Expression and characterisation of equine interleukin 2 and interleukin 4

Karen Dohmann; Bettina Wagner; David W. Horohov; Wolfgang Leibold

doi:10.1016/S0165-2427(00)00249-X

Expression and characterisation of equine interleukin 2 and interleukin 4

Karen Dohmann, Bettina Wagner, David W. Horohov, Wolfgang Leibold

Veterinary Science

Research output: Contribution to journal › Article › peer-review

26 Scopus citations

Abstract

In the present study, we describe the expression of equine IL2 and IL4. The cDNA of equine IL2 or IL4 was cloned in a mammalian expression vector, containing c-terminal myc- and six histidines His₆-epitopes for recognition and purification of equine cytokines. The vector constructs were used for transfection of chinese hamster ovary (CHO) cells. Purified equine cytokines were characterised by western blotting. Equine IL2 was secreted with a molecular weight of approximately 17.1 kDa, whereas IL4 was expressed in three different sizes of 17.1, 19.6 and 22.1 kDa, probably due to different glycosylation modifications. The biological activities of both cytokines were tested by proliferation assays using leukoagglutinin (LAG) prestimulated equine PBMC. Both, equine IL2 and IL4 induced dose-dependent lymphocyte proliferation. In contrast to IL4, IL2 supported the proliferation of B cells.

Original language	English
Pages (from-to)	243-256
Number of pages	14
Journal	Veterinary Immunology and Immunopathology
Volume	77
Issue number	3-4
DOIs	https://doi.org/10.1016/S0165-2427(00)00249-X
State	Published - Dec 29 2000

Bibliographical note

Funding Information:
We thank Dr. D.P. Lunn for the gift of the monoclonal antibodies CVS4 and CVS8. This work was supported by the DFG Graduiertenkolleg: Zell- und Molekularbiologie in der Tiermedizin in Hannover.

Keywords

Biochemical characterisation
Horse IL2 and IL4
Interleukins
Mammalian expression
Proliferations assays

ASJC Scopus subject areas

Immunology
General Veterinary

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10.1016/S0165-2427(00)00249-X

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title = "Expression and characterisation of equine interleukin 2 and interleukin 4",

abstract = "In the present study, we describe the expression of equine IL2 and IL4. The cDNA of equine IL2 or IL4 was cloned in a mammalian expression vector, containing c-terminal myc- and six histidines His6-epitopes for recognition and purification of equine cytokines. The vector constructs were used for transfection of chinese hamster ovary (CHO) cells. Purified equine cytokines were characterised by western blotting. Equine IL2 was secreted with a molecular weight of approximately 17.1 kDa, whereas IL4 was expressed in three different sizes of 17.1, 19.6 and 22.1 kDa, probably due to different glycosylation modifications. The biological activities of both cytokines were tested by proliferation assays using leukoagglutinin (LAG) prestimulated equine PBMC. Both, equine IL2 and IL4 induced dose-dependent lymphocyte proliferation. In contrast to IL4, IL2 supported the proliferation of B cells.",

keywords = "Biochemical characterisation, Horse IL2 and IL4, Interleukins, Mammalian expression, Proliferations assays",

author = "Karen Dohmann and Bettina Wagner and Horohov, {David W.} and Wolfgang Leibold",

note = "Funding Information: We thank Dr. D.P. Lunn for the gift of the monoclonal antibodies CVS4 and CVS8. This work was supported by the DFG Graduiertenkolleg: Zell- und Molekularbiologie in der Tiermedizin in Hannover.",

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AU - Dohmann, Karen

AU - Wagner, Bettina

AU - Horohov, David W.

AU - Leibold, Wolfgang

N1 - Funding Information: We thank Dr. D.P. Lunn for the gift of the monoclonal antibodies CVS4 and CVS8. This work was supported by the DFG Graduiertenkolleg: Zell- und Molekularbiologie in der Tiermedizin in Hannover.

PY - 2000/12/29

Y1 - 2000/12/29

N2 - In the present study, we describe the expression of equine IL2 and IL4. The cDNA of equine IL2 or IL4 was cloned in a mammalian expression vector, containing c-terminal myc- and six histidines His6-epitopes for recognition and purification of equine cytokines. The vector constructs were used for transfection of chinese hamster ovary (CHO) cells. Purified equine cytokines were characterised by western blotting. Equine IL2 was secreted with a molecular weight of approximately 17.1 kDa, whereas IL4 was expressed in three different sizes of 17.1, 19.6 and 22.1 kDa, probably due to different glycosylation modifications. The biological activities of both cytokines were tested by proliferation assays using leukoagglutinin (LAG) prestimulated equine PBMC. Both, equine IL2 and IL4 induced dose-dependent lymphocyte proliferation. In contrast to IL4, IL2 supported the proliferation of B cells.

AB - In the present study, we describe the expression of equine IL2 and IL4. The cDNA of equine IL2 or IL4 was cloned in a mammalian expression vector, containing c-terminal myc- and six histidines His6-epitopes for recognition and purification of equine cytokines. The vector constructs were used for transfection of chinese hamster ovary (CHO) cells. Purified equine cytokines were characterised by western blotting. Equine IL2 was secreted with a molecular weight of approximately 17.1 kDa, whereas IL4 was expressed in three different sizes of 17.1, 19.6 and 22.1 kDa, probably due to different glycosylation modifications. The biological activities of both cytokines were tested by proliferation assays using leukoagglutinin (LAG) prestimulated equine PBMC. Both, equine IL2 and IL4 induced dose-dependent lymphocyte proliferation. In contrast to IL4, IL2 supported the proliferation of B cells.

KW - Biochemical characterisation

KW - Horse IL2 and IL4

KW - Interleukins

KW - Mammalian expression

KW - Proliferations assays

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IS - 3-4

ER -

Expression and characterisation of equine interleukin 2 and interleukin 4

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

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