Expression and distribution of the Kit receptor in bovine bone marrow cells

Hirokazu Hikono National Institute of Animal Health, Tsukuba, Ibaraki 305-0856, Japan.

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 DVM
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Masato Ohta National Institute of Animal Health, Tsukuba, Ibaraki 305-0856, Japan.

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 PhD
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Jian-Hua Zhou Institute of Society for Techno-innovation of Agriculture, Forestry, and Fisheries, Tsukuba, Ibaraki 305-0854, Japan.
Present address is the Edward G. Madigan Laboratory, University of Illinois, Urbana, IL 61801.

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Michiharu Sakurai National Institute of Animal Health, Tsukuba, Ibaraki 305-0856, Japan.

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Abstract

Objective—To characterize the expression and distribution of the Kit receptor in bovine bone marrow cells (BMC) and to define the function of its ligand, stem cell factor (SCF).

Animals—Six 7- to 70-day-old healthy male Holstein-Friesian calves.

Procedures—Expression and distribution of the Kit receptor were assessed by use of flow cytometry with monoclonal antibodies (mAb) against the bovine Kit protein. Using Giemsa-stained centrifuged preparations, the histologic appearance of Kit receptor positive (Kit+) BMC were evaluated. Semisolid cultures supplemented with granulocyte colony-stimulating factor (G-CSF) and SCF were used to measure the colony formation capacity of Kit+ BMC.

Results—The Kit receptor was expressed on approximately 18% of total BMC. Most of Kit+ BMC did not coexpress lineage markers, but a small subset of this population did coexpress CD3. The Kit+CD3– BMC were a heterogeneous cell population comprising blast-like cells such as myeloblasts, promyelocytes, rubriblasts, and prorubricytes. Conversely, Kit+CD3+ BMC had a lymphocyte-like appearance. Kit+ BMC formed colonies in semisolid culture with G-CSF, whereas Kit– BMC failed to grow. Addition of SCF to G-CSF resulted in superadditive enhancement in colony numbers and size.

Conclusions—The Kit receptor is expressed primarily on immature blood cells in bovine bone marrow, and Kit+ BMC contain hematopoietic progenitor cells that are reactive to G-CSF. In addition, SCF synergizes with G-CSF to stimulate colony formation by these cells. Our results suggest that the Kit receptor and its ligand, SCF, are involved in early stages of granulopoiesis in calves. ( Am J Vet Res 2001;62:974–977)

Abstract

Objective—To characterize the expression and distribution of the Kit receptor in bovine bone marrow cells (BMC) and to define the function of its ligand, stem cell factor (SCF).

Animals—Six 7- to 70-day-old healthy male Holstein-Friesian calves.

Procedures—Expression and distribution of the Kit receptor were assessed by use of flow cytometry with monoclonal antibodies (mAb) against the bovine Kit protein. Using Giemsa-stained centrifuged preparations, the histologic appearance of Kit receptor positive (Kit+) BMC were evaluated. Semisolid cultures supplemented with granulocyte colony-stimulating factor (G-CSF) and SCF were used to measure the colony formation capacity of Kit+ BMC.

Results—The Kit receptor was expressed on approximately 18% of total BMC. Most of Kit+ BMC did not coexpress lineage markers, but a small subset of this population did coexpress CD3. The Kit+CD3– BMC were a heterogeneous cell population comprising blast-like cells such as myeloblasts, promyelocytes, rubriblasts, and prorubricytes. Conversely, Kit+CD3+ BMC had a lymphocyte-like appearance. Kit+ BMC formed colonies in semisolid culture with G-CSF, whereas Kit– BMC failed to grow. Addition of SCF to G-CSF resulted in superadditive enhancement in colony numbers and size.

Conclusions—The Kit receptor is expressed primarily on immature blood cells in bovine bone marrow, and Kit+ BMC contain hematopoietic progenitor cells that are reactive to G-CSF. In addition, SCF synergizes with G-CSF to stimulate colony formation by these cells. Our results suggest that the Kit receptor and its ligand, SCF, are involved in early stages of granulopoiesis in calves. ( Am J Vet Res 2001;62:974–977)

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