Amplification of immunological functions by subcutaneous injection of intermediate-high dose interleukin-2 for 2 years after autologous stem cell transplantation in children with stage IV neuroblastoma.

BACKGROUND: Immunotherapy given post-autologous stem cell transplantation may eliminate residual tumor cells escaping the conditioning protocol. METHODS: Five children suffering from stage IV neuroblastoma were treated by recombinant interleukin-2 (IL-2) post-autologous peripheral blood stem cell transplantation. The patients' peripheral mononuclear cells were monitored for CD3+ and CD56+ levels, their proliferative response and killing of various cell lines targets. RESULTS: An increase in the level of total lymphocytes, mainly due to expansion of T cells, and enhanced proliferative response to phytohemaglutinin were observed. Elevated cytotoxicity against K562 and neuroblastoma target cells was detected in four patients and against K562 targets in one patient. Toxicity included mild thrombocytopenia, and fever in four patients and mild to moderate encephalopathy which necessitated withdrawing one patient from the protocol. Three of five patients studied are alive today, one of them whose IL-2 was stopped, is in relapse. Two patients have died. CONCLUSIONS: Immunotherapy with s.c. intermediate-high dose IL-2 is feasible and results in expansion of T cells and in stimulation of killing activity against several targets including in some cases, neuroblastoma tumor cells.

Successful human umbilical cord blood stem cell transplantation without conditioning in severe combined immune deficiency.

A 2-month-old girl with severe combined immunodeficiency (SCID), presented with mild staphylococcal skin infection, lymphopenia, low T cell number, absence of B cells, high number of NK cells, and a negligible response to mitogens. Since her older brother died as a result of SCID 2 years earlier, cord blood was harvested from a sister born 2 1/2 years earlier, who was normal and fully matched both by serology and molecular typing. In view of her clinical condition and in spite of a high number of NK cells with normal activity, HUCBT without preparative conditioning was performed. No G-CSF was administered. Engraftment with mixed chimerism was evident 3 weeks post transplantation. There were no peritransplantation complications. Eighteen months post transplantation, the girl is in excellent condition, blood counts are normal, T cell engraftment is complete, B cell engraftment is proceeding gradually, and the mitogen stimulation tests are normal. Due to the unique nature of HUCB hematopoietic cells, engraftment without conditioning may be possible in patients with SCID with fully matched donors. This is the first HUCBT performed without conditioning.

Stratified horizontal flow in vertically vibrated granular layers.

A layer of granular material on a vertically vibrating sawtooth-shaped base exhibits horizontal flow whose speed and direction depend on the parameters specifying the system in a complex manner. Discrete-particle simulations reveal that the induced flow rate varies with height within the granular layer and oppositely directed flows can occur at different levels. The behavior of the overall flow is readily understood once this feature is taken into account.

Interleukin 2 induces human acute lymphocytic leukemia cells to manifest lymphokine-activated-killer (LAK) cytotoxicity.

Lymphokine-activated killer cells (LAK) were originally distinguished from natural killers (NK) and cytotoxic T lymphocytes. Recently, however, IL 2-activated NK cells were suggested as the major source of LAK reactivity in human peripheral blood (PBL). Because certain T cell acute lymphoblastic leukemia (T-ALL) cells are phenotypically similar to LAK precursors, we have asked whether these leukemic cells can be induced toward LAK-cytotoxicity and express NK reactivity before stimulation. Five out of seven T-ALL preparations were induced by IL 2 to kill target cells. The cytotoxicity of the leukemic-LAK cells resembled that of normal LAK effectors as they lysed efficiently the NK-resistant target Daudi, as well as fresh human sarcoma, carcinoma, and renal cancer cells but not normal PBL. The ALL-LAK precursors phenotype was T3-, T4-, T8-, and T11+, similar to most normal LAK precursors. In contrast to normal PBL that generated LAK effectors when their proliferation was inhibited, the irradiated, nonproliferating T-ALL leukemic cells did not respond to IL 2. Therefore, the T-ALL LAK cytotoxicity was attributed to the leukemic cells rather than to residual normal lymphocytes. The IL 2-responding T-ALL cells did not express autonomous NK type cytotoxicity, suggesting that they reflect LAK precursors of non-NK origin. The homogeneous leukemic preparations with inducible LAK cytotoxicity described herein provide a model system for studying normal LAK cells.

Monocyte PGE2 secretion in Hodgkin's disease and its relation to decreased cellular immunity.

The secretion of PGE2 from monocytes of newly diagnosed patients with Hodgkin's disease (HD) was compared to that of patients in remission, who were not receiving either chemotherapy or radiotherapy, and normal controls. We found that monocyte monolayers of some patients, both newly diagnosed and those in remission, secreted markedly elevated levels of PGE2. The lymphocyte proliferative response to PHA was increased to a similar extent in both newly diagnosed patients and those in remission when cultured in the presence of indomethacin. PGE2 concentrations in the medium of mononuclear cultures correlated with the lymphocyte proliferative response to PHA (P less than 0.05). However, no correlation of monocyte PGE2 production with decreased E rosette forming lymphocytes, anergy or clinical stage could be demonstrated. We suggest that PGE2 secretion by monocytes is indicative of an 'activated' state of these cells. It is, however, unlikely that PGE2 is the only molecular species responsible for the decreased cellular immune function in HD. 'Activated' monocytes may be part of the immune response in this disease and may be responsible for the decreased cellular immunity.

Lymphokine-activated killer (LAK) cells: interferon-gamma synergizes with interleukin-2 to induce LAK cytotoxicity in homogeneous leukemic preparations.

Lymphokine-activated killer (LAK) cells are generated by the incubation of lymphocytes with high levels of interleukin-2 (IL-2). We report here that interferon-gamma (IFN-gamma) acts synergistically with low levels of IL-2 to promote LAK differentiation in peripheral blood lymphocytes as well as in homogeneous T acute lymphocytic leukemic cells exhibiting LAK precursor reactivity. No augmentation of LAK response was observed with IFN-alpha-2, IFN-beta-1, and IFN-beta-2/IL-6. The synergism between IL-2 and IFN-gamma was expressed in the ability of activated lymphocytes to lyse natural killer resistant cell line targets and surgically removed melanoma cells. The augmented LAK response due to IFN-gamma does not reflect up-regulation of the high-affinity IL-2 receptors consisting both of alpha and beta subunits, since expression of the alpha (Tac) subunit on the responding leukemic cells was not increased by IFN-gamma. The observed IFN-gamma/IL-2 synergism in the induction of monoclonal LAK precursors suggests that a single precursor cell responds to both IFN-gamma and IL-2 and that different mechanisms underlie the basal IL-2-mediated LAK response and its enhancement by IFN-gamma.

Studies on human milk macrophages: effect of activation on phagocytosis and secretion of prostaglandin E2 and lysozyme.

Breast milk macrophages cultured in vitro synthesized and secreted increasing amounts of protein, lysozyme, and prostaglandin E2(PGE2) into the extracellular medium. These cells were also shown to actively phagocytose labeled zymosan particles in culture. Morphologic characteristics, phagocytosis, and secretory responses of the macrophages were altered depending on the presence of various stimuli in the culture. Concanavalin A, endotoxin and zymosan particles, but not latex particles, all resulted in an increased PGE2 secretion into the medium. Although total protein synthesis was not altered by any of these stimuli, Concanavalin A and endotoxin resulted in a decreased lysozyme concentration in the extracellular medium. Concanavalin A enhanced, whereas endotoxin and prior phagocytosis of latex particles inhibited phagocytosis of labeled zymosan particles. These findings indicate that phagocytosis and secretions of milk macrophages may be altered depending on the nature of the stimulating agent.

IgE induces secretion of prostaglandin E2 by human monocytes.

IgE was isolated from a patient with the hyper IgE, recurrent infection syndrome by immunoadsorption on sepharose bound goat anti-human IgE. Addition of this IgE to a monolayer culture of human monocytes resulted in a dose-dependent increase in PGE2 secretion. The addition of F(ab')2 fraction of goat anti-human IgE in the presence of sub-stimulating doses of IgE markedly increased PGE2 secretion; whereas addition of F(ab')2 fragment of irrelevant goat IgG had no effect. Similar activation of monocytes which could be enhanced by anti IgE was observed in the presence of the patient's serum. No such effect was seen in the presence of normal human serum. These results indicate that IgE may activate human monocytes and induce PGE secretion.

The effect of human monocytes and macrophages on lymphocyte proliferation.

Human monocytes suppressed both the phytohaemagglutin (PHA) or antigen-induced lymphocyte proliferative response, when the monocyte: lymphocyte ratio was increased or when the monocytes were stimulated with zymosan or endotoxin. The effect of monocytes on autologous lymphocyte proliferation was compared with that of macrophages obtained by culturing monocytes in vitro for 7 days. The lymphocyte proliferative responses were increased in the presence of macrophages, however, neither increasing their number nor stimulation by zymosan or endotoxin altered the autologous lymphocyte proliferative response to PHA or purified protein derivative (PPD). The PGE2 concentration in the medium of both the cultured monocytes or macrophages activated by zymosan and endotoxin rose markedly without a corresponding suppressor effect on lymphocyte proliferation in the presence of macrophages in the culture. Thus it seems that while PGE2 is a useful marker of mononuclear phagocyte activation, other molecular species are of importance in determining the lymphocyte proliferative response to mitogens and antigens.

The mutual clonal origin of the lymphoplasmocytic and lymphoma cell in alpha-heavy chain disease.

Biosynthetic studies in alpha-heavy chain disease were performed on the gut tumour which was composed mainly of lymphoplasmocytic cells and on the mesenteric lymph node tumour composed mainly of immunoblasts. The gut tumour cells synthesised alpha-heavy chains and secreted them during 2-5 hr culture, whereas the lymph node tumour cells synthesized alpha-heavy chains which were shed into the culture medium only after 20 hr. These chains were shown to be present on the surface of the immunoblastic tumour cells by enzymatic radioiodination. Both the surface and the secreted alpha-heavy chain of the lymph node and gut tumour were found to be smaller than the alpha-heavy chain of myeloma proteins. These results suggest that the lymphoblasmocytic and the immunoblastic tumour cells originate from the same defective clone.

Studies on the effect of intralipid on human monocyte functions in vitro.

Intralipid (IL) particles were ingested by human monocytes in culture. These particles remained within the cells for periods of up to 3 weeks in culture. The presence of IL particles did not alter the normal secretion of lysozyme or prostaglandin E2 (PGE2), nor was the normal process of biochemical activation altered, as evidenced by the expected increase in PGE2 secretion and superoxide anion production by stimulated monocytes. Phagocytosis of zymosan particles was increased in monocytes that had ingested IL. These results indicate that in this in vitro system essential monocyte functions were not altered following ingestion of IL.

Spectrum of clinical manifestations of antiphospholipid antibodies in childhood and adolescence.

Nineteen children with circulating anticoagulants or anticardiolipin antibodies and negative or only weakly positive antinuclear factors were reviewed for their clinical manifestations. The relationship between these antibodies and the morbidity in the pediatric age group will be discussed.