Biomedical subjects
Publications and source records attributed to Y Ohtsuki.
A 22-year-old female with a thymic carcinoma is reported. The tumor was refractory to both chemotherapy and irradiation. The patient died with an aggressive clinical course. Cytogenetic study showed that the tumor cells had a chromosome translocation, t(15;19)(q15;p13), which was not identified previously in human cancer.
The immunohistochemical distribution of alpha and beta subunits of S-100 protein (S-100 alpha, S-100 beta, respectively) in 138 cases of human brain tumors was investigated by the avidin-biotin immunoperoxidase method. Brain tumors can be divided into four groups: group 1 [S-100 alpha (+) and/or S-100 beta (+)]; astrocytoma, glioblastoma, ependymoma, subependymoma, oligodendroglioma, choroid plexus papilloma, gangliocytoma, meningioma, chordoma, malignant melanoma. Group 2 [S-100 alpha (+) and S-100 beta (-)]; pineoblastoma, pituitary adenoma, craniopharyngioma, rhabdomyosarcoma. Group 3 [S-100 alpha (-) and S-100 beta (+)]; acoustic Schwannoma. Group 4 [S-100 alpha (-) and S-100 beta (-)]; medulloblastoma malignant lymphoma, germinoma. The S-100 beta immunoreactivity pattern in brain tumors was similar to those obtained using conventional anti-S-100 protein sera. In the first group of brain tumors both the number of positively stained tumor cells and the staining intensity were generally greater for S-100 beta than for S-100 alpha with a few exceptions including one gemistocytic astrocytoma, one subependymoma, one malignant melanoma, and some cases of glioblastomas. As to the relationship between malignancy and S-100 protein in glioma, S-100 beta immunoreactivity decreased according to degree of malignancy, while that of S-100 alpha varied, suggesting a heterogeneity of tumor cells in glioblastomas. Immunostaining for S-100 alpha and S-100 beta might become a useful diagnostic procedure in brain tumors and may give us more detailed and precise data of S-100 protein in brain tumors.
Two cell lines, CNS-5 and CNS-6, were established by cocultivation of sedimented cells in cerebrospinal fluid (CSF) from two anti-human T-lymphotropic virus type I (HTLV-I) antibody-positive male patients with encephalopathy and HTLV-I-associated myelopathy, respectively, with peripheral blood mononuclear cells from a healthy seronegative female. These cell lines, possessing a normal female karyotype, revealed similar characteristics as follows; they expressed HTLV-I-related antigens, they produced C-type retrovirus particles, HTLV-I provirus genomes were integrated into their DNAs, and they had CD4+ activated T-cell markers. In addition, immunocytochemical and immunoelectron microscopic studies showed peculiar immunoreactivity of these cell lines with anti-alpha/beta T-cell antigen receptor (TCR) antibodies; beta Fl, defining beta chain epitope, was only positive in the perinuclear spaces and rough endoplasmic reticulum in some cells, and WT31, recognizing alpha/beta framework, was mostly negative, while CD3 was expressed in the majority of the cells. These facts indicate that HTLV-I-infected cells were present in CSF of these two patients, and suggest that neurological disorders associated with HTLV-I may not be restricted to myelopathy and may include brain abnormalities.
A monoclonal antibody, termed K-20, was generated against an anaplastic thymic carcinoma cell line, Ty-82. Subcapsular thymic epithelial cells of the thymus and blood vessels in various organs were shown to react with the K-20 monoclonal antibody by immunohistochemical staining. Immunofluorescent study revealed that various haematopoietic fresh cells and cell lines did not show any significant reactivity with K-20, except for one Epstein-Barr-virus-carrying lymphoma cell line (SP-50B). Western immunoblotting and affinity purification procedure revealed that K-20 was directed to a protein with a molecular weight of 28 kDa. K-20 is unique in its restrictive reactivity with human subcapsular thymic epithelial cells.
Twelve cases of phaeochromocytoma (PCC) and four cases of paraganglioma (PGG) were studied by immunohistochemistry and immunoelectron microscopy in order to demonstrate HLA-DR (Ia)-antigen-positive dendritic cells (IaDCs). Dense infiltration of IaDCs was detected in the majority of PCCs revealing high urinary or serum catecholamine levels, but in aggressively growing PCCs, a familial PCC and all PGGs, few IaDCs were demonstrated. Interestingly, these IaDCs were negative for S-100 protein. Although S-100-protein-positive sustentacular-like cells (SCs), morphologically similar to IaDCs, were also present, these were clearly distinguished from IaDCs by our double immunostaining method. Ultrastructurally, IaDCs had smooth or slightly indented nuclei and contained a moderate amount of endoplasmic reticulum, small mitochondria and vacuoles, extending elongated cytoplasmic processes. These results suggest that determination of the quantity of IaDCs is a highly effective method of assessing the character of PCCs, in particular, their prognosis.
A newly established human osteosarcoma cell line, HS-Os-1, from an osteoblastic tumor arising in the left humerus of an 11-year-old girl was morphologically characterized in vitro and in vivo. HS-Os-1 cells in a monolayer have been maintained for more than 2 years since the initial cultivation, and were round or polygonal in shape with marked pleomorphism. Their cytoplasm was strongly positive for specific markers of osteoblasts, such as alkaline phosphatase and osteocalcin. Tumors induced in nude mice by HS-Os-1 cell inoculation at passage 12 or 23 revealed typical histological features of osteoblastic osteosarcoma, similar to those observed in the original tumor, producing prominent osteoid matrix with calcification. Ultrastructurally, HS-Os-1 cells in vitro and tumor cells in vivo showed similar well-developed, markedly dilated rough endoplasmic reticulum, polysomes and microfilaments in their cytoplasm. Additionally, many collagen fibers associated with deposition of electron-dense material were detected in the stroma featuring osteoid matrix. Thus, the HS-Os-1 cell line was shown to exhibit its osteoblastic nature in vitro and in vivo, and therefore might become an extremely useful tool for various pathomorphological investigations on human osteosarcomas.
Single cilium formation was studied in two human hematopoietic cell lines, KCL-22 and MT-2, KCL-22, established from a patient with chronic myelogenous leukemia in blastic crisis, and MT-2, a human cord T cell line carrying human T-lymphotropic virus type I, were transplanted into hamsters. Ciliogenesis was observed in both cell lines, only after transplantation into hamsters.
We present a case of pre-elastofibroma-like lesion, a kind of elastic-producing fibrous tumor. The small colonic polyp, which was found in a 49-year-old asymptomatic man in association with a large colonic adenoma, showed submucosal nodular deposits of fine granular or fibrillar eosinophilic materials with interspersed fibroblastic cells. Elastic stain revealed these deposits to consist mainly of dark gray granular or partially fibrillar dense elastinophilic materials, most of which were digested with elastase. This stromal lesion somewhat resembled a pre-elastofibroma. Therefore, pre-elastofibroma-like lesions should be kept in mind as a possible origin of colonic polyp.
A case of pancreatic pseudocyst complicating with pregnancy and delivery was reported. This primipara patient with pancreatic pseudocyst was able to achieve a full-term, transvaginal delivery of a mature baby. The usefulness of ultrasonic monitoring of the pseudocyst during labor was clinically demonstrated. When intra-abdominal pressure increased, pancreatic pseudocyst was deformed.
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A new human lymphoma cell line, designated DL-40, was established from the peripheral blood of a 64-year-old woman with leukemic conversion of aggressive large cell lymphoma. The cell line grew in suspension with or without forming clumps of cells and exhibited large, round, or multiple nuclei in the relatively abundant cytoplasm that was positive for acid phosphatase. The cells expressed a Ki-1 antigen (CD30), E+, CD2+, CD4+, CD45+, Ia+ phenotype and had rearranged T-cell receptor beta chain but were negative for CD15, HTLV-I, and Epstein-Barr virus nuclear antigen. Chromosome analysis of this cell line showed a human female karyotype with complex hyperdiploid abnormalities. DL-40 cells produced tumors histologically similar to the original lymphoma when transplanted into nude mice and immunosuppressed hamsters. The DL-40 cell line could provide a useful tool for the understanding of biology of the Ki-1-positive non-Hodgkin's lymphoma.
To determine the minimum volume of blood required to transmit human T-cell leukemia virus type I (HTLV-I), heparinized blood was collected from a virus-infected female rabbit and aliquots of 10, 5, 1, 0.5, 0.1, and 0.01 mL were transfused into groups of two male rabbits each. All 10 rabbits transfused with 10 to 0.1 mL and 1 of 2 rabbits transfused with 0.01 mL seroconverted for HTLV-I after 2 to 4 weeks. HTLV-I-producing lymphoid cell lines of recipient origin were established from one seroconverted rabbit of each aliquot group. To determine the ability of passive immunization to protect against HTLV-I infection, two groups of three rabbits were first transfused with 5 mL of blood from the same virus-infected rabbit and then infused after 24 or 48 hours with 10 mL of HTLV-I immune globulin (77 mg/mL of IgG) prepared from seropositive healthy persons. None of the 24-hour immunization group seroconverted for HTLV-I during the observation period of six months; however, all of the 48-hour immunization group became seropositive after 2 to 4 weeks. These results indicate that HTLV-I can be transmitted with as little as 0.01 mL of virus-infected blood, and that passive immunization is effective in preventing cell-to-cell infection of HTLV-I when given within 24 hours of transfusion of virus-infected blood.
An isochromosome 11q in a patient with M5a type acute monoblastic leukemia is reported. The leukemic cells had a few azurophilic and alpha-naphthyl butyrate esterase-positive granules in the cytoplasm. Electron microscopy showed indented nuclei, abundant perinuclear fibrous bundles, and small lysosomes which were characteristic of monocytoid cells. A strong inhibitory activity against urokinase was detected in the cell lysates as compared with other leukemic cells. Cytogenetic analysis of the leukemic cells showed the karyotype 47, XY, +i(11q), which has not been observed hitherto in acute monocytic leukemia.
Four rabbits inoculated intravenously with milk cells from 4 post-partum women seropositive for human T-cell leukemia virus type I (HTLV-I) and one rabbit inoculated with semen cells from a seropositive healthy man seroconverted for HTLV-I after 3-5 weeks but no seroconversion occurred in 2 rabbits inoculated with milk cells from a seronegative mother or with heated (56 degrees C, 30 min) milk cells from a seropositive mother. Attempts were made to isolate HTLV-I from peripheral blood lymphocytes harvested 5-15 weeks after cell inoculation and cultured in the presence of interleukin-2. An HTLV-I-carrying lymphoid cell line of rabbit origin was established from a rabbit inoculated with milk cells. Another long-term culture, derived from a rabbit inoculated with semen cells, also expressed HTLV-I antigens and harbored virus particles. Furthermore, transfusion of 20 ml of blood from all 5 seroconverted rabbits, but not from the 2 seronegative ones, caused seroconversion in normal recipient rabbits after 4-6 weeks.
Type-C virus-like particles (VLPs) were found in an Epstein-Barr (EB) virus-infected human B-cell lymphoma cell line, SP-50B, that was established from a patient with non-Hodgkin lymphoma. The cell line continuously produces a small number of type-C VLPs, 150-200 nm in diameter, over 1 year. SP-50B cells were negative for HTLV-I and HTLV-II antigens and did not contain the HTLV-I genome. In addition, two EB virus nuclear antigen (EBNA)-positive B-cell lines, SP-54-Cord and SP-57-CLL, were established from human cord blood and chronic lymphocytic leukemia (CLL), respectively, by coculture with lethally irradiated SP-50B cells. Type-C VLPs with the same morphology were also found in both cell lines.
Co-cultivation of peripheral blood mononuclear cells, from a 4-year-old boy with infectious mononucleosis, with a human T lymphotrophic virus type I (HTLV-I)-producing T-cell line resulted in the establishment of an HTLV-I infected Epstein-Barr virus (EBV)-producing B-cell line. Ultrastructural examination revealed the presence of ribosome-lamella complexes (RLCs), in a small proportion of long-term cultured cells. Evidence is presented of continuity between RLC and the rough endoplasmic reticulum, and in some instances there was also an intimate relationship with tubuloreticular structures. The findings support the view that RLC is derived from the rough endoplasmic reticulum and also provide further evidence of the non-specific nature of RLC in relation to cell type.
The effects of human alpha-, beta-, or gamma-interferon (IFN) on the replication and production of human T-lymphotrophic virus type-I (HTLV-I) were investigated in a human T-cell line, MT-2. Virus transmission and production estimated by syncytium formation and HTLV-I-associated reverse transcriptase (RT) activity were strongly suppressed in the presence of alpha- and beta-IFN, but not gamma-IFN. However, the expression of virus specific proteins gp46 but not p19, p24, p28, p36, and gp68 was affected with IFNs as revealed by Western blotting analysis. Electron microscopic observations showed that some of the HTLV-I particles were trapped in the intracellular vacuoles in the presence of high doses of alpha- or beta-IFN. Continuous supply of IFNs appeared to be essential for the constant suppression of RT activity. These results suggest that alpha- and beta-IFN do not inhibit HTLV-I gene expression strikingly but suppress processing or assembly of virus proteins and/or releasing of virions in the late phase of maturation.
In order to clarify the function of human S100 beta-positive T-cells, S100 beta-positive T-leukemia cells (S100 beta TLC) were examined in vitro. S100 beta TLC were obtained from the peripheral blood of a patient with S100 beta-positive T-cell leukemia and enriched by an E-rosetting method. Two dimensional flow cytometric analysis indicated that the vast majority of the E-positive fraction were S100 beta TLC expressing CD3 and CD8 antigens. Although S100 beta TLC expressed CD3 antigen, they were negative for the alpha/beta and gamma/delta T-cell antigen receptor (TCR) defined by monoclonal antibodies (mabs) WT-31 and delta TCS-1, respectively. It was speculated that S100 beta TLC initially expressed alpha/beta TCR but lost it during malignant transformation. When S100 beta TLC were cultured for 24 h, they acquired cytotoxic activity towards various NK-sensitive cell lines including K-562, Molt-3 and CEM-CCLF, but did not exhibit lysing activity towards NK-resistant cell lines including Raji, Daudi and MT-1. Despite the NK-activity of cultured S100 beta TLC, they lacked the morphological features of large granular lymphocytes (LGL). S100 beta TLC did not exhibit lymphokine-activated killer (LAK) activity. When S100 beta TLC were cocultivated with NK-sensitive cells or NK-resistant cells, they selectively bound to NK-sensitive cells, indicating that they lysed target cells by cell-to-cell contact. The finding that S100 beta TLC lacked TCR molecules and their NK activity was not inhibited by mabs reactive with the CD3-TCR complex indicated that the CD3-TCR complex was not involved in their target recognition.(ABSTRACT TRUNCATED AT 250 WORDS)