cholecalciferol and 4-phenylbutyric-acid

Successful treatment of acute promyelocytic leukemia (APL) has identified several novel approaches to induce leukemic cell differentiation and selective apoptosis by overcoming the site-specific transcriptional repression by dominant fusion leukemogenic proteins characteristic of APL and other forms of acute myelogenous leukemia (AML). These therapeutic approaches include the use of site-specific ligands, receptors and cytokines, disruption of dominant fusion leukemogenic proteins, chromatin remodeling and combining the above with cytotoxic chemotherapy. With the exception of cytotoxic chemotherapy, the above therapeutic strategies do not significantly affect normal hematopoiesis and their combinations have been shown to be synergistic in inducing myeloid differentiation and apoptosis in several AML cell lines and in patients with APL. These approaches are, in general, non-cross resistant and should be well tolerated particularly in elderly patients with AML. Clinical studies which include biologic end points for differentiation induction, histone acetylation and selective apoptosis are presently in development to evaluate these strategies in the treatment of AML.

Topics: Acute Disease; Adult; Aged; Animals; Antineoplastic Agents; Apoptosis; Butyrates; Cell Differentiation; Cholecalciferol; Chromatin; Drug Design; Drug Synergism; Gene Expression Regulation, Leukemic; Granulocyte Colony-Stimulating Factor; Humans; Leukemia, Myeloid; Ligands; Mice; Middle Aged; Oncogene Proteins, Fusion; Phenylbutyrates; Transcription, Genetic; Translocation, Genetic; Tretinoin

Development of new tuberculosis (TB) drugs and alternative treatment strategies are urgently required to control the global spread of TB. Previous results have shown that vitamin D3 (vitD3) and 4-phenyl butyrate (PBA) are potent inducers of the host defense peptide LL-37 that possess anti-mycobacterial effects.. To examine if oral adjunctive therapy with 5,000IU vitD3 or 2x500 mg PBA or PBA+vitD3 to standard chemotherapy would lead to enhanced recovery in sputum smear-positive pulmonary TB patients.. Adult TB patients (n = 288) were enrolled in a randomized, double-blind, placebo-controlled trial conducted in Bangladesh. Primary endpoints included proportions of patients with a negative sputum culture at week 4 and reduction in clinical symptoms at week 8. Clinical assessments and sputum smear microscopy were performed weekly up to week 4, fortnightly up to week 12 and at week 24; TB culture was performed at week 0, 4 and 8; concentrations of LL-37 in cells, 25-hydroxyvitamin D3 (25(OH)D3) in plasma and ex vivo bactericidal function of monocyte-derived macrophages (MDM) were determined at week 0, 4, 8, 12 and additionally at week 24 for plasma 25(OH)D3.. At week 4, 71% (46/65) of the patients in the PBA+vitD3-group (p = 0.001) and 61.3% (38/62) in the vitD3-group (p = 0.032) were culture negative compared to 42.2% (27/64) in the placebo-group. The odds of sputum culture being negative at week 4 was 3.42 times higher in the PBA+vitD3-group (p = 0.001) and 2.2 times higher in vitD3-group (p = 0.032) compared to placebo. The concentration of LL-37 in MDM was significantly higher in the PBA-group compared to placebo at week 12 (p = 0.034). Decline in intracellular Mtb growth in MDM was earlier in the PBA-group compared to placebo (log rank 11.38, p = 0.01).. Adjunct therapy with PBA+vitD3 or vitD3 or PBA to standard short-course therapy demonstrated beneficial effects towards clinical recovery and holds potential for host-directed-therapy in the treatment of TB.. clinicaltrials.gov NCT01580007.

Topics: Administration, Oral; Adult; Antimicrobial Cationic Peptides; Antitubercular Agents; C-Reactive Protein; Calcifediol; Calcium; Cathelicidins; Cholecalciferol; Double-Blind Method; Drug Therapy, Combination; Female; Humans; Leukocytes, Mononuclear; Logistic Models; Macrophages; Phenylbutyrates; Placebo Effect; RNA, Messenger; Sputum; Treatment Outcome; Tuberculosis, Pulmonary

We earlier showed that 4-phenylbutyrate (PB) can induce cathelicidin LL-37 expression synergistically with 1,25-dihydroxyvitamin D3 in a lung epithelial cell line. We aimed to evaluate a therapeutic dose of PB alone or in combination with vitamin D3 for induction of LL-37 expression in immune cells and enhancement of antimycobacterial activity in monocyte-derived macrophages (MDM).. Healthy volunteers were enrolled in an 8-days open trial with three doses of PB [250 mg (Group-I), 500 mg (Group-II) or 1000 mg (Group-III)] twice daily (b.d.) together with vitamin D3 {5000 IU once daily (o.d.)}, PB (500 mg b.d.) (Group-IV) or vitamin D3 (5000 IU o.d.) (Group-V), given orally for 4 days. Blood was collected on day-0, day-4 and day-8; plasma was separated, peripheral blood mononuclear cells (PBMC), non-adherent lymphocytes (NAL) and MDM were cultured. LL-37 transcript in cells and peptide concentrations in supernatant were determined by qPCR and ELISA, respectively. In plasma, 25-hydorxyvitamin D3 levels were determined by ELISA. MDM-mediated killing of Mycobacterium tuberculosis (Mtb) (H37Rv) was performed by conventional culture method.. MDM from Group-II had increased concentration of LL-37 peptide and transcript at day-4, while Group-I showed increased transcript at day-4 and day-8 compared to day-0 (p < 0.05). Both Group-I and -II exhibited higher levels of transcript on day-4 compared to Group-III and Group-V (p < 0.035). Increased induction of peptide was observed in lymphocytes from Group-II on day-4 compared to Group-I and Group-IV (p < 0.05), while Group-IV showed increased levels on day-8 compared to Group-I and Group-III (p < 0.04). Intracellular killing of Mtb on day-4 was significantly increased compared to day-0 in Group-I, -II and -V (p < 0.05).. The results demonstrate that 500 mg b.d. PB with 5000 IU o.d. vitamin D3 is the optimal dose for the induction of LL-37 in macrophages and lymphocytes and intracellular killing of Mtb by macrophages. Hence, this dose has potential application in the treatment of TB and is now being used in a clinical trial of adults with active pulmonary TB (NCT01580007).

Topics: Administration, Oral; Adolescent; Adult; Antimicrobial Cationic Peptides; Antineoplastic Agents; Calcium; Cathelicidins; Cells, Cultured; Cholecalciferol; Dose-Response Relationship, Drug; Drug Therapy, Combination; Female; Humans; Immunity, Innate; Macrophages; Male; Middle Aged; Mycobacterium tuberculosis; Phenylbutyrates; RNA, Messenger; Tuberculosis; Up-Regulation; Vitamins; Young Adult

Maintaining fish health is one of the most important aims in aquaculture. Prevention of fish diseases therefore is crucial and can be achieved by various different strategies, including most often a combination of different methods such as optimal feed and fish density, as well as strengthening the immune system. Understanding the fish innate immune system and developing methods to activate it, in an effort to prevent infections in the first place, has been a goal in recent years. In this study we choose different inducers of the innate immune system and examined their effects in vitro on the salmon cell line CHSE-214. We found that the butyrate derivatives 4-phenyl butyrate (PBA) and β-hydroxy-β-methyl butyrate (HMB) induce the expression of various innate immune genes differentially over 24-72 h. Similarly, lipids generated from fish oils were found to have an effect on the expression of the antimicrobial peptides cathelicidin and hepcidin, as well as iNOS and the viral receptor RIG-1. Interestingly we found that vitamin D3, similar as in mammals, was able to increase cathelicidin expression in fish cells. The observed induction of these different innate immune factors correlated with antibacterial activity against Aeromonas salmonicida and antiviral activity against IPNV and ISAV in vitro. To relate this data to the in vivo situation we examined cathelicidin expression in juvenile salmon and found that salmon families vary greatly in their basal cathelicidin levels. Examining cathelicidin levels in families known to be resistant to IPNV showed that these QTL-families had lower basal levels of cathelicidin in gills, than non QTL-families. Feeding fish with HMB caused a robust increase in cathelicidin expression in gills, but not skin and this was independent of the fish being resistant to IPNV. These findings support the use of fish cell lines as a tool to develop new inducers of the fish innate immune system, but also highlight the importance of the tissue studied in vivo. Understanding the response of the innate immune system in different tissues and what effect this might have on infections and downstream cellular pathways is an interesting research topic for the future.

Topics: Aeromonas salmonicida; Animals; Birnaviridae Infections; Cell Line; Cholecalciferol; Fish Diseases; Fish Proteins; Furunculosis; Gene Expression; Gram-Negative Bacterial Infections; Immunity, Innate; Infectious pancreatic necrosis virus; Lipids; Phenylbutyrates; Salmo salar; Valerates

Antimicrobial peptides (AMPs) constitute an indispensable arm of innate immunity against infectious microbes in humans. Induction of endogenous AMPs may become an alternative therapy against infections. Our previous studies have demonstrated phenylbutyrate (PBA) as a novel inducer of the AMPs cathelicidin (encoded by the CAMP gene) and human beta-defensin-1 in the human bronchial epithelial cell line VA10. In this work, we have continued by studying molecular mechanisms of PBA mediated induction of LL-37 expression and associated pathways in the human bronchial epithelial cell line VA10. In this study we demonstrate vitamin D receptor (VDR) as a key transcription factor required for PBA mediated up-regulation of the CAMP gene expression. PBA also increases mRNA expression of the vitamin D3 regulated genes CYP24A1 and CD14. The siRNA knockdown of VDR reduced PBA mediated increase in CAMP, CYP24A1 and CD14 expression. Furthermore, we demonstrate that PBA enhances Toll-Like Receptor 5 ligand flagellin regulated mRNA expression of the inflammatory cytokine TNFα and chemokine CXCL8. PBA also up-regulates the expression of the genes encoding the growth factor cytokines transforming growth factor (TGF) α, TGFβ1 and TGFβ2. Our results indicate that TGFβ type I receptor and epidermal growth factor receptor are involved in PBA mediated CAMP regulation. Finally, we show that co-treatment with PBA and vitamin D3 reduces Pseudomonas aeruginosa growth in vitro.

Topics: Antimicrobial Cationic Peptides; beta-Defensins; Cathelicidins; Cell Line; Cholecalciferol; Cytokines; Flagellin; Gene Silencing; Humans; Inflammation; Intercellular Signaling Peptides and Proteins; Ligands; Microbial Sensitivity Tests; Phenylbutyrates; Pseudomonas aeruginosa; Receptors, Calcitriol; RNA, Messenger; Signal Transduction; Toll-Like Receptor 5; Up-Regulation; Vitamin D3 24-Hydroxylase

A promising strategy in the fight against multidrug-resistant pathogens is the induction of endogenous AMPs, with compounds such as VitD₃ and PBA. These compounds display an array of immunomodulatory effects that remain to be investigated in further detail to establish their role in the clearance of infection and possible modulation of AMP expression. Here, we have investigated the effects of VitD₃ and PBA on human monocyte-DC differentiation and found that VitD₃ and PBA promote the development of a stretched CD14⁺/CD1a⁻ DC subset. This subset produced enhanced levels of ROS and human cathelicidin; furthermore, it displayed enhanced killing capacity of Staphylococcus aureus compared with control DCs. When experiments were performed in WT and cathelicidin-deficient mice, we established that a ROS-producing, stretched DC subset was also induced in mouse-derived cells, independent of cathelicidin expression. However, in contrast to the human DCs, enhanced cathelicidin expression and enhanced antimicrobial activities were not found in the murine VitD₃/PBA DC subset. In conclusion, the results of this study show that VitD₃ and PBA induce a human DC subset that is effective against infection. These results promote further research into the use of these compounds as an antimicrobial treatment strategy.

Topics: Animals; Antimicrobial Cationic Peptides; Cathelicidins; Cell Differentiation; Cells, Cultured; Cholecalciferol; Cytokines; Dendritic Cells; Humans; Mice; Mice, Inbred C57BL; Monocytes; Phenylbutyrates; Reactive Oxygen Species