Effect of somatic antigens of Dirofilaria repens adult worms on angiogenesis, cell proliferation and migration and pseudo-capillary formation in human endothelial cells

María Del Pilar Pérez Rodríguez; Claudia Alarcón-Torrecillas; Miguel Pericacho; Iván Rodríguez-Escolar; Elena Carretón; Rodrigo Morchón

doi:10.1186/s13071-023-05726-z

Effect of somatic antigens of Dirofilaria repens adult worms on angiogenesis, cell proliferation and migration and pseudo-capillary formation in human endothelial cells

Parasit Vectors. 2023 Mar 16;16(1):105. doi: 10.1186/s13071-023-05726-z.

Authors

María Del Pilar Pérez Rodríguez¹, Claudia Alarcón-Torrecillas², Miguel Pericacho², Iván Rodríguez-Escolar¹, Elena Carretón³, Rodrigo Morchón^{4

5}

Affiliations

¹ Zoonotic Diseases and One Health Group, IBSAL-CIETUS (Biomedical Research Institute of Salamanca Research Centre for Tropical Diseases), Faculty of Pharmacy, University of Salamanca, 37007, Salamanca, Spain.
² Department of Physiology and Pharmacology, Biomedical Research Institute of Salamanca (IBSAL), University of Salamanca, Salamanca, Spain.
³ Faculty of Veterinary Medicine, Research Institute of Biomedical and Health Sciences (IUIBS), University of Las Palmas de Gran Canaria, Arucas, 35413, Las Palmas, Spain.
⁴ Zoonotic Diseases and One Health Group, IBSAL-CIETUS (Biomedical Research Institute of Salamanca Research Centre for Tropical Diseases), Faculty of Pharmacy, University of Salamanca, 37007, Salamanca, Spain. rmorgar@usal.es.
⁵ Faculty of Veterinary Medicine, Research Institute of Biomedical and Health Sciences (IUIBS), University of Las Palmas de Gran Canaria, Arucas, 35413, Las Palmas, Spain. rmorgar@usal.es.

Abstract

Background: Angiogenesis is defined as the formation of new vessels by sprouting of endothelial cells from pre-existing vessels in response to stimuli, such as hypoxia or inflammation. Subcutaneous dirofilariasis, caused by Dirofilaria repens, is a zoonotic disease characterized by the formation of subcutaneous nodules with the presence of at least one encapsulated worm, showing perivascular vascularization around it. The aim of this study is to analyze whether the somatic antigen of adult D. repens worms interacts with and modulates the angiogenic mechanism, cell proliferation and migration, and formation of pseudo-capillaries.

Methods: The expression of VEGF-A, VEGFR-1/sFlt, VEGFR-2, mEnd and sEnd in cultures of human vascular endothelial cells stimulated with somatic antigen of adult worms of D. repens (DrSA), vascular endothelial growth factor (VEGF) and DrSA + VEGF were evaluated by using ELISA commercial kits. Cellular viability was analyzed by live cell count, cytotoxicity assays by using a commercial kit, cell proliferation by MTT-based assay, cell migration by wound-healing assay carried out by scratching wounds and capacity of formation of pseudo-capillaries analyzing cell connections and cell groups in Matrigel cell cultures. In all cases unstimulated cultures were used as controls.

Results: DrSA + VEGF significantly increased the expression of VEGF-A, VEGFR-2 and mEndoglin compared to other groups and unstimulated cultures. Moreover, DrSA + VEGF produced cell proliferation and migration and increased the formation of pseudo-capillaries.

Conclusions: Somatic antigen of adult D. repens worms activated the proangiogenic mechanism, cell proliferation and cell migration as well as formation of pseudo-capillaries in this in vitro human endothelial cell model. These processes could be related to the survival of adult D. repens in subcutaneous nodules in infected hosts.

Keywords: Angiogenesis; Cell migration; Cell proliferation; Dirofilaria repens adult worms; Pseudo-capillary formation; Somatic antigens.

MeSH terms

Adult
Animals
Capillaries
Cell Proliferation
Dirofilaria repens*
Dirofilariasis*
Endothelial Cells
Humans
Vascular Endothelial Growth Factor A
Vascular Endothelial Growth Factor Receptor-2

Substances

Vascular Endothelial Growth Factor A
Vascular Endothelial Growth Factor Receptor-2