Biomedical subjects

L Lu

Publications and source records attributed to L Lu.

At least 199 records · Page 11Linked to original sources

PubMed2000

[Study on normal pregnancy and recurrent spontaneous abortions from helper T cell 1/helper T cell 2 cytokines balance].

OBJECTIVE: To investigate the function of Th1 (IFN-gamma, TNF-alpha) and Th2 (IL-4, IL-10) cytokine balance in the maternal-fetal tolerance. METHODS: The IFN-gamma, TNF-alpha, IL-4 and IL-10 mRNA were studied from PBMCs of 11 normal non-pregnancy control, 10 women in first trimester of normal pregnancy, 11 women in last trimester of normal pregnancy by semi-quantitative reverse transcriptional-polymerase chain reaction (RT-PCR) method. RESULTS: (1) The IFN-gamma mRNA decreased [(24.68 +/- 5.55)%, P < 0.05], while the IL-10 mRNA rose [(20.72 +/- 5.36)%, P < 0.01] in normal pregnancy women compared with the non-pregnancy control [IFN-gamma(30.76 +/- 5.35)%, IL-10 (13.58 +/- 3.91)%]. (2) IL-10 mRNA decreased [(9.48 +/- 2.84)%, P < 0.05], while the IFN-gamma mRNA rose [(36.08 +/- 5.75)%, P < 0.05] in RSA women compared with the non-pregnancy control. (3) The IFN-gamma mRNA rose (P < 0.01), while the IL-10 mRNA decreased (P < 0.01) in RSA women compared with women in their first trimester of normal pregnancy [IFN-gamma(27.23 +/- 5.74)%, IL-10 (18.61 +/- 5.43)%]. CONCLUSIONS: The results suggest that women in normal pregnancy had mainly the Th2 cytokines. The breakage of the Th1/Th2 balance may result in RSA. The results provide clue that Th1/Th2 balance plays a role in maternal-fetal tolerance.

Abortion, Habitual↗

PubMed2000

[The prevention of infection complicating acute necrotizing pancreatitis:an experimental study].

OBJECTIVE: To observe the effects of 5 intervention measures on infection complicating acute necrotizing pancreatitis (ANP) in dogs and rats. METHODS: A lethal model of ANP was reproduced by infusion of artificial bile into the biliopancreatic duct. Animals were divided randomly into: ANP group (no treatment); Chinese medicine group ("Qing Yi Tang"); Bifidobacterium mixture group; purgation group (MgSO(4)); selective decontamination of the digestive tract (SDD) group; and somatostatin group. The pancreas and intestine were observed morphologically and tight junction on ileum epithelia was assessed on cryofracture replicas. Blood and/or tissue levels of DAO and D-lactic acid and uric contents of lactulose/mannitol (L/M), served as indicators of gut barrier function, were measured at various time points. Intestinal flora and incidence of bacterial translocation (BT) to organs were examined. RESULTS: In early stage of ANP, mucosal and epithelial tight junction damage and flora disturbance occurred in the gut. In addition, the gut barrier function indicators deteriorated. The BT rates were as high as 78.6% (canine) and 59.5% (rat). Treatment with Chinese medicine markedly improved gut barrier function and reduced BT rate (32.1% - 37.0%). Having similar purgative response as Chinese medicine, MgSO4 did not show any beneficial effect on gut barrier protection and the incidence of BT was not reduced. In bifidobacterium mixture and SDD group, flora balance was preserved well and similar results as Chinese medicine were obtained. The BT rates of these two groups decreased to 33.9% and 33.3%, respectively. Somatostatin markedly blunted pancreatic tissue injury and ameliorated gut barrier damage during early phase of ANP. All intervention measures except for purgation also decreased mortalities (14.3% - 35.3%, compared with 58.8% of the no treatment group). CONCLUSIONS: Treatment with Chinese medicine, bifidobacterium mixture, SDD and somatostatin attenuated gut barrier damage and BT after ANP, and could be used to prevent secondary infection after ANP, but purgation alone is not effective.

Animals↗

PubMed2000

[The effect of adrenalectomy on morphine dependence behavior of rats with forced-swimming].

In this study, the possible action of stress and glucocorticoids on the drug-seeking behavior was explored in the rats with morphine dependence. Forty rats were assigned into four groups randomly and received different treatments as follows: adrenalectomy; adrenalectomy + hydrocortisone 20 mg/kg S.C.; adrenalectomy + hydrocortisone 40 mg/kg S.C.; saline S.C.; The effect of adrenalectomy and hydrocortisone on morphine conditioned place preference (CPP) in rats with forced-swimming was observed. The results showed: 1. there was no significant difference for the adrenalectomized rats' spending time between the morphine and saline-paired side during the test session (t = 1.84, P > 0.05), but obvious differences of time spent between the morphine and saline-paired side were observed among the other three groups (P < 0.05 or P < 0.01); 2. the time spent in the morphine-paired side of the adrenalectomized group significantly decreased, compared with those of the other three groups, and there was no difference for the time spent in the morphine-paired side among the groups of adrenalectomy + hydrocortisone 20 mg/kg S.C., adrenalectomy + hydrocortisone 40 mg/kg S.C., and saline S.C.. These data suggested that adrenalectomy could decrease the facilitation action of morphine CPP induced by forced swimming in rats, whereas administration with glucocorticoids should reverse this effect.

Adrenalectomy↗

PubMed2000

[The different effects of CaM inhibitors of phenothiazines on the proliferation of Saccharomyces cerevisiae and Schizosaccharomyces pombe].

Low concentration of phenothiazines apparently stimulated the proliferation of S. pombe, the cell density incubated for 54 hours by preincubating the cells with 20 mumol/L trifluoperazine (TFP) in the EMM-Ca medium was two times more than the control. The stimulation was more obvious with lowing the concentration of calcium in the culture medium, TFP cooperated and complemented with calcium in stimulating the proliferation of S. pombe. When the original inoculated cell density was 5 x 10(6) cells/ml or during the logarithm period of growth curve, the proliferation of S. pombe wasn't affected by the low concentration of TFP. While when the concentration of TFP was increased to 100 mumol/L, the promotion effect of TFP on proliferation of S. pombe declined obviously and the proliferation of S. pombe was inhibited completely when TFP up to 200 mumol/L. The cell proliferation also could be inhibited by CaM antagonist W7 and W7-agarose, the inhibition was increased with increasing the concentration of antagonist. On the other hand, 20 mumol/L TFP used by the same method as above arrested the cell division cycle of Saccharomyces cerevisiae at a single G2 + M nuclei stage, the cells was penetrated easily by TFP, the fluorescence in cells was very obvious when TFP was 20 mumol/L, but it was difficult to penetrat by TFP in the cells of S. pombe and the Ca2+ influx of S. pombe could be induced rapidly by 20 mumol/L TFP. In this article, the cause of different effects of TFP on cell proliferation of S. pombe and S. cerevisiae was discussed, it was due to the difference of penetration of TFP and stimulation by calcium in the two kinds of cells.

Calcium↗

PubMed2000

[Audiological findings and mitochondrial DNA mutation in a large family with matrilineal sensorineural hearing loss].

OBJECTIVE: To explore audiological features of matrilineal non-syndromic deafness and its molecular mechanism. METHODS: A large family with 41 members having inherited deafness was studied. Complete history and the data of general and otolaryngological examinations were collected. All subjects were screened for mitochondrial DNA A1555G mutation by molecular analysis. Audiological evaluation included puretone audiometry, auditory brainstem responses and transiently evoked otoacoustic emissions. RESULTS: All subjects were in good health generally. Molecular analysis showed that all maternal relatives with or without hearing loss harbored the A1555G mitochondrial mutation. No mutation was found among spouses and paternal relatives. Audiological results showed notable symmetric bilateral sensorineural hearing loss in 17 of 20 maternal relatives, in which 5 cases had a progressive hearing loss in the recent 11 years. The age of appearance of hearing loss ranged from 1 to 50 years. CONCLUSION: All hearing-impaired subjects of this family had late-onset sensorineural hearing loss. Most of which were progressive. The A1555G mitochondrial mutation in the 12S rRNA gene is responsible for the disorder. Other factors, such as nuclear genes or environmental determinants, may influence the clinical expression of mutant mtDNA.

Acoustic Impedance Tests↗

PubMed2000

[An experiment of gut barrier function damage following multiple firearm injuries in a porcine model].

OBJECTIVE: To study the characteristics and pathogenesis of gut barrier damage following multiple firearm injuries in porcine models. METHODS: 24 small pigs were divided into 4 groups (n = 6 each): control group(group C), group H (gunshot-induced tangential fracture of parietal bone), group L (gunshot-induced comminuted fracture of bilateral femora) and group M (combined group H + L). Gastric intramucosal pH (pHi), plasma endotoxin in portal vein, and plasma D-lactic acid were measured and blood samples were cultured at different time intervals after trauma. The animals were sacrificed at 72 h and ileum tissues were harvested for pathological examination and DAO measurement. RESULT: In group M at 72 h, pHi was significantly lower than that of group H and L(P < 0.01), plasma endotoxin level was significantly higher than that of group H(P < 0.01) and group L(P < 0.05), D-lactic acid level was significantly higher than that of group H(P < 0.01), and bacterial isolation rate in blood was much higher than that of group H and L(P < 0.05). Necrosis and peeling off were revealed at ileum villus top in all trauma groups, especially in group M, in which ileum DAO content also declined in most cases. CONCLUSION: Multiple traumas were prone to gastrointestinal ischemia even when hemorrhagic shock did not take place. The damage of gut barriers in multiple traumas were more severe than that in one-site trauma, which enhanced the incidence of intestinal endotoxin and bacterial translocation and trend to cause enterogenous infection.

Amine Oxidase (Copper-Containing)↗

PubMed2000

[Spectra on the influence of grinding to kaolinite structure].

Kaolinite clay is one of important ceramic materials. In this paper, the kaolinite clay was ground by dry grinding and wet grinding. The powder samples ground for different time were measured by FTIR spectra and XRD, and the changing rules of the IR absorption frequency and XRD were respectively discussed. The results indicated that wet grinding was superior to dry one. Kaolinite structure began to change while it was ground by dry mothed for 12 h, and the layer structure was destroyed completely till 42 h. However, the day structure didn't change much until 54 h at the condition of wet grinding. The mechanism of the kaolinite structure changed were also discussed in detail.

English Abstract↗

PubMed1999

Protection from cell death by mcl-1 is mediated by membrane hyperpolarization induced by K(+) channel activation.

Mcl-1, a member of the Bcl-2 family, has been identified as an inhibitor of apoptosis induced by anticancer agents and radiation in myeloblastic leukemia cells. The molecular mechanism underlying this phenomenon, however, is not yet understood. In the present study, we report that hyperpolarization of the membrane potential is required for prevention of mcl-1 mediated cell death in murine myeloblastic FDC-P1 cells. In cells transfected with mcl-1, the membrane potential, measured by the whole-cell patch clamp, was hyperpolarized more than -30 mV compared with control cells. The membrane potential was repolarized by increased extracellular K(+) concentration (56 mV per 10-fold change in K(+) concentration). Using the cell-attached patch-clamp technique, K(+) channel activity was 1.7 times higher in mcl-1 transfected cells (NP(o) = 22.7 +/- 3. 3%) than control cells (NP(o) = 13.2 +/- 1.9%). Viabilities of control and mcl-1 transfected cells after treatment with the cytotoxin etoposide (20 microgram/ml), were 37.9 +/- 3.9% and 78.2 +/- 2.0%, respectively. Suppression of K(+) channel activity by 4-aminopyridine (4-AP) before etoposide treatment significantly reduced the viability of mcl-1 transfected cells to 49.0 +/- 4.6%. These results indicate that as part of the prevention of cell death, mcl-1 causes a hyperpolarization of membrane potential through activation of K(+) channel activity.

4-Aminopyridine↗

PubMed1999

Cyclosporine A inhibits the expression of costimulatory molecules on in vitro-generated dendritic cells: association with reduced nuclear translocation of nuclear factor kappa B.

BACKGROUND: The maturation of dendritic cells (DC) is influenced by various factors, in particular cytokine-mediated signaling events. These include modulation of the activation of nuclear factor kappa B (NF-kappaB), which controls the transcription of genes encoding major histocompatibility complex (MHC) antigens, and costimulatory/accessory molecules for T-cell activation. Here, we investigated the influence of cyclosporine A (CsA) on the in vitro maturation of DC, and on the nuclear translocation and DNA binding of NF-kappaB. METHODS: DC progenitors were propagated from mouse bone marrow in granulocyte-macrophage colony-stimulating factor (GM-CSF) or in GM-CSF plus either transforming growth factor (TGF)-beta or interleukin (IL)-4, in the presence or absence of CsA (1 microg/ml). After 5 days of culture, cell surface expression of MHC class I/II, CD40, CD80, and CD86 was analyzed by flow cytometry, and nuclear NF-kappaB proteins by electrophoretic mobility shift, antibody supershift, and Western blot assays. The antigen-presenting function of DC was determined in one-way mixed leukocyte reactions. RESULTS: Exposure of replicating DC progenitors propagated in GM-CSF or GM-CSF+TGF-beta to CsA reduced costimulatory molecule expression, without affecting MHC antigen expression. Nuclear extracts from the CsA-treated DC revealed a decrease in nuclear translocation of NF-kappaB (p50). Mixed leukocyte reaction data were consistent with the flow cytometry and gel shift assay results, and showed reduced allostimulatory ability of the CsA-treated cells compared with untreated controls. Addition of IL-4 from the start of DC cultures conferred resistance to CsA-induced inhibition of NF-kappaB nuclear translocation and DC maturation. CONCLUSIONS: CsA differentially inhibits the expression of key cell surface costimulatory molecules by in vitro-generated DC. This effect can be overcome, at least in part, by IL-4 and augmented by TGF-beta. The inhibition is linked to a decrease in nuclear translocation/DNA binding of NF-kappaB. Thus, CsA can alter the antigen-presenting function of DC for T-cell activation.

Animals↗

PubMed1999

Differential effects of exogenous interleukin-10 on cardiac allograft survival: inhibition of rejection by recipient pretreatment reflects impaired host accessory cell function.

BACKGROUND: There have been conflicting reports of the influence of exogenous mammalian interleukin (IL)-10 on immune reactivity. These findings may reflect the pleiotropic effects of IL-10 on the functions of antigen-presenting cells and immune effector cells. The purpose of this study was to extend observations of the influence of the cytokine on organ allograft survival and to investigate its effects on the function of accessory and immune effector cells in a mouse cardiac transplant model. METHODS: C3H (H2k) recipients of heterotopic vascularized B10 (H-2b) heart allografts were treated with recombinant (r) mouse IL-10 over a wide range of doses (0.2-200 microg/day), either before the transplant (days -3, -2, -1), peri-operatively (days -1, 0, 1), or after the transplant (days 0-6). Anti-donor cytotoxic T lymphocyte activity of host spleen and graft-infiltrating cells, and circulating complement-dependent cytotoxic antibody titers were determined by isotope release assays. Mixed leukocyte reactions were used to determine the influence of IL-10 on the function of antigen-presenting cells and allogeneic responder T cells. RESULTS: Recipient pre-transplant administration of IL-10 (days -3, -2, -1) prolonged graft survival at all doses tested. Donor pretreatment with IL-10 (25 microg/day; days -3, -2, -1) was also effective, but less. A pre-transplant or perioperative course of IL-10, however, did not significantly affect the immunosuppressive action of tacrolimus given on days 0-6. If given only after the transplant, IL-10 either had no effect on graft survival or (at high dosage) accelerated rejection and prevented the immunosuppressive effect of cyclosporine. Pretransplant treatment of graft recipients with IL-10 reduced splenic anti-donor cytotoxic T lymphocyte activity and the incidence of graft-infiltrating CD8+ cells. There was no significant effect on circulating alloantibody titers. MLR assays revealed that preincubation of responder cells, but not stimulator spleen cells with IL-10, inhibited T cell proliferation, whereas addition of IL-10 after the start of culture modestly enhanced proliferation. Preincubation of purified T responders with IL-10 showed no inhibitory effect. CONCLUSION: The modest and opposing effects of exogenous IL-10 on organ allograft survival are dependent on timing and dosage. Recipient pretreatment prolongs graft survival. This finding, together with the MLR results, suggest that IL-10 inhibits the function of host immune accessory cells and that the direct pathway of alloantigen presentation may be less susceptible to inhibition by IL-10.

Animals↗

PubMed1999

Retroviral-mediated gene transduction of c-kit into single hematopoietic progenitor cells from cord blood enhances erythroid colony formation and decreases sensitivity to inhibition by tumor necrosis factor-alpha and transforming growth factor-beta1.

The c-kit receptor and its ligand, steel factor (SLF), are critical for optimal hematopoiesis. We evaluated effects of transducing cord blood (CB) progenitor cells with a retrovirus encoding human c-kit cDNA. CD34(+) cells were sorted as a population or as 1 cell/well for cells expressing high levels of CD34 and different levels of c-kit (++,+,Lo/-), transduced and then cultured in the presence of granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), IL-6, erythropoietin (Epo) +/- SLF in the absence of serum. At a single-cell level, transduction with c-kit, but not with control (neo only), virus significantly increased colony formation, especially by erythroid and multipotential progenitors. The enhancing effect of c-kit transduction was inversely correlated with expression of c-kit protein before transduction. The greatest enhancing effects were noted in CD34KitLo+/- cells transduced with c-kit. The stimulating effect was apparent even in the absence of exogenously added SLF, but in the presence of GM-CSF, IL-3, IL-6, and Epo. Enzyme-linked immunosorbent assay (ELISA) of SLF protein, reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of SLF mRNA expression in CD34+ cells, and use of neutralizing antibodies to SLF and/or c-kit suggested the presence of endogenous, although probably very low level, expression of SLF by these progenitor cells. Transduction of c-kit significantly decreased sensitivity of progenitor cells to the inhibitory effects of transforming growth factor-beta1 and tumor necrosis factor-alpha. c-kit-transduced cells had increased expression of c-kit protein and decreased spontaneous or cytokine-induced apoptosis. Our results suggest that transduced c-kit into selected progenitor cells can enhance proliferation and decrease apoptosis and that endogenous SLF may mediate this effect.

Antigens, CD↗

PubMed1999

Genomic sequence, structural organization, molecular evolution, and aberrant rearrangement of promyelocytic leukemia zinc finger gene.

The promyelocytic leukemia zinc finger gene (PLZF) is involved in chromosomal translocation t(11;17) associated with acute promyelocytic leukemia. In this work, a 201-kilobase genomic DNA region containing the entire PLZF gene was sequenced. Repeated elements account for 19.83%, and no obvious coding information other than PLZF is present over this region. PLZF contains six exons and five introns, and the exon organization corresponds well with protein domains. There are at least four alternative splicings (AS-I, -II, -III, and -IV) within exon 1. AS-I could be detected in most tissues tested whereas AS-II, -III, and -IV were present in the stomach, testis, and heart, respectively. Although splicing donor and acceptor signals at exon-intron boundaries for AS-I and exons 1-6 were classical (gt-ag), AS-II, -III, and -IV had atypical splicing sites. These alternative splicings, nevertheless, maintained the ORF and may encode isoforms with absence of important functional domains. In mRNA species without AS-I, there is a relatively long 5' UTR of 6.0 kilobases. A TATA box and several transcription factor binding sites were found in the putative promoter region upstream of the transcription start site. PLZF is a well conserved gene from Caenorhabditis elegans to human. PLZF paralogous sequences are found in human genome. The presence of two MLL/PLZF-like alignments on human chromosome 11q23 and 19 suggests a syntenic replication during evolution. The chromosomal breakpoints and joining sites in the index acute promyelocytic leukemia case with t(11;17) also were characterized, which suggests the involvement of DNA damage-repair mechanism.

Alternative Splicing↗

PubMed1999

Increased apoptosis of immunoreactive host cells and augmented donor leukocyte chimerism, not sustained inhibition of B7 molecule expression are associated with prolonged cardiac allograft survival in mice preconditioned with immature donor dendritic cells plus anti-CD40L mAb.

BACKGROUND: We previously reported the association among donor leukocyte chimerism, apoptosis of presumedly IL-2-deficient graft-infiltrating host cells, and the spontaneous donor-specific tolerance induced by liver but not heart allografts in mice. Survival of the rejection-prone heart allografts in the same strain combination is modestly prolonged by the pretransplant infusion of immature, costimulatory molecule-(CM) deficient donor dendritic cells (DC), an effect that is markedly potentiated by concomitant CM blockade with anti-CD40L (CD154) monoclonal antibody (mAb). We investigated whether the long survival of the heart allografts in the pretreated mice was associated with donor leukocyte chimerism and apoptosis of graft-infiltrating cells, if these end points were similar to those in the spontaneously tolerant liver transplant model, and whether the pretreatment effect was dependent on sustained inhibition of CM expression of the infused immature donor DC. In addition, apoptosis was assessed in the host spleen and lymph nodes, a critical determination not reported in previous studies of either spontaneous or "treatment-aided" organ tolerance models. METHODS: Seven days before transplantation of hearts from B10 (H-2b) donors, 2x10(6) donor-derived immature DC were infused i.v. into C3H (H-2k) recipient mice with or without a concomitant i.p. injection of anti-CD40L mAb. Donor cells were detected posttransplantation by immunohistochemical staining for major histocompatibility complex class II (I-Ab) in the cells of recipient lymphoid tissue. CM expression was determined by two-color labeling. Host responses to donor alloantigen were quantified by mixed leukocyte reaction, and cytotoxic T lymphocyte (CTL) assays. Apoptotic death in graft-infiltrating cells and in areas of T-dependent lymphoid tissue was visualized by terminal deoxynucleotidyltransferase-catalyzed dUTP-digoxigenin nick-end labeling and quantitative spectrofluorometry. Interleukin-2 production and localization were estimated by immunohistochemistry. RESULTS: Compared with control heart transplantation or heart transplantation after only DC administration, concomitant pretreatment with immature donor DC and anti-CD40L mAb caused sustained elevation of donor (I-Ab+) cells (microchimerism) in the spleen including T cell areas. More than 80% of the I-Ab+ cells in combined treatment animals also were CD86+, reflecting failure of the mAb to inhibit CD40/ CD80/CD86 up-regulation on immature DC in vitro after their interaction with host T cells. Donor-specific CTL activity in graft-infiltrating cells and spleen cell populations of these animals was present on day 8, but decreased strikingly to normal control levels by day 14. The decrease was associated with enhanced apoptosis of graft-infiltrating cells and of cells in the spleen where interleukin-2 production was inhibited. The highest levels of splenic microchimerism were found in mice with long surviving grafts (>100 days). In contrast, CTL activity was persistently elevated in control heart graft recipients with comparatively low levels of apoptotic activity and high levels of interleukin-2. CONCLUSION: The donor-specific acceptance of rejection-prone heart allografts by recipients pretreated with immature donor DC and anti-CD40L mAb is not dependent on sustained inhibition of donor DC CM (CD86) expression. Instead, the pretreatment facilitates a tolerogenic cascade similar to that in spontaneously tolerant liver recipients that involves: (1) chimerism-driven immune activation, succeeded by deletion of host immune responder cells by apoptosis in the spleen and allograft that is linked to interleukin-2 deficiency in both locations and (2) persistence of comparatively large numbers of donor-derived leukocytes. These tolerogenic mechanisms are thought to be generic, explaining the tolerance induced by allografts spontaneously, or with the aid of various kinds of immunosuppression.

Animals↗

PubMed1999

Modulation of marrow stromal cell function using poly(D,L-lactic acid)-block-poly(ethylene glycol)-monomethyl ether surfaces.

The adhesion of marrow stromal osteoblasts and the adsorption of fetal bovine serum (FBS) proteins to end-capped poly(D,L-lactic acid) 50:50 (PLA50) of molecular weight 17,000 (PLA5017), non-end-capped PLA50 of molecular weight 11,000 (PLA5011h), and a diblock copolymer made of poly(ethylene glycol)-monomethyl ether of molecular weight 5,000 and PLA50 of molecular weight 20,000 (Me. PEG5-PLA20) were investigated. Cell attachment and proliferation on both PLA50 polymers were equally good. The block copolymer did not allow the proliferation of cells. However, the attached cells were highly differentiated and metabolically active in contrast to the cells on PLA50. Moreover, surface analysis studies using electron spectroscopy revealed that FBS proteins adsorbed well from aqueous solutions to the PLA50 surfaces while they adsorbed substantially less to the block copolymer. These results suggest that Me.PEG-PLA block copolymers may be used to regulate protein adsorption and, therefore, cell adhesion by varying the block composition of the copolymer.

Adsorption↗

PubMed1999

Dendritic cells as regulators of immune reactivity: implications for transplantation.

Dendritic cells are now regarded not only as the initiators but also as regulators of immune responses. They are potentially powerful tools for the therapeutic manipulation of immune reactivity in cancer, infectious disease, and allograft rejection. We provide a brief overview of the properties of dendritic cells, with emphasis on recently acquired information, then focus attention on their capacity to modulate immune reactivity, and its relevance to transplantation.

Animals↗

PubMed1999

Targeted mutagenesis of the POU-domain gene Brn4/Pou3f4 causes developmental defects in the inner ear.

Targeted mutagenesis in mice demonstrates that the POU-domain gene Brn4/Pou3f4 plays a crucial role in the patterning of the mesenchymal compartment of the inner ear. Brn4 is expressed extensively throughout the condensing mesenchyme of the developing inner ear. Mutant animals displayed behavioral anomalies that resulted from functional deficits in both the auditory and vestibular systems, including vertical head bobbing, changes in gait, and hearing loss. Anatomical analyses of the temporal bone, which is derived in part from the otic mesenchyme, demonstrated several dysplastic features in the mutant animals, including enlargement of the internal auditory meatus. Many phenotypic features of the mutant animals resulted from the reduction or thinning of the bony compartment of the inner ear. Histological analyses demonstrated a hypoplasia of those regions of the cochlea derived from otic mesenchyme, including the spiral limbus, the scala tympani, and strial fibrocytes. Interestingly, we observed a reduction in the coiling of the cochlea, which suggests that Brn-4 plays a role in the epithelial-mesenchymal communication necessary for the cochlear anlage to develop correctly. Finally, the stapes demonstrated several malformations, including changes in the size and morphology of its footplate. Because the stapes anlage does not express the Brn4 gene, stapes malformations suggest that the Brn4 gene also plays a role in mesenchymal-mesenchymal signaling. On the basis of these data, we suggest that Brn-4 enhances the survival of mesodermal cells during the mesenchymal remodeling that forms the mature bony labyrinth and regulates inductive signaling mechanisms in the otic mesenchyme.

Acoustic Stimulation↗

PubMed1999

A requirement for K+-channel activity in growth factor-mediated extracellular signal-regulated kinase activation in human myeloblastic leukemia ML-1 cells.

Voltage-gated K+ channels have been shown to be required for proliferation of various types of cells. Much evidence indicates that K+-channel activity is required for G1 progression of the cell cycle in different cell backgrounds, suggesting that K+-channel activity is required for early-stage cell proliferation in these cells. However, little is known about the molecular mechanisms that underlie this phenomenon. We have shown in human myeloblastic leukemia ML-1 cells that K+ channels are activated by epidermal growth factor (EGF), whereas serum starvation deprivation suppressed their activity. In addition, voltage-gated K+ channels are required for G1/S-phase transition of the cell cycle. We report here that suppression of K+ channels prevented the activation of extracellular signal-regulated protein kinase 2 (ERK-2) in response to EGF and serum. However, blockade of K+ channels did not prevent ERK-2 activation induced by 12-O-tetradecanoyl-phorbol 13-acetate (TPA). Elimination of extracellular Ca2+ did not alter either ERK-2 activation or the effect of K+-channel blockade on ERK-2 activation. Our data demonstrate that the K+ channel is a part of the EGF-mediated mitogenic signal-transduction process and is required for initiation of the EGF-mediated mitogen-activated protein kinase (MAPK) pathways. Our findings may thus explain why an increase in K+-channel activity is associated with cell proliferation in many types of cells, including ML-1 cells.

Calcium-Calmodulin-Dependent Protein Kinases↗

PubMed1999

A novel polymorphism in the MCP-1 gene regulatory region that influences MCP-1 expression.

Two novel polymorphisms in the distal regulatory region of the MCP-1 gene were identified by directly sequencing PCR amplified genomic DNA. These polymorphisms are located at positions -2518 (G or A) and -2076 (A or T) relative to the major transcriptional start site of the gene. To examine the effect of these polymorphisms on MCP-1 transcription, polymorphic variants of the MCP-1 distal regulatory region were placed upstream of a luciferase reporter gene and transfected into A172 cells. IL-1beta-induced luciferase activity was significantly greater from cells transfected with constructs containing G at position -2518. The polymorphism at -2076 did not affect MCP-1 transcription. IL-1beta-treated peripheral blood mononuclear cells from individuals heterozygous or homozygous for G at -2518 produced more MCP-1 than cells from individuals homozygous for A at -2518. These data identify a polymorphism in the MCP-1 distal regulatory region that affects the level of MCP-1 expression in response to an inflammatory stimulus.

Alleles↗