Takahisa Yoshida

Lymph nodes in incipient adult T-cell leukemia-lymphoma with Hodgkin's disease-like histologic features

Lymph nodes were examined from four patients with incipient adult T‐cell leukemia‐lymphoma (ATLL) who had mild lymphadenopathy, fatigue, no or a few atypical lymphocytes in their peripheral blood, and integrated proviral human T‐cell lymphotrophic virus type I (HTLV‐I) DNA in the nodes. The HTLV‐I DNA was detected by southern blot analysis and/or polymerase chain reaction in the lymph nodes of all cases. The nodal architecture was preserved. Some scattered or aggregated highly lobular, cerebriform, or Reed‐Sternberg‐like giant cells were observed, with occasional mitoses and diffuse infiltration of small to medium‐sized lymphocytes, with no or minimal nuclear abnormalies in the enlarged paracortex. The giant cells were usually positive for Ki‐1 and also for UCHL‐1 and other T‐cell markers but negative for Ber‐H2. Rearrangement and/or deletion of T‐cell receptors were found in three of four patients. All patients died within 2 years, with transformation to overt leukemia‐lymphoma occurring in three patients, and pulmonary carcinoma in one. The incipient or prelymphomatous phase of ATLL should be differentiated from Hodgkins disease because of the distinctly different prognoses of these two diseases.

Clinicopathological study of Ki-1-positive lymphomas.

We examined an antibody against Ki-1 antigen in 161 cases of malignant lymphoma, four of histiocytic sarcoma, and six of nonspecific lymphadenitis, using monoclonal antibody Ki-1, which is known to react selectively with activated lymphocytes, Reed-Sternberg cells, and Hodgkins cells. Among them, 12 cases of malignant lymphoma demonstrated a diffuse positive cell membrane and/or cytoplasmic reaction of tumor cells and were categorized as Ki-1-positive lymphoma. Nine of these cases exhibited large cells with indented nuclei, distinct nucleoli, and abundant basophilic or amphophilic cytoplasm. Of the remaining three cases, two were of medium-sized and one of small-cell type. Immunologically, the 12 cases of malignant lymphoma demonstrated T-helper/inducer phenotype in six cases, B-cell in two case, and non-T, non-B in four cases. Tac and HLADR were positive in 9/12 and 4/5, respectively, and markers for histiocytes (lysozyme, alpha-1 anti-chymotrypsin, and OK-M1) were usually negative. Clinically, T-cell Ki-1-positive lymphoma was most likely to occur in the elderly, at extranodal sites, and had a rather poor prognosis (mean survival 35.5 months) as compared with B-cell and non-T, non-B lymphoma (7-52 months survival).

Anti-human T-cell lymphotrophic virus type I antibody-positive adult T-cell leukemia/lymphoma with no monoclonal proviral DNA. A clinicopathologic and immunologic study

Proviral DNA of adult T‐cell leukemia virus (HTLV‐I) was examined by the standard Southern blotting method in lymph nodes of 45 patients with anti‐HTLV‐I antibody (ATLA)‐positive adult T‐cell leukemia/lymphoma (ATLL). Six of these patients revealed no monoclonal proviral HTLV‐I DNA in tumor cells. These six patients showed typical flower cells in peripheral blood; they comprised five cases of the smoldering type and one of lymphoma type. They showed a longer clinical course than ATLL patients with integrated proviral HTLV‐I DNA. Five of the six patients were alive from 8 to 36 months after onset; the other patient died 9 months after onset. Histologically, they exhibited features of T‐cell malignancy but with absence of the typical cerebriform giant cells that are usually present in ATLL. The tumor cells represented T‐cell markers, usually CD4, but CD25 was negative. Rearrangement of the T‐cell receptor gene Cβ was found in four of the six cases. On the basis of these results, cases of ATLL with no monoclonal proviral HTLV‐I DNA should be clinicopathologically differentiated from those with integrated proviral DNA. Cancer 64:2515–2524, 1989.

Genotypic and Immunophenotypic Analysis of Anaplastic Large Cell Lymphoma (Ki-1 Lymphoma)*

Genotypic and immunological analysis was performed in 10 patients with anaplastic large cell lymphoma (Ki-1 lymphoma), 4 were male and 6 female. Immunophenotypically, 9 of these patients expressed some T cell markers; CD 2 in 9, CD 4 in 9, CD 3 in 6, CD 8 in 4, and UCHL-1 in 8. For B cell markers, 4 patients expressed B 1 and one expressed L 26; these patients also expressed T cell markers. In genotypic analysis, 9 of 10 cases displayed some incidence of T cell receptor gene (TCR) rearrangement or deletion, which was found in 3 patients for C beta 1, 7 for C beta 2, 8 for J gamma, 5 for C gamma (Hind III), 5 for C gamma (EcoRI), 3 for J delta 1 (Hind III), 3 for J delta 1 (BamHI), 3 for J delta 2, and 2 for C delta. One patient with rearrangement of TCR also showed rearrangement of immunoglobulin heavy chain. Another patient exhibited rearrangement of both TCR and immunoglobulin chain. These results indicate a common T cell lineage for this type of lymphoma.

A T‐cell neoplasia showing clinicopathologic features of malignant histiocytosis with novel chromosomal abnormalities and N‐ras mutation

Malignant histiocytosis (MH) is a distinct disease entity defined clinically and morphologically. However, the neoplastic origin of MH is not well established. The authors report a 26‐year‐old woman who showed the typical clinicopathologic features of so‐called MH. Cytogenetic and molecular genetic examinations were performed in addition to the morphologic and immunologic approach. The expression of CD2 and T‐cell receptor gene rearrangements indicated the T‐cell origin of this case. CD30, which is positive for anaplastic large cell lymphoma (Ki‐1 lymphoma), was not expressed. The cytogenetic study revealed a clonal chromosome abnormality involving 3q25, 6p21, 11p15, and 11q21. An N‐ras point mutation within codon 12 (GGT→GCT) was also detected. These findings indicate that MH defined clinically and morphologically is not a tumor of true histiocytic origin and that it should be reclassified on the basis of immunologic, cytogenetic, and molecular genetic data.

Establishment of hybridomas producing cancer specific human antibodies from B cell line derived from PBL of a patient with adult T cell leukemia

Adult T cell leukemia (ATL) is a malignant disease characterized by tumorous proliferation of CD4+ T cells infected with retrovirus human T cell leukemia virus Type-I (HTLV-I) and concurs with an autoimmune disease and cancer due to attenuated immune response. In this study, we established ATL patient derived B-cell line TM-1 producing cancer-specific IgM antibodies, and further characterized its antigen specificity by establishing hybridomas fused with human-mouse origin hetero-myeloma cell line RF-S1. We established three hybridoma cell lines termed 2E12, 3E9, and 3E10, which continuously secreted human IgM antibodies. Immunohistochemical staining of formalin-fixed tissue section using antibodies secreted from these hybridomas showed that these antibodies specifically recognized tumor sites of human colon adenocarcinomas. Antibody produced from hybridoma 3E9 bound to some of leukemic cell lines, but not to normal human PBL, which was evidenced by the flow cytometric analysis, indicating that antibody produced from 3E9 recognizes cell surface antigen specifically expressed in the leukemic cells.

Limited TCR Vβ Usage of Infiltrating T cells in Synovial Tissues from Patients with HTLV-I Associated Arthropathy

Human T cell lymphotropic virus type-I (HTLV-I) is the etiologic agent of adult T cell leukemia/lymphoma and recently has also been suggested to be involved in chronic arthritis. The synovia of patients with rheumatoid arthritis (RA) contains activated T lymphocytes, with a restricted expression of T cell receptor (TCR) variable (V) beta gene segments. To characterize the T-cell populations of RA among HTLV-I carriers and noncarriers, we performed the immunohistochemical staining of CD4 and CDB, as well as a reverse transcription polymerase chain reaction (RT-PCR) to estimate the proportion of TCR beta RNA containing any particular V elements on the synovial specimens. In all but one HTLV-I carrier, the proviral DNA and/or RNA expression of HTLV-I was detected in the synovium. The CD4-positive cells proliferated markedly in the HTLV-I carriers compared with the noncarriers. In contrast to mononuclear cells in the peripheral blood, synovial T cells expressed only a few V beta transcripts, and no definite difference was observed between the carriers and the noncarriers. These results suggest that a common major antigen associated with the pathogenesis of RA may thus selectively interact with the V beta component of the TCR. Using RT-PCR, we studied the expression of the recombination-activating gene-1 (RAG-1), which was used in the V(D)J recombination of immunoglobulin and TCR genes. In all cases, RAG-1 was transcripted. The results supported the possibility that the extrathymic development of the selected TCR V beta T cells occurred in the synovia.

Diagnosis of high risk lymphoma by molecular genotypic analysis: Bigenotype and multiple rearranged bands as risk factor.

We examined 132 non-Hodgkins lymphomas for rearrangements of TcR β and Ig genes. These histologic subclassification was based on the modified International Working Formulation (WF). Of these, 58 were derived from T-cell and 74 were from B-cell due to the results of immunophenotype. In high grade group of T-cell lymphomas, bigenotypic pattern with Cβ and JH probe was 14% and multiple rearranged bands pattern was 13%. On B-cell lymphomas, the bigenotypic pattern was 12% in high grade, 4.7% in intermediate grade and nothing in low grade. The multiband pattern was observed in only two cases (one is lymphoblastic subtype and the other one large). However, there is no corellation between the frequency of biphenotypic or multibands pattern, and WF subclassification in B-cell lymphoma. The presence of these specific rearrangement patterns in T-cell lymphoma (except for AILD) was associated with a poor prognosis. These findings indicate that the presence of bigenotypic and/or multibands pattern indentifies a high-risk group of patients with T-cell lymphoma.