Characterisation of bovine transferrin receptor on normal activated and Theileria parva-transformed lymphocytes by a new monoclonal antibody

Jan Naessens, Dennis J. Grab, Gerhard Fritsch

Research output: Contribution to journalArticle

Abstract

A murine IgM monoclonal antibody (mAb), IL-A77, has been generated that recognises the bovine transferrin receptor (TfR) and will be a useful tool to measure the activation state of bovine lymphocytes and macrophages. The antigen is detected on immature erythroid cells and proliferating lymphocytes. It is undetectable on resting lymphocytes, but appears within 24 h after stimulation with concanavalin A (ConA) or pokeweed mitogen (PWM). Immune precipitations of lysates of both labeled activated lymphocytes and bone marrow erythroid cells showed that, similar to human TfR, to bovine receptor is a disulfide-bonded dimer of two identical chains of M(r) 970000. A similar 97000 M(r) protein was eluted from a column containing immobilised bovine transferrin (Tf) using conditions known to elute the human TfR, and this protein was recognised by mAb IL-A77, proving that it detected bovine TfR. Although the mAb inhibited binding of transferrin to its receptor, it did not block proliferation of Theileria parva-transformed or ConA-stimulated lymphocytes. When cells were metabolically labeled with 35S-methionine, a second 90000-M(r) TfR band was detected in Theileria parva-transformed cells, but not in stimulated lymphocytes. This form of the TfR was not expressed on the cell surface. It may be an unusual precursor of the receptor, a parasite-modified receptor or it may be of parasite origin and necessary for transfer of iron into the intracellular parasite.

Original languageEnglish (US)
Pages (from-to)65-76
Number of pages12
JournalVeterinary Immunology and Immunopathology
Volume52
Issue number1-2
DOIs
StatePublished - Jun 15 1996
Externally publishedYes

Fingerprint

Theileria parva
Transferrin Receptors
transferrin
monoclonal antibodies
lymphocytes
Monoclonal Antibodies
Lymphocytes
receptors
cattle
Erythroid Cells
Parasites
Transferrin
Concanavalin A
Pokeweed Mitogens
concanavalin A
parasites
Immunoprecipitation
Bone Marrow Cells
Disulfides
Methionine

Keywords

  • bovine
  • monoclonal antibody
  • Theileria parva
  • transferrin receptor

ASJC Scopus subject areas

  • Animal Science and Zoology
  • Immunology
  • veterinary(all)

Cite this

Characterisation of bovine transferrin receptor on normal activated and Theileria parva-transformed lymphocytes by a new monoclonal antibody. / Naessens, Jan; Grab, Dennis J.; Fritsch, Gerhard.

In: Veterinary Immunology and Immunopathology, Vol. 52, No. 1-2, 15.06.1996, p. 65-76.

Research output: Contribution to journalArticle

@article{b594272c3def4c708b5a7ae8f62c975f,
title = "Characterisation of bovine transferrin receptor on normal activated and Theileria parva-transformed lymphocytes by a new monoclonal antibody",
abstract = "A murine IgM monoclonal antibody (mAb), IL-A77, has been generated that recognises the bovine transferrin receptor (TfR) and will be a useful tool to measure the activation state of bovine lymphocytes and macrophages. The antigen is detected on immature erythroid cells and proliferating lymphocytes. It is undetectable on resting lymphocytes, but appears within 24 h after stimulation with concanavalin A (ConA) or pokeweed mitogen (PWM). Immune precipitations of lysates of both labeled activated lymphocytes and bone marrow erythroid cells showed that, similar to human TfR, to bovine receptor is a disulfide-bonded dimer of two identical chains of M(r) 970000. A similar 97000 M(r) protein was eluted from a column containing immobilised bovine transferrin (Tf) using conditions known to elute the human TfR, and this protein was recognised by mAb IL-A77, proving that it detected bovine TfR. Although the mAb inhibited binding of transferrin to its receptor, it did not block proliferation of Theileria parva-transformed or ConA-stimulated lymphocytes. When cells were metabolically labeled with 35S-methionine, a second 90000-M(r) TfR band was detected in Theileria parva-transformed cells, but not in stimulated lymphocytes. This form of the TfR was not expressed on the cell surface. It may be an unusual precursor of the receptor, a parasite-modified receptor or it may be of parasite origin and necessary for transfer of iron into the intracellular parasite.",
keywords = "bovine, monoclonal antibody, Theileria parva, transferrin receptor",
author = "Jan Naessens and Grab, {Dennis J.} and Gerhard Fritsch",
year = "1996",
month = "6",
day = "15",
doi = "10.1016/0165-2427(95)05537-1",
language = "English (US)",
volume = "52",
pages = "65--76",
journal = "Veterinary Immunology and Immunopathology",
issn = "0165-2427",
publisher = "Elsevier",
number = "1-2",

}

TY - JOUR

T1 - Characterisation of bovine transferrin receptor on normal activated and Theileria parva-transformed lymphocytes by a new monoclonal antibody

AU - Naessens, Jan

AU - Grab, Dennis J.

AU - Fritsch, Gerhard

PY - 1996/6/15

Y1 - 1996/6/15

N2 - A murine IgM monoclonal antibody (mAb), IL-A77, has been generated that recognises the bovine transferrin receptor (TfR) and will be a useful tool to measure the activation state of bovine lymphocytes and macrophages. The antigen is detected on immature erythroid cells and proliferating lymphocytes. It is undetectable on resting lymphocytes, but appears within 24 h after stimulation with concanavalin A (ConA) or pokeweed mitogen (PWM). Immune precipitations of lysates of both labeled activated lymphocytes and bone marrow erythroid cells showed that, similar to human TfR, to bovine receptor is a disulfide-bonded dimer of two identical chains of M(r) 970000. A similar 97000 M(r) protein was eluted from a column containing immobilised bovine transferrin (Tf) using conditions known to elute the human TfR, and this protein was recognised by mAb IL-A77, proving that it detected bovine TfR. Although the mAb inhibited binding of transferrin to its receptor, it did not block proliferation of Theileria parva-transformed or ConA-stimulated lymphocytes. When cells were metabolically labeled with 35S-methionine, a second 90000-M(r) TfR band was detected in Theileria parva-transformed cells, but not in stimulated lymphocytes. This form of the TfR was not expressed on the cell surface. It may be an unusual precursor of the receptor, a parasite-modified receptor or it may be of parasite origin and necessary for transfer of iron into the intracellular parasite.

AB - A murine IgM monoclonal antibody (mAb), IL-A77, has been generated that recognises the bovine transferrin receptor (TfR) and will be a useful tool to measure the activation state of bovine lymphocytes and macrophages. The antigen is detected on immature erythroid cells and proliferating lymphocytes. It is undetectable on resting lymphocytes, but appears within 24 h after stimulation with concanavalin A (ConA) or pokeweed mitogen (PWM). Immune precipitations of lysates of both labeled activated lymphocytes and bone marrow erythroid cells showed that, similar to human TfR, to bovine receptor is a disulfide-bonded dimer of two identical chains of M(r) 970000. A similar 97000 M(r) protein was eluted from a column containing immobilised bovine transferrin (Tf) using conditions known to elute the human TfR, and this protein was recognised by mAb IL-A77, proving that it detected bovine TfR. Although the mAb inhibited binding of transferrin to its receptor, it did not block proliferation of Theileria parva-transformed or ConA-stimulated lymphocytes. When cells were metabolically labeled with 35S-methionine, a second 90000-M(r) TfR band was detected in Theileria parva-transformed cells, but not in stimulated lymphocytes. This form of the TfR was not expressed on the cell surface. It may be an unusual precursor of the receptor, a parasite-modified receptor or it may be of parasite origin and necessary for transfer of iron into the intracellular parasite.

KW - bovine

KW - monoclonal antibody

KW - Theileria parva

KW - transferrin receptor

UR - http://www.scopus.com/inward/record.url?scp=0029905476&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029905476&partnerID=8YFLogxK

U2 - 10.1016/0165-2427(95)05537-1

DO - 10.1016/0165-2427(95)05537-1

M3 - Article

C2 - 8807777

AN - SCOPUS:0029905476

VL - 52

SP - 65

EP - 76

JO - Veterinary Immunology and Immunopathology

JF - Veterinary Immunology and Immunopathology

SN - 0165-2427

IS - 1-2

ER -