Proliferation Of Human Mononuclear Cells Research Articles

Plant extracts of three species of the genus Baccharis induce human mononuclear cells proliferation

Las plantas de uso en medicina tradicional constituyen una fuente importante de compuestos con actividad inmunomoduladora; entre ellas las especies del genero Baccharis, conocidas popularmente como “Jaguareteka´a” en nuestro pais, son ampliamente empleadas. En este estudio se evaluo la actividad inmunomoduladora de extractos metanolicos de tres especies del genero Baccharis (B. trimera, B. notosergilay B. punctulata) sobre la proliferacion de celulas mononucleares humanas de sangre periferica. Los extractos de las tres especies estudiadas estimularon la proliferacion de las celulas mononucleares. Especificamente, el extracto de B. notosergila estimulo la proliferacion celular a todas las concentraciones probadas (5, 10, 25 y 50 μg/mL), mientras que los extractos de B. trimera y B. punctulata mostraron este efecto a 5 y 10 μg/mL. Ademas, por presentar mayor induccion de la proliferacion, se realizo un fraccionamiento con diferentes solventes del extracto metanolico de B. notosergila y B. punctulata. La fraccion de acetato de etilo de ambos extractos vegetales aumento la proliferacion celular, sugiriendo que compuestos de polaridad media son los responsables de esta actividad. Estos resultados demuestran que los extractos de B. trimera, B. notosergila y B. punctulata poseen actividad inmunomoduladora sobre celulas mononucleares humanas y serviran de base a otros estudios para determinar el o los componentes activos de los extractos sobre el sistema inmune.

Human Umbilical Cord Blood Mesenchymal Stem Cells Reduce Colitis in Mice by Activating NOD2 Signaling to COX2

Decreased levels or function of nucleotide-binding oligomerization domain 2 (NOD2) are associated with Crohn's disease. NOD2 regulates intestinal inflammation, and also is expressed by human umbilical cord blood-derived mesenchymal stem cells (hUCB-MSCs), to regulate their differentiation. We investigated whether NOD2 is required for the anti-inflammatory activities of MSCs in mice with colitis. Colitis was induced in mice by administration of dextran sulfate sodium or trinitrobenzene sulfonic acid. Mice then were given intraperitoneal injections of NOD2-activated hUCB-MSCs; colon tissues and mesenteric lymph nodes were collected for histologic analyses. A bromodeoxyuridine assay was used to determine the ability of hUCB-MSCs to inhibit proliferation of human mononuclear cells in culture. Administration of hUCB-MSCs reduced the severity of colitis in mice. The anti-inflammatory effects of hUCB-MSCs were greatly increased by activation of NOD2 by its ligand, muramyl dipeptide (MDP). Administration of NOD2-activated hUCB-MSCs increased anti-inflammatory responses in colons of mice, such as production of interleukin (IL)-10 and infiltration by T regulatory cells, and reduced production of inflammatory cytokines. Proliferation of mononuclear cells was inhibited significantly by co-culture with hUCB-MSCs that had been stimulated with MDP. MDP induced prolonged production of prostaglandin (PG)E2 in hUCB-MSCs via the NOD2-RIP2 pathway, which suppressed proliferation of mononuclear cells derived from hUCB. PGE2 produced by hUCB-MSCs in response to MDP increased production of IL-10 and T regulatory cells. In mice, production of PGE2 by MSCs and subsequent production of IL-10 were required to reduce the severity of colitis. Activation of NOD2 is required for the ability of hUCB-MSCs to reduce the severity of colitis in mice. NOD2 signaling increases the ability of these cells to suppress mononuclear cell proliferation by inducing production of PGE2.

A new chloroquinolinyl chalcone derivative as inhibitor of inflammatory and immune response in mice and rats.

The synthetic chalcone derivative 1-(2,4-dichlorophenyl)-3-(3-(6,7-dimethoxy-2-chloroquinolinyl))-2-propen-1-one (ClDQ) was evaluated for its anti-inflammatory, analgesic and immunomodulatory efficacy in-vitro and in-vivo. ClDQ concentration-dependently inhibited the production of nitric oxide (NO) (IC50 4.3 microM) and prostaglandin E(2) (PGE(2)) (IC50 1.8 microM) in RAW 264.7 macrophages stimulated with lipopolysaccharide. Human mononuclear cell proliferation was significantly inhibited by 10 microM ClDQ. Oral administration of ClDQ (10-30 mg kg(-1)) in the 24-h zymosan-stimulated mouse air-pouch model produced a dose-dependent reduction of cell migration as well as NO and PGE(2) levels in exudates. ClDQ (20 mg kg(-1), p.o.) inhibited ear swelling and leucocyte infiltration in the delayed-type hypersensitivity response to 2,4-dinitrofluorobenzene in mice. In the rat adjuvant-arthritis model, this compound reduced joint inflammation as well as PGE(2) and cytokine levels. In addition, ClDQ displayed analgesic effects in the phenylbenzoquinone-induced abdominal constriction model in mice and in the late phase of the nociceptive response to formalin. Our findings indicated the potential interest of ClDQ in the modulation of some immune and inflammatory conditions.

The Pharmacology of Two Novel Long-Acting Phosphodiesterase 3/4 Inhibitors, RPL554 [9,10-Dimethoxy-2(2,4,6-trimethylphenylimino)-3-(N-carbamoyl-2-aminoethyl)-3,4,6,7-tetrahydro-2H-pyrimido[6,1-a]isoquinolin-4-one] and RPL565 [6,7-Dihydro-2-(2,6-diisopropylphenoxy)-9,10-dimethoxy-4H-pyrimido[6,1-a]isoquinolin-4-one

The pharmacology of two novel, trequinsin-like PDE3/4 inhibitors, RPL554 [9,10-dimethoxy-2(2,4,6-trimethylphenylimino)-3-(N-carbamoyl-2-aminoethyl)-3,4,6,7-tetrahydro-2H-pyrimido-[6,1-a]isoquinolin-4-one] and RPL565 [6,7-dihydro-2-(2,6-diisopropylphenoxy)-9,10-dimethoxy-4H-pyrimido[6,1-a]isoquinolin-4-one], has been investigated in a number of in vitro and in vivo assays. Electrical field stimulation-induced contraction of guinea pig superfused isolated tracheal preparations was significantly inhibited by RPL554 (10 microM) and RPL565 (10 microM) (percentage control; 93 +/- 1.2 and 84.4 +/- 2.7, respectively). Contractile responses were suppressed for up to 12 h after termination of superfusion with RPL554 demonstrating a long duration of action. RPL554 and RPL565 inhibited, in a concentration-dependent manner, lipopolysaccharide-induced tumor necrosis factor alpha release from human monocytes [IC50; 0.52 microM (0.38-0.69) and 0.25 microM (0.18-0.35), respectively] and proliferation of human mononuclear cells to phytohemagglutinin [IC50; 0.46 microM (0.24-0.9) and 2.90 microM (1.6-5.4), respectively]. The inhibitory effect of these drugs in vitro was translated into anti-inflammatory activity in vivo. RPL554 (10 mg/kg) and RPL565 (10 mg/kg) administered orally significantly inhibited eosinophil recruitment following antigen challenge in ovalbumin-sensitized guinea pigs. Likewise, inhalation of dry powder containing RPL554 by conscious guinea pigs (25% in micronized lactose) 1.5 h before antigen exposure significantly inhibited the recruitment of eosinophils to the airways. Exposure of conscious guinea pigs to inhalation of dry powder containing RPL554 (2.5%) and RPL565 (25%) in micronized lactose significantly inhibited histamine-induced plasma protein extravasation in the trachea and histamine-induced bronchoconstriction over a 5.5-h period. Thus, RPL554 and RPL565 are novel, long-acting PDE 3/4 inhibitors exhibiting a broad range of both bronchoprotective and anti-inflammatory activities.

Immunomodulatory functions of extracts from the Chinese medicinal fungus Cordyceps cicadae

The effects of Cordyceps cicadae extracted fractions on human mononuclear cells (HMNC) proliferation were determined by tritiated thymidine uptake. The results indicated that aqueous methanol (50%) extracts of C. cicadae ascocarps portion (CC-1-2) enhanced HMNC proliferation activated with phytohemagglutinin (PHA) with an EC 50 of 13.8±4.6 μg/ml. By contrast, the methanol (100%) extracts of C. cicadae insect-body portion (CC-2-1) suppressed HMNC proliferation stimulated by PHA with an IC 50 of 32.5±5.2 μg/ml. Cell viability test indicated that inhibitory effects of CC-2-1 on HMNC proliferation were not through direct cytotoxicity. The action mechanisms of CC-1-2 and CC-2-1 may involve the regulation of interleukin-2 (IL-2) and interferon-γ (IFN-γ) production in HMNC. Since CC-2-1 suppressed IL-2 and IFN-γ production in HMNC induced with PHA. The CC-1-2 enhanced IL-2 and IFN-γ production of HMNC stimulated with PHA in a concentration dependent manner. Therefore, the results demonstrated that C. cicadae contained growth modulators for HMNC.

Epitope mapping of neutralizing TSST-1 specific antibodies induced by immunization with toxin or toxoids

Toxic shock syndrome toxin-1 (TSST-1), a superantigen produced by Staphylococcus aureus, is a potent stimulator of the immune system. T-cells are activated by crosslinking of MHC class II molecules on antigen presenting cells with T-cell receptors (TCR). TSST-1 is associated with the majority of the cases of menstrual staphylococcal toxic shock, a severe and life-threatening multisystem disorder. Even though antibody mediated protection has been studied, information on antibody specificity directed to individual antigenic determinants of the protein is incomplete. To obtain immunogens with low toxicity, we generated a double-site mutant ( dmTSST-1), modified at solvent-exposed residues predicted to be important for both MHC class II and TCR binding, and detoxified recombinantly expressed TSST-1 ( rTSST-1) as well as native TSST-1 ( nTSST-1) isolated from Staphylococcus aureus by treatment with formaldehyde. Rabbits were immunized with rTSST-1, nTSST-1, dmTSST-1, and formaldehyde inactivated toxoids. The sera obtained were used to map the antigen-reactive regions of the molecule and to identify specificities of antibodies induced by immunization with the different antigens. To detect linear antigenic epitopes of TSST-1 the reactivity of the sera with 11-meric peptides having an overhang of four residues, covering the entire molecule of TSST-1, have been studied. We found that sera of TSST-1 immunized rabbits predominantly reacted with N-terminal residues 1–15, while sera generated with formaldehyde inactivated toxoid recognized a total of 7 regions located at the N- and C-terminus and internal sites of TSST-1. Despite different specificities all sera were able to inhibit TSST-1 induced proliferation of human mononuclear cells.

MOSQUITO LARVAE SOLUBLE FRACTIONS INDUCE DNA SYNTHESIS ALTERATIONS ON HUMAN MONONUCLEAR CELLS

Mosquito larvae soluble fractions obtained by molecular exclusion chromatography altered the mitotic rate of several epithelial cell populations in hepatectomised mice, as well as the proliferation of human mononuclear cells (MNC), stimulating or inhibiting them depending on the fraction and dose applied. The effect was also thermolabile, suggesting a proteic nature of the compounds involved. Analysis of cell viability after culture indicated that the extract did not have lethal toxic effects. One fraction with a molecular weight ranging between 12–80kDa caused only an inhibitory effect. In the present study, we performed further characterisation of this fraction by assaying the effect of new fractions obtained from this one, by the use of a column with a lower molecular weight exclusion range. Assays were performed on the proliferation of adult human MNCs. Our results showed that two out of four of the sub-fractions analysed, with a MW of about 70 and 17kDa, caused a dose-dependent response, either inhibiting or stimulating MNC proliferation respectively.

Differential antigen-stimulated proliferation of human mononuclear cells by recombinant Schistosoma japonicum antigens in a Chinese population.

Peripheral blood mononuclear cells (PBMC) from 117 individuals living on two islands in an area (Dongting Lake) endemic for schistosomiasis japonica in China, and 15 control individuals from a non-endemic area of China, were assessed for antigen-stimulated proliferation against five recombinant Schistosoma japonicum antigens of recognized interest in the development of immunity to schistosomiasis. Two recombinant antigens, paramyosin and 28-kD glutathione-S-transferase, stimulated cellular proliferation (stimulation index > or = 3.0) in 38.5% and 42.5% of subjects, respectively, a level similar to that induced by a soluble whole parasite extract (51.3%). In contrast, three other recombinant antigens tested--a fatty acid binding protein, 22-kD tegumental membrane-associated antigen, and glyceraldehyde-3-phosphate dehydrogenase--stimulated PBMC proliferation in only 3-8% of subjects. Moreover, we also identified a positive association between the degree of exposure, and cellular proliferation following stimulation with recombinant paramyosin or whole parasite extract. Highly significant differences in antigen-stimulated proliferation were also observed between the two islands, Niangashan and Qingshan. The whole parasite extract stimulated proliferation in 90% of subjects from Niangashan island compared with only 42.1% of subjects from Qingshan island (chi2 = 16.88, P = 0.00004), while glutathione-S-transferase stimulated proliferation in 77.3% of subjects from Niangashan island compared with only 34.7% of subjects from Qingshan island (chi2 = 13.09, P = 0.003). A similar, but not significant, trend was observed for paramyosin and the fatty-acid binding protein. The identification of differential cellular proliferative responses to specific schistosome antigens within an infected human population may have important practical implications for vaccine development against schistosomiasis japonica.

MOSQUITO LARVAE PROTEINS MODULATE LUTEINIZING HORMONE AND PROLACTIN RELEASE IN PITUITARY CELLS OF NORMAL AND ESTROGENIZED RATS

Whilst looking for vertebrate growth factor homologues in insects, we found that a soluble fraction of a 12–80kDa molecular weight band peaking at 25kDa, isolated from mosquito larvae extracts by gel permeation chromatography, had a modulatory effect on mouse hepatocytes and adult human mononuclear cell proliferation. The effect disappeared after heating the extract at 90°C for 30min, suggesting that the active factor may be a protein. In order to determine the activity of the extract on cell function, we assessed the effect of the extract on pituitary hormone secretion in vitro. We assayed a dialyzed fraction (MW greater than 12kDa) of mosquito larvae for its effect on the release of luteinizing hormone (LH) and prolactin (PRL) from dispersed rat pituitary cells. In normal anterior pituitary (AP) cells we found that the extract had a stimulatory effect on LH release but an inhibitory action on prolactin secretion. In AP cells obtained from estrogen-induced hyperplasia, the extract had an inhibitory effect on prolactin secretion. In all cases the effects were time- and dose-dependent. Interference of the mosquito proteins with the radioimmunoassay was checked and found to be negligible. After a 60min incubation, cell viability was comparable in control and treated cells. Furthermore, the biological effect of the extract was thermally unstable. Our results suggest that mosquito larvae may share common factors with mammals, probably peptidic in nature, which are able to modulate cell function.

PARTIAL CHARACTERIZATION OF MOSQUITO LARVAE EXTRACT INDUCING DNA SYNTHESIS ALTERATIONS ON HUMAN MONONUCLEAR CELLS

A crude mosquito larvae and dialysed extract alters the mitotic rate of several epithelial cell populations in normal young and adult hepatectomized mice. A crude extract also showed a biphasic effect on the proliferation of human mononuclear cells (MNCs), either stimulating or inhibiting them depending on the dose applied. In the present paper, we assayed the effect of the dialysed mosquito larvae extract and two different protein fractions on human MNCs. Analysis of cell viability after culture indicated that the extract did not have toxic effects. Our results show a dual response of the MNCs to the dialysed, as well as to the protein fraction, with the highest molecular weight inhibiting or stimulating proliferation, depending on the dose applied. The protein fraction with the lowest molecular weight (range between 12–80kDa) showed only an inhibitory effect on cell proliferation.

Vandellia cordifolia regulated cell proliferation and cytokines production in human mononuclear cells.

Vandellia cordifolia (V. cordifolia) used for treatment inflammation in traditional Chinese medicine was selected for immunopharmacological activity test. The effects of V. cordifolia extracted fractions on human mononuclear cells (HMNC) proliferation were determined by tritiated thymidine uptake. The results indicated that VC-ME fraction suppressed HMNC proliferation activated with phytohemagglutinin (PHA) and stimulated cell cycle progression was arrested at the G0/G1 stage. The inhibitory mechanisms may involve the blocking of interleukin-2 (IL-2) and interferon-gamma (IFN-gamma) production, since VC-ME suppressed IL-2 and IFN-gamma production of HMNC in a dose-dependent manner. Therefore, it is suggested that immunomodulatory agents are contained in V. cordifolia.

Immunomodulatory principles of Dichrocephala bicolor.

From the ethanolic extract of Dichrocephala bicolor eight compounds-4,5-dicaffeoyl quinic acid (1); 3,4-dicaffeoyl quinic acid (2); 3,5-dicaffeoyl quinic acid (3); ethyl 4,5-dicaffeoyl quinate (4); methyl 3,5-dicaffeoyl quinate (5); 5-caffeoyl quinic acid (6); caffeic acid (7); and quercetin-3-O-rutinoside (8)-were isolated and identified. All of them were selected for immunopharmacological activity testing. Human mononuclear cells (HMNC) were used as target cells. Cell proliferation was determined by 3H-thymidine uptake. Compounds 2 and 6 potently enhanced HMNC proliferation and interferon-gamma production. Enhancement mechanisms may involve the increase of cytokines production.

Thalidomide and its metabolites have no effect on human lymphocyte proliferation.

Thalidomide is a drug that is being used in several diseases with an immunological component, but the effects on the different immune functions have only been studied partially. Therefore, we studied the effect of thalidomide on PPD-or Con-A-induced proliferation of human mononuclear cells. We found no direct effect of thalidomide at up to 50 micrograms/ml on the cultures. Cells taken from subjects 6 h after ingestion of 200 mg of thalidomide proliferated equally well to PPD and Con-A than cells taken prior to drug administration. Plasma taken from subjects that ingested 200 mg of thalidomide 6 h before did not affect the proliferative response of their own cells when added to the cultures. Plasma from rabbits that were injected with doses 5 or 15 times higher than the dose given to humans did not diminish the proliferative response of human mononuclear cells to PPD. We conclude that neither thalidomide nor its metabolites affect the proliferative response of human mononuclear cells.

Inhibition of human mononuclear cell proliferation, interleukin synthesis, mRNA for IL-2, IL-6, and leukotriene B4 synthesis by a lipoxygenase inhibitor

Human peripheral blood mononuclear cells (PBMCs) were cultured in vitro with various stimuli and in the presence or absence of a 5-lipoxygenase (5-LO) inhibitor, A-63162, to measure its effects on PBMC proliferation, interleukin-2 receptor (IL-2R) expression, interleukin-2 (IL-2) synthesis, interleukin-6 (IL-6) synthesis, and accumulation of messenger RNA for IL-2 or IL-6. A-63162 inhibited PBMC proliferation stimulated by phytohemagglutinin (PHA) and phorbol myristate acetate (PMA) plus A23187, IL-2 receptor expression stimulated by PHA, and IL-2 or IL-6 synthesis induced by PHA plus PMA or PMA plus A23187. At the same concentration, A-63162 inhibited accumulation of mRNA for IL-2 or IL-6 and also inhibited leukotriene B4 (LTB4) synthesis. Our data indicate that the 5-LO inhibitor A-63162 has immunosuppressive activity that may be due to inhibition of LTB4 production or to direct inhibitory actions of A-63162 on IL-2 and IL-6 synthesis.

The immunosuppressive effect of ursodeoxycholic acid: A comparativein vitro study on human peripheral blood mononuclear cells

Ursodeoxycholic acid is an efficient treatment for putatively immune-mediated liver diseases, but its mechanism of action is unknown. We studied human mononuclear cell proliferation as an in vitro model for cell-mediated immunity in the presence of ursodeoxycholic acid, its glycoconjugate and tauroconjugate and chenodeoxycholic acid at concentrations of 5, 25 and 50 mumol/L. Proliferation was inhibited in a dose-dependent manner compared with control values (15% to 54% depending on the bile acid, concentration and mitogen used), except at 5 mumol/L where inhibition was significant with only one mitogenic stimulus of the three used. With one mitogen (phorbolester) the inhibition was additive with that of cyclosporine. The number of cell-surface receptors studied was not modified by bile acids. Interleukin-2 production was decreased 35% to 60% by ursodeoxycholic acid and its conjugates. The proliferation of the interleukin-2-dependent cell line CTLL-2 was also inhibited. The immunosuppression was reversible except at a chenodeoxycholic acid concentration of 50 mumol/L. Because bile acids are able to partition into membranes and change their properties, we speculate that this allows them to interact with cell-surface receptors or signaling systems within the membrane or on its inner face, thus impairing their function. This would inhibit the numerous extracellular messages that lymphocytes need to proliferate.

Selective inhibition of T cell activation by an inhibitor of S-adenosyl-L-homocysteine hydrolase.

Defects in the enzymes involved in the pathway of S-adenosylmethionine (AdoMet) metabolism, or inhibition of those enzymes, results in profound immunodeficiency. We have examined MDL 28,842, a novel irreversible inhibitor of S-adenosyl-L-homocysteine hydrolase (AdoHcyase), an enzyme involved in AdoMet metabolism, to determine its effect on the immune system and to investigate its potential as an immunosuppressive agent. The stimulation of human mononuclear cell proliferation in vitro with Con A, a T cell mitogen, and PWM, a T-dependent B cell mitogen, were inhibited by MDL 28,842. The 50% inhibitory concentration for both were 0.33 microM. In murine spleen cells, MDL 28,842 was a potent, nontoxic, inhibitor of Con A-stimulated T cell proliferation (IC50 = 0.19 microM) but did not affect LPS-induced B cell proliferation. This selective suppression was also observed when enriched murine T and B cells were stimulated with mitogens, although S-adenosyl-L-homocysteine (AdoHcy), the substrate of AdoHcyase, was similarly elevated in both populations. In addition to proliferation in response to a number of stimuli, IL-2 production and the expression of IL-2R by mitogen-stimulated T cells were inhibited by MDL 28,842. These results suggest a direct effect of MDL 28,842 on T cells. In vivo, the antibody response to a T cell-dependent Ag, OVA, was inhibited by MDL 28,842. The response of splenic T cells from these animals to OVA in vitro were similarly depressed compared with controls. The results demonstrate that MDL 28,842 is a potent nontoxic immunosuppressive agent, which has selectivity for T cells and therefore may be useful in the treatment of T cell-mediated disorders, such as autoimmune disease and tissue transplantation.

Cytotoxicity of mebendazole against established cell lines from the human, rat, and mouse liver.

The direct cytotoxicity of mebendazole (MBZ) was investigated by using cell lines derived from human, mouse and rat liver. It was demonstrated that Chang liver cells (derived from human liver) were more sensitive to the cytotoxic effects of MBZ than the other two cell lines. Longer incubation of the cells with MBZ resulted in stronger toxicity, and the cytotoxicity was dependent on the MBZ concentration above a certain threshold value (0.25-0.50 mg/l in a 42-h culture). Inhibition of the proliferation of Chang liver cells by MBZ was detected at a concentration of 0.008 mg/l, a lower concentration than that having a cytotoxic effect. The other two cell lines were less sensitive to the inhibitory effect of MBZ. Proliferation of human mononuclear cells following stimulation by phytohemagglutinin (PHA) was inhibited by MBZ, and this inhibition was more extensive than that of cells stimulated with whole formalin-treated Pseudomonas aeruginosa. It is suggested that dividing cells may be more sensitive to MBZ cytotoxicity. This anti-proliferative effect may be related to its clinically known side effects, such as hepatotoxicity and bone marrow suppression.

IgG Anti-IgE Autoantibodies in Immunoregulation

Human sera contain anti-IgE autoantibodies with diverse biological functions in vitro. Opposite functions can be shown for triggering of IgE-mediated histamine release from human basophils in terms of anaphylactogenic or nonanaphylactogenic autoantibodies. Furthermore, autoantibodies are either capable of removing IgE from the surface of CD23-positive cells of binding more IgE to such cells. A similar dichotomy seems also to exist for the effect of autoantibodies on human IgE mRNA synthesis as well as IL-4-induced proliferation of human mononuclear cells. Thus, anti-IgE autoantibodies may represent a key factor in the manifestation and the development of allergic disease.

Parasite antigens are present in breast milk of women infected with Onchocerca volvulus.

We used a noncompetitive two-site ELISA with 5 monoclonal antibodies to determine whether parasite antigens are present in breast milk from women infected with Onchocerca volvulus. Seven out of 13 available milk samples contained significant amounts of filarial antigens. Antigen indices in milk correlated with levels of microfilarodermia (Rs = 0.74, P less than 0.005). Antigen-containing milk samples markedly inhibited mitogen-induced proliferation of human mononuclear cells and activated cells within this population that suppressed the proliferative response of autologous lymphocytes to mitogens and antigens. These findings indicate that parasite products are present in breast milk of O. volvulus-infected women and suggest that these may induce immune tolerance and/or suppression in infants born of infected mothers.

Aryl-fluoroquinolone derivatives A-56619 (difloxacin) and A-56620 inhibit mitogen-induced human mononuclear cell proliferation.

Aryl-fluoroquinolone derivatives A-56619 (difloxacin) and A-56620 were found to inhibit human peripheral blood mononuclear cell (MNC) proliferation (measured by [3H]thymidine uptake) that was induced by concanavalin A or monoclonal antibody OKT3. These antimicrobial agents exert their maximum suppressive effect when added within the first 24 h after the onset of culture with concanavalin A. No increase in the concentration of mitogen or the duration of incubation of MNC cultures reversed this inhibitory effect, but the removal of the drug from cultures reversed the suppression of DNA synthesis. A-56619 appeared not to interfere with the triggering of MNC activation by mitogen because it did not inhibit mitogen-induced increase in protein synthesis (measured by [3H]leucine incorporation), interleukin-2 receptor expression (measured by the binding of fluorescein-conjugated monoclonal antibody against interleukin-2 receptor), and cell volume. These findings are considered in terms of possible interference of aryl-fluoroquinolones with mammalian topoisomerase and DNA polymerases.