Identification of cell surface receptors for murine macrophage inflammatory protein-1 alpha.
We have produced recombinant proteins for a cytokine, L2G25BP (macrophage inflammatory protein-1 alpha) (MIP-1 alpha). By using the recombinant protein (rMIP-1 alpha), receptors for MIP-1 alpha were identified on Con A-stimulated and unstimulated CTLL-R8, a T cell line, and LPS-stimulated RAW 264.7, a macrophage cell line. The 125I-rMIP-1 alpha binds to the receptor in a specific and saturable manner. Scatchard analysis indicated a single class of high affinity receptor, with a Kd of approximately 1.5 x 10(-9) M and approximately 1200 binding sites/Con A-stimulated CTLL-R8 cell and a Kd of 0.9 x 10(-9) M and approximately 380 binding sites/RAW 264.7 cell. 125I-rMIP-1 alpha binding was inhibited by unlabeled rMIP-1 alpha in a dose-dependent manner, but not by IL-1 alpha or IL-2. rMIP-1 alpha inhibited the proliferation of unstimulated CTLL-R8 cells. Rabbit anti-rMIP-1 alpha antibodies blocked the growth-inhibitory effect of the rMIP-1 alpha on CTLL-R8 cells.
Oh
K O
KO
Department of Microbiology and Immunology, Walther Oncology Center, Indiana University School of Medicine, Indianapolis 46202.
Zhou
Z
Z
Kim
K K
KK
Samanta
H
H
Fraser
M
M
Kim
Y J
YJ
Broxmeyer
H E
HE
Kwon
B S
BS
D000818:Animals; D015152:Blotting, Northern; D002455:Cell Division; D002460:Cell Line; D054407:Chemokine CCL4; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D003602:Cytotoxicity, Immunologic; D015870:Gene Expression; D007111:Immunity, Cellular; D066298:In Vitro Techniques; D019402:Macrophage Inflammatory Proteins; D008264:Macrophages / Q000166:cytology / Q000502:physiology*; D051379:Mice; D008970:Molecular Weight; D015846:Monokines / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics; D011956:Receptors, Cell Surface / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000166:cytology / Q000502:physiology*
D054407:Chemokine CCL4; D016207:Cytokines; D019402:Macrophage Inflammatory Proteins; D015846:Monokines; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins