Restoration by Levamisole of Rosette-Forming Cells Inhibited by Histamine, DB-Camp and Adenosine

Db-CAMP and adenosine in vitro markedly inhibited the E-rosette formation of peripheral T-cells of healthy subjects. Histamine in vitro also inhibited the E-rosette formation, but only in patients with allergic disorders.

Levamisole in vitro significantly restored the E-rosette formation of T-cells inhibited by db-CAMP, adenosine and histamine.     

Differntiation and Regulation of Lymphocyte Populations: Evidence for Immunopotentiator-Induced T Cell Recruitment

Isoprinosine, the pacetaminobenzoic acid salt of inosine dimethylaminoisopropanol (1:3 molar ratio) and sodium diethyldithiocaxbamate are two immunopotentiators which share an ability to induce in vivo acquisition of a specific T-cell marker by undifferentiated precursor lymphoid cells of healthy nu/nu mice, without affecting the B-cell lineage.

Serum from treated nu/nu mice tested in dual assays, contains a selective inducer of prothymocytes.     

Modification of actin in peritoneal macrophages after diazepam treatment

Abstract

We have investigated the effect of therapeutic doses of diazepam (7 μg/mouse) on the association of actin with the macrophage cytoskeleton using cytochemical and morphological methods.

Results obtained indicated that diazepam was able to modulate the content of actin in macrophages; such an effect proved to be time-dependent. After fixation and staining for indirect immunofluorescence with actin antibody, peritoneal macrophages from mice treated for short time with diazepam, showed a fluorescent intensity increase compared to control mice. The fluorescent intensity augmented reaching peak value within 14 days of treatment. Afterwards, this value dropped below control value for mice that underwent longer treatments. In the in vitro experiments concentrations of 10^?5 M, diazepam inhibited a well cell spread and a lower amount of actin after 15 min of incubation was also revealed.

These results suggest that administration of diazepam in vivo plays a role in both the nonspecific and specific immune response, producing in the macrophages a reorganization process of microfilaments.     

Effect of L-Alanosine on Immune Response

L-alanosine is an antitumor compound which has recently entered clinical trials. Single i.p. doses of this drug inhibit antibody production to thymus-dependent and thymus-independent antigens as well as delayed hypersensitivity to SRBC in mice. Moreover, the in vitro lymphoproliferative responses of splenocytes to Concanaval in A and bacterial Lipopolysaccharides are also affected when the drug is added upon setting up the cultures. On the other hand, in vivo treatment with L-alanosine does not impair spleen natural killer (NK) activity.

The results are discussed in an effort to characterize the immuno-suppressive activity of L-alanosine.     

Action of Levamisole on E-Rosette Forming Cells and Serum Adenosine Deaminase in Hodgkin's Disease

The interaction of levamisole and/or adenosine in vitro an E-rosette forming cells (RFC) of healthy subjects and on low RFC of patients affected with various diseases was investigated. Levamisole did not enhance normal RFC. The drug enhanced low RFC in some patients (responders) but not in others (non-responders). Adenosine inhibited normal and low RFC and this inhibition could be reversed by levamisole.

Further, levamisole 150 mg 2 days a week was given to 12 Hodgkin's disease (HD) responders for 2 weeks, then 150 mg 1 day weekly for 2 1/2 months After one week of treatment RFC increased and the enhancement could be maintained during the whole treatment period.

Lastly levamisole in vitro and in vivo diminished serum adenosine deaminase activity in normal subjects and HD patients.     

Mrphological and Functional Changes in Mouse Splenic Lymphocytes Following in Vivo and in Vitro Exposure to Chlorphentermine

With repeated administration to animals, the cationic, amphiphilic drug, cnlorphentermine (CP), has been shown by others to induce a phospholipidosis in lymphocytes. In the present study mouse splenic lymphocytes, exposed to CP, either in vivo or in vitro, developed morphological changes consistant With the induction of phospholipidosis. In addition, CP induced functional changes in lymphocytes. Mice, treated with CP in vivo, demonstrated a significantly depressed ability to generate a delayed hypersensitivity response or to produce antibody-secreting cells against de novo antigens. Mouse splenic lymphocytes, exposed to 10^-7 M CP for 3 days in vitro, demonstrated a signficantly depressed blastogenic response to the mitogens phytohemagglutinin, concanavalin A and lipopolysaccharide.

CP inhibited an event that occurred early during lymphocyte activation, but was subsequent to mitogen/receptor coupling. In addition, CP significantly depressed the increased uptake of choline that occurs in lymphocytes following cellular activation. Since the presence of phospholipidosis is indicative of an impairment in phospho lipid metabolism, these results taken together provide evidence for a relationship between this phenomenon and a ltered immune function.     

The Effect of the Non-Steroidal Anti-Inflammatory Drug Etodolac on Macrophage Migration In Vitro and In Vivo

The possible mode of action of Etodolac, a potent anti-inflammatory drug, has been investigated. The effect of Etodolac on macrophage inflammatory response was studied in vivo and in vitro. Etodolac significantly depressed the influx of inflammatory macrophages into peritoneal cavity following stimulation with a sterile irritant. This decrease in macrophage accumulation in vivo correlated with the effect of Etodolac on the macrophage chemotaxis in vitro. Etodolac was also capable of reducing the mecrophage ability to migrate towards a chemoattractant. In vivo Etodolac should reduce the amount of damage produced at the site of chronic inflammation since fewer macrophages would migrate to the inflammatory sites.     

In Vitro Lymphocyte Sensitivity Test to Methisoprinol in Different Pathological Conditions

The present paper describes an in vitro study with Methisoprinol, carried out by means of various lymphocyte sensitivity tests in different pathological conditions.

Methisoprinol does not exert any effect - neither on responsiveness nor on lymphocyte function - in healthy subjects, while in subjects with immunodepressive pathology the drug is capable of increasing these parameters either alone (rosettes) or in conjunction with mitogens (rosetting) at doses between 50 and 100 µ;g/ml.     

Modulatory Effect of Isoprinosine on Lymphocyte Proliferative Response Under Immunodeppressive Conditions

In the present work the effect of Isoprinosine on the mitogenic responses of T and B lymphocytes has been studied. We have found that Isoprinosine can enhance in vitro the response to Concanavalin A. This enhancement was more apparent in cell cultures showing an initially low blastogenic response. In low responses artificially induced by treatments in vivo with cyclophosphamide, our results indicate that Isoprinosine, administered in vivo, does not enhance the response to Con A of treated mice. However, addition of Isoprinosine (75 μg/ml) to cultures of spleen cells from mice previously treated with cyclophosphamide enhanced the suppressed response up to normal levels. Neither in vivo nor in vitro Isoprinosine treatments increased the response of lymphocytes to lipopolysaccharide, but usually inhibited the blastogenesis of B cells.     

Modulation of Inflammatory Peritoneal Cell Function and Metabolism by Methotrexate

Methotrexate (MTX) is widely used in cancer chemotherapy, altnough the effects of MTX on cellular antitumor defense mechanisms are poorly understood. To evaluate the effect of MTX on the cellular inflammatory response, male Sprague-Dawley rats were treated with four daily i.p. injections of MTX or a control vehicle. Kats treated with daily doses of 1.2 mg/kg MTX demonstrated a significant reduction in number of peritoneal exudate cells, specifically macrophages, collected 96 hours following the inflammatory stimulus. To determine if metabolic perturbations also occur upon exposure to LTX, glucose oxidation and protein synthesis by inflammatory cells were monitored in vitro. At a MTX concentration of 10^-3M, peritoneal exudate cell ¹⁴C-1-glucose and ¹⁴C-6-glucose oxidation was significantly depressed.

¹⁴C-1-leucine incorporation into TCA precipitable protein was inhibited at 4 × 10^-3M MTX. Peritoneal exudate cell viability was not altered at these concentrations of MTX. These results demonstrate that MTX, at therapeutic concentrations, can depress the influx of macrophages to an inflammatory site and also diminish energy metabolism and protein synthesis by inflammatory cells.     

Enhancement of Antibody-Independent Phagocytosis by N-(6-Aminohexyl)-5-Chloro-1-Naphthalenesulfonamide (W-7)

A low-molecular weight compound N-(6-aminohexyl)-5-chloro-1-naphthalenesulfonamide (W-7) resulted in strongly enhanced phagocytosis of polystyrene latex beads by peritoneal macrophages of BALB/c mice after intraperitoneal administration. Binding of C3 cleavage product (C3b) to peritoneal macrophages after intraperitoneal injection of W-7 was also enhanced as shown by fluorescent antibody technique. Antibody against W-7 was not detected in the sera of these mice. Therefore, close association of enhanced phagocytosis of polystyrene latex beads with increased binding of C3 cleavage product to macrophages was demonstrated through induction by a low-molecular weight compound in vivo.     

Inhibition of Yeast Phagocytosis by Dexamethasone in Macrophage Cultures: Reversibility of the Effect and Enhanced Suppression in Cultures of Stimulated Macrophages

This investigation was initiated to characterize further the ability of 1μM dexamethasone to suppress the ingestion of heat-killed Saccharomyces cerevisiae particles in cultures of murine resident peritoneal macrophages. Time course studies revealed that the inhibitory response required the continual presence of the steroid in the culture medium. In addition, increased inhibitory responses occurred after dexamethasone was supplied to previously untreated cultures of resident macrophages that became stimulated by differentiating in vitro. These findings indicate that glucocorticoids act directly on macrophages to decrease their phagocytic capacity, which in vivo would reduce host resistance.     

In Vivo or in Vitro Modulating Effects of Vincristine on the Generation of Allogeneic Cytotoxic Lymphocytes in Vitro

Cytotoxic lymphocytes carrying the H-2ⁱ⁵ haplotype were generated in vitro against alloantigens specified by the IC-S-G-D regions of H-2.

Responder lymphocytes were collected from donors B10.A(5R) non-treated or treated with 3 or 0.6 mg/Kg, ip of Vincristine (VCR) and incubated with irradiated B10 spleen cells. High doses of VCR produced depression of the cytotoxic response (CR), whereas 0.6 mg/Kg of the drug produced substantial increase of the response.

Further experiments were conducted adding VCR in vitro to the tissue culture during the generation of cytotoxic lymphocytes. Depression of CR occurred following treatment with high concentrations of VCR. On the other hand, marked increase of CR was found using responder lymphocytes treated in vitro with low concentrations (i.e. 0.016-0.0032 μq/ml) of the drug.

The mechanism of VCR-induced enhancement of CR has not been elucidated. However, the present data show the possibility of increasing host immune responses by treatment of donor mice or of responder lymphocytes with VCR at selected doses or concentrations.     

Book Reviews

MOLECULAR APPROACHES TO IMMUNOLOGY Eds. E.E. Smith and D.W. Ribbons Academic Press, Inc., New York, 1975 354 pages, hard bound, $15.00

IMMUNOBIOLCGY OF THE MACROPHAGE D. S. Nelson, Editor Academic Press, New York, 1976; xiii + 633 pages, hardbound, $39.00

INFECTION AND THE COMPROMISED HOST J.C. Allen, Ed. Williams & Wilkins, Baltimore, 1976 Softbound, 176 pages, $10.95

TRANSMISSIBLE DISEASE AND BLOOD TRANSFUSION T.J. Greenwalt and G.A. Jamieson, eds.: Grune and Stratton, New York, 1975; 298 pages, hardbound, $26.50

IMMUNOADSORBENTS IN PROTEIN PURIFICATION E. Ruoslahti, Ed. University Park Press, Baltimore, 1976; Hardbound, 86 pages, $16.50 (Supplement No. 2 of the Scandinavian Journal of Immunology).     

Effect of a Traditional Chinese Medicine, Bu-Zhong-Yi-Qi-Tang (Japanese Name: Hochu-Ekki-To) on the Protection Against Listeria Monocytogenes Infection in Mice

ABSTRCTS

Effects of Bu-Zhong-Yi-Qi-Tang (Japanese name : Hochu-ekki-to) on the resistance against Listeria monocytogenes were observed in ICR mice orally administered this medicine daily for 10 days. Survival rates were increased by the pretreatment in mice inoculated i.v. with bacteria 1 day after the last administration and in mice inoculated i.p. 4 days after the last administration. After an i.v. inoculation of L. monocytogenes, the numbers of bacteria in the spleen and liver increased gradually to kill mice by day 5 in untreated group but the bacterial numbers increased slightly by day 3 and decreased from day 3 to day 8 in Hochu-ekki-to pretreated group. After an i.p. inoculation, the number of bacteria in the peritoneal cavity decreased very rapidly within 6h in Hochu-ekki-to treated group compared to that in untreated group. After the administration, number of polymorphonuclear cells increased in the peripheral blood, peritoneal cavity and spleen. In treated mice, macrophages increased in number in the peritoneal cavity and the spleen but decreased in the peripheral blood. Peritoneal macrophages from treated mice showed an enhanced activity to kill L. monocytogenes in vitro within 60 min after ingestion of bacteria. Hochu-ekki-to may augment the host defense against L. monocytogenes through the activation of macrophage series during an early phase of infection.     

A Role for Anti-Inflammatory Agents and Cyclic Amp in Regulating Fibronectin-Mediated Phagocytosis

The present study deals with the effects of anti-inflammatory drugs and agents known to elevate intracellular levels of cyclic AMP (cAMP) on plasma fibronectin-mediated (PFn) phagocytosis of radiolabeled, gelatin-coated latex particles (g-Ltx*) by inflammatory macrophages. Monolayers of casein-elicited peritoneal macrophages were preincubated with the specified agents for either 1 or 24 hrs at 37°C prior to the measurements of phagocytosis in the presence of human plasma fibronectin (47 ug/ml) and heparin (6.7 U/ml).

Under these conditions, prostaglandin E₁, colchicine, vincristine, and cytochalasin B were all effective in inhibiting g-Ltx* phagocytosis by macrophages in a dose-dependent fashion. More potent inhibition of phagocytosis was manifested by agents known to increase intracellular levels of cAMP in phagocytic cells. Dibutyryl cyclic AMP (dbcAMP), d, 1-isoproterenol and aminophylline (10^--⁵ to 10^--³M) were all effective in reducing the uptake of g-Ltx* by macrophages. The combination of dbcAMP and aminophylline acted additively. These studies demonstrate that anti-inflammatory drugs and cAMP-elevating agents exert potent inhibitory effects on fibronectin-mediated phagocytosis of gelatin-coated particles by macrophages. Thus, our system provides a suitable in vitro model for further investigations into the humoral regulation of pnagocytosis of denatured collagen-coated particles and tissue debris by inflammatory phagocytic cells.     

Interaction of Macrophages and Lymphocytes in Rat Skin Allografts

Light microscopy of 2μm sections of rejecting rat skin allografts, embedded in hydroxyethyl methacrylate, revealed among the cells infiltrating the graft base extravascular macrophages containing a small lymphocyte. Toluidine blue staining indicated DNA degradation in some of these phagocytosed lymphocytes. More frequently small lymphocytes were in intimate contact with the surface of the macrophages, resembling 'Periopolesis', which others have previously observed in vitro. These macrophage-lymphocyte interactions were not seen in sections of autografts. Despite a previous report that diphenylhydantoin (phenytoin) impairs macrophage function, these macrophage-lymphocyte interactions were present in grafts placed in rats receiving this drug. This treatment did not hasten or delay the onset of graft rejection. These in vivo findings both accord with recent in vitro studies on the mechanisms of phagocytosis and with reports that phagocytosis is one of the effector mechanisms in allograft rejection. However, macrophage phagocytosis of lymphocytes has also been observed in testicular lymph collected from conscious normal sheep.     

In Vitro and in Vivo Activity Profile of RHC 2851: A New Orally Effective Antiallergic Agent

RHC 2851 has been investigated for its antiallergic activity in three in vitro and two in vivo models of anaphylaxis. We have also compared its activity profile in these models with that of disodium cromoglycate (DSCG), doxantrazole, ketotifen and oxatomide. RHC 2851, given i.p., was 6 times more potent than DSCG, and given orally it was 3 times more potent than doxantrazole. As an inhibitor of mediator release, the activity profile of RHC 2851 was identical to that of DSCG in the following respects: inhibition of IgE-mediated in vitro release of histamine from rat mast cells (RMC) but not human basophils (HUB), possession of tachyphylactic properties and demonstration of rapid loss of inhibitory activity as a function of time before antigen challenge as well as inability to inhibit IgG₁-mediated release of histamine, both in vitro and in vivo, and lack of mediator antagonist activity. Ketotifen and oxatomide did not inhibit either IgE or IgG₁-mediated release of histamine in vitro or in vivo, and were potent mediator antagonists in vivo. We conclude that RHC 2851 is an orally effective inhibitor of mediator release with a mechanism of action similar to that of DSCG and different from that of ketotifen and oxatomide.     

Flow Cytometric Analysis of in Vivo Activation of Murine Spleen Cells

The in vivo effects of two polyclonal immune stimulators were studied in several strains of mice by analyzing the percentages of cells in various phases of the cell cycle by flow cytometry. Both lipopolysaccharide (LPS) and polyriboinosinic, cytidylic acid (rI.rC) were capable of inducing an increase in the percentage of cells undergoing DNA synthesis (S phase) in the spleens of several mouse strains. The response to both LPS and rI.rC was maximal between days 3 and 5 following injection. Optimal in vivo responses to LPS occurred at 3-30 μg, and to rI.rC at 100 μg; however, responses were observed over a broad dose range. No similar increase in S-phase cells was observed following injection of non-mitogenic T-independent antigens. Specific antibody was also measured after in vivo administration of rI.rC. There was a dissociation between the ability of an injection to induce specific antibody and to induce proliferation. These studies extend our knowledge of in vivo lymphocyte activation, and provide a basis for a detailed anafisis of lymphocyte activation following a variety of immune modulators in vivo.     

Diazepam Inhibits Phagocytosis and Killing Exerted by Polymorphonuclear Cells and Monocytes from Healthy Donors. In Vitro Studies

The effect of a benzodiazepine (BDZ), diazepam on human polymorphonuclear cell (PMN) and monocyte pha gocytosis and killing from healthy volunteers has been evaluated. Diazepam is able to inhibit in vitro both functions exerted by PMN and monocytes at 10^-5 and 10^-6 M concentrations/ 4 × 10⁶ phagocytes. 10^-7 M con centration was not effective in all the instances.

These results are discussed for their possible clinical implications, since previous studies have shown that in patients with phobic disorder there is evidence for reduced phagocytosis and killing capacities.