Human monocytes were isolated from the peripheral blood of normal donors and allowed to differentiate in vitro into macrophages. The susceptibility of these cells to infection with a virulent Mycobacterium avium and its modulation by some soluble factors was monitored. The virulent strain of Mycobacterium avium grew progressively in untreated macrophage monolayers. Interleukin-6 (IL-6) was tested for its ability to modulate the macrophage-mycobacteria interaction. Surprisingly, IL-6 was shown to increase M. avium growth in macrophage monolayers by twofold as compared with untreated cells, when added before or after infection. Moreover, addition of rIL-6 to replicating mycobacteria in vitro enhanced their growth two- to three-fold as compared with cultures treated with rIL-6 and a rabbit antiserum to rIL-6. Treatment with IL-6 and interferon-gamma (IFN-gamma) or IL-4 did not modify the growth promoting effect of IL-6 in human macrophages. Overall, our results suggest that IL-6 may contribute significantly to the pathogenesis of infections with M. avium by promoting mycobacterial growth.

Resident peritoneal cells were obtained from BALB/c mice and enriched for cells of the macrophage lineage by adherence onto 96 well tissue culture plates. Adherent cells were then exposed to various recombinant cytokines or supernatants from cell cultures, for 24 h. The ability of such adherent antigen presenting cells (APC) to support proliferation and development of helper function in T-lymphocyte populations, primed with sheep erythrocytes (SRBC), was examined. The addition of cytokines to the APC population did not enhance either proliferation of the T-cells nor helper function, assessed by assay of polyclonal IgG secretion in second cultures, beyond that obtained with control APC. The potent macrophage activators interferon-gamma and lipopolysaccharide caused a significant decrease in both parameters of T-cell activity. This effect was caused by a prostaglandin-mediated pathway inasmuch as indomethacin (1-5 microM) prevented it. Further analysis showed that this negative signal predominated until macrophages were diluted below 5% of the total cell population. At 0.5% macrophages, interferon-gamma stimulated APC function of these cells compared with untreated macrophages. Despite the relative difficulty in manipulating the T-cell response by attempted modulation of the APC with cytokines, the simple manoeuvre of incubation of otherwise responsive, primed T-cells with a high dose (10%) of SRBC during in vitro restimulation, caused the proliferation and helper function of these T-cells to be markedly decreased. This phenomenon was seen regardless of the cytokine used to stimulate the APC population. These studies further clarify the dual role the macrophage in regulation of T-cell responses.

Les cytokines sont des glycoprotéines importantes secrétées par les leucocytes. Ces facteurs sont impliqués dans la réponse immune au niveau cellulaire et humoral. Des résultats sont présentés ici, qui suggèrent que certaines cytokines peuvent agir comme facteurs de croissance pour certaines bactéries. L'interleukine-2 stimule la multiplication d'une souche virulente de Escherichia coli tandis que l'interleukine-6 stimule la multiplication d'une souche de Mycobacterium avium. Ces résultats suggèrent que les bactéries virulentes ont développé la capacité d'utiliser les cytokines comme facteurs de croissance.

The effect of human recombinant interleukin-2 (IL-2) and human recombinant granulocyte-macrophage colony-stimulating factor on the growth of a virulent strain of Escherichia coli in tissue culture medium and in untreated, normal mouse serum was investigated. Both of these cytokines enhanced the growth of the microorganism two- to threefold in tissue culture medium with or without additional fetal calf serum and in untreated mouse serum. IL-4 did not have any effect on the growth of this microbe under the conditions tested. That the enhancement of growth seen with recombinant IL-2 was due to the active cytokine was shown by the following data: (i) addition of an antibody to IL-2 abrogated the growth-promoting effect; (ii) the excipient buffer, which contained everything except the active cytokine, was inactive in modifying bacterial growth; and (iii) heat-inactivated recombinant IL-2 did not promote enhanced microbial growth. The enhancement of growth with IL-2 was significant with concentrations as low as 1 U/ml. Growth of an avirulent strain of E. coli was not stimulated by IL-2. Moreover, addition of IL-2 to growth virulent E. coli in tissue culture medium led to rapid removal of the cytokine from the medium. Collectively, these data suggest that cytokines may act as growth factors for some virulent bacteria.

Murine peritoneal macrophages were isolated by adherence and their listericidal activity assessed in the presence or absence of selected cytokines. Untreated macrophages were not highly listericidal, showing moderate killing in the first 2 h after infection, and allowed progressive microbial growth thereafter (up to 9 h). Pre-treatment of cells with 10 to 100 U/ml of IFN-gamma allowed macrophages to develop sustained listericidal activity for the 9-h observation period, with a 2-log reduction of Listeria CFU per monolayer. Pulsing of cells with TNF-alpha alone did not result in enhanced microbicidal activity but TNF-alpha potentiated IFN-gamma-induced listericidal activity, resulting in high levels of killing when both cytokines were present. Conversely, macrophages pre-treated with interleukin-3 (IL-3) or colony-stimulating factor-1 (CSF-1) were found to be much more permissive for Listeria growth. Neither IL-3 nor CSF-1 abrogated IFN-gamma-induced listericidal activity. Moreover, neither IL-3 nor CSF-1 had any effect on the ability of macrophages to develop a respiratory burst following Listeria infection, as judged by H2O2 release following in vitro infection. Overall, these results suggest that different cytokines may have opposing effects on intracellular microbial growth, and that the balance of cytokine production in vivo may determine the resistance or susceptibility of the infected host.

The ability of soluble factors to modulate the growth of a virulent strain of Mycobacterium avium in murine peritoneal macrophages was studied. The virulent strain, TMC 702, grew progressively in the organs of susceptible BALB/C mice. In addition, this strain of M. avium grew progressively in untreated peritoneal macrophages. Treatment of macrophage monolayers with interferon-gamma (IFN-gamma) did not change significantly the intracellular growth of M. avium. Addition of indomethacin to IFN-gamma-treated macrophage monolayers rendered them significantly more bacteriostatic than macrophages treated with interferon alone, suggesting a role for prostaglandins in inducing unresponsiveness to IFN-gamma in infected cells. Additionally, treatment with tumour necrosis factor-alpha led to a modest increase in bacteriostasis, as compared to untreated monolayers. Further experiments with recombinant interleukins showed that interleukin-4 (IL-4), on its own, could increase bacteriostatic activity against M. avium in a reproducible fashion. Experiments with interleukin combinations showed that IFN-gamma and IL-4 treatment of macrophages rendered these cells almost fully bacteriostatic against M. avium, inclusion of scavengers of reactive oxygen species did not modify the beneficial effect of IFN-gamma and IL-4. Overall, our results suggest an important role for interleukins in modulating the interaction between virulent mycobacteria and murine macrophages.

The ability of a variety of soluble factors, alone or in combination, to endow murine resident peritoneal macrophages with listericidal activity was assessed. Inhibition of growth and (or) killing of Listeria in infected macrophages was determined by the uptake of [3H]uracil following lysis of the infected macrophage monolayers. Interferon-gamma was shown to induce modest listericidal activity in murine resident macrophages as compared with untreated monolayers. Treatment with tumour necrosis factor alpha also induced significant listericidal activity in this system. Among other cytokines tested, IL-4 induced an ability to inhibit growth of Listeria in resident macrophages. The ability of cytokines tested, IL-4 induced an ability to inhibit growth of Listeria in resident macrophages. The ability of cytokines to act in an additive or synergistic fashion with IFN-gamma was also investigated. Combinations of IFN-gamma and IL-4 and IFN-gamma and IL-2 induced listericidal activity not greater than that seen with IFN-gamma alone. IFN-gamma and TNF-alpha were shown to increase bactericidal activity in an additive fashion. However, elicited macrophages were shown to spontaneously exert a significant listericidal activity that was not enhanced by cytokine treatment. Collectively, these findings show that cytokine treatment induced rather modest enhancement in listericidal activity in murine resident peritoneal macrophages and no enhancement whatsoever in elicited macrophages. Thus, in in vivo situations where Listeria organisms are completely cleared from the infected organs, mechanisms other than lymphokine-induced listericidal activity of resident macrophages would seem to be operating.

This study was concerned with the handling of ingested tubercle bacilli by normal human macrophages. Intracellular growth was determined after exposure of macrophages to viable bacilli in vitro and the effect of various cytokines, alone or in combination, on bacilli growth/survival was determined. It was found that Mycobacterium tuberculosis (M.tb) grew quite readily in untreated cultured human macrophages. Treatment with soluble factors showed that a crude lymphokine containing supernatant elicited with Concanavalin A (Con A) was ineffective at reducing growth of M.tb in vitro; similarly a crude lymphokine preparation from M.tb lysate-stimulated mononuclear cells failed to induce any mycobacteriostatic activity in human monocyte-derived macrophages. Recombinant cytokines were then evaluated for their ability to modulate growth of the tubercle bacilli in human macrophages. Recombinant interferon-gamma (IFN-gamma), interleukin-2 (IL-2) and recombinant interleukin 4 (IL-4) were all ineffective at modifying M. tuberculosis growth in human macrophages. Recombinant tumour necrosis-alpha (TNF-alpha) curbed the growth of the bacilli in human macrophages in a reproducible fashion. No cytokine combination was more efficient than TNF-alpha alone. These studies thus highlight the resistance of virulent mycobacteria against different mechanisms of cytokine-induced macrophage bactericidal activity.

The ability of a virulent strain of Mycobacterium avium to infect and replicate within human monocyte-derived macrophages of normal donors was assessed. Moreover, the ability of selected cytokines to modulate the intracellular growth of M. avium was investigated. Our virulent strain of M. avium grew progressively in human macrophages. Treatment of macrophage monolayers with interferon-gamma (IFN-gamma) did not lead to any significant change in the infection pattern. Conversely, treatment with tumour necrosis factor-alpha (TNF-alpha) led to a significant reduction in the growth of M. avium in the macrophages. In contrast, treatment of macrophages with interleukin-6 (IL-6) enhanced their susceptibility to M. avium significantly. This finding was substantiated by other results which showed that IL-6 increased the growth of M. avium in tissue culture medium. These results suggest that cytokines may influence the M. avium-macrophage interaction, in a positive or negative manner.

The immunomodulatory properties of platelet factor 4 (PF4) have been examined in vitro and in vivo. This agent prevented the induction of concanavalin A (Con A)-induced suppressor cells in vitro in a dose-dependent manner but it did not affect the function of established Con A suppressor cells. This effect was not due to an enhanced production of interleukin-2 (IL-2) by lymphocytes exposed to PF4. The delayed-type hypersensitivity (DTH) reaction to sheep erythrocytes (SRBC) was used as a model for the generation of antigen-specific suppression in vivo. PF4 enhanced the magnitude of the swelling following SRBC challenge 10 days after sensitization by the i.p. route or following sensitization by both the s.c. and i.p. routes. These studies show that PF4 has immunomodulatory activities in well-defined models of cell-mediated immunity and suggest that this agent has a potential use in the dissection of events in antigen-specific suppression.

An investigation has been made into the ability of neoplastic B lymphocytes obtained from lymphoid tissue of patients with non-Hodgkin's lymphoma (NHL) to secrete immunoglobulin (Ig) in vitro. The majority of the cell populations secreted IgM (17/24 patients), identified as pentameric in three cases examined, and free monotypic light chains (23/24 patients) of the same type as the surface Ig. Secretion of IgD (6/21 patients) and IgG (3/21 patients) was found less frequently. The amounts of Ig secreted were variable and there was no significant difference in the patterns of secretion of cells from NHL patients when compared to previous studies of chronic lymphocytic leukaemia (CLL), nor was there any clear correlation with the histological type. For four of the patients, anti-idiotypic antibody was produced and was used to demonstrate the idiotypic nature of the secreted Ig, and also to show its presence in the serum. The level of idiotypic IgM was measured in one patient during chemotherapy and appeared to correlate well with disease. Such idiotypic Ig must be taken into account when planning treatment of B cell neoplasms with antiidiotypic antibody since it could act as a block to antibody attack. Assessment of the ability of tumour cells to secrete Ig in vitro provides a useful preliminary screen when choosing such patients since a high secretion rate together with extensive disease could lead to unacceptable levels of serum idiotypic Ig.

Immunoglobulin (Ig) could not be detected on the surface or in the cytoplasm of neoplastic cells from five cases of follicle centre cell lymphoma with centroblastic/centrocytic follicular histology when examined by immunohistology of frozen or wax embedded sections. Examination by fluorescein labelled antibodies of cells in suspensions prepared from the biopsies revealed a monotypic surface Ig positive population in one case and a surface or cytoplasmic Ig kappa:lambda light chain imbalance in a further two cases consistent with neoplastic B cell involvement: in all cases the proportion of cells failing to express Ig or T cell markers ranged from 24 to 75%. The monoclonal antibodies B1 (Pan B cell), FMC4 (HLA class II) and J5 (cALL antigen) stained the majority of cells in suspension with residual cells staining with UCHT1 or OKT11 (T cell monoclonal antibodies). In frozen sections, neoplastic follicular cells did not stain with UCHT1. However, in the one case tested these cells stained with the antibodies B1 and FMC4. In paraffin sections J chain could be demonstrated in the cytoplasm of three out of five cases. Cells from four cases were cultured in vitro for Ig production: two failed to produce Ig and monotypic light chains were the sole Ig product of the remaining two cases. The failure to express Ig by the majority of the neoplastic cells from the cases described in this report is at variance with the follicular histology of these neoplasms. Mechanisms responsible for this failure are discussed with reference to current models of B cell differentiation.

Summary This report describes 17 cases of chlormethiazole abuse or dependence. These include one case with symptoms and signs of withdrawal and two other similar cases where undoubted dependence was combined with excessive alcohol intake. Seven other patients with alcoholism who indulged in drug-seeking behaviour involving chlormethiazole are also reported, together with a further seven abusers of various other drugs who were also discovered to be taking chlormethiazole.

A method of analyzing the data obtained from a Compton electron spectrometer in order to obtain the incident photon spectrum is described. Practical considerations limit the application of this analysis to photon energies above about 5 Mev. Application of this analysis to a measurement of the spectrum of the Case betatron is described.

A 60\ifmmode^\circ\else\textdegree\fi{} magnetic spectrometer is used to obtain a relative measurement of the integrated energy spectrum of secondary electrons produced at depths of one-half inch and four inches in aluminum by 17.8-Mev bremsstrahlung. The experimental spectra were found to agree with theoretical spectra calculated by the method due to Brysk to within the experimental error of 5%.

The scattering of 16-Mev bremsstrahlung by 3, 6, and 9 in. of iron was measured for small angles using a thallium-activated sodium-iodide scintillation counter. The experimental results closely fitted a theoretical curve calculated on the basis of single scattering.