Angiogenic response in an in vitro model of dog microvascular endothelial cells stimulated with antigenic extracts from Dirofilaria immitis adult worms.
Animals
Antigens, Bacterial
/ pharmacology
Antigens, Helminth
/ pharmacology
Capillaries
/ drug effects
Cell Survival
/ drug effects
Cells, Cultured
Dirofilaria immitis
/ chemistry
Dogs
Endothelial Cells
/ drug effects
Inflammation
Neovascularization, Physiologic
/ drug effects
Wolbachia
/ chemistry
Angiogenic factors
Canine microvascular endothelial cells
Dirofilaria immitis antigenic extracts
Pseudocapillaries formation
Wolbachia amount
Journal
Parasites & vectors
ISSN: 1756-3305
Titre abrégé: Parasit Vectors
Pays: England
ID NLM: 101462774
Informations de publication
Date de publication:
24 Jun 2019
24 Jun 2019
Historique:
received:
27
02
2019
accepted:
17
06
2019
entrez:
26
6
2019
pubmed:
27
6
2019
medline:
14
8
2019
Statut:
epublish
Résumé
Angiogenesis can occur under pathological conditions when stimuli such as inflammation, vascular obstruction or hypoxia exist. These stimuli are present in cardiopulmonary dirofilariosis (Dirofilaria immitis). The aim of this study was to analyze the capacity of D. immitis antigens to modify the expression of angiogenic factors and trigger the formation of pseudocapillaries (tube-like structures) in an in vitro model of endothelial cells. The expression of VEGF-A, sFlt, mEndoglin and sEndoglin in cultures of canine microvascular endothelial cells stimulated with extract of adult worms of D. immitis obtained from an untreated dog (DiSA) and from a dog treated for 15 days with doxycycline (tDiSA), was determined by using commercial kits. The capacity of pseudocapillary formation was evaluated analyzing cell connections and cell groups in Matrigel cell cultures stimulated with DiSA and tDiSA. In both cases non-stimulated cultures were used as controls. First, we demonstrated that worms obtained from the dog treated with doxycycline showed a significantly lower amount of Wolbachia (less than 60%) than worms removed from the untreated dog. Only DiSA was able to significantly increase the expression of the proangiogenic factor VEGF-A in the endotelial cells cultures. None of the D. immitis extracts modified the expression of sFlt. tDiSA extract was able to modify the expression of the endoglins, significantly decreasing the expression of the pro-angiogenic mEndoglin and increasing the anti-angiogenic sEndoglin. The formation of pseudocapillaries was negatively influenced by tDiSA, which reduced the organization and number of cellular connections. The ability of antigens from adult D. immitis worms to modify the expression of pro and anti-angiogenic factors in endotelial cell cultures was demonstrated, as well as the trend to form pseudocapillaries in vitro. The capacity of stimulation may be linked to the amount of Wolbachia present in the antigenic extracts.
Sections du résumé
BACKGROUND
BACKGROUND
Angiogenesis can occur under pathological conditions when stimuli such as inflammation, vascular obstruction or hypoxia exist. These stimuli are present in cardiopulmonary dirofilariosis (Dirofilaria immitis). The aim of this study was to analyze the capacity of D. immitis antigens to modify the expression of angiogenic factors and trigger the formation of pseudocapillaries (tube-like structures) in an in vitro model of endothelial cells.
METHODS
METHODS
The expression of VEGF-A, sFlt, mEndoglin and sEndoglin in cultures of canine microvascular endothelial cells stimulated with extract of adult worms of D. immitis obtained from an untreated dog (DiSA) and from a dog treated for 15 days with doxycycline (tDiSA), was determined by using commercial kits. The capacity of pseudocapillary formation was evaluated analyzing cell connections and cell groups in Matrigel cell cultures stimulated with DiSA and tDiSA. In both cases non-stimulated cultures were used as controls.
RESULTS
RESULTS
First, we demonstrated that worms obtained from the dog treated with doxycycline showed a significantly lower amount of Wolbachia (less than 60%) than worms removed from the untreated dog. Only DiSA was able to significantly increase the expression of the proangiogenic factor VEGF-A in the endotelial cells cultures. None of the D. immitis extracts modified the expression of sFlt. tDiSA extract was able to modify the expression of the endoglins, significantly decreasing the expression of the pro-angiogenic mEndoglin and increasing the anti-angiogenic sEndoglin. The formation of pseudocapillaries was negatively influenced by tDiSA, which reduced the organization and number of cellular connections.
CONCLUSIONS
CONCLUSIONS
The ability of antigens from adult D. immitis worms to modify the expression of pro and anti-angiogenic factors in endotelial cell cultures was demonstrated, as well as the trend to form pseudocapillaries in vitro. The capacity of stimulation may be linked to the amount of Wolbachia present in the antigenic extracts.
Identifiants
pubmed: 31234915
doi: 10.1186/s13071-019-3570-0
pii: 10.1186/s13071-019-3570-0
pmc: PMC6591997
doi:
Substances chimiques
Antigens, Bacterial
0
Antigens, Helminth
0
Types de publication
Journal Article
Langues
eng
Pagination
315Subventions
Organisme : Agencia de Desarrollo Económico
ID : Castilla y León
Organisme : Agencia de Desarrollo Económico
ID : Spain (cofinanced with FEDER funds)
Organisme : RICET Red de Investigación de Centros de Enfermedades Tropicales
ID : Project No. RD16/0027/0018
Organisme : Financial support for Valencia team obtained by RICET
ID : Project No. RD16/0027/0023
Références
Cancer Cell. 2002 Apr;1(3):219-27
pubmed: 12086857
Parassitologia. 2001 Dec;43(4):173-8
pubmed: 12402526
Am J Pathol. 2003 Oct;163(4):1321-7
pubmed: 14507641
Circ Res. 2006 Aug 4;99(3):248-56
pubmed: 16840721
Circ Res. 2007 Oct 26;101(9):948-56
pubmed: 17823371
Adv Parasitol. 2008;66:193-285
pubmed: 18486691
Parasitol Int. 2008 Dec;57(4):441-6
pubmed: 18603468
Vet Parasitol. 2008 Dec 10;158(3):223-31
pubmed: 18922634
Vet Parasitol. 2008 Dec 10;158(3):191-5
pubmed: 18947926
PLoS Pathog. 2009 Dec;5(12):e1000688
pubmed: 20011114
Lymphat Res Biol. 2009 Dec;7(4):215-9
pubmed: 20143920
Anticancer Res. 2010 Mar;30(3):805-10
pubmed: 20393000
J Vis Exp. 2011 Sep 07;(55):null
pubmed: 21931289
Clin Microbiol Rev. 2012 Jul;25(3):507-44
pubmed: 22763636
Circulation. 2012 Nov 27;126(22):2612-24
pubmed: 23110859
Nihon Juigaku Zasshi. 1990 Feb;52(1):159-61
pubmed: 2313946
Vet Parasitol. 2014 Jun 16;203(1-2):241-6
pubmed: 24566125
Enferm Infecc Microbiol Clin. 2015 Apr;33(4):233-7
pubmed: 24948573
Exp Parasitol. 1989 Jul;69(1):9-15
pubmed: 2499488
PLoS One. 2015 Apr 13;10(4):e0124445
pubmed: 25875022
Parasit Vectors. 2015 Apr 01;8:194
pubmed: 25888952
Trends Parasitol. 2016 Apr;32(4):325-335
pubmed: 26775037
Angiogenesis. 2017 Feb;20(1):1-24
pubmed: 27943030
Actas Dermosifiliogr. 2017 Jul - Aug;108(6):515-523
pubmed: 28162227
Life Sci. 2017 Apr 15;175:52-60
pubmed: 28336397
Expert Opin Biol Ther. 2017 Sep;17(9):1053-1063
pubmed: 28656781
Int J Mol Sci. 2018 Apr 27;19(5):null
pubmed: 29702562
Mol Biochem Parasitol. 1995 Nov;74(2):223-7
pubmed: 8719164