Blood dendritic cells in cattle infected with bovine leukemia virus (BLV): isolation and phenotyping

Szczotka, M.; Kuźmak, Jacek; Kostro, K.; Bednarek, D.; Purzycka, M.

doi:10.2478/v10181-012-0095-2

Cited by 3 publications

(5 citation statements)

References 42 publications

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“…Interleukin 6 induces haematopoietic cell proliferation, functional maturation and differentiation, and deficiency of IL-6 can affect BMSC function and impair haematopoietic differentiation and supportive activity. In our investigations, low levels of IL-6 secretion were found in cell cultures of bone marrow-and spleen-originated BLV-infected stem cells, which is in agreement with other authors' findings and indicative of the stemness of these bone marrow and spleen cells when infected with BLV (31). Contrastingly, IL-6 secretion was higher in the blood and lymph node stem cells.…”

Section: Discussionsupporting

confidence: 93%

“…The data reported by others indicated an increase in the secretion of IL-10 during the progression of BLV infection. Elevated concentrations of this cytokine were noted in experimentally infected sheep, and this was associated with progression of the disease (31). Interleukin 10 actively blocked BLV replication in adherent cultures of monocytes/macrophages, which suggested involvement of this cytokine in mechanisms of viral latency (11).…”

Section: Discussionmentioning

confidence: 97%

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Cytokine secretion in stem cells of cattle infected with bovine leukaemia virus

Szczotka,

Kuźmak

2024

Journal of Veterinary Research

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Introduction Bovine leukaemia virus (BLV) is a Deltaretrovirus responsible for enzootic bovine leukosis, the most common neoplastic disease of cattle. It deregulates the immune system, favouring secondary infections and changes in the blood and lymphatic tissues. Blood homeostasis depends on functional haematopoietic stem cells (HSCs). Bone marrow is populated by these cells, which express CD34+ and CD35+ surface antigens and produce and release cytokines involved in the maintenance of haematopoiesis. The aim of the study was determination of the profile of cytokine production by CD34+ stem cells of cattle naturally infected with BLV. Material and Methods The HSCs were generated from the blood and lymphoid organs of cows infected with BLV and healthy control cows with immunomagnetic separation and anti-CD34+ monoclonal antibodies. Isolated CD34+ cells were cultivated for two weeks with interleukin (IL)-4 and granulocyte-macrophage colony-stimulating factor. The levels of IL-6, IL-10, IL-12p40, IL-12p70, interferon gamma (IFN-γ) and tumour necrosis factor alpha (TNF-α) were determined in culture fluid by flow cytometry. Results The expression of IL-6, IL-12p70 and TNF-α in blood HSCs was higher in BLV+ cows than in the control animals. In bone marrow HSCs of infected cows, IL-12, TNF-α and IFN-γ were more concentrated, but in these cows’ spleen HSCs only expression of IL-10 was elevated. In HSCs isolated from the lymph nodes of leukaemic cows, only TNF-α secretion was lower than in control cows, the other cytokines being more potently secreted. Conclusion Infection with BLV caused statistically significant differences in cytokine expression by HSC CD34+ cells.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 97%

Cytokine secretion in stem cells of cattle infected with bovine leukaemia virus

Szczotka,

Kuźmak

2024

Journal of Veterinary Research

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“…The methods of DC generation and cell culture were adapted from those described by Szczotka et al ( 23 ). Briefly, DCs were generated from the bovine blood monocytes.…”

Section: Methodsmentioning

confidence: 99%

“…The viraemic state is rather short and can be detected during the first days after viral infection. The detection of viral antigens is difficult because viral protein production is suppressed by cytokines, especially by interleukin (IL)-6 and IL-10 ( 20 , 23 ). These investigations indicated that BLV infection caused disease when changes occurred in host factors.…”

Section: Introductionmentioning

confidence: 99%

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

Szczotka,

Wasiak,

Kuźmak

2024

Journal of Veterinary Research

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Introduction Exosomes are nanosized lipid bilayer membranous microvesicles, extracellularly released from a variety of mammalian cells. They mediate intercellular signalling by transporting several types of RNA, lipids and proteins and participate in the intercellular exchange of DNA, RNA, micro RNA, proteins and other components. These microvesicles are present in all body fluids in physiological and pathological conditions and reflect the state of the host organism. The aim of the study was the isolation and molecular determination of exosomes in blood and supernatant fluids of bovine dendritic cell cultures infected with bovine leukaemia virus (BLV). Material and Methods Exosomes were isolated by ultracentrifugation from the blood sera, plasma and supernatant of bovine BLV-infected and uninfected control dendritic cell cultures and their presence was confirmed with scanning electron and transmission electron microscopy. Western blot analysis of the structural BLV glycoprotein 51 (Env) and protein 24 (Gag) and of the tetraspanin exosomal markers CD9, CD63 and flotillin-1 was undertaken in BLV+ and control BLV− cattle. Results In exosomes of leukaemic cattle both BLV proteins and exosomal markers were detected. In healthy control animals only exosomal markers were determined. Conclusion Proteins of BLV were released with exosomes and could be transferred into recipient cells as an alternative propagation route not requiring virus infection.

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“…Reactivation of this enzyme in tumour cells has been observed in humans (Allsopp et al 1992, Hoos et al 1998 and in some animal species (pig, dog, and cat), so the telomerase activity has been proposed as a marker of tumour in these mammals (Varon et al 1997, Biller et al 1998,Yazawa et al 1999, Pathak et al 2000, Biller et al 2001, Nasir et al 2001, Argyle et al 2003. The high level of telomerase activity was determined in cattle infected with bovine leukaemia virus (BLV) in blood lymphocytes, sera, and supernatants of in vitro cultured dendritic cell (Szczotka et al 2009(Szczotka et al , 2011. Telomerase activity in tumour tissue and blood samples in BLV-infected cows was investigated by Suzuki et al (2008).…”

Section: Introductionmentioning

confidence: 99%

Polish Journal of Veterinary Sciences

Szczotka

Kocki

Iwan

et al. 2019

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Telomeres are repetitive sequence structures at the ends of chromosomes. They consist of the double stranded DNA repeats followed by the short single stranded DNA. In humans and other verterbrates the telomeric sequence is composed of tandem of TTAGGG repeats. With each cells division telomeres shorten by up to 200 base pairs. Telomerase is an enzyme responsible for continuous cell growth and is repressed in most somatic cells, except proliferating progenitor cells, but in more than 85% of cancer cells telomerase expression is observed. Tumour cells with metastatic potential may demonstrate a high telomerase activity, allowing cells to escape from the inhibition of cell proliferation due to shortened telomeres. Determination of telomerase expression was performed with the use of PCR ELISA in samples isolated from bovine leukaemia virus (BLV) infected cows. Telomerase activity was found in almost all investigated samples. The relative telomerase activity (RTA) was higher in infected cows than in healthy animals and the differences were statistically significant (α=0.05). In blood lymphocytes of BLV-infected cows the mean values of telomerase expression determined in real-time PCR were 3534.12 copies, in the healthy group there were 1010.10 copies and these differences were also statistically significant. For telomere length evaluation the Telomere PNA/FITC FISH and Telomere PNA/FITC FISH for flow cytometry were used. The mean fluorescence intensity of telomere sequences calculated on the surface of interphase nuclei of leukaemic blood lymphocytes was lower than that in the control animals and the difference was statistically significant. The mean length of telomeres in BLV-infected and healthy cows was 31. 63 ± 12.62 and 38.4 ± 4.03, (p=0.112), respectively.

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Blood dendritic cells in cattle infected with bovine leukemia virus (BLV): isolation and phenotyping

Cited by 3 publications

References 42 publications

Cytokine secretion in stem cells of cattle infected with bovine leukaemia virus

Cytokine secretion in stem cells of cattle infected with bovine leukaemia virus

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

Polish Journal of Veterinary Sciences

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