Biomedical subjects

X Cao

Publications and source records attributed to X Cao.

At least 55 records · Page 3Linked to original sources

PubMed2001

Interleukin 18 transfection enhances antitumor immunity induced by dendritic cell-tumor cell conjugates.

Dendritic cell (DC)-based tumor vaccine represents a promising approach to the immunotherapy of malignant tumors. We prepared a novel type of DC-based vaccine, stable conjugates of DCs and EL4 cells transduced with cDNA of OVA (E.G7). Immunization with DC-E.G7 conjugates led to generation of T helper (Th) 1 cytokine-producing cells, antigen-specific CD8(+) T cells, and strong antitumor immunity that is dependent on both CD4(+) T cells and CD8(+) T cells. To further increase the potency of the vaccine, interleukin 18-transfected DCs were used to prepare the IL18DC-E.G7 conjugates. Immunization with such conjugates significantly increased the production of Th1 cytokine-producing cells and the number of antigen-specific CD8(+) T cells, as well as stronger antitumor immunity. Furthermore, the increased Th1 cytokine production and stronger antitumor effect were not observed in mice depleted of IFN-gamma. These data indicated that DC-tumor cell conjugates are a potent tumor vaccine. Interleukin 18 can be administrated using gene-transfected cells and enhances antitumor immunity, which is mainly mediated by IFN-gamma.

Animals↗

PubMed2001

Identification and characterization of DPZF, a novel human BTB/POZ zinc finger protein sharing homology to BCL-6.

The C2H2 zinc finger protein family is one of the largest families of transcription factors. We identified a novel BTB/POZ zinc finger gene from human dendritic cells (DC), which encodes a 733-residue protein with a BTB/POZ domain at the N-terminal and 4 C2H2 zinc fingers at C-terminal. It was designated dendritic cell-derived BTB/POZ zinc finger (DPZF). DPZF protein shares closest homology to BCL-6, with the highest homology present in the BTB/POZ and zinc finger domains. Like BCL-6, DPZF gene is localized on chromosome 3. It is widely expressed in hematopoietic tissues, including DC, monocytes, B cells, and T cells. DPZF protein expression is detectable in lymphoid neoplasm with a molecular mass of 100 kD, especially in B lymphoma. These indicate that DPZF may be a transcription factor closely related to BCL-6, and may be involved in hematopoiesis, oncogenesis, and immune responses.

Amino Acid Sequence↗

PubMed2001

Dexamethasone inhibits the antigen presentation of dendritic cells in MHC class II pathway.

Glucocorticoids (GC) are physiological inhibitors of inflammatory responses and are widely used as anti-inflammatory and immunosuppressive agents in treatment of many autoimmune and allergic diseases. In the present study, we demonstrated that one of the mechanisms by which GC can suppress the immune responses is to inhibit the differentiation and antigen presentation of dendritic cells (DC). DC were differentiated from murine bone marrow hematopoietic progenitor cells by culture with GM-CSF and IL-4 with or without dexamethasone (Dex). Our data showed that Dex, in a dose dependent manner, down-regulated surface expression of CD86, CD40, CD54 and MHC class II molecules by DC, but the expression of MHC class I, CD80, CD95 and CD95L were not affected. In addition, Dex-treated DC showed an impaired function to activate alloreactive T cells and to secrete IL-Ibeta and IL-12p70. Moreover, Dex inhibited DC to present antigen by MHC class II pathway. However, the endocytotic activity of DC was not affected. The inhibitory effect of Dex on the expression of costimulatory molecules and the antigen-presenting capacity of DC could be blocked by the addition of RU486, a potent steroid hormone antagonist, suggesting the requirement of binding to cytosolic receptors in the above-described action of Dex. Since DC have the unique property to present antigen to responding naive T cells and are required in the induction of a primary response, the functional suppression of DC by Dex may be one of the mechanisms by which GC regulate immune responses in vivo.

Animals↗

PubMed2001

Regulation of Stat3 activation by MEK kinase 1.

Stat3 is a latent transcription factor activated by various cytokines and growth factors. Phosphorylation on Tyr-705 is a prerequisite for dimer formation, nuclear translocation, binding to its cognate DNA sequences, and regulation of the target gene transcription. Ser-727 phosphorylation of Stat3 plays an additional role in the regulation of transcription. MEK kinase 1 (MEKK1) is a mitogen-activated protein kinase (MAPK) kinase kinase (MAPKKK) that activates the c-Jun NH(2)-terminal kinase signaling pathway. Here we report that MEKK1 is involved in the regulation of Stat3 activation by growth factors. Kinase-inactive MEKK1 inhibits Stat3 phosphorylation on tyrosine and serine, and its transcriptional activity stimulated by epidermal growth factor and platelet-derived growth factor in different cell types. In contrast, active MEKK1 induces Stat3 tyrosine and serine phosphorylation leading to a functionally active Stat3 capable of binding DNA and enhancing transcription. Ser-727 is phosphorylated by MEKK1 in vitro, whereas Tyr-705 phosphorylation induced by MEKK1 involves Src and Janus kinases in vivo. These data demonstrate for the first time a novel role of MEKK1 to modulate tyrosine kinases that results in the activation of specific members of STAT family.

Acute-Phase Proteins↗

PubMed2001

Identification of a novel type I cytokine receptor CRL2 preferentially expressed by human dendritic cells and activated monocytes.

From a human dendritic cell (DC) cDNA library, we identified a novel type I cytokine receptor, designated as cytokine receptor-like molecule 2 (CRL2). CRL2 cDNA encoded a 371-residue type I transmembrane protein with an extracellular domain of 210 residues and an intracellular domain of 119 residues. Its extracellular domain contains conserved cysteine residues and WAS-like motif in place of the hallmark of WSXWS motif present in other type I cytokine receptors. The intracellular domain contained a membrane-proximal "box 1" motif and conserved tyrosine residue potentially as a binding site for signal transducing molecules. CRL2 protein shares significant homology with common cytokine receptor (gammac) and interleukin-13 receptor alpha1 chain. Northern blot analysis showed that CRL2 was restrictedly expressed by spleen and peripheral blood leukocytes, and abundantly expressed by HL-60 cells. RT-PCR analysis demonstrated that CRL2 was preferentially expressed by DC and monocytes. Interestingly, CRL2 expression was up-regulated when monocytes were activated by LPS. These indicate that CRL2 may be involved in the biological functions of DC and monocytes. The Ba/F3 transfectants of CRL2 was retrovirally established with the expressed FLAG-tagged CRL2 in the size of approximately 48 kD, which could be efficiently immunoprecipitated. We also prepared a CRL2Ig fusion protein. The identification of its ligand and involvement of signal transduction will help to elucidate its potential function.

Amino Acid Sequence↗

PubMed2001

SeO(2) induces apoptosis with down-regulation of Bcl-2 and up-regulation of P53 expression in both immortal human hepatic cell line and hepatoma cell line.

An immortal human hepatic cell line HL-7702 and human hepatoma cell line SMMC-7721 were treated with 3-30 microM SeO(2). SeO(2) at 30 microM markedly inhibited cell proliferation and viability, and prompted apoptosis of both normal hepatic and hepatoma cells after 48h treatment. SeO(2) could also down-regulate the Bcl-2 level, greatly in HL-7702 and slightly in SMMC-7721 cells, but up-regulate wild type P53 level a little in HL-7702 and significantly in SMMC-7721 cells. The Bcl-2/P53 value was closely correlated with the apoptotic rate as well as SeO(2) concentrations.

Apoptosis↗

PubMed2001

Cloning and characterization of a novel ITIM containing lectin-like immunoreceptor LLIR and its two transmembrane region deletion variants.

A novel full-length cDNA was cloned from human dendritic cells (DC) by subtractive cloning and RACE. The deduced protein is a type II lectin-like membrane protein that contains an ITIM proximal to N terminal and is designated as lectin-like immunoreceptor (LLIR). The gene of LLIR is located in a region of chromosomal 12p13 and shows highest homologous with ASGPR. Two alternatively spliced transmembraneless variants of LLIR were identified by RT-PCR and named as LLIRv1 and LLIRv2. RT-PCR and immunoblotting analysis revealed that LLIR was expressed with much higher level in immature DC than in mature DC. The ITIM in LLIR was demonstrated to bind SHP-1 in HL-60 cell after the tyrosine had been phosphorylated. In addition, the mRNA expression level of LLIRv2 was raised when leukemia cells were induced to differentiate by PMA.

3T3 Cells↗

PubMed2001

Hoxa-9 represses transforming growth factor-beta-induced osteopontin gene transcription.

Smad2 and Smad3 are downstream transforming growth factor-beta (TGF-beta) signaling molecules. Upon phosphorylation by its type I receptor, Smad2 or Smad3 forms a complex with Smad4 and translocates to the nucleus where the complex activates target gene transcription. In the present study, we report that Smad3 binds directly to the osteopontin (OPN) promoter and that Smad4 interacts with the Hox protein and displaces it from its cognate DNA binding site in response to TGF-beta stimulation. In gel shift assays, the glutathione S-transferase-Smad3 fusion protein was found to bind to a 50-base pair DNA element (-179 to -229) from the OPN promoter. Also, we found that both Hoxc-8 and Hoxa-9 bound to a Hox binding site adjacent to Smad3 binding sequence. Interestingly, Smad4, the common partner for both bone morphogenic protein and TGF-beta signaling pathways, inhibited the binding of Hox protein to DNA. FLAG-tagged Smad4 coimmunoprecipitated with HA-tagged Hoxa-9 from cotransfected COS-1 cells, demonstrating an interaction between Smad4 and Hoxa-9. Transfection studies showed that Hoxa-9 is a strong transcriptional repressor; it suppresses the transcription of the luciferase reporter gene driven by a 124-base pair OPN promoter fragment containing both Smad3 and Hox binding sites. Taken together, these data demonstrate a unique TGF-beta-induced transcription mechanism. Smad3 and Smad4 exhibit different functions in activation of OPN transcription. Smad3 binds directly to the OPN promoter as a sequence-specific activator, and Smad4 displaces the transcription repressor, Hoxa-9, by formation of Smad4/Hox complex as part of the transcription mechanism in response to TGF-beta stimulation.

Animals↗

PubMed2001

Transcriptional mechanisms of bone morphogenetic protein-induced osteoprotegrin gene expression.

Osteoprotegerin (OPG), an osteoblast-secreted decoy receptor, specifically binds to osteoclast differentiation factor and inhibits osteoclast maturation. Members of the transforming growth factor-beta superfamily including bone morphogenetic proteins (BMPs) stimulate OPG mRNA expression. In this study, we have characterized the transcription mechanism of BMP-induced OPG gene expression. Transfection of Smad1 and a constitutively active BMP type IA receptor ALK3 (Q233) stimulated the OPG promoter. Deletion analysis of the OPG promoter identified two Hoxc-8 binding sites that respond to BMP stimulation. Glutathione S-transferase-Hoxc-8 protein binds to these two Hox sites specifically. Consistent with the transfection results of the native promoter, ALK3 or Smad1 linker region, which interacts with Hoxc-8, stimulated the activation of the reporter construct with the two Hox sites. Overexpression of Hoxc-8 inhibited the induced promoter activity. When the two Hox binding sites were mutated, ALK3 or Smad1 linker region no longer activated the transcription. Importantly, Smad1 linker region induced both OPG promoter activity and endogenous OPG protein expression in 2T3 osteoblastic cells. The medium from cells transfected with Smad1 linker region expression plasmid effectively inhibited osteoclastogenesis. Collectively, our data indicate that Hox sites mediate both OPG promoter construct activity and endogenous OPG gene expression in response to BMP stimulation.

Animals↗

PubMed2001

Depletion of taurine in experimental diabetic neuropathy: implications for nerve metabolic, vascular, and functional deficits.

In diabetes, increased oxidative stress, disruption of signal transduction pathways, and endothelial dysfunction have been critically implicated in the pathogenesis of experimental diabetic neuropathy (EDN). The development of nerve conduction slowing in diabetes is accompanied by depletion of the beta-amino acid taurine. Since taurine functions as an antioxidant, calcium modulator, and vasodilator, taurine depletion may provide a pathogenetic link between nerve metabolic, vascular, and functional deficits complicating diabetes. The mechanism(s) of nerve taurine depletion, the localization of critical taurine deficits, and its pathophysiological significance in EDN are however unknown. This study explored the pathophysiological effects of selective nerve taurine replacement in streptozotocin-diabetic (STZ-D) rats. A polyclonal human taurine transporter (TT) antibody was also generated in order to determine potential loci of critical taurine depletion. Two weeks of STZ-D reduced sciatic motor nerve conduction velocity (NCV) by 23% (P < 0.01), decreased composite nerve blood flow by 38% (P < 0.01), and reduced nerve taurine content by 29% (P < 0.05). In STZ-D rats, a 1% taurine diet corrected nerve taurine depletion, prevented motor NCV slowing, and partially attenuated composite nerve blood flow deficits. After 6 weeks of STZ-D, a 1% taurine diet ameliorated motor NCV slowing and endoneurial nutritive blood flow deficits, prevented digital sensory NCV slowing, and reduced ouabain-sensitive nerve (Na,K)-ATPase activity. Immunohistochemical studies localized taurine and the TT to the vascular endothelium and Schwann cells of the sciatic nerve. In conclusion, taurine depletion in the vascular endothelium and Schwann cells of the sciatic nerve may contribute to the neurovascular and metabolic deficits in EDN.

Animals↗

PubMed2001

Experimental study of serum substantia nigra neuron autoantibody and its effect in Parkinson disease patients.

To investigate the serum substantia nigra neuron autoantibody and its effect in the patients with Parkinson disease (PD), substantia nigra slices and a rat model of injection of serum from PD patients in unilateral side substantia nigra were applied. The results showed that the positive rate of substantia nigra neuron autoantibody in PD patients was significantly higher than in the healthy control group (36.67% vs 6.67%, P < 0.01), but no significant difference was found between PD group and myasthenia gravis (MG) group (26.67%, P > 0.05). The sera from PD patients positive for substantia nigra neuron autoantibody could decrease the number of the dopaminergic neurons more seriously than those from MG and the healthy once respectively (both P < 0.01). The results suggested that the immunological mechanism might partly play a role in the development of PD.

Adult↗

PubMed2001

The potent antitumor effects of combined p16 gene and GM-CSF gene therapy through efficient induction of antitumor immunity.

PURPOSE: Tumor suppressor gene therapy and cytokine gene therapy have limited antitumor effects when used alone. Thus, in the present study, we investigated the antitumor potentials of the combined transfer of the p16 tumor suppressor gene and the murine granulocyte-macrophage colony-stimulating factor (GM-CSF) gene. METHODS: The adenovirus-harboring p16 gene (Adp16) and adenovirus-harboring GM-CSF (AdGMCSF) gene were utilized for the treatment of established tumors in vivo. The mice were inoculated s.c. with Renca renal carcinoma cells and 3 days later received an intratumoral injection of Adp16 in combination with AdGMCSF. RESULTS: The results demonstrated that tumor-bearing mice treated with Adp16 and Ad-GMCSF showed more potent inhibition of tumor growth and a prolonged survival period than mice treated with Adp16. AdGMCSF, adenovirus-expressing beta-galactosidase or PBS (P<0.01). Treatments of the mice with Adp16 alone or AdGMCSF alone also showed obvious antitumor effects as compared with those mice treated with PBS (P<0.05). After combined p16 and AdGMCSF gene therapy, the expression of H2Kd and Fas molecules on freshly isolated tumor cells increased markedly, and more CD(4)+ T cells and CD(8)+ T cells infiltrated in the tumor sites. The cytotoxicity of natural killer cells and specific cytotoxic T lymphocytes increased more significantly after the combined therapy. CONCLUSIONS: Our results demonstrated that combination p16 gene and GM-CSF gene therapy could inhibit the growth of established tumors in mice more significantly through efficient induction of antitumor immunity.

Adenoviridae↗

PubMed2001

Cloning and characterization of a novel zinc finger protein (MDZF) that is associated with monocytic differentiation of acute promyelocytic leukemia cells.

AIM: To study the characteristics of a novel zinc finger protein designated as monocytic differentiation-associated zinc finger protein (MDZF) and its role in the differentiation of leukemia cells. METHODS: The mRNA expression of MDZF in tissues and cells was analyzed by Northern blot and RT-PCR. Polyclonal antibodies against the N terminus of MDZF were used to analyze protein expression in hematopoietic cell lines and subcellular location of MDZF in promyelocytic NB4 cells. The NB4 cells treated with TPA or ATRA at different intervals were harvested and then the expression level of MDZF protein was determined by Western blot. RESULT: A full-length cDNA was successfully identified from a human monocyte-derived dendritic cell cDNA library which encodes 610 amino acids with eight C2H2 zinc finger motifs and one POZ domain. It was located on chromosome 3 according to the genome database. This novel zinc finger protein was designated as MDZF. One transcript isoform of MDZF was also cloned by RT-PCR. Northern blot showed that MDZF mRNA was restrictedly expressed in heart, skeleton muscle, kidney, liver, and placenta. MDZF protein was expressed in all hematopoietic cell lines examined. Immunoblotting and confocal analysis indicated a dominant cytoplasmic location of MDZF in NB4 cells. Furthermore, after NB4 cells were treated with TPA for 48 h and differentiated into monocytes, MDZF expression increased fivefold in the NB4 cells, but no effect was observed in NB4 cells treated with ATRA. CONCLUSION: A novel zinc finger protein MDZF was cloned. MDZF is upregulated in monocytic, but not granulocytic, differentiation of NB4 acute promyelocytic leukemia cells. MDZF may be a candidate regulator of monocytic differentiation.

Amino Acid Sequence↗

PubMed2001

Vaccination with IL-18 gene-modified, superantigen-coated tumor cells elicits potent antitumor immune response.

OBJECTIVE: To investigate the induction of antitumor immune response by vaccination with interleukin-18 (IL-18) gene-modified, C215Fab-SEA-coated tumor cells. MATERIALS: A B16-C215 cell clone stably expressing C215 antigen was established by transfecting the gene-encoding C215 antigen into B16 melanoma cells. The manipulated tumor cell vaccine was prepared with B16-C215 cells genetically modified with the IL-18 gene, coated with the fusion protein of SEA and the Fab region of C215 mAb (C215Fab-SEA) which specifically binds to the C215 antigen and then irradiated. C57BL/6 mice were vaccinated with IL-18 gene-modified, C215Fab-SEA-coated B16-C215 cells followed by tumor challenge. Tumor growth and survival time were observed. The expansion of CD4+, CD8+ cells in lymphocytes derived from draining lymph node was detected by FACS. Induction of CTL activity by vaccination was measured by 51Cr release assay. RESULTS: IL-18 gene-modified, C215Fab-SEA-coated B16-C215 cell vaccine effectively stimulated lymphocyte proliferation and CD4+, CD8+ cell expansion in vitro. It was more immunogenic than B16-C215 cells genetically modified with IL-18 gene alone or B16-C215 cells coated with C215Fab-SEA alone. Immunization of the mice with the manipulated vaccine elicited protective immunity against the following tumor challenge of parental B16-C215 and wild-type B16 cells. Significant expansion of CD4+, CD8+ T cells was observed in the draining lymph node of the immunized mice when compared with that in unvaccinated mice. Higher CTL activity was induced in vaccinated mice than that in unvaccinated mice. CONCLUSION: Vaccination with IL-18 gene-modified, C215Fab-SEA-coated tumor cells elicited potent antitumor response through induction of tumor-specific immune response.

Animals↗

PubMed2001

Topoisomerase-I- and Alu-mediated genomic deletions of the APC gene in familial adenomatous polyposis.

Germline mutation in the adenomatous polyposis coli (APC) gene results in familial adenomatous polyposis (FAP), a heritable form of colorectal cancer. We have previously reported two novel mutations that delete exons 11 and 14 of the APC gene, respectively, at the cDNA level without any splice junction defects at the genomic level. We describe here the precise breakpoints of the two mutations and the possible mechanisms leading to the genomic rearrangement. The first rearrangement is most likely a topoisomerase-I-mediated non-homologous recombination resulting in a 2-kb deletion that deletes exon 11 of the APC gene. Both 5' and 3' breakpoints have two topoisomerase I recognition sites and runs of pyrimidines within the 10-bp sequences in their vicinity. Further, the 3' breakpoint has an adenine-thymidine-rich region. This is probably the first report of a topoisomerase-I-mediated germline mutation in a tumor suppressor gene. The second rearrangement is most likely an Alu-Alu homologous recombination resulting in a 6-kb deletion encompassing exon 14. The Alu elements at the 5' and 3' breakpoints include the 26-bp core sequence thought to stimulate recombination. In both rearrangements, partial sequences from the long interspersed nuclear element family are in the vicinity of the breakpoints. Other than serving as markers for regions of DNA damage, their precise role in the recombination events, if any, is unclear. Both deletions result in truncated APC proteins missing the beta-catenin- and axin-binding domains, resulting in severe polyposis and cancer.

Adenomatous Polyposis Coli↗

PubMed2001

Effects of redox potential and pH value on the release of rare earth elements from soil.

Equilibrium release experiments were conducted under three different pH values of 3.5, 5.5 and 7.5 as well as three redox potentials of 400, 0 and -100 Mv to investigate the influence of redox potential and pH value on the La, Ce, Gd and Y release of from the simulated-REEs-accumulation (SRA) soil. Oxygen and nitrogen were allowed to flow into soil suspension to adjust redox potential to a preset value, and 1 mol/l HCl or 1 mol/l NaOH solutions were added into the soil suspension to keep pH at a preset value. Results indicated that La, Ce, Gd and Y release increased gradually with the decrease of pH value or Eh, and the influence of redox potential on Ce was more remarkable than on La, Gd and Y. At the same time. It was observed that La, Ce, Gd and Y releases were positively correlated with the release of Fe and Mn, indicating that La, Ce, Gd and Y releases might originate from dissolution of Fe-Mn oxyhydroxides under reduction and low pH conditions. Moreover, it was found that alteration of pH value and redox potential might affect the change of La, Ce, Gd and Y species in the soil. The contents of La, Ce, Gd and Y in exchangeable fraction and Fe-Mn oxide fraction in the solid phase from soil suspension separation decreased with the decline of pH value and redox potential. Multiple stepwise regression analysis showed that exchangeable fraction and Fe-Mn oxide fraction predominately contributed to the La, Ce, Gd and Y release. Low pH value and redox potential were more favorable to La, Ce, Gd and Y releases following the change of their species. The La, Ce, Gd and Y contents in exchangeable fraction and Fe-Mn oxide fraction are the main contributors to their release.

Biological Availability↗

PubMed2001

Defining the concentration gradient of nerve growth factor for guided neurite outgrowth.

The developing axon is believed to navigate towards its target tissue in response to a concentration gradient of neurotrophic factors, among other diffusible and surface-bound stimuli. However, the minimum concentration gradient required for guidance over the maximum distance is still unknown, largely because well-defined systems have not been utilized to address this question. In this study, a linear concentration gradient of nerve growth factor was achieved across a 5-mm agarose membrane that separated a nerve growth factor source compartment from a sink compartment. The concentrations in both compartments were maintained constant (and different). Both concentration and concentration gradient were well defined across the membrane, allowing us to study the relative importance of concentration gradient vs concentration for neurite guidance. The orientation of PC12 cell neurites was studied in response to a series of nerve growth factor concentration gradients in vitro. For effective guidance of PC12 cell neurite outgrowth, a minimum concentration gradient of 133ng/ml per mm was required, below which guidance was ineffective. Higher gradients were effective for guidance yet were limited by the concentration of nerve growth factor in the source compartment. At a nerve growth factor concentration of 995ng/ml, the PC12 cells' receptors were saturated, thereby limiting the maximum effective distance for guidance to less than 7.5mm in response to a diffusible nerve growth factor cue. This distance exceeds the 0.5-2mm distance observed by others for effective neurite guidance. Using this model system, we propose that the minimum concentration gradient can be defined for other cells and growth factors. Ultimately, it is anticipated that such concentration gradients could be included in a device to promote regeneration.

Animals↗

PubMed2001

The electrochemical and spectroelectrochemical behaviors of SOD at cysteine modified gold electrode.

Different promoters were used to enhance the rate of the electron transfer process of superoxide dismutase (SOD). A quasi-reversible cyclic voltammogram (CV) of SOD was observed on the cysteine-modified gold wire electrode. The coupled adsorption/desorption processes and the characteristic Raman spectra of different states of SOD molecules on the electrode surface were studied with electrochemical quartz crystal microbalance (EQCM) and in-situ Raman spectroelectrochemical techniques. The possible promotion mechanism was discussed.

Adsorption↗