Wharton's jelly cells from sheep umbilical cord maintained with different culture media are permissive to in vitro infection by Small Ruminant Lentiviruses

Abstract

This study aimed to isolate cells from the Wharton's jelly of umbilical cord (WJUC) of sheep collected during natural parturition using different culture media, in addition to reporting for the first time the permissiveness of these cells to in vitro infection by small ruminant lentiviruses. Ten umbilical cords were collected from healthy sheep. Each cord explants were grown in different media consisting of MEM, low glucose DMEM, M199, and RPMI-1640. The permissiveness of infection of sheep cells from WJUC was tested with CAEV-Cork and MVV-K1514 strains, inoculating 0.1 MOI of each viral strain. Four supernatants from each strain were obtained from WJUC sheep cell cultures infected in different media. The results demonstrated the presence of cytopathic effect after the in vitro infection by CAEV-Cork and MVV-K1514 with all of the tested culture media. Nested-PCR detected proviral DNA in all supernatants. Supernatants containing CAEV-Cork viruses had TCID50/ml titres of 105.5 in MEM, 104.0 in low glucose DMEM, 105.0 in M199, and 105.7 in RPMI-1640. Supernatants containing the MVV-K1514 virus had TCID50/ml titres of 104.3 in MEM, 103.5 in low-glucose DMEM, 104.7 in M199, and 103.5 in RPMI-1640. Sheep cells from WJUC are permissive to in vitro infection by small ruminant lentivirus.

Vertical transmissibility of small ruminant lentivirus

“…In the present study, results from nPCR and Western Blot demonstrated the occurrence of SRLV-positive neonates from mothers infected by these viral agents. These results were confirmed via cell culture, and observation of syncytia and cell destruction, which are some of the main cytopathic effects characteristic of SRLV evidenced in the literature [20,26,32,33].…”

“…The maternal-filial transmission of SRLV over time has shown strong evidence of its occurrence with tropism to the organs of the reproductive system of goats and umbilical cord cells to this group of retroviruses [19,20]. In the present study, results from nPCR and Western Blot demonstrated the occurrence of SRLV-positive neonates from mothers infected by these viral agents.…”

“…In another study [46] that evaluated the transmission of the Maedi-Visna virus through the colostrum of infected sheep, it was found that one of the offspring, which was separated immediately after birth and, therefore, before the ingestion of colostrum, was seropositive in the commercial enzyme immunoassay (ELISA). Although the placenta of ruminants is syndesmocorial [47], a fact that prevents the passage of antibodies from the mother to the young, it is not known, to date, if, due to this characteristic, it would also prevent the passage of SRLV to the fetus, or whether fetal infection can occur due to the presence of the virus in other reproductive organs, whose isolation of the virus in the female reproductive system is already reported in the literature [19][20][21].…”

“…In addition, the hypothesis has been raised that intrauterine transmission is likely to occur, since there are reports of the detection of pro-viral DNA in tissues of the uterus, oviduct and ovary, demonstrating tropism of the SRLV in these regions [19]. Associated with this, there is also the fact that umbilical cord cells from small ruminants have been shown to be permissible to infection in vitro by these viral agents [20]. Thus, it is possible to deduce that a similar situation may occur in vivo, since uterine goat epithelial cells have already shown susceptibility to infection by SRLV in in vivo tests [21].…”

In vitro antiviral effect of ethanolic extracts from Azadirachta indica and Melia azedarach against goat lentivirus in colostrum and milk

“…The MTT assay was based on the methodology described by Dias et al 43 A cell suspension was prepared at a concentration of 2.0 × 10 5 cells/mL in 12 96-well plates. The samples were then incubated in a CO 2 oven for 24 h. Afterward, the wells were washed with 200 µl sterile PBS-1X at 37 °C.…”