Inhibition effect of shengma-gegen-tang on measles virus in Vero cells and human peripheral blood mononuclear cells

The nucleotide sequence of gene 6 encoding the rotavirus major capsid protein VP6 of EDIM strain (EW) was determined and compared to that of 20 previously reported strains with known subgroup specificities. Multiple alignments of amino acid sequences exhibited a high level of sequence conservation (87 to 99.2%). Site-specific mutagenesis experiments were undertaken to localize regions involved in subgroup specificity. Amino acid positions 305, 315, and a region 296-299 (or 301 for equine strain H-2) were identified as contributing to subgroup epitopes. A single amino acid mutation at position 305 or 315 was sufficient to change the subgroup specificity of EW VP6 protein from non I/II to subgroup I- or subgroup II-like, respectively. Mutation at these sites may be another important mechanism for subgroup variation, along with gene reassortment.     

Inhibition of muscarinic receptor-stimulated glial cell proliferation by ethanol

Acetylcholine and other muscarinic agonists stimulate the proliferation of rat cortical astrocytes and 132 1N1 human astrocytoma cells by activating muscarinic m3 cholinergic receptors. Ethanol was a potent inhibitor of carbachol-stimulated proliferation, measured by [3H]thymidine incorporation, with an IC50 of 10 mM. On the other hand, basal and serum-stimulated proliferation of astrocytes and astrocytoma cells was inhibited by ethanol with lower potency (IC50 = 200-250 mM). Concentration-response experiments with carbachol, in the presence of 10 mM ethanol, suggested that inhibition of proliferation by the alcohol was of the noncompetitive type. Experiments with acetaldehyde and with the alcohol dehydrogenase inhibitor 4-methylpyrazole suggested that the inhibitory effect of alcohol was due to ethanol itself and not to its metabolite acetaldehyde. Proliferation of astrocytoma cells induced by carbachol and the inhibitory effects of ethanol were also confirmed by flow cytometry using the 5-bromodeoxyuridine-Hoechst 33258 method. Ethanol (10 mM) had no effect on proliferation induced by 50 micrograms/ml insulin and 100 ng/ml platelet-derived growth factor BB; on the other hand, the mitogenic effect of 1 mM histamine, 100 U/ml interleukin-1, and 100 ng/ml 12-O-tetradecanoylphorbol 13-acetate were inhibited by approximately 50%. These results indicate that proliferation of glial cells induced by muscarinic agonists is especially sensitive to the inhibitory effect of ethanol. This action of ethanol may be relevant to its developmental neurotoxicity, particularly microencephaly, which is one of the common features of the fetal alcohol syndrome.     

Suppression of HIV-1 replication in peripheral blood mononuclear cells by fasudil

Fasudil is a potent inhibitor for various protein kinases such as myosin light chain kinase and protein kinase C. It has been used as a drug for improvement of intracranial vasospasm and following ischaemic diseases. In this report, we demonstrate that fasudil suppressed the replication of human immunodeficiency virus type 1 (HIV-1) in mitogen-activated peripheral blood mononuclear cells. Our finding shows that fasudil may be useful as a new and distinct chemotherapeutic agent against HIV-1 infection.     

Immunoglobulin-secreting cells in healthy peripheral blood mononuclear cells

Immunoglobulin-secreting cells were measured in healthy uncultured peripheral blood mononuclear cells (PBMCs) in vitro. The percentage of IgM-, IgG- and IgA-secreting cells in adult PBMCs was 0.053, 0.099 and 0.065%, respectively. The percentage of IgM-, IgG- and IgA-secreting cells was 0.73, 5.2 and 3.8% of surface IgM-, IgG- and IgA-bearing cells, respectively. The numbers of IgM-, IgG- and IgA-secreting cells increased with age in childhood. However, the numbers of all three classes were slightly decreased in adults compared with children aged 9-15 years. These results may explain the difference in immunity in vivo.     

Immunological status of patients subjected to cardiac surgery: effect of lactoferrin on proliferation and production of interleukin 6 and tumor necrosis factor alpha by peripheral blood mononuclear cells in vitro

The aim of this investigation was to monitor proliferation and cytokine production by peripheral blood mononuclear cells (PBMC) of patients subjected to cardiac surgery. Another goal was to establish regulatory effects of lactoferrin (LF) on these immune reactions in vitro. PBMC were tested before, during surgery and on day 1 and day 8-10 following operation. In control donors, low spontaneous and phytohemagglutinin (PHA)-induced proliferation of PBMC, as well as lipopolysaccharide (LPS)-induced TNF-alpha secretion was stimulated by LF, but high production of this cytokine was inhibited. In patients, the proliferation of PBMC and the ability to produce IL-6 TNF-alpha by these cells underwent characteristic changes depending on preoperative immune reactivity of patients. In general, low, preoperative reactivity of PBMC showed a tendency to increase within the monitoring period whereas moderate/high responsiveness was diminishing. LF had, in majority of cases, a down-regulatory effect on the proliferative response, best pronounced in patients of high/moderate preoperative response. Similarly, LF exhibited, in general, an inhibitory effect on LPS-induced IL-6 production. In terms of TNF-alpha production, a considerable up-regulatory effect of LF, particularly in low responding patients was of a special interest. In summary, we suggest that LF may play a role in lowering the immune response of patients to surgery and promoting tissue regeneration.     

Phenotype and engraftment potential of cytokine-mobilized peripheral blood mononuclear cells

Atypical fibroxanthoma is a superficial variant of pleomorphic malignant fibrous histiocytoma. Histopathologically, it is characterized by a dermal nodule composed of bizarre cells arranged in a haphazard-to-fascicular pattern. These cells are spindle or rounded, pleomorphic and with numerous atypical mitotic figures. Some cells appear polygonal with ample and foamy cytoplasm. We recently encountered two elderly patients with atypical fibroxanthoma on their face. Histopathologically, one of the lesions was composed, almost entirely, of clear cells, whereas in the other one aggregations of clear cells constituted a half of the neoplasm. Atypical multinucleated cells with a Touton-like appearance were present. In addition to clear cells, areas of more conventional atypical spindle cells arranged in fascicles were seen, supporting the diagnosis of atypical fibroxanthoma. PAS staining failed to demonstrate glycogen in neoplastic cells. Immunohistochemistry revealed that neoplastic cells expressed positivity for vimentin, muscle-specific actin, and alpha smooth muscle actin, whereas cytokeratin, S-100 protein, EMA, CEA, and desmin were negative. Ultrastructural studies showed that neoplastic cells contained abundant rough endoplasmic reticulum, mitochondria, and numerous lipid vacuoles within the cytoplasm. Clear-cell atypical fibroxanthoma is a rare variant of atypical fibroxanthoma that should be differentiated from other clear-cell neoplasms of the skin.     

IL-10 synthesis and secretion by peripheral blood mononuclear cells in haemodialysis patients

PURPOSE OF STUDY: IL-10 may explain the paradox between immunodeficiency and oversecretion of cytokines in chronic haemodialysis (HD) patients. We analysed the secretion of IL-10 by PBMC and the expression of IL-10 mRNA in 10 long-term HD patients (108-276 months), 10 short-term HD patients (3-18 months), and 10 healthy controls. RESULTS: Spontaneous IL-10 secretion was higher in HD patients than in controls (15 pg/ml vs 2 pg/ml, P = 0.004). It was detected in 13 of 20 patients and in 1 of 10 controls (P = 0.01). IL-10 mRNA expression was also higher in HD patients than in controls. Spontaneous secretions of IL-10 and IL-6 were positively correlated in patients. IL-10 secretion in response to LPS was higher than the upper limit of control range in 4 of 10 long-term HD patients and in no short-term HD patients (P = 0.04). IL-10 mRNA expression was also higher in long-term than in short-term HD patients. CONCLUSIONS: This study demonstrates that IL-10 is spontaneously synthesized and secreted in HD patients, supporting an immunomodulating role in this setting. The greater IL-10-producing capacity in long-term HD patients indicates a chronic effect of haemodialysis on PBMC responsiveness.     

Interleukin-1 receptor antagonist synthesis by peripheral blood mononuclear cells in hemodialysis patients

BACKGROUND: Pro-inflammatory cytokines like interleukin (IL)-1 beta and tumor necrosis factor-alpha (TANF-alpha) are believed to play a significant role in dialysis-related morbidity. It has been previously demonstrated that the endogenous synthesis of interleukin-1 receptor antagonist (IL-1Ra) is a reliable marker of the level of IL-1 beta synthesis in hemodialysis (HD) patients. In this study, we assessed the impact of clinical and laboratory variables on IL-1Ra synthesis by peripheral blood mononuclear cells (PBMC) in patients on HD with unsubstituted cellulose dialyzers. METHODS: IL-1Ra by PBMC was measured by a specific non-cross-reactive radioimmunoassay. Day to day variation in cytokine synthesis, the correlation between cytokine synthesis under different in vitro stimulatory conditions, and the influence of clinical and laboratory variables on cytokine synthesis were studied. RESULTS: Although there was a trend towards greater IL-1Ra synthesis by unstimulated, endotoxin-stimulated and IgG-stimulated PBMC drawn before the second and third dialysis sessions of the week when compared to the first dialysis treatment, this was not statistically significant. There was a strong correlation between IL-1Ra synthesis by PBMC cultured under different stimulatory conditions that was best observed between IL-1Ra cell content and from endotoxin-stimulated PBMC (r = 0.51, P = 0.0001), and endotoxin- and IgG-stimulated PBMC (r = 0.44, P = 0.0001). In addition, there was a close correlation between total synthesis (cell associated and secreted) and secreted levels of IL-1Ra in unstimulated (r = 0.59, P = 0.0001) and endotoxin-stimulated PBMC (r = 0.69, P = 0.0001). Interestingly, there was an inverse correlation between IL-1Ra synthesis and duration of dialysis that was strongest for secreted IL-1Ra from unstimulated (r = -0.50, P = 0.002) and endotoxin-stimulated PBMC (r = -0.34, P = 0.04). There was no significant correlation between IL-1Ra synthesis by PBMC and other clinical and laboratory indices. CONCLUSIONS: The observations from this study indicate that: (1) in HD patients, there were no significant differences in cytokine synthesis by PBMC drawn before the three different dialysis treatments during the week; (2) there is a close relationship between IL-1Ra synthesis from PBMC cultured under different stimulatory conditions; (3) the secreted levels of IL-1Ra correlate directly with total synthesis (cell-associated and secreted); (4) with the exception of duration of dialysis, none of the other clinical or laboratory parameters correlated with cytokine synthesis; and (5) the diminished endotoxin- or IgG-stimulated IL-1Ra synthesis with increasing time on dialysis is possibly another sign of the impaired host-defense system in patients on long-term hemodialysis.     

Effect of two-chambered bicarbonate lactate-buffered peritoneal dialysis fluids on peripheral blood mononuclear cell and polymorphonuclear cell function in vitro

Low pH, high osmolality, increasing glucose concentration, and glucose degradation products (GDP) formed during heat sterilization of conventional peritoneal dialysis (PD) fluids have been shown to have a detrimental effect on cells involved in peritoneal host defense. The two-chambered PD fluid bag in which glucose at pH approximately 3 is separated from a bicarbonate (25 mmol/L)-lactate (15 mmol/L) buffer during heat sterilization permits PD fluids with lower GDP to be delivered to the patient at neutral pH. To establish the possible benefit of two-chambered bag PD fluids on peripheral blood mononuclear cell (PBMC) and polymorphonuclear (PMN) cell function, we compared conventional 1.5% Dianeal (1.5%D) with 1.5% two-chambered bag bicarbonate-lactate (1.5%D-B), and conventional 4.25% Dianeal (4.25%D) with 4.25% two-chambered bag bicarbonate-lactate (4.25%D-B). Furthermore, to study the effect of the sterilization process on PBMC and PMN function, we compared filter-sterilized 4.25%D (4.25%D-F) with 4.25%D and 4.25%D-B. PBMC were harvested by Ficoll-Hypaque separation, and 2.5 x 10(6) cells in RPMI were incubated with an equal volume of the test fluids for 4 hours, pelleted, and resuspended in RPMI containing 10 ng endotoxin for a further 20 hours. Tumor necrosis factor alpha (TNF-alpha) production by endotoxin-stimulated PBMC was not significantly different (P = 0.10) between 1.5%D-B and 1.5%D, but was significantly higher (P = 0.01) with 4.25%D-B compared with 4.25%D. PBMC exposed to filter-sterilized fluid (4.25%D-F) showed significantly higher endotoxin-stimulated TNF-alpha production compared with 4.25%D (P = 0.02), but was not significantly different from 4.25%D-B (P = 0.40). PMN were harvested by Ficoll-Hypaque separation and 10 x 10(6) cells incubated with test fluids for 30 minutes. After incubation, phagocytosis (phagocytosis index) was determined by the uptake of 14C-labeled Staphylococcus aureus, oxidative burst by reduction of ferricytochrome C to ferrocytochrome C on stimulation with PMA, and enzyme release by measurement of endotoxin-stimulated bactericidal/permeability increasing protein (BPI). Bicarbonate-lactate two-chambered fluids of similar osmolality and glucose concentration conferred a significant improvement in phagocytosis (P = 0.02 for 1.5%D-B and P < 0.001 for 4.25%D-B). Oxidative burst and BPI release were significantly higher in 4.25%D-B compared with 4.25%D (P < 0.001). Filter-sterilized 4.25%D-F conferred a significant improvement in phagocytosis and oxidative burst compared with 4.25%D (P < 0.001) or 4.25%D-B (P < 0.001). Furthermore, conventional 4.25%D was associated with significantly lower BPI release compared with 4.25%D-F (P = 0.01). GDP's acetaldehyde and 5-HMF were analyzed in 4.25%D-B, 4.25%D, and 4.25%D-F. Acetaldehyde was below the lower limit (0.79 ppm) of the standard curve in 4.25%D-B and 4.25%D-F fluids but was detected (3.76 to 5.12 ppm) in all of the 4.25%D fluids. Relative levels of 5-HMF in the 4.25%D-B (0.032 to 0.041 Abs @ 284 nm) and 4.25%D (0.031 to 0.036 Abs @ 284 nm) were similar. The lowest levels (0.001 Abs @ 284 nm) were observed in the filter-sterilized 4.25%D-F. The beneficial effects of two-chambered bicarbonate lactate-buffered PD fluids on PBMC and PMN function are probably related to reduction of GDP from heat sterilization of glucose in a separate chamber at a lower pH. This improvement in biocompatibility could have a beneficial affect on peritoneal defenses.     

Inhibition of rabbit lens epithelial cell proliferation

OBJECTIVE: To study the ability of homoharringtonine, 5-fluorouracil and adriamycin on inhibiting the proliferation of rabbit lens epithelial cells (RLEC) and the prevention of after cataract by using homoharringtonine. METHODS: RLEC were isolated and cultured. (1) The passage RLEC were placed in 24-well tissue culture plates and incubated for 48 hours, then exposed to different concentrations of homoharringtonine, 5-fluorouracil and adriamycin for 24 and 72 hours; (2) The passage RLEC and homoharringtonine, 5-fluorouracil, adriamycin were placed and cultured for 24 hours to investigate the rate of attached cells; (3) The morphological changes of RLEC were studied under light microscope. RESULTS: The ID50 of homoharringtonine, 5-fluorouracil and adriamycin exposed to RLEC for 24 hours were 0.84 micrograms, 0.58 micrograms and 4.50 ng/ml and those for 72 hours were 0.49 micrograms, 0.33 micrograms and 3.85 ng/ml respectively. In the homoharringtonine group, the rate of attached cells was less than that of 5-fluorouracil and adriamycin groups. The study of the morphological changes showed that the different concentrations of antiproliferative drugs affected on RLEC at different regions. CONCLUSION: The authors consider that homoharringtonine may be more effective for the prevention of after cataract than 5-fluorouracil and adriamycin.     

Effects of diheptyldiselenide (DDS) on human tumor cell lines and on peripheral blood mononuclear cells

In vitro effects of graded concentrations of diheptyldiselenide (DDS) on human tumor cell proliferation, and on the proliferative responses and immunological functions of peripheral blood mononuclear cells (MNC) were investigated. The agent significantly decreased tumor cell proliferation in a dose and time dependent manner. Proliferative responses of MNC to phytohemagglutinin (PHA) and interleukin-2 (IL-2) were also significantly depressed when MNCs were exposed to DDS (250 microM for 18 h) led to a significant increase in NK activity only in MNC samples showing very limited baseline NK function. On the other hand, generation of LAK cells was significantly inhibited by DDS. However, when the agent was added to the effector and target cell mixture during the 4 h 51Cr release cytotoxicity assay, no influence was found on NK and LAK-mediated target cell lysis. These studies show that high concentrations of DDS inhibit tumor cell proliferation and could also impair certain proliferative-dependent immune functions, without directly affecting cell-mediated cytolytic activity of effector cells.     

Cytokine production by human peripheral blood mononuclear cells: differential senstivity to glutamine availability

Human peripheral blood mononuclear cells (PBMCs) were cultured in the presence of different glutamine concentrations (0, 0.1, 0.4, 0.6, 2 mM) and stimulated with concanavalin A (Con A) or bacterial lipopolysaccharide (LPS). The concentrations of T lymphocyte- and monocyte-derived cytokines were measured in the culture medium 24 h later. The availability of glutamine significantly increased the production of interleukin (IL)-2 (2-fold), IL-10 (4-fold) and interferon (IFN)-gamma (4.5-fold) by Con A-stimulated PBMCs. Maximal production of these cytokines occurred at 0.1 mM glutamine and increasing the concentration of glutamine above that did not lead to a further increase in cytokine production. Glutamine availability resulted in small increases (24 to 35%) in the production of IL-1alpha, IL-1beta, IL-6 and tumour necrosis factor (TNF)-alpha by Con A-stimulated PBMCs; again maximal production occurred at a glutamine concentration of 0.1 mM. Glutamine availability did not influence the production of IL-1beta or TNF-alpha by LPS-stimulated PBMCs, while there was a small increase (17 to 32%) in the production of IL-1alpha, IL-6 and IL-10 by these cells at a glutamine concentration of 0.1 mM. It is concluded that glutamine enhances the production of T lymphocyte-derived cytokines with only minimal effects on production of cytokines by monocytes.     

The effects of retinol on in vitro immunoglobulin synthesis by cord blood and adult peripheral blood mononuclear cells

In this study we examined the effects of retinol (ROH), a metabolic precursor of retinoic acid (RA), on Staphylococcus aureus Cowan I (SAC)-induced immunoglobulin synthesis of cord blood mononuclear cells (CBMC) and adult peripheral blood mononuclear cells (PBMC). ROH augmented SAC-induced IgM synthesis of CBMC by 5.9 +/- 1.5-fold (n = 7, mean +/- s.d.), and IgG synthesis of adult PBMC by 16.3 +/- 5.1-fold (n = 3) at optimal concentrations of 10(-6) M and 10(-11) M, respectively. No augmenting effects could be demonstrated for the other immunoglobulin isotypes. Time-course studies showed that the synthesis of IgM by CBMC was accelerated with detectable immunoglobulin in supernatant fluids starting on day 3. ROH augmented immunoglobulin synthesis of CBMC stimulated by Epstein-Barr virus (EBV), a T cell-independent polyclonal activator, and of EBV-transformed B cell clones (2.5 +/- 0.2 and 4.1 +/- 1.5-fold increase, respectively), which suggests that ROH can act directly on B cells to enhance immunoglobulin synthesis. In contrast, when ROH was preincubated with cord blood T cells, washed and added to the B cell-enriched fraction with SAC, no increase (0.9-1.8-fold) in IgM synthesis was obtained. Thus, the principal mechanism(s) by which ROH augments immunoglobulin synthesis is by acting on B cells. This is in contrast to the immunoglobulin-enhancing effects of RA which is mediated by T cells, or T cell products, e.g. cytokine. Our studies suggest that RA and ROH may have different pathways of immunoglobulin-enhancing effects, perhaps mediated by different retinoid binding proteins resulting in gene activation and immunoglobulin synthesis.     

Proliferation of human peripheral blood mononuclear cells during calcium channel blockade

To evaluate the role of intracellular calcium and particularly Ca2+ uptake in the initiation of lymphocyte mitogenesis, the effect of mibefradil--which blocks both L- and T-type calcium channels with a more selective blockade of T-type channels--on the proliferation of human peripheral blood mononuclear cells (PBMC) is compared with the effect of nifedipine, which blocks only the L-type calcium channel. The rate of 3H-thymidine, 3H-uridine, and 3H-leucine incorporation into control and concanavalin A-stimulated PBMC in the presence or absence of the calcium channel blockers mibefradil or nifedipine (1, 10, or 50 micromol/L), and of the intracellular calcium antagonist TMB-8 or the calmodulin antagonist W-7 (1, 10, 25, or 50 micromol/L) was assayed in cells cultured for 3 days. The cellular cytotoxicity and the cell number in growing cultures was also determined in mibefradil- or nifedipine-treated control or stimulated cells. Mibefradil and nifedipine reduced the cell number and the 3H-thymidine, 3H-uridine, or 3H-leucine incorporation or the de novo DNA, RNA, or protein synthesis in control and concanavalin A-stimulated human PBMC in a concentration-dependent manner. Mibefradil exhibited a more pronounced inhibition than nifedipine. The inhibitory effect of mibefradil or nifedipine on DNA synthesis was dependent upon the timing of treatment with the drugs. The inhibitory effect of mibefradil or nifedipine on the lymphoproliferative response was nearly abolished if the drugs were added 20 h after cell stimulation. A markedly reduced inhibitory effect was found when mibefradil or nifedipine were added 1 to 7 h after cell stimulation. However, regardless of time of addition, TMB-8 and W-7 caused a persistent inhibition of the proliferation of human PBMC. Our data show that mibefradil had a more pronounced inhibitory effect on the proliferation of human PBMC than nifedipine and that this inhibitory effect on de novo DNA synthesis was dependent upon the timing of treatment with both drugs. Mibefradil and nifedipine also reduce RNA and protein synthesis in human PBMC. Therefore, administration of these calcium channel blockers to inhibit cellular proliferation might be most beneficial at anatomic sites where cellular proliferation is not already an active process, while being ineffective in the presence of ongoing active proliferation, as suggested by some prospective studies.     

Interleukin 1 production by peripheral blood mononuclear cells of children with bronchial asthma in remission

To analyse the change of the immunological response in the remission state of children with bronchial asthma, we studied the interleukin 1 (IL-1) production of peripheral blood mononuclear cells (PBMC) obtained from children with bronchial asthma sensitized by mite antigen. After PBMC were cultured for 24 hours with Dermatophagoides farinae (Df) or lipopolysaccharide (LPS), the PBMC-derived culture supernatant was estimated for IL-1 alpha and beta by enzyme-linked immunosorbent assay (ELISA). PBMC from some of subjects with active asthma produced IL-1 alpha and beta without any stimulation, but not those from controls or subjects in remission. IL-1 alpha and beta production of PBMC stimulated with Df was observed in all three groups, but IL-1 produced by subjects with active asthma was higher than that produced by subjects in the other two groups. Moreover, when PBMC were incubated with LPS, the secretion of both IL-1 alpha and beta was enhanced. PBMC from patients with active asthma produced both IL-1 alpha and beta in amounts comparable to those produced by PBMC from control subjects, but IL-1 production of PBMC from patients in remission was lower than in the other two groups. IL-1 beta production was about ten times as much as IL-1 alpha. Df-induced IL-1 production of PBMC from asthmatic patients sensitized by mite antigen, which was increased in the active state, was down-regulated in the remission state. Moreover, nonspecific stimuli such as LPS may induce the suppressive factors which down-regulate IL-1 production.(ABSTRACT TRUNCATED AT 250 WORDS)     

Inhibition of glomerular mesangial cell proliferation and extracellular matrix deposition by halofuginone

Mesangial cell proliferation, increased deposition of collagen, and expansion of the mesangial extracellular matrix (ECM) are key features in the development of mesangioproliferative diseases. Halofuginone, a low molecular weight anti-coccidial quinoazolinone derivative, inhibits collagen type alpha 1(I) gene expression and synthesis. We investigated the effect of halofuginone on both normal and SV40 transformed mesangial cell proliferation, collagen synthesis, and ECM deposition. Proliferation of both cell types was almost completely inhibited in the presence of 50 ng/ml halofuginone. The cells were arrested in the late G1 phase of the cell cycle and resumed their normal growth rate following removal of the compound from the culture medium. The antiproliferative effect of halofuginone was associated with inhibition of tyrosine phosphorylation of cellular proteins. Similar results were obtained whether the mesangial cells were seeded on regular tissue culture plastic or in close contact with a naturally produced ECM resembling their local environment in vivo. Halofuginone also inhibited synthesis and deposition of ECM by mesangial cells as indicated by a substantial reduction in 14C-glycine and Na2(35)SO4 incorporation into the ECM, and by the inhibition of collagen type I synthesis and gene expression. It is proposed that by inhibiting collagen type I synthesis and matrix deposition, halofuginone exerts a potent antiproliferative effect that may be applied to inhibit mesangial cell proliferation and matrix expansion in a variety of chronic progressive glomerular diseases.     

Inhibition of smooth muscle cell proliferation and neointimal growth by low-anticoagulant heparin

Low-anticoagulant heparin (LA-heparin) obtained by affinity chromatography on antithrombin III Sepharose inhibits the proliferation of cultured arterial smooth muscle cells in an in vitro bioassay system as effectively as standard heparin. A growth inhibition of smooth muscle cells of about 60% is achieved when LA-heparin or heparin is added to the culture medium to a concentration of 50 micrograms/ml. In normolipemic rats LA-heparin suppresses the formation of neointimal thickenings and stenosis after balloon catheter-induced deendothelialization of the carotid artery. In terms of mass a dose of 5 mg/kg body weight/d given subcutaneously twice daily one week before and 2 weeks after balloon injury the cross sectional area of the neointima is reduced to 36% as compared with the nontreated control group (100%). This 64% reduction is statistically highly significant (p < 0.001). After treatment with 0.5 mg LA-heparin/kg/d the reduction of the neointima was 11% (p < 0.05). At a dose of 5 mg/kg body weight single or repeated administrations of LA-heparin caused only a small and transient increase in activated partial thromboplastin time values. The results show that subcutaneous administration of LA-heparin very effectively prevents smooth muscle cell proliferation and balloon catheter-induced neointimal growth. The well tolerated systemic application of this chemically non-modified LA-heparin might justify clinical trials for prevention of restenosis after percutaneous transluminal coronary angioplasty or other invasive cardiovascular interventions without complications of bleeding.     

HIV type 1 Tat protein inhibits interleukin 12 production by human peripheral blood mononuclear cells

HIV-1 Tat protein, which trans-activates HIV-1 expression, exerts many effects on host immune function. Meanwhile, PBMCs and pulmonary macrophages from HIV-1-infected patients produce only a small amount of IL-12, which plays an essential role in the development of helper T type 1 (Th1) cells, and in the generation of cytotoxic T lymphocytes. We examined the possibility that Tat suppresses IL-12 production by PBMCs from healthy donors. Tat significantly inhibited IL-12 production by human PBMCs stimulated with Staphylococcus aureus Cowan 1 strain (SAC) at concentrations between 5 and 40 ng/ml. Immunoabsorption by using polyclonal antibody to Tat abolished the suppression of the IL-12 production by Tat. Tat at the same concentrations did not affect IL-10, IL-6, or TNF-alpha production. Other HIV-1 proteins (Nef and gp120) did not influence IL-12 production. Tat also suppressed the expression of mRNA encoding the p40 chain of IL-12, whereas it did not affect the expression of mRNA encoding IL-10 and beta-actin. IL-12 production by monocytes, separated from PBMCs by the adhesion method, was also inhibited by Tat. These results suggest that Tat protein is one of the main causes of decreased IL-12 production by PBMCs (mostly by monocytes) from HIV-1-infected individuals.