Macrophage-like Cell Line (HS-P) from a Rat Histiocytic Sarcoma

doi:10.1053/jcpa.2000.0452

Journal of Comparative Pathology

Volume 124, Issues 2–3, February 2001, Pages 183-191

https://doi.org/10.1053/jcpa.2000.0452 Get rights and content

Abstract

With future exploration of macrophage properties in mind, we established a novel cell line (HS-P) from a transplantable histiocytic sarcoma, derived originally from a tumour in an aged F344 rat. HS-P was subjected to 70 serial passages, in which the mean doubling time was 15·7 h. The cells, which were round, oval or polygonal in shape, were arranged in a compact sheet. They reacted to varying degrees for lysosomal enzymes (acid phosphatase and non-specific esterase) and with the following antibodies: ED1/ED2 (rat macrophage/histiocyte-specific), OX6 (rat MHC class II-specific), lysozyme antibody and α1-antichymotrypsin antibody. Electron microscopically, HS-P cells showed lysosomes and prominent cell projections. These findings indicated that the cultured cells were macrophage-like. Syngeneic rats inoculated subcutaneously or intraperitoneally with HS-P cells invariably developed sarcomatous tumours consisting of monomorphic mononuclear cells, which exhibited cytochemical properties similar to those of cultured HS-P cells. Bioassay and reverse transcription-polymerase chain reaction methods revealed that tumour necrosis factor-α increased on addition of lipopolysaccharide (LPS), indicating that HS-P cells remained LPS-responsive. HS-P cells may prove to be a useful tool for in-vitro studies of macrophage function.

References (33)

H.T. Cook et al.
Interleukin-4 ameliorates crescentic glomerulonephritis in Wistar Kyoto rats
Kidney International
((1999))
B.E. Drysdale et al.
Identification of a lipopolysaccharide inducible transcription factor in murine macrophages
Molecular Immunology
((1996))
T.W. Jungi et al.
Human macrophages respond to LPS in a serum-independent, CD14-dependent manner
Immunology Letters
((1996))
J. Yamate et al.
Morphological characteristics of a transplantable Íhistiocytic sarcoma (HS-J) in F344 rats and appearance of renal tubular hyaline droplets in HS-J-bearing rats
Journal of Comparative Pathology
((1997))
S.J. Baker et al.
The Salmonella typhi melittin resistance gene pqaB affects intracellular growth in PMA-differentiated U937 cells, polymyxin B resistance and lipopolysaccharide
Microbiology
((1999))
M. Benahmed et al.
LPS increases biomedical degradation by human monocytes in vitro
Journal of Biomedical Materials Research
((1997))
S.J. Benn et al.
Effect of extracellular matrix on gene expression and mRNA stability in primary rat hepatocytes
Journal of Toxicologic Pathology
((1998))
J.G. Damoiseaux et al.
Rat macrophage lysosomal membrane antigen recognized by monoclonal antibody ED1
Immunology
((1994))
J. Delabie et al.
True histiocytic neoplasm of Langerhan's cell type
Journal of Pathology
((1991))
A.J. Demetris et al.
Analysis of chronic rejection and obliterative arteriopathy: possible contributions of donor-antigen-presenting cells and lymphatic disruption
American Journal of Pathology
((1997))

C.D. Dijkstra et al.

The heterogeneity of mononuclear phagocytes in lymphoid organs: distinct macrophage subpopulations in the rat recognized by monoclonal antibodies ED1, ED2 and ED3

Immunology

((1985))

S. Eustis et al.

Liver

In Pathology of the Fischer Rat G. A. BoorÍman, S. L. Eustis, M. R. Elwell, C. A. Montgomery Jr and W. F. MacKenzie, Eds, Academic Press, San Diego

((1990))

J.E. Hines et al.

In vivo responses of macrophages and perisinusoidal cells to cholestatic liver injury

American Journal of Pathology

((1993))

S.M. Hsu et al.

Lymphomas of true histiocytic origin. Expression of different phenotypes in so-called true histiocytic lymphoma and malignant histiocytosis

American Journal of Pathology

((1991))

K. Iwabuchi et al.

Csk overexpression reduces several monokines and nitric oxide production but enhances prostaglandin E2 production in response to lipopolysaccharide in the macrophage cell line J774A·1

European Journal of Immunology

((1997))

J.E. Laithwaite et al.

Enhanced macrophage resistance to Pseudomonas exotoxin A is correlated with decreased LRP expression

Infection and Immunity

((1999))

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Regular ArticleMacrophage-like Cell Line (HS-P) from a Rat Histiocytic Sarcoma

Abstract

Kidney International

Molecular Immunology

Immunology Letters

Journal of Comparative Pathology

The Salmonella typhi melittin resistance gene pqaB affects intracellular growth in PMA-differentiated U937 cells, polymyxin B resistance and lipopolysaccharide

Microbiology

LPS increases biomedical degradation by human monocytes in vitro

Journal of Biomedical Materials Research

Effect of extracellular matrix on gene expression and mRNA stability in primary rat hepatocytes

Journal of Toxicologic Pathology

Rat macrophage lysosomal membrane antigen recognized by monoclonal antibody ED1

Immunology

True histiocytic neoplasm of Langerhan's cell type

Journal of Pathology

Analysis of chronic rejection and obliterative arteriopathy: possible contributions of donor-antigen-presenting cells and lymphatic disruption

American Journal of Pathology

The heterogeneity of mononuclear phagocytes in lymphoid organs: distinct macrophage subpopulations in the rat recognized by monoclonal antibodies ED1, ED2 and ED3

Immunology

Liver

In Pathology of the Fischer Rat G. A. BoorÍman, S. L. Eustis, M. R. Elwell, C. A. Montgomery Jr and W. F. MacKenzie, Eds, Academic Press, San Diego

In vivo responses of macrophages and perisinusoidal cells to cholestatic liver injury

American Journal of Pathology

Lymphomas of true histiocytic origin. Expression of different phenotypes in so-called true histiocytic lymphoma and malignant histiocytosis

American Journal of Pathology

Csk overexpression reduces several monokines and nitric oxide production but enhances prostaglandin E2 production in response to lipopolysaccharide in the macrophage cell line J774A·1

European Journal of Immunology

Enhanced macrophage resistance to Pseudomonas exotoxin A is correlated with decreased LRP expression

Infection and Immunity

Regular Article
Macrophage-like Cell Line (HS-P) from a Rat Histiocytic Sarcoma