<abstract xmlns="http://www.w3.org/1999/xhtml">

<sec id="j_jvetres-2021-0043_s_005">
<h3>Introduction</h3>
<p>Granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) are cytokines widely used in <italic>ex vivo</italic> monocyte differentiation experiments, vaccine formulations and disease treatment. The aim of this study was to produce recombinant bovine GM-CSF and IL-4 in an episomal expression system that conserves the postransductional modification of the native proteins and to use the products to differentiate bovine monocytes into dendritic cells.</p></sec>
<sec id="j_jvetres-2021-0043_s_006">
<h3>Material and Methods</h3>
<p>The recombinant proteins rGM-CSF and rIL-4 were expressed in PEAKrapid CRL-2828 human kidney cells, ATCC CRL-2828. The functional activity of the recombinant cytokines was monitored by registering morphological changes in bovine monocytes and assessing the expression of CD14 upon incubation with them.</p></sec>
<sec id="j_jvetres-2021-0043_s_007">
<h3>Results</h3>
<p>Both recombinant proteins were detected in the cell culture supernatant of transfected cells. Culture supernatants of transfected cells induced in bovine monocytes morphological changes that resemble macrophages or dendritic cells. In addition, bovine cells treated with rGM-CSF and rIL-4 showed reduced expression of the macrophage surface marker CD14 compared with untreated cells. This effect indicates the expected differentiation. The expression of the cytokines was stable after many successive cell passages and a freeze/thaw cycle.</p></sec>
<sec id="j_jvetres-2021-0043_s_008">
<h3>Conclusions</h3>
<p>The semi-stable mammalian episomal expression system used in this study allowed us to easily produce functional bovine rGM-CSF and rIL-4 without the need for protein purification steps.</p></sec></abstract>

Introduction
Granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) are cytokines widely used in ex vivo monocyte differentiation experiments, vaccine formulations and disease treatment. The aim of this study was to produce recombinant bovine GM-CSF and IL-4 in an episomal expression system that conserves the postransductional modification of the native proteins and to use the products to differentiate bovine monocytes into dendritic cells.

Material and Methods
The recombinant proteins rGM-CSF and rIL-4 were expressed in PEAKrapid CRL-2828 human kidney cells, ATCC CRL-2828. The functional activity of the recombinant cytokines was monitored by registering morphological changes in bovine monocytes and assessing the expression of CD14 upon incubation with them.

Results
Both recombinant proteins were detected in the cell culture supernatant of transfected cells. Culture supernatants of transfected cells induced in bovine monocytes morphological changes that resemble macrophages or dendritic cells. In addition, bovine cells treated with rGM-CSF and rIL-4 showed reduced expression of the macrophage surface marker CD14 compared with untreated cells. This effect indicates the expected differentiation. The expression of the cytokines was stable after many successive cell passages and a freeze/thaw cycle.

Conclusions
The semi-stable mammalian episomal expression system used in this study allowed us to easily produce functional bovine rGM-CSF and rIL-4 without the need for protein purification steps.

Semi-stable production of bovine IL-4 and GM-CSF in the mammalian episomal expression system

Journal of Veterinary Research

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Introduction
Granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) are cytokines widely used in ex vivo monocyte differentiation...

Semi-stable production of bovine IL-4 and GM-CSF in the mammalian episomal expression system

Pubblicato online: 19 ago 2021

Pagine: 315 - 321

Ricevuto: 22 gen 2021

Accettato: 26 lug 2021

DOI: https://doi.org/10.2478/jvetres-2021-0043

Parole chiave
bovine GM-CSF, bovine IL-4, dendritic cells, macrophages, mammalian episomal expression

© 2021 F.C. Blanco et al. published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1

Fig. 2

Fig. 3

Semi-stable production of bovine IL-4 and GM-CSF in the mammalian episomal expression system

Pubblicato online: 19 ago 2021

Pagine: 315 - 321

Ricevuto: 22 gen 2021

Accettato: 26 lug 2021

DOI: https://doi.org/10.2478/jvetres-2021-0043

Parole chiavebovine GM-CSF, bovine IL-4, dendritic cells, macrophages, mammalian episomal expression

© 2021 F.C. Blanco et al. published by Sciendo

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1

Fig. 2

Fig. 3

Parole chiave
bovine GM-CSF, bovine IL-4, dendritic cells, macrophages, mammalian episomal expression