells, but only in one case (case 2) was IL-10 expression observed. Despite the variable expression of CDw150, the clonal cells of our patients did not express IFN-γ, usually upregulated by CDw150 (Cocks et al., 1995Cocks B.G. Chang C.-C.J. Carballido J.M. Yssel H. de Vries J.E. Aversa G. A novel receptor involved in T-cell activation.Nature. 1995; 376: 260-263Crossref PubMed Scopus (414) Google Scholar). Taken together, the phenotype CD45RA+, CD45RO+, CDw150±, IL-4+, IL-2, IFN-γ, and with the variation in IL-10 expression between the patients, the clonal cells seem to be intermediate forms between naive CD45RA+ and CD45RO+ Th2 cells. Possibly, they might represent cells that have not attained complete maturity, or they could be mature Th2 cells partly reverted towards a more naive or resting T cell type (Hamann et al., 1996Hamann D. Baars P.A. Hooibrink B. van Lier R.A.W. Heterogeneity of the human CD4+ T-cell population: two distinct CD4+ T-cell subsets characterized by coexpression of CD4RA and CD45RO isoforms.Blood. 1996; 88: 3513-3521PubMed Google Scholar). The differences in the immunohistologic staining intensity of IL-4-positive cells must be interpreted with caution, and the interactions with other cells of the microenvironment were not studied. Our finding of malignant cells with the phenotype CD45RA+, CD45RO+ is in concordance with the heterogeneity of RA+, RA+/RO+, and RO+ expression between four SS patients studied with flow cytometry by Urban et al. 1 Interestingly, the IL-4 positivity may contribute to the low frequency of apoptotic cells found in skin lesions of CTCL patients,2 as IL-4 blocks the action of caspase 3 (Manna and Aggarwal, 1998Manna S.K. Aggarwal B. Interleukin-4 down-regulates both forms of tumor necrosis factor receptor and receptor-mediated apoptosis, NK-κB, AP-1, and c-June N-terminal kinase.J Biol Chem. 1998; 273: 33333-33341Crossref PubMed Scopus (66) Google Scholar), and IL-4-producing T cells have been shown to be resistant to activation-induced apoptosis (Carbonari et al., 2000Carbonari M. Tedesco T. Del Porto P. Paganelli R. Fiorilli M. Human Tcells with a type-2 cytokine profile are resistant to apoptosis induced by primary activation: consequences for immunopathogenesis.Clin Exp Immunol. 2000; 120: 454-462https://doi.org/10.1046/j.1365-2249.2000.01243.xCrossref PubMed Scopus (18) Google Scholar). In contrast toVermeer et al., 1999Vermeer M.H. Geelen F.A.M.J. Kummer J.A. Meijer C.J.L.M. Willemze R. Expression of cytotoxic proteins by neoplastic T cells in mycosis fungoides increases with progression from plaque stage to tumor stage disease.Am J Pathol. 1999; 154: 1203-1210Abstract Full Text Full Text PDF PubMed Scopus (61) Google Scholar, who showed cytotoxic T-lymphocyte-associated granzyme B and cytotoxic-granule-associated TIA protein expression in morphologically identified malignant cells in MF patients, we did not find notable granzyme B expression in our SS patients in this or our previous study, 2Nevala H, Karenko L, Ranki A: Pro- and antiapoptotic markers in cutaneous T-cell lymphoma (CTCL) skin infiltrates and in comparison to lymphomatoid papulosis. Submitted. although the sample of lymphomatoid papulosis showed clear positive staining. Our finding of abundant amounts of malignant chromosomal clone cells in lymph nodes with dermatopathic lymphonoditis histology, and obtained even several months prior to malignant infiltrate in the skin, suggests that malignant clones may rise extracutaneously and they may invade lymph nodes early in the course of the disease. It is noteworthy that the SS of one patient evolved from MF. The concept of an early systemic nature of CTCL has gained evidence also from some previous studies (Whang-Peng et al., 1982Whang-Peng J. Bunn P.A. Knutsen T. Matthews M.J. Schechter G. Minna J.D. Clinical implication of cytogenetic studies in cutaneous T-cell lymphoma (CTCL).Cancer. 1982; 50: 1539-1553Crossref PubMed Scopus (83) Google Scholar;Veelken et al., 1995Veelken H. Wood G.S. Sklar J. Molecular staging of cutaneous T-cell lymphoma: evidence for systemic involvement in early disease.J Invest Dermatol. 1995; 104: 889-894Crossref PubMed Scopus (38) Google Scholar;Dommann et al., 1996Dommann S.N.W. Dommann-Scherrer C.C. Dours-Zimmermann M.-T. Zimmermann D.R. Kural-Serbes B. Burg G. Clonal disease in extracutaneous compartments in cutaneous T-cell lymphomas. A comparative study between cutaneous T-cell lymphomas and pseudo lymphomas.Arch Dermatol Res. 1996; 288: 163-167https://doi.org/10.1007/s004030050040Crossref PubMed Google Scholar;Trotter et al., 1997Trotter M.J. Whittaker S.J. Orchard G.E. Smith N.P. Cutaneous histopathology of Sèzary syndrome: a study of 41 cases with proven circulating T-cell clone.J Cutan Pathol. 1997; 24: 286-291Crossref PubMed Scopus (75) Google Scholar;Tok et al., 1998Tok J. Szabolcs M.J. Silvers D.N. Zhong J. Matsushima A.Y. Detection of clonal T-cell receptor gamma chain gene rearrangements by polymerase chain reaction and denaturing gradient gel electrophoresis (PCR/DGGE) in archival specimens from patients with early cutaneous T-cell lymphoma: correlation of histologic findings with PCR/DGGE.J Am Acad Dermatol. 1998; 38: 453-460Abstract Full Text Full Text PDF PubMed Scopus (50) Google Scholar), and early extracutaneous T cell chromosomal aberrations or T cell clonality have been verified even in large or small plaque parapsoriasis more recently (Karenko et al., 1997Karenko L. Hyytinen E. Sarna S. Ranki A. Chromosomal abnormalities in cutaneous T-cell lymphoma (CTCL) and its premalignant conditions as detected by G-banding and interphase cytogenetic methods.J Invest Dermatol. 1997; 108: 22-29Crossref PubMed Scopus (57) Google Scholar;Muche et al., 1999Muche J.M. Lugowsky A. Heim J. Friedrich M. Audring H. Sterry W. Demonstrating of frequent occurrence of clonal T-cell in the peripheral blood but not in the skin of patients with small plaque parapsoriasis.Blood. 1999; 94: 1409-1417PubMed Google Scholar). Thus, lymph node biopsies should be obtained during the early phases of CTCL, possibly even prior to clinical enlargement, to demonstrate the presence of malignant cells. A prerequisite for the use of FICTION in the routine study of lymph nodes or other tissues of CTCL patients, however, is the definition of clonal chromosomal or gene level aberrations suitable for in situ hybridization for identification of the malignant clone. The authors wish to thank Dr. Riitta Kauppila (Institute of Forensic Medicine, Helsinki University, Helsinki, Finland) for the postmortem samples, Dr. Tapio Visakorpi (Institute of Medical Technology, Tampere University, Finland) for the centromere-specific probes, Dr. Marketta Kähkönen, Ph.D., lecturer in genetics, for TCR analysis of the lymph node, and Mrs. Kaija Järvinen and Mrs. Marianne Karlsberg for technical assistance." @default.
• W2023862568 created "2016-06-24" @default.
• W2023862568 creator A5009120400 @default.
• W2023862568 creator A5010386138 @default.
• W2023862568 creator A5043654576 @default.
• W2023862568 creator A5069645530 @default.
• W2023862568 creator A5001298683 @default.
• W2023862568 date "2001-01-01" @default.
• W2023862568 modified "2023-10-14" @default.
• W2023862568 title "Chromosomally Clonal T Cells in the Skin, Blood, or Lymph Nodes of Two Sezary Syndrome Patients Express CD45RA, CD45RO, CDw150, and Interleukin-4, but no Interleukin-2 or Interferon-γ" @default.
• W2023862568 cites W1508488789 @default.
• W2023862568 cites W1525894265 @default.
• W2023862568 cites W1734655579 @default.
• W2023862568 cites W1963656304 @default.
• W2023862568 cites W1970221999 @default.
• W2023862568 cites W1970401456 @default.
• W2023862568 cites W1977834275 @default.
• W2023862568 cites W1979841260 @default.
• W2023862568 cites W1994390440 @default.
• W2023862568 cites W1996448581 @default.
• W2023862568 cites W2001045543 @default.
• W2023862568 cites W2005862831 @default.
• W2023862568 cites W2006159237 @default.
• W2023862568 cites W2031254710 @default.
• W2023862568 cites W2033626681 @default.
• W2023862568 cites W2046544122 @default.
• W2023862568 cites W2055970395 @default.
• W2023862568 cites W2062567403 @default.
• W2023862568 cites W2075685998 @default.
• W2023862568 cites W2087117578 @default.
• W2023862568 cites W2088256979 @default.
• W2023862568 cites W2093518407 @default.
• W2023862568 cites W2129663642 @default.
• W2023862568 cites W2165721674 @default.
• W2023862568 cites W2274031442 @default.
• W2023862568 cites W2337543356 @default.
• W2023862568 cites W2379022034 @default.
• W2023862568 cites W2473215883 @default.
• W2023862568 doi "https://doi.org/10.1046/j.1523-1747.2001.00207.x" @default.
• W2023862568 hasPubMedId "https://pubmed.ncbi.nlm.nih.gov/11168816" @default.
• W2023862568 hasPublicationYear "2001" @default.
• W2023862568 type Work @default.
• W2023862568 sameAs 2023862568 @default.
• W2023862568 citedByCount "15" @default.
• W2023862568 countsByYear W20238625682016 @default.
• W2023862568 countsByYear W20238625682022 @default.
• W2023862568 crossrefType "journal-article" @default.
• W2023862568 hasAuthorship W2023862568A5001298683 @default.
• W2023862568 hasAuthorship W2023862568A5009120400 @default.
• W2023862568 hasAuthorship W2023862568A5010386138 @default.
• W2023862568 hasAuthorship W2023862568A5043654576 @default.
• W2023862568 hasAuthorship W2023862568A5069645530 @default.
• W2023862568 hasBestOaLocation W20238625681 @default.
• W2023862568 hasConcept C142724271 @default.
• W2023862568 hasConcept C191732599 @default.
• W2023862568 hasConcept C203014093 @default.
• W2023862568 hasConcept C2776178377 @default.
• W2023862568 hasConcept C2777371288 @default.
• W2023862568 hasConcept C2778690821 @default.
• W2023862568 hasConcept C2779720271 @default.
• W2023862568 hasConcept C2780841837 @default.
• W2023862568 hasConcept C3020380090 @default.
• W2023862568 hasConcept C47348012 @default.
• W2023862568 hasConcept C71924100 @default.
• W2023862568 hasConcept C74172505 @default.
• W2023862568 hasConcept C86803240 @default.
• W2023862568 hasConcept C98119934 @default.
• W2023862568 hasConceptScore W2023862568C142724271 @default.
• W2023862568 hasConceptScore W2023862568C191732599 @default.
• W2023862568 hasConceptScore W2023862568C203014093 @default.
• W2023862568 hasConceptScore W2023862568C2776178377 @default.
• W2023862568 hasConceptScore W2023862568C2777371288 @default.
• W2023862568 hasConceptScore W2023862568C2778690821 @default.
• W2023862568 hasConceptScore W2023862568C2779720271 @default.
• W2023862568 hasConceptScore W2023862568C2780841837 @default.
• W2023862568 hasConceptScore W2023862568C3020380090 @default.
• W2023862568 hasConceptScore W2023862568C47348012 @default.
• W2023862568 hasConceptScore W2023862568C71924100 @default.
• W2023862568 hasConceptScore W2023862568C74172505 @default.
• W2023862568 hasConceptScore W2023862568C86803240 @default.
• W2023862568 hasConceptScore W2023862568C98119934 @default.
• W2023862568 hasIssue "1" @default.
• W2023862568 hasLocation W20238625681 @default.
• W2023862568 hasLocation W20238625682 @default.
• W2023862568 hasOpenAccess W2023862568 @default.
• W2023862568 hasPrimaryLocation W20238625681 @default.
• W2023862568 hasRelatedWork W1997441608 @default.
• W2023862568 hasRelatedWork W2054642945 @default.
• W2023862568 hasRelatedWork W2065367149 @default.
• W2023862568 hasRelatedWork W2076599494 @default.
• W2023862568 hasRelatedWork W2144596714 @default.
• W2023862568 hasRelatedWork W2153579277 @default.
• W2023862568 hasRelatedWork W2160715338 @default.
• W2023862568 hasRelatedWork W2462542604 @default.
• W2023862568 hasRelatedWork W3134597572 @default.
• W2023862568 hasRelatedWork W4361263315 @default.
• W2023862568 hasVolume "116" @default.
• W2023862568 isParatext "false" @default.

• next