RESUMEN
The urothelium of the urinary bladder represents the first line of defense. However, uropathogenic E. coli (UPEC) damage the urothelium and cause acute bacterial infection. Here, we demonstrate the crosstalk between macrophages and the urothelium stimulating macrophage migration into the urothelium. Using spatial proteomics by MALDI-MSI and LC-MS/MS, a novel algorithm revealed the spatial activation and migration of macrophages. Analysis of the spatial proteome unravelled the coexpression of Myo9b and F4/80 in the infected urothelium, indicating that macrophages have entered the urothelium upon infection. Immunofluorescence microscopy additionally indicated that intraurothelial macrophages phagocytosed UPEC and eliminated neutrophils. Further analysis of the spatial proteome by MALDI-MSI showed strong expression of IL-6 in the urothelium and local inhibition of this molecule reduced macrophage migration into the urothelium and aggravated the infection. After IL-6 inhibition, the expression of matrix metalloproteinases and chemokines, such as CX3CL1 was reduced in the urothelium. Accordingly, macrophage migration into the urothelium was diminished in the absence of CX3CL1 signaling in Cx3cr1gfp/gfp mice. Conclusively, this study describes the crosstalk between the infected urothelium and macrophages through IL-6-induced CX3CL1 expression. Such crosstalk facilitates the relocation of macrophages into the urothelium and reduces bacterial burden in the urinary bladder.
Asunto(s)
Comunicación Celular , Quimiocina CX3CL1/metabolismo , Interleucina-6/metabolismo , Macrófagos/metabolismo , Proteómica , Urotelio/inmunología , Urotelio/metabolismo , Animales , Modelos Animales de Enfermedad , Susceptibilidad a Enfermedades , Técnica del Anticuerpo Fluorescente , Inmunohistoquímica , Macrófagos/inmunología , Ratones , Proteómica/métodos , Vejiga Urinaria/inmunología , Vejiga Urinaria/metabolismo , Vejiga Urinaria/microbiología , Infecciones Urinarias/etiología , Infecciones Urinarias/metabolismo , Infecciones Urinarias/patología , Urotelio/microbiologíaRESUMEN
Cis-dichloro-oxovanadium(IV) complexes [VO(L1/L2)Cl2], where L1 is N-(4-(5,5-difluoro-1,3,7,9-tetramethyl-5H-4Ê4,5Ê4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborinin-10-yl)benzyl)-1-(pyridin-2-yl)-N-(pyridin-2-ylmethyl)methanamine in 1 and L2 is N-(4-(5,5-difluoro-2,8-diiodo-1,3,7,9-tetramethyl-5H-4Ê4,5Ê4-dipyrrolo[1,2-c:2',1'-f][1,3,2]diazaborinin-10-yl)benzyl)-1-(pyridin-2-yl)-N-(pyridin-2-ylmethyl)methanamine in 2) having 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene as boron-dipyrromethene (BODIPY) appended dipicolylamine bases were prepared, characterized and their photocytotoxicity studied. X-ray crystal structure of 1 showed distorted octahedral geometry with a VIVON3Cl2 core having Cl-V-Cl angle of 91.93(4)°. The complexes showed variable solution conductivity properties. They were non-electrolytes in dry DMF at 25⯰C but showed 1:1 electrolytic behavior in an aqueous medium due to dissociation of one chloride ligand as evidenced from the mass spectral study. Complexes 1 and 2 showed absorption bands at 500 and 535â¯nm, respectively. The calf thymus DNA melting study revealed their interaction through DNA crosslinking on exposure to light which was further confirmed from the alkaline agarose gel electrophoresis using plasmid supercoiled pUC19 DNA. Complex 2 showed disruption of the mitochondrial membrane potential in the JC-1 (1,1',3,3'-tetraethyl-5,5',6,6'-tetrachloroimidacarbocyanine iodide) assay. The complexes were photocytotoxic in visible light (400-700â¯nm, power: 10â¯Jâ¯cm-2) in cervical cancer HeLa and breast cancer MCF-7 cells. Complex 2 having a photoactive diiodoboron-dipyrromethene moiety gave a singlet oxygen quantum yield (ΦΔ) value of ~0.6. It showed singlet oxygen mediated apoptotic photodynamic therapy activity with remarkably low IC50 (half maximal inhibitory concentration) value of ~0.15⯵M. The cis-disposition of chlorides gave a cis-divacant 4-coordinate intermediate structure from the density functional theory (DFT) study thus mimicking the DNA crosslinking property of cisplatin.
Asunto(s)
Compuestos de Boro , Citotoxinas , ADN , Fármacos Fotosensibilizantes , Porfobilinógeno/análogos & derivados , Vanadatos , Boro/química , Boro/farmacología , Compuestos de Boro/síntesis química , Compuestos de Boro/química , Compuestos de Boro/farmacología , Cristalografía por Rayos X , Citotoxinas/síntesis química , Citotoxinas/química , Citotoxinas/farmacología , ADN/química , ADN/metabolismo , Células HeLa , Humanos , Células MCF-7 , Fármacos Fotosensibilizantes/síntesis química , Fármacos Fotosensibilizantes/química , Fármacos Fotosensibilizantes/farmacología , Porfobilinógeno/síntesis química , Porfobilinógeno/química , Porfobilinógeno/farmacología , Vanadatos/química , Vanadatos/farmacologíaRESUMEN
Natural killer (NK) cells comprise of â¼70% of the immune cell population in the maternal decidua and â¼15% of the mononuclear cells in the peripheral blood. The decidual NK cells capable of producing high levels of cytokines are functionally distinct from the peripheral NK cells that exhibit high cytotoxicity. The numbers of peripheral NK cells and their cytotoxicity potential have been correlated with pregnancy outcome. In the same context, glycodelin, an immunomodulatory protein, has been recognized to be essential for the establishment and maintenance of pregnancy, and its' reduced levels are associated with recurrent spontaneous abortions. We investigated the effect of glycodelin on the peripheral NK cells. Our results reveal that glycodelin suppresses the cytotoxicity of peripheral NK cells via downregulating perforin, granzyme B and IFNγ. Glycodelin also induces caspase-dependent death in only activated peripheral NK cells, the effect suggested to be mediated by glycodelin upon engaging with the CD7 cell surface receptor. Thus, during pregnancy, glycodelin modulates the function and the number of cytotoxic NK cells that pose a deleterious effect on the fetus, a semi-allograft. This study provides insights into the mechanism of the regulatory effect of glycodelin on NK cells and could possibly be exploited for the management of miscarriages.
Asunto(s)
Glicodelina/metabolismo , Células Asesinas Naturales/inmunología , Células T Asesinas Naturales/inmunología , Aborto Habitual/inmunología , Antígenos CD7/metabolismo , Comunicación Celular/inmunología , Línea Celular Tumoral , Regulación hacia Abajo/inmunología , Implantación del Embrión/inmunología , Femenino , Granzimas/metabolismo , Humanos , Interferón gamma/metabolismo , Células Asesinas Naturales/metabolismo , Leucocitos Mononucleares , Recuento de Linfocitos , Células T Asesinas Naturales/metabolismo , Perforina/metabolismo , Cultivo Primario de Células , Trofoblastos/inmunologíaRESUMEN
Correction for 'Endoplasmic reticulum targeting tumour selective photocytotoxic oxovanadium(iv) complexes having vitamin-B6 and acridinyl moieties' by Samya Banerjee et al., Dalton Trans., 2016, 45, 783-796.
RESUMEN
Correction for 'Endoplasmic reticulum targeted chemotherapeutics: the remarkable photo-cytotoxicity of an oxovanadium(iv) vitamin-B6 complex in visible light' by Samya Banerjee et al., Chem. Commun., 2014, 50, 5590-5592.
RESUMEN
Herein, siRNA transfection efficiency of a unique set of α-tocopherylated gemini lipids has been established in vitro and in vivo. High efficacy of oncogene silencing achieved using the biomacromolecular assembly, formed from siRNA complexes of co-liposomes containing an α-tocopherylated gemini lipid, has been utilized for tumor regression via chemosensitization. Delivery studies with the gemini bearing hydroxyethyl headgroup with octamethylene spacer (TH8S) pointed to a higher siRNA transfection efficacy than its analog without hydroxyethyl group (T8S). Owing to p53 upregulation, transfected cells showed enhanced sensitivity to the chemotherapeutic agent, doxorubicin. Studies in murine model revealed significantly low levels of survivin mRNA in xenograft tumors injected with siRNA lipoplexes, leading to effective inhibition of tumor growth and an increase in sensitivity of the tumors toward doxorubicin. These findings enable us to propose the anti-survivin siRNA carrying TH8S co-liposomes as a potent member of cancer management strategies using suicide gene therapy.
Asunto(s)
Doxorrubicina , Técnicas de Silenciamiento del Gen , Lípidos , Neoplasias , ARN Interferente Pequeño , Transfección , Proteína p53 Supresora de Tumor/genética , alfa-Tocoferol , Animales , Doxorrubicina/química , Doxorrubicina/farmacocinética , Doxorrubicina/farmacología , Células HEK293 , Células Hep G2 , Humanos , Lípidos/química , Lípidos/farmacocinética , Lípidos/farmacología , Liposomas , Ratones , Neoplasias/tratamiento farmacológico , Neoplasias/genética , Neoplasias/metabolismo , Neoplasias/patología , ARN Interferente Pequeño/química , ARN Interferente Pequeño/farmacocinética , ARN Interferente Pequeño/farmacología , Proteína p53 Supresora de Tumor/deficiencia , Proteína p53 Supresora de Tumor/metabolismo , alfa-Tocoferol/química , alfa-Tocoferol/farmacocinética , alfa-Tocoferol/farmacologíaRESUMEN
Ly6C+ monocytes are important components of the innate immune defense against infections. These cells have been shown to proliferate in the bone marrow of mice with systemic infections. However, the proliferative capacity of Ly6C+ monocytes in infected peripheral tissues as well as the associated regulatory mechanisms remain unclear. In this study, we analyzed the proliferative capacity of Ly6C+ monocytes in the urinary bladder after infection with uropathogenic E. coli, one of the most prevalent pathogen worldwide, and in LPS-induced peritonitis. We show that Ly6C+ monocytes proliferated in the bladder after infection with uropathogenic E. coli and in the peritoneum after intraperitoneal injection of LPS. We identified IL-6, a molecule that is highly expressed in infections, as a crucial regulator of Ly6C+ monocyte proliferation. Inhibition of IL-6 via administration of antibodies against IL-6 or gp130 impeded Ly6C+ monocyte proliferation. Furthermore, repression of IL-6 trans-signaling via administration of soluble gp130 markedly reduced the proliferation of Ly6C+ monocytes. Overall, this study describes the proliferation of Ly6C+ monocytes using models of urinary tract infection and LPS-induced peritonitis. IL-6 trans-signaling was identified as the regulator of Ly6C+ monocyte proliferation.
Asunto(s)
Antígenos Ly/metabolismo , Proliferación Celular , Infecciones por Escherichia coli/microbiología , Interleucina-6/metabolismo , Monocitos/inmunología , Infecciones Urinarias/inmunología , Animales , Antígenos Ly/inmunología , Diferenciación Celular , Células Cultivadas , Escherichia coli/patogenicidad , Infecciones por Escherichia coli/complicaciones , Femenino , Interleucina-6/genética , Ratones , Ratones Endogámicos C57BL , Monocitos/metabolismo , Transducción de Señal , Infecciones Urinarias/metabolismo , Infecciones Urinarias/microbiologíaRESUMEN
Oxovanadium(IV) complexes [VO(L1/L2)Cl2]n+ (1,2) of (anthracenyl)terpyridine (An-tpy as L1 in 1, n=0) and triphenylphosphonium-appended (anthracenyl)terpyridine (An-tpy-TPP+ as L2 in 2, n=1) were synthesized, characterized and their DNA crosslinking ability, photocytotoxicity in visible light and cellular localization in cancer cells studied. The bromide derivative of 2, viz. [VO(An-tpy-TPP)Br2]Br (3) is structurally characterized. The structure showed trans disposition of two halides in the coordination sphere and the TPP+ unit is a pendant to the terpyridyl ligand. The DNA melting and comet assay studies on the complexes suggest the formation of DNA crosslinks. Complexes 1 and 2 displayed ~10 fold increase in cytotoxicity on exposure to visible light (400-700nm) when compared to those in dark in HeLa and MCF-7 cells. FACScan (Fluorescence Associated Cell Sorter Scan) analysis showed cellular apoptosis when treated with the complex in visible light in comparison to their dark controls. Fluorescence microscopic studies using complex 2 revealed its mitochondrial localization within the cancer cells.
Asunto(s)
Antracenos , Reactivos de Enlaces Cruzados , ADN de Neoplasias/metabolismo , Neoplasias/tratamiento farmacológico , Fototerapia , Vanadatos , Antracenos/síntesis química , Antracenos/química , Antracenos/farmacología , Apoptosis/efectos de los fármacos , Reactivos de Enlaces Cruzados/síntesis química , Reactivos de Enlaces Cruzados/química , Reactivos de Enlaces Cruzados/farmacología , Células HeLa , Humanos , Mitocondrias/metabolismo , Mitocondrias/patología , Neoplasias/metabolismo , Neoplasias/patología , Vanadatos/síntesis química , Vanadatos/química , Vanadatos/farmacologíaRESUMEN
Oxovanadium(iv) complexes of vitamin-B6 Schiff base, viz., [VO(HL(1)/L(2)/L(3))(B)]Cl (), where B is 2,2'-bipyridine (bpy in and ), 11-(9-acridinyl)dipyrido[3,2-a:2',3'-c]phenazine (acdppz in and ), H2L(1)·HCl is 3-hydroxy-5-(hydroxymethyl)-4-(((2-hydroxyphenyl)imino)methyl)-2-methylpyridin-1-ium chloride (in and ), HL(2) is 2-(((2-(1H-imidazol-4-yl)ethyl)imino)methyl)phenol (in ) and HL(3) is 4-(((2-(1H-imidazol-4-yl)ethyl)imino)methyl)-5-(hydroxymethyl)-2-methylpyridin-3-ol (in ) were synthesized, characterized and their cellular uptake, photo-activated cytotoxicity and intracellular localization were studied. Complexes , as the perchlorate salt of , and , as the hexafluorophosphate salt of , were structurally characterized. Vitamin-B6 transporting membrane carrier (VTC) mediated entry into tumour cells in preference to the normal ones seems to be responsible for the higher cellular uptake of the complexes into HeLa and MCF-7 cells over MCF-10A cells. Complexes and having acdppz as the photosensitizer exhibit remarkable photocytotoxicity in these cancer cells giving IC50 of <0.9 µM. The complexes remain non-toxic in the dark. The complexes show photo-induced apoptotic cell death via singlet oxygen ((1)O2) generation. Fluorescence microscopy reveals specific localization of complex to endoplasmic reticulum (ER) and generation of (1)O2 possibly leads to apoptotic cell death by triggering ER stress response (ERSR).
Asunto(s)
Complejos de Coordinación/química , Retículo Endoplásmico/metabolismo , Fármacos Fotosensibilizantes/química , Vanadatos/química , Vitamina B 6/química , 2,2'-Dipiridil/química , Apoptosis/efectos de los fármacos , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Complejos de Coordinación/síntesis química , Complejos de Coordinación/farmacología , Cristalografía por Rayos X , Retículo Endoplásmico/efectos de los fármacos , Células HeLa , Humanos , Luz , Células MCF-7 , Microscopía Fluorescente , Conformación Molecular , Fármacos Fotosensibilizantes/síntesis química , Fármacos Fotosensibilizantes/farmacología , Teoría Cuántica , Bases de Schiff/química , Oxígeno Singlete/metabolismo , SolubilidadRESUMEN
An oxovanadium(IV) vitamin-B6 Schiff base complex, viz. [VO(HL)(acdppz)]Cl, having (acridinyl)dipyridophenazine (acdppz) shows specific localization to endoplasmic reticulum (ER) and remarkable apoptotic photocytotoxicity in visible light (400-700 nm) in HeLa and MCF-7 cancer cells (IC50 < 0.6 µM) while being non-toxic in the dark and to MCF-10A normal cells (IC50 > 40 µM).
Asunto(s)
Retículo Endoplásmico/efectos de los fármacos , Retículo Endoplásmico/efectos de la radiación , Luz , Compuestos Organometálicos/farmacología , Fotoquimioterapia , Vanadatos/química , Vitamina B 6/química , Células HeLa , Humanos , Células MCF-7 , Modelos Moleculares , Conformación Molecular , Compuestos Organometálicos/química , Fármacos Fotosensibilizantes/química , Fármacos Fotosensibilizantes/farmacología , Bases de Schiff/químicaRESUMEN
The phagocytes of the innate immune system, macrophages and neutrophils, contribute to antibacterial defense, but their functional specialization and cooperation is unclear. Here, we report that three distinct phagocyte subsets play highly coordinated roles in bacterial urinary tract infection. Ly6C(-) macrophages acted as tissue-resident sentinels that attracted circulating neutrophils and Ly6C(+) macrophages. Such Ly6C(+) macrophages played a previously undescribed helper role: once recruited to the site of infection, they produced the cytokine TNF, which caused Ly6C(-) macrophages to secrete CXCL2. This chemokine activated matrix metalloproteinase-9 in neutrophils, allowing their entry into the uroepithelium to combat the bacteria. In summary, the sentinel macrophages elicit the powerful antibacterial functions of neutrophils only after confirmation by the helper macrophages, reminiscent of the licensing role of helper T cells in antiviral adaptive immunity. These findings identify helper macrophages and TNF as critical regulators in innate immunity against bacterial infections in epithelia.