Lewis Glasser

Experimental studies on the role of alkyl lysophospholipids in autologous bone marrow transplatation

The selective cytocidal effect of alkyl lysophospholipids against neoplastic cells while sparing normal cells make these ideal candidates for purging leukemic cells from bone marrows obtained during remission. To test the feasibility of such an approach, a murine model and an in vitro human cell model were developed. In the murine system a mixture of normal bone marrow cells and WEHI IIIB myelomonocytic leukemic cells was incubated with varying doses of 1-O-octadecyl-2-O-methyl-rac-glycero-3-phosphocholine (ET-Me) for 24 hr before being injected into tail veins of lethally irradiated Balb/c mice. At doses of 20 and 100 μg/ml, long-term survivors were noted. The additional steps of freezing and thawing following incubation resulted in significantly longer survival with doses of 10 to 50 μg/ml, but were toxic to marrow stem cells at 100 μg/ml.In the in vitro model, normal marrow progenitor cells and leukemic cells (the promyelocytic cell line HL60) were exposed to varying concentrations of ET-Me for 1 and 4 hr alone or mixed, and clonogenicity was assayed by colony formation in semisolid medium during 7–14 days’ incubation. At doses up to 100 μg/ml exposed for 4 hr normal progenitor cells were spared and HL60 colonies eliminated. Other phospholipids analogues were less effective in eliminating leukemic cells, but spared normal progenitor cells.A survey of fresh leukemic cells found varying degrees of sensitivity to ET-Me, indicating the need for testing a variety of compounds.These studies clearly indicated the potential usefulness of alkyl lysophospholipid compounds in selectively purging leukemic cells from remission marrows for autologous bone marrow transplantation.

The effect of in vivo dexamethasone on lymphocyte subpopulations: differential response of EAhu rosette-forming cells.

The purpose of this study was to compare the differential effect of dexamethasone on “third population” lymphocytes with its effect on B and T lymphocytes. Five human volunteers were given a single oral dose of dexamethasone (6–8 mg). Samples were evaluated prior to the drug, 5 hr after drug ingestion and 24 hr following the poststeroid sample. Surface marker studies included SIg, EAC rosettes, mouse rosettes, EAhu (Ripley) rosettes, total E-rosette-forming lymphocytes, Tμ lymphocytes, and Tγ lymphocytes. Functional studies included mitogen stimulation with phytohemagglutinin, concanavalin A, and pokeweed and antigen stimulation with herpes simplex virus and Candida albicans. At the nadir of the steroid-induced lymphopenia absolute counts for all B-cell markers were decreased. No differential effects were noted in SIg using polyvalent, IgM, IgG, κ, or λ antisera. Total T lymphocytes decreased 55% (P < 0.05). There was a differential effect on Tμ and Tγ subsets and the former were significantly decreased. Unlike other lymphocyte subpopulations, cells with high avidity Fc receptors for IgG molecules, i.e., EAhu (Ripley)-rosette-forming and Tγ lymphocytes, were unresponsive to steroid treatment. EAhu (Ripley)-rosette-forming lymphocytes increased from a baseline value of 15 to 28% after dexamethasone ingestion but absolute cell counts remained essentially unchanged with a mean baseline value of 287/μl compared with a poststeroid count of 250/μl. A rebound phenomenon was noted for B, T, Tμ, and Tγ lymphocytes (P < 0.05). Functional studies of the peripheral blood mixture of lymphocytes at the nadir of the lymphopenia showed significantly suppressed responses to both mitogens and antigens.

New ultrastructural observations: Parallel tubular arrays in human Tγ lymphoid cells

Abstract T γ cells are E-rosetting cells bearing Fc receptors for IgG (E + , Fc γ + cells). Third population (non-T, non-B) lymphoid cells are also Fc γ + cells and contain unique inclusions called parallel tubular arrays (PTA). Although T γ cells and third population lymphoid cells should belong to a similar population of cells, previous ultrastructural studies on purified T γ cells have failed to reveal the presence of PTA. In this study, we have unequivocally demonstrated PTA in the majority of T γ cells using simple rosetting techniques. A total of 76 EA hu -rosettes and 108 EA ox -rosettes prepared from an E + enriched fraction (using sheep erythrocytes as marker particles) were directly examined by electron microscopy. PTA were found in 87% of the EA hu -rosettes and 82% of the EA ox -rosettes. Ammonium chloride, commonly used in other laboratories to lyse erythrocytes during the purification procedure was found to cause a marked decrease in the number of ultrastructurally distinct PTA profiles. In contrast, hypotonic lysis had no effect on cellular ultrastructure. This study showed for the first time that T γ cells are ultrastructurally similar to other Fc γ + lymphoid cells and contain PTA as a distinct marker. The significance of our findings to the basic function of this E + Fc γ + lymphoid population is discussed.

The in vivo development of plasma cells A morphologic study of human cerebrospinal fluid

This is a case study of a patient with the clinical diagnosis of meningoencephalitis. It demonstrates the in vivo development of plasma cells in the central nervous system. All the stages of in vitro mitogen-induced lymphocyte transformation previously described by light and electron microscopy are recapitulated in this analysis of cerebrospinal fluid. It is postulated that this represents a local humoral immune reaction in the central nervous system and provides morphologic support for the concept that the gamma globulin in cerebrospinal fluid is produced locally in part, in addition to its origin from the plasma proteins.

Basophilic meningitis secondary to lymphoma

In this case report of basophilic meningitis, the patient was a 5-year-old girl under treatment for a diffuse lymphocytic lymphoma. She presented with headache and bilateral papilledema. Cerebrospinal fluid examination showed 60 percent basophils. Subsequent specimens showed a rare blast. It is postulated that after the lymphoma cells had spread to the meninges, a cell-mediated immune reaction was initiated with the appearance of basophils in the exudate.

Phagocytosis in acute leukemia

The phagocytic potential of leukemic cells in various types of acute leukemia was studied. Cases included lymphoblastic leukemia, myeloblastic leukemia, myelomonocytic leukemia, monocytic leukemia, progranulocytic leukemia, blast transformation of chronic myelocytic leukemia, and unclassified leukemias. Cytochemical stains were used as an aid in classification. These included Sudan black B, naphthol AS‐D chloroacetate esterase, α‐naphthyl butyrate esterase, acid phosphatase, and periodic acid‐Schiff. Phagocytosis was evaluated after incubation of leukemic cells with Candida albicans. Rare phogocytic activity was seen in lymphoblastic leukemia, unclassified leukemias, blast crises in chronic myelocytic leukemia, and progranulocytic leukemia. Myeloblastic leukemias were feebly phagocytic. Myelomonocytic leukemia and monocytic leukemia both exhibited marked phagocytosis which distinguished them from the other acute leukemias. Myelomonocytic leukemia could be differentiated from acute monocytic leukemia by its greater phagocytic capacity.

Dose in Granulocyte Transfusion

Excerpt To the editor: In a recent article, Winston and associates (1) presented data that led them to conclude that therapeutic granulocyte transfusions offered no substantial benefit over optimal...

The molecular pathobiology of stored neutrophils

Abstract The neutrophil is a short-lived cell because it has limited biosynthetic capacity to repair and maintain its component parts. Studies on neutrophil storage show generalized defects involving energy metabolism, phagocytosis, microbial killing, chemotaxis, membrane receptors, microtubules, microfilaments, and stimulus—response coupling. Some storage conditions are more detrimental to certain functions than others, e.g. cold induces a severe chemotactic defect. Thus, room temperature storage is preferable to refrigerated storage. For optimal storage, it is necessary to have sufficient glucose in the medium and to maintain an adequate pH. During storage, a dismantling of the functional integrity of the cell occurs with the most highly integrated functions being lost initially, while primitive functions are retained the longest. With minor exceptions, the generalized metabolic, structural, and functional defects noted in stored neutrophils are not peculiar to storage but most likely mirror the inexorable in vivo programmed senescence of the neutrophil.