Carthamus tinctorius Enhances the Antitumor Activity of Dendritic Cell Vaccines via Polarization toward Th1 Cytokines and Increase of Cytotoxic T Lymphocytes

Carthamus tinctorius (CT), also named safflower, is a traditional Chinese medicine widely used to improve blood circulation. CT also has been studied for its antitumor activity in certain cancers. To investigate the effects of CT on the dendritic cell (DC)-based vaccine in cancer treatment, cytokine secretion of mouse splenic T lymphocytes and the maturation of DCs in response to CT were analyzed. To assess the antitumor activity of CT extract on mouse CD117+ (c-kit)-derived DCs pulsed with JC mammal tumor antigens, the JC tumor was challenged by the CT-treated DC vaccine in vivo. CT stimulated IFN-γ and IL-10 secretion of splenic T lymphocytes and enhanced the maturation of DCs by enhancing immunological molecule expression. When DC vaccine was pulsed with tumor antigens along with CT extract, the levels of TNF-α and IL-1β were dramatically increased with a dose-dependent response and more immunologic and co-stimulatory molecules were expressed on the DC surface. In addition, CT-treated tumor lysate-pulsed DC vaccine reduced the tumor weight in tumor-bearing mice by 15.3% more than tumor lysate-pulsed DC vaccine without CT treatment. CT polarized cytokine secretion toward the Th1 pathway and also increased the population of cytotoxic T lymphocytes ex vivo. In conclusion, CT activates DCs might promote the recognition of antigens and facilitate antigen presentation to Th1 immune responses

Analysis of maturation-specific genes in soybean seeds: Gene structure, desiccation induction and abscisic acid responsiveness

This study describes two maturation (Mat) genes which share some structural similarities with a special group of genes, whose expression can be induced by water stress and also by exogenous abscisic acid (ABA) application. Previous studies have identified a 31-kDa maturation polypeptide (MAT) which accumulates during normal and precocious maturation of soybean (Glycine max L. Merr) seeds, and disappears rapidly during germination. The corresponding mRNA can also be induced in germinating seeds with ABA treatment, but not in normally developing seeds. By screening a 65 day-after-flower (DAF) seed cDNA library with a partial cDNA clone corresponding to the 31 kDa MAT protein, we were able to isolate and characterize two closely-related gene sequences represented by the pMat1 and pMat9 clones. Primer extension studies indicated that these two clones were full length cDNA clones. In the soybean genome, we identified at least two copies of the Mat1 genes and one Mat1-related gene, probably the Mat9 gene, in both the Williams'82 and T157 varieties. The nucleotide sequence comparison of cDNA and genomic clones showed these Mat genes do not contain any introns, which was confirmed by polymerase-chain-reaction (PCR) analysis of soybean seed mRNA and genomic DNA. The pMat 1 and pMat9 clones not only differ in size (51 nucleotides less in the coding region of pMat9 than in the coding region of pMat1), but the corresponding genes also differ in their expression patterns during normal and precocious maturation stages, and also by differential response to exogenous abscisic acid during germination. The protein sequence analysis showed that MAT proteins contain a highly conserved C-terminal region, which is also present in rice RAB (ABA-responsive) proteins and also in the cotton LEA and barley dehydrin proteins. The N-termini of MAT proteins also contains a short 7-amino acid stretch which might be related to the tissue-specificity expression or activity of these proteins. Possible roles of MAT proteins in seed maturation processes are discussed.U of I OnlyETDs are only available to UIUC Users without author permissio

Dipeptidyl Peptidase-Iv Inhibitors: An Evolving Treatment for Type 2 Diabetes from the Incretin Concept

Glucose-dependent insulinotropic polypeptide (GIP) and glucagon-like peptide (GLP-1) are the 2 major incretin hormones released after meals to enhance glucose-stimulated insulin secretion. In patients with type 2 diabetes, a loss of activity of GIP for insulinotropic function and a reduced secretion of GLP-1 exist in response to oral glucose while GLP-1 action is preserved. GLP-1 is therefore an attractive avenue for treating type 2 diabetes. Due to the short circulating half-life of GLP-1, which is degraded by dipeptidyl peptidase IV (DPP-IV), 2 approaches have been undertaken. One is to develop long-acting GLP-1 analogs, such as exendin-4 that is resistant to degradation. Here we review another approach for developing DPP-IV inhibitors. This group of potential drugs covers several major chemical classes and their derivatives, such as amino acid amide, carbocyclic, alkylamine, and heterocyclic compounds. More than 100 patents have been issued for DPP-IV inhibitors to be used either as a monotherapy or in combination with other antidiabetic agents for the treatment of type 2 diabetes, as well as metabolic syndrome, osteoporosis, and arthritis. Structure-based drug design is currently under intensive investigation for future development of more selective therapeutic agents

Specific Medicinal Plant Polysaccharides Effectively Enhance the Potency of a DC-Based Vaccine against Mouse Mammary Tumor Metastasis

<div><p>Dendritic cell (DC) vaccines are a newly emerging immunotherapeutic approach for the treatment and prevention of cancer, but major challenges still remain particularly with respect to clinical efficacy. Engineering and optimization of adjuvant formulations for DC-based vaccines is one strategy through which more efficacious treatments may be obtained. In this study, we developed a new <i>ex vivo</i> approach for DC vaccine preparation. We evaluated two highly purified mixed polysaccharide fractions from the root of <i>Astragalus membranaceus</i> and <i>Codonopsis pilosulae</i>, named Am and Cp, for their use in enhancing the efficiency of a DC-based cancer vaccine against metastasis of 4T1 mammary carcinoma in mice. Mixed lymphocyte reaction showed all Am-, Cp- and [Am+Cp]-treated DCs enhanced mouse CD4⁺ and CD8⁺ T-cell proliferation. [Am+Cp]-treated DCs exhibited the strongest anti-4T1 metastasis activity in test mice. Treatments with Am, Cp and [Am+Cp] also resulted in augmented expression of CD40, CD80 and CD86 markers in test DCs. Bioinformatics analysis of the cytokine array data from treated DCs identified that [Am+Cp] is efficacious in activation of specific immune functions via mediating the expression of cytokines/chemokines involved in the recruitment and differentiation of defined immune cells. Biochemical analysis revealed that Am and Cp are composed mainly of polysaccharides containing a high level (70–95%) glucose residues, but few or no (< 1%) mannose residues. In summary, our findings suggest that the specific plant polysaccharides Am and Cp extracted from traditional Chinese medicines can be effectively used instead of bacterial LPS as a potent adjuvant in the formulation of a DC-based vaccine for cancer immunotherapies.</p></div

Adjuvant effect of [Am+Cp] phytoextract on DC vaccine against <i>in vivo</i> metastasis of 4T1 mammary tumors.

<p>(A) Treatment schema used for the cancer vaccine experiment. (B) Percentage of whole body organs free from metastasis. (C) Survival rate of treated 4T1 tumor-bearing mice after resection of primary tumor(s). The free from tumor metastasis rate and mouse survival rate were statistically analyzed at day 51 after resection of the primary tumor. (D[a]) Metastatic pulmonary foci of 4T1 (shown as arrows) in lung were detected in 4T1 tumor-resected mice. (D[b]) Histological staining of tumor-bearing lung tissue sections with H&E. Arrows indicate metastatic tumors (MT: metastatic tumors; A: alveolus). (E) Bioluminescence imaging of the whole mouse body obtained with an in vivo imaging system (IVIS). (F) Immunohistochemistry staining for CD8⁺ T cells in lung tissue section. Arrows indicate the infiltrating CD8⁺ T cells in the tumor site of the lung tissues (MT: metastatic tumors).</p

Cytokine array/profiling analysis of the alteration of multiple cytokines/chemokines in conditioned culture media of Cp-, Am-, [Am+Cp]- and LPS-treated DCs.

<p>Cytokine array membranes were incubated with cultured media from DCs that were treated with TCL for 2 h, and then with Cp, Am, [Am+Cp] or LPS for another 22 h. Squares mark the cytokines and chemokines secreted from DCs that were increased in Cp, Am, [Am+Cp] or LPS treatments: <b>1.</b> CSF3, <b>2.</b> CSF2, <b>3.</b> CCL1, <b>4.</b> IL-1α, <b>5.</b> IL-1β, <b>6.</b> IL-5, <b>7.</b> IL-6, <b>8.</b> IL-7, <b>9.</b> IL-10, <b>10.</b> CXCL10, <b>11.</b> CXCL11, <b>12.</b> CXCL1, <b>13.</b> M-CSF, <b>14.</b> CCL2, <b>15.</b> CXCL9, <b>16.</b> CCL3, <b>17.</b> CCL4, <b>18.</b> CXCL2, <b>19.</b> TNF-α, <b>20.</b> TREM-1.</p

Flow cytometric analysis of expression of surface markers on DCs with different treatments.

<p>(A) Phenotypic changes in expression of CD40, CD80 and CD86 maturation markers in TCL-loaded DCs as a response to treatment with Cp, Am or [Am+Cp] phytoextracts, all at a concentration of 200 μg/ml. Anti-CD40, CD80 or CD86 antibodies were conjugated with FITC. (B) The mean fluorescence intensity (MFI) of (B[a]) CD40, (B[b]) CD80, or (B[c]) CD86 in DCs from different groups were calculated and are presented as a bar chart. Data represent the mean ± SD obtained from three independent experiments. A <i>P</i> value of less than 0.05 was considered significant (*, <i>P</i> < 0.05; **, <i>P</i> < 0.01; ***, <i>P</i> < 0.001; n.s, no significance).</p

Stimulation (in fold change) of cytokines and chemokines in DCs-treated with Cp, Am, [Am+Cp] or LPS groups compared to control group.

<p>* Fold change = treatment group/control untreated group</p><p>** Control group: DC+TCL</p><p>Stimulation (in fold change) of cytokines and chemokines in DCs-treated with Cp, Am, [Am+Cp] or LPS groups compared to control group.</p

<i>Graptopetalum paraguayense</i> Inhibits Liver Fibrosis by Blocking TGF-β Signaling In Vivo and In Vitro

Background and Aims: Liver fibrosis is the excessive accumulation of extracellular matrix proteins, including collagen, which occurs in most types of chronic liver diseases. Advanced liver fibrosis results in cirrhosis, liver failure, and portal hypertension. Activated hepatic perivascular stellate cells, portal fibroblasts, and myofibroblasts of bone marrow origin have been identified as major collagen-producing cells in the injured liver. These cells are activated by fibrogenic cytokines, such as TGF-&#946;1. The inhibition of TGF-&#946;1 function or synthesis is a major target for the development of antifibrotic therapies. Our previous study showed that the water and ethanol extracts of Graptopetalum paraguayense (GP), a Chinese herbal medicine, can prevent dimethylnitrosamine (DMN)-induced hepatic inflammation and fibrosis in rats. Methods: We used rat hepatic stellate HSC-T6 cells and a diethylnitrosamine (DEN)-induced rat liver injury model to test the potential mechanism of GP extracts and its fraction, HH-F3. Results: We demonstrated that GP extracts and HH-F3 downregulated the expression levels of extracellular matrix (ECM) proteins and inhibited the proliferation and migration via suppression of the TGF-&#946;1 pathway in rat hepatic stellate HSC-T6 cells. Moreover, the HH-F3 fraction decreased hepatic collagen content and reduced plasma AST, ALT, and &#947;-GT activities in a DEN-induced rat liver injury model, suggesting that GP/HH-F3 has hepatoprotective effects against DEN-induced liver fibrosis. Conclusion: These findings indicate that GP/HH-F3 may be a potential therapeutic agent for the treatment of liver fibrosis. The inhibition of TGF-&#946;-mediated fibrogenesis may be a central mechanism by which GP/HH-F3 protects the liver from injury