Kunihiko Tominaga

Analysis of the immunoglobulin heavy chain gene of secondary diffuse large B-cell lymphoma that subsequently developed in four cases with B-cell chronic lymphocytic leukemia or lymphoplasmacytoid lymphoma (Richter syndrome)

The immunoglobulin heavy chain gene (IgH gene) was analysed in four cases of B‐cell Richter syndrome, in order to determine whether a secondary diffuse large B‐cell lymphoma (DLBCL) could arise from the same clone as the initial B‐cell chronic lymphocytic leukemia (B‐CLL) and lymphoplasmacytoid lymphoma (LPL) or be a de novo event, and whether secondary DLBCL shows an intraclonal microheterogeneity. Both the initial B‐CLL and secondary DLBCL in two cases expressed CD5 antigen. Both samples of the initial B‐CLL or LPL and the secondary DLBCL in three cases were examined for comparison. The polymerase chain reaction‐amplified IgH gene of secondary DLBCL in two cases (CD5+ case and CD5− case) were different from those of the initial B‐CLL, revealing a new malignant clone. The other case (CD5−) showed that secondary DLBCL had a sequence identical to the initial LPL, indicating the same clonal origin. The variable region of the IgH gene of secondary DLBCL (CD5+ two cases and CD5− two cases) exhibited a 0.5–9.0% somatic mutation range and no intraclonal microheterogeneity.

Histiocytic sarcoma of the spleen associated with hypoalbuminemia, hypo gamma-globulinemia and thrombocytopenia as a possibly unique clinical entity--report of three cases.

We report three patients with histiocytic sarcoma of the spleen associated with severe hypoalbuminemia, hypo gamma-globulinemia and thrombocytopenia. After the clinical diagnosis of splenic tumor of unknown origin was made, all three patients underwent splenectomy. The histiocytic origin of the tumor was confirmed histopathologically and immunohistochemically using a panel of antibodies. In contrast to malignant histiocytosis (MH), which typically reveals severe generalized clinical manifestations and a rapidly fatal course caused by the disseminated proliferation of neoplastic histiocytes, these patients were asymptomatic or showed only mild clinical symptoms for a long period of time until the recurrence was detected by which time the tumor cells had already spread to other organs. All three cases were characteristically associated with hypoalbuminemia, hypo gamma-globulinemia and thrombocytopenia, which returned to normal after splenectomy. Splenic histiocytic sarcoma with the features described here may represent a unique clinical entity, distinct from MH.

Analysis of lectin binding properties on human Burkitt's lymphoma cell lines that show high spontaneous metastasis to distant organs in SCID mice : The binding sites for soybean agglutinin lectin masked by sialylation are closely associated with metastatic lymphoma cells

Alterations in cell surface carbohydrates on human lymphoma cell lines with different spontaneous metastatic potential in the severe combined immunodeficiency (SCID) mouse model were analyzed. A difference In cell surface carbohydrates between high‐ (HBL‐2, HBL‐7 and HBL‐8) and no‐ or low‐(HBL‐4, HBL‐6, Daudi and Raji) spontaneous metastatic human lymphoma cell lines were analyzed on a FACScan using fluorescein‐isothiocyanate (FITC)‐conjugated lectins. The most consistent difference in lectin binding properties was found with soybean agglutinin (SBA) lectin. High‐metastatic lymphoma cells (HBL‐7 and HBL‐8 cells) in vitro were found to bind the SBA lectin, but the cells in vivo (in primary tumors and metastatic tumors of SCID mice) showed considerably reduced SBA lectin binding. In addition, HBL‐2 cells that almost did not bind SBA lectin in vitro and in vivo showed high spontaneous metastasis. Neuraminidase treatment revealed that SBA lectin binding sites were masked by sialic acid. On the other hand, no‐ or low‐metastatlc lymphoma cells in vitro and in vivo were found to bind SBA lectin. HBL‐8 cell clones without SBA lectin binding showed high spontaneous metastasis to distant organs in SCID mice but HBL‐8 cell clones with SBA lectin binding showed very low spontaneous metastasis. Sophora Japonica agglutinin (SJA) lectin is able to recognize the carbohydrates in common with SBA lectin, but it does not appear to be associated with metastatic capacity. These results suggest that the sialylation of particular carbohydrate residues on human lymphoma cells that are recognized by SBA lectin may be associated with the spontaneously metastatic capacity of human lymphoma cell lines in our SCID mouse model.

NECROTIZING LYMPHADENITIS Electron Microscopical and Immunohistochemical Study

Seven cases of necrotizing lymphadenitis (NEL) including a pair of male siblings, a female suffering from non‐bacterial meningitis, and two cases with the proliferation of monocytes and/or macrophages in the bone marrow are reported. This disease is clinically documented by the occurrence in young adults usually accompanied by painful cervical lymphadenopathy with fever and leukopenia (below 4,000/mm3). Morphological features were characterized by nuclear debris from degenerated lymphocytes and the appearance of blastoid cells and/or immunoblasts and macrophages. Ultrastructurally, tubular Inclusions with a close relation to the endoplasmic reticulum were observed in various kinds of cells in the lesion. Immunohistochemical studies revealed that the ratio of helper/suppressor T‐lymphocytes became low at the active stage and returned to normal range at the recovery stage. By immunohistochemical study it was confirmed that suppressor cells mainly corresponded to large transformed lymphocytes and/or immunoblasts and helper cells were degenerated by an unknown agent. Though the pathogenesis of NEL is still uncertain, it is suggested that T‐lymphocytes are mainly involved during the course of the disease and lymphocytes show cellular debris or blastoid transformation. ACTA PATHOL. JPN. 37:1071–1084, 1987.

Clinicopathological and immunophenotypic studies on 12 cases with B cell chronic lymphocytic leukemia

To clarify the histogenesis of B cell chronic lymphocytic leukemia (BCLL), clinicopathological and immunophenotypic studies were performed using a large panel of monoclonal antibodies on 12 cases with BCLL including three caes with prolymphocytic/chronic lymphocytic leukemia (CLL/PL). Immunophenotypically, CD19 and CD20 were positive for all cases of this series and CD5, CD21, CD22, CD23, CD25, CD38, Leu‐8, KB‐61, and bcl‐2 protein were expressed in variable proportion from case to case. CD10, however, did not react. No alkaline phosphatase (ALP) positive cases were found. The phenotype of BCLL was similar to that of B cells of the mantle zone (MZ) of secondary follicle in the lymph node. It is therefore postulated that the neoplastic cells of BCLL in these cases might be derived from B cells of the MZ. Moreover, the cells possibly originated from the lymphocytes located in the inner layer of the MZ, since ALP+ B cells are usually observed in the outer layer of the MZ. The pseudofollicular (PF) pattern was observed in four biopsied lymph nodes among five cases tested, but no such a pattern in an aspiration clot of bone marrow. These four cases consisted of three cases with CLL and a case with CLL/PL. The immunohistochemical study showed that there were many proliferating cells showing Ki‐67+ in the PF area of the lymph nodes. In these cases, leukemic cells might have developed from the PF area of the lymph node.

A Case of Lymphoblastic Lymphoma with Pre-B Cell Phenotype

Naoya Nakamura, MD, Department of Pathology, Fukushima Medical College, 1 Hikarigaoka, Fukushima 960-12 (Japan) Lymphoblastic lymphoma usually shows a T cell phenotype [1], but rarely some lymphoblastic lymphoma shows a pre-B cell phenotype. We present an uncommon case of lymphoblastic lymphoma expressing a pre-B cell phenotype and review the literture. An 11-year-old Japanese boy complained of right cervical and bilateral inguinal lymphadenopathy in April 1986. Hepatosplenomegaly was not found. A surgical specimen obtained from the right cervical lymph node showed malignant lymphoma, lymphoblastic type. A chest X ray and computed tomographic scan of the thorax and abdomen revealed no abnormal finding. A hemogram of peripheral blood did not show leukemic picture. Bone marrow aspiration showed normocellularity with no leukemic cell. The patient received a combination chemotherapy and prophylactic cranial irradiation (2,400 rad), and went into complete remission. He is now receiving maintenance chemotherapy. Microscopically, normal architecture of the cervical lymph node was almost effaced by a diffuse infiltration of atypical lymphoid cells consistent with lym-phoblasts, although a few atrophic follicles were observed. The neoplastic cells had convoluted nuclei with dusty chromatin and small prominent nucleoli, and scanty basophilic cytoplasm. Electron microscopy revealed that the neoplastic cells had indented or convoluted nuclei with small prominent nucleoli. The chromatin was evenly distributed in the nucleus with peripheral condensation of heterochromatin along the nuclear membrane. The cytoplasm possessed numerous ribosomes and polyribosomes, several mitochondria and a few short rough endoplasmic cister-nae. The neoplastic cells reacted with anti-HLA-DR, anti-CD19(B4), and antiCDlO(CALLA), but not with anti-CD20(Bl), anti-CD21(B2), anti-T-cell-associated antigens and anti-myeloid-associated antigens. The neoplastic cells did not express any surface immunoglobulins. Immunoelectron microscopy demonstrated that the neoplastic cells bored the cytoplasmic μ-chain (fig. 1.). In addition, the neoplastic cells were positive for TdT activity. These findings indicate that the present case represents a neoplastic counterpart of pre-B cell. Non-T ALL with a pre-B cell phenotype is not uncommon. However, pre-B cell lymphoblastic lymphoma without leukemic blood picture is very rare; to our knowledge several cases have been reported [2–7] (table 1). These reported cases displayed the common characteristics of indented or convoluted nuclei as well as the preferential involvement of skin and lymph nodes. In addition, these cases often showed lytic bone lesions. The present case is somewhat different

Alkaline phosphatase-positive B cell lymphomas

Alkaline phosphatase (ALP) activity of 70 cases of non‐Hodgkins lymphomas of the B‐cell type was studied. ALP activity was found in malignant lymphoma (ML), follicular, small cleaved cell (1/5 cases); ML, diffuse, small cleaved cell (3/13 cases); and mantle zone lymphoma (intermediate lymphocytic lymphoma) (2/2 cases). The ALP‐positive neoplastic cells simultaneously displayed the characteristic immunophenotype of mantle zone (MZ) B lymphocytes of secondary follicle (SIg D +, BA‐1 +, IL‐2R+ and Leu‐1+). All other B‐cell lymphomas, including ML, follicular, mixed small cleaved and large cell (9 cases); ML, follicular, large cell (4 cases); ML, mixed small and large cell (7 cases); ML, diffuse, large cell (27 cases); and ML, small noncleaved cell (3 cases), were consistently negative for ALP. The present study indicates that ALP‐positive lymphomas including follicular lymphomas and diffuse lymphomas may be neoplastic counterparts of ALP‐positive MZ B lymphocytes.

Characterization of a newly established human acinic cell adenocarcinoma cell line (HACC) originating from the salivary gland : Morphological features and role of various growth factors on the growth of the HACC cell line

Human acinic cell adenocarcinoma cell (HACC) line was established from the pleural effusion that contains meta‐static tumor cells of acinic cell adenocarcinoma of papillary and microcystic type originating from the parotid gland. The HACC cells grew in an adherent monolayer with a doubling time of 66 h. Implanted tumor of SCID mice revealed similar histologlcal findings to that of the primary tumor. The HACC cells produced mucin and expressed epithelial markers as well as α1‐antitrypsin and lysozyme, whereas salivary peptide P‐C was expressed in cultured HACC cells but not In the primary and Implanted HACC cell tumors. S‐100 protein was also expressed in both the primary tumor and HACC cell line. Neither amplification of common oncogenes nor expression of p53 was observed. The receptor for epidermal growth factor (EGF) was expressed, indicating EGF and transforming growth factor‐α (TGF‐α) enhanced the growth of the HACC line. Unexpectedly, tumor necrosis factor‐α (TNF‐α) also enhanced the growth of the HACC line significantly. However, there was no evidence of autocrine growth using these growth factors. In contrast, TGF‐β1 inhibited the growth of the HACC cell line through apoptosis. The HACC cell line has features similar to both acinar and intercalated ductal cells of the salivary gland. Epidermal growth factor, TGF‐α and TNF‐α are potential growth factors for the HACC cell line. The HACC cell line may be a good model for studying the biological behavior of salivary gland neoplasms.