Interferon-Induced Surface Alterations in Hairy Cells. A Review

Hairy cells (HCs), derived from the peripheral blood and spleen of hairy cell leukemia (HCL) patients, constantly displayed both ruffles and microvilli. HCs which were kept in culture for up to three days exhibited extremely polarized and active surfaces with elongated microvilli and exaggerated spiked ruffles. Cells derived from 11 cases of HCL were treated with alpha-interferon (IFN) in-vitro and examined by immuno -scanning electron microscopy (immuno-SEM). In 8 cases, up to one-third of the IFN-treated hairy cells displayed deformed surfaces with bubbling membrane and markedly villous bud-like formations. Monoclonal antibodies (MoAb), used in conjunction with immuno-qold labeling, facilitated better correlation between these morphological changes and the immunological profiles of the cells before and after interferon treatment in-vitro. Immuno-SEM analyses revealed no remarkable changes in the labeling of HCs with Leu-14 and Leu-MS MoAbs before and after IFN treatment, even in cases showing membrane changes. However, a significant increase in the labeling intensity for HLA-DR and HLA-DQ was noticed in HCs from cases where IFN-induced membrane changes were evident. A review of the literature on membrane changes in IFN-treated cells proposes that such immuno-ultrastructural alterations might reflect unique interferon-induced membrane reorganization in the target malignant cells

Phorbol Ester (TPA)-Induced Surface Membrane Alterations in B-Type Hairy Cell and Lymphocytic Leukemia Cells

This report documents phorbol ester (TPA)-induced changes in cell morphology, and in vitro growth patterns in 9 patients with hairy cell leukemia (HCL), 21 with B-type CLL and non-Hodgkin\u27s lymphoma in leukemic phase (NHL), and 10 with acute non lymphoblastic leukemia (ANLL). TPA caused cells from HCL to adhere strongly and produce elongated cytoplasmic extensions. Many of these cells had an appearance resembling fibroblasts, while others showed marked surface ruffling and spreading containing increased dense bodies, and phagolysosomal vacuoles as seen by transmission electron microscopy. This HCL in vitro growth pattern after TPA exposure differed from that seen in B-CLL and NHL cells, which only adhered moderately after 72 hours and readily detached in clumps. CLL and NHL-cells did not show ultrastructural features of macrophages but had either plasmacytic or HCL features. It is suggested that these different growth patterns may aid in distinguishing HCL from other B-cell neoplasias. The expression of surface markers, tartrate resistant acid phosphatase (TRAP) and Ig secretion were studied in some B-CLL, NHL and HCL cells after exposure to different concentrations of TPA for up to 6 days. Results showed that the documented changes were frequently both dose and time dependent and the most striking HCL-features were encountered after 6 days incubation with higher concentrations of TPA. However, individual variation from case to case was noted. Nevertheless, it seems that TPA induces neoplastic B-cells to mature into secreting plasmacytoid lymphocytes, and cells with features of HCL with variable expression of surface markers and TRAP

Topical Modes in the Preparation of Human Spleen Specimens for Routine Scanning Electron Microscopy Studies

Various preparatory techniques were used to improve scanning electron microscopy images of the fine structure of vascular, cellular, and cordal-reticular components of normal human spleens. The progressive method of fixation (GTGO) applied in the present study, allowed air drying of the tissues and rendered the specimens conductive even in newly fractured surfaces. Vascular perfusion proved necessary only in studies of the splenic blood vessels, while a simple immersion of tissue blocks in the washing solution resulted in better images of the white pulps. Interstitial (trans-splenic) perfusion was found to be superior to vascular perfusion for routine preparation of spleen tissues, and freeze-cracking did not necessarily lead to improved images of the specimen\u27s surfaces. Combined with proper washing and shaping protocols, the GTGO procedure is shown to be a superior mode of specimen preparation, abolishing most traditional artifacts and obtaining clear images of the complex splenic tissue