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Extracellular vesicles in cattle infected with bovine leukaemia virus: isolation and molecular analysis

Maria Szczotka

Maria Szczotka

Department of BiochemistryPuławy, Poland
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Szczotka, Maria
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Magdalena Wasiak

Magdalena Wasiak

Department of Pathology National Veterinary Research InstitutePuławy, Poland
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Wasiak, Magdalena
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Jacek Kuźmak

Jacek Kuźmak

Department of BiochemistryPuławy, Poland
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Kuźmak, Jacek
  
May 27, 2024
Extracellular vesicles in cattle infected with bovine leukaemia virus: isolation and molecular analysis's Cover Image
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Published Online: May 27, 2024
Page range: 189 - 198
Received: Dec 15, 2023
Accepted: May 21, 2024
DOI: https://doi.org/10.2478/jvetres-2024-0031
Keywords
© 2024 Maria Szczotka et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1.

The presence of CD9, CD63 and flotillin-1 exosomal markers in exosomes isolated from the blood plasma of bovine leukaemia virus–infected cows as detected by Western blot M – Marker ladder
The presence of CD9, CD63 and flotillin-1 exosomal markers in exosomes isolated from the blood plasma of bovine leukaemia virus–infected cows as detected by Western blot M – Marker ladder

Fig. 2.

The presence of glycoprotein (gp)51 and core protein (p)24 viral markers in exosomes isolated from the blood plasma of bovine leukaemia virus–infected cows as detected by Western blot M – Marker ladder; K+ – exosomes in supernatant of permanently bovine leukaemia virus–infected foetal lamb kidney cell culture; 1–4 – exosomes isolated from the blood plasma of bovine leukaemia virus–infected cows
The presence of glycoprotein (gp)51 and core protein (p)24 viral markers in exosomes isolated from the blood plasma of bovine leukaemia virus–infected cows as detected by Western blot M – Marker ladder; K+ – exosomes in supernatant of permanently bovine leukaemia virus–infected foetal lamb kidney cell culture; 1–4 – exosomes isolated from the blood plasma of bovine leukaemia virus–infected cows

Fig. 3.

Expression of bovine leukaemia virus glycoprotein 51 in dendritic cells generated from blood samples of bovine leukaemia virus–infected cattle, as visualised by an immunofluorescence reaction
Expression of bovine leukaemia virus glycoprotein 51 in dendritic cells generated from blood samples of bovine leukaemia virus–infected cattle, as visualised by an immunofluorescence reaction

Fig. 5.

Extracellular vesicles in in vitro culture of dendritic cells isolated from the blood of bovine leukaemia virus–infected cows seen in scanning electron microscopy Mag – magnification; EHT – electron high tension; WD – working distance; SE1 – secondary electron 1
Extracellular vesicles in in vitro culture of dendritic cells isolated from the blood of bovine leukaemia virus–infected cows seen in scanning electron microscopy Mag – magnification; EHT – electron high tension; WD – working distance; SE1 – secondary electron 1

Fig. 6.

Extracellular vesicles in in vitro culture of dendritic cells isolated from the blood of bovine leukaemia virus–infected cows seen in scanning electron microscopy Mag – magnification; EHT – electron high tension; WD – working distance; SE1 – secondary electron 1
Extracellular vesicles in in vitro culture of dendritic cells isolated from the blood of bovine leukaemia virus–infected cows seen in scanning electron microscopy Mag – magnification; EHT – electron high tension; WD – working distance; SE1 – secondary electron 1

Fig. 7.

Extracellular vesicles in in vitro culture of dendritic cells isolated from the blood of bovine leukaemia virus–infected cows seen in scanning electron microscopy. Magnification – 5.00 KX
Extracellular vesicles in in vitro culture of dendritic cells isolated from the blood of bovine leukaemia virus–infected cows seen in scanning electron microscopy. Magnification – 5.00 KX

Fig. 9.

Exosomes in in vitro culture of HeLa cells seen in scanning electron microscopy Mag – magnification; EHT – electron high tension; WD – working distance; SE1 – secondary electron 1
Exosomes in in vitro culture of HeLa cells seen in scanning electron microscopy Mag – magnification; EHT – electron high tension; WD – working distance; SE1 – secondary electron 1

Fig. 8.

Electron micrograph of exosomes in in vitro culture of dendritic cells seen in transmission electron microscopy Len – size
Electron micrograph of exosomes in in vitro culture of dendritic cells seen in transmission electron microscopy Len – size

Fig. 10.

Electron micrograph of exosomes in in vitro culture of dendritic cells isolated from the blood of bovine leukaemia virus–infected cattle seen in transmission electron microscopy
Electron micrograph of exosomes in in vitro culture of dendritic cells isolated from the blood of bovine leukaemia virus–infected cattle seen in transmission electron microscopy

Fig. 4.

Expression of bovine leukaemia virus glycoprotein 51 in a permanently bovine leukaemia virus–infected foetal lamb kidney cell line, as visualised by an immunofluorescence reaction
Expression of bovine leukaemia virus glycoprotein 51 in a permanently bovine leukaemia virus–infected foetal lamb kidney cell line, as visualised by an immunofluorescence reaction

The viral and exosomal markers detected by Western blot in bovine leukaemia virus (BLV)-infected cows

Origin of exosomes BLV infection BLV markers Cellular markers
gp51 p24 CD63 CD9 flotillin-1
Positive control – supernatant of FLK-BLV culture + + + + + +
Supernatant of cultured in vitro BLV-infected bovine DCs 1 + + + + + +
2 + + + + + +
3 + + + + + +
4 + + + + + +
5 + + + + + +
Negative control – plasma BLV− − − − + + +
Positive control – lysate of FLK–BLV cells + + + + + +
Bovine BLV+ sera (1–7) and BLV− sera (8–11) 1 + + + + + +
2 + + + + + +
3 + + + + + +
4 + + + + + +
5 + + + + + +
6 + + + + + +
7 + + + + + +
8 − − − + + +
9 − − − + + +
10 − − − + + +
11 − − − + + +
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