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The changes occurring in surface morphology during the transformation of normal peripheral blood lymphocytes by phytohaemagglutinin (PHA) are described and the surface ultrastructure of the PHA-induced blasts is compared with that of pokeweed mitogen (PWM)-and rabbit anti-beta2-microglobulin antiserum-induced blasts. Both mitogen-specific and non-specific changes were observed and their possible relationship to the activation of lymphocyte subpopulations is discussed. Similar surface characteristics found in various tissue lymphoid cells are also described.

The properties have been studied of a haemic cell line established from a patient with longstanding non-Hodgkin's lymphoma after transformation to acute leukaemia. The cells are EBNA, Fc, C3 and SmIg and Ia-like antigen negative. The continuous in vitro proliferation and karyotype abnormalities suggest malignancy. The cells contain only the lymphoid type of alkaline phosphatase and a high level of terminal deoxynucleotidyl transferase and as they formed E-rosettes during the first year in culture they may be of T-cell lineage. Since the patients' leukaemic cells have not been studied it is impossible to know whether the cultured cells represent an in vitro proliferation of the patient's malignant cells or an outgrowth of a subpopulation of cells. In the cytoplasm of many cells inclusion bodies containing virus-like particles, similar to those observed in fresh human leukaemia cells, were often seen. As this new virus is biologically active, infection of the cells by virus may explain their continuous proliferation in vitro.

A detailed study is described of a case of hairy cell leukaemia, presenting with a serum paraprotein of an immunoglobulin (Ig) class different from that synthesised by the neoplastic cells. The case was unusual in its association with leukaemic arthropathy but ultrastructurally the hairy cells were typical. By immunofluorenscence and immuno!electron microscopy the neoplastic cells expressed IgAh both on the cell surface and intracellularly in the rough endoplasmic reticulum, perinuclear space and Golgi apparatus. No Ig was observed in the ribosomal-lamellae complexes. These cells also synthesised and secreted Ig of class Ah in culture. However the serum paraprotein was of class IgAx and could not be attributed to an abnormal population of plasma cells in the bone marrow. There was no other evidence for myeloma and the IgAx paraproteinaemia appeared to be benign, apparently unrelated to the neoplastic proliferation of hairy cells.

The ultrastructural localization of immunoglobulin heavy and light chains has been investigated in nine haematopoietic cell lines, using a technique which involves the treatment of lightly prefixed cells with saponin to allow penetration of the antibody-peroxidase conjugate. The synthesis and secretion of immunoglobulin was also studied in these cell lines. Immunoglobulin was found to be localized in the cisternae and on the membranes of the rough endoplasmic reticulum, perinuclear space and/or Golgi apparatus. In each case staining for heavy chains was weak or absent in the perinuclear space while staining for light chains was usually strong. Additionally in three cell lines immunoperoxidase staining indicated that heavy chains were absent from the Golgi apparatus despite the observed presence of light chains in the Golgi apparatus and the secretion of combined immunoglobulin into the supernatant. The results obtained suggest compartmentalization of the synthesis of light and heavy chains and indicate that the technique of immunoelectron microscopy may significantly contribute to an understanding of the mechanisms involved in immunoglobulin synthesis, intracellular transport and secretion.

The ultrastructural localization of immunoglobulin heavy and light chains has been investigated in nine haematopoietic cell lines, using a technique which involves the treatment of lightly prefixed cells with saponin to allow penetration of the antibody‐peroxidase conjugate. The synthesis and secretion of immunoglobulin was also studied in these cell lines. Immunoglobulin was found to be localized in the cisternae and on the membranes of the rough endoplasmic reticulum, perinuclear space and/or Golgi apparatus. In each case staining for heavy chains was weak or absent in the perinuclear space while staining for light chains was usually strong. Additionally in three cell lines immunoperoxidase staining indicated that heavy chains were absent from the Golgi apparatus despite the observed presence of light chains in the Golgi apparatus and the secretion of combined immunoglobulin into the supernatant. The results obtained suggest compartmentalization of the synthesis of light and heavy chains and indicate that the technique of immunoelectron microscopy may significantly contribute to an understanding of the mechanisms involved in immunoglobulin synthesis, intracellular transport and secretion.

The distribution of light and heavy immunoglobulin chains in chronic lymphocytic leukemia (CLL) cells has been investigated at the ultrastructural level using an immunoperoxidase technique. Light chains were localized in the lumens of the perinuclear space and rough endoplasmic reticulum, while staining for heavy chains was weak or negative and generally confined to the membranes of the rough endoplasmic reticulum. This pattern is consistent with immunoglobulin biosynthetic studies on CLL cells in which light chains are synthesized in excess over heavy chains and secreted as the exclusive immunoglobulin product. The pokeweed mitogen stimulation of two populations of CLL cells for 6 days resulted in a balanced synthesis and secretion of light and heavy chains that was reflected in concomitant change in light and heavy chain distribution and intensity of staining using the immunoperoxidase technique.