Characterization of lymphotoxin-alpha beta complexes on the surface of mouse lymphocytes.
The lymphotoxin-alpha beta complex (LT alpha beta) is found on the surface of activated lymphocytes and binds to a specific receptor called the LT beta receptor (LT beta R). In the mouse, signaling through this pathway is important for lymph node development and splenic organization, yet the biochemical properties of murine LT alpha and LT beta are essentially unknown. Here we have used soluble receptor-Ig forms of LT beta R and TNF-R55 and mAbs specific for murine LT alpha, LT beta, and LT beta R to characterize the appearance of surface LT alpha beta complexes and LT beta R on several common murine cell lines. Cells that bound LT beta R also bound anti-LT alpha and anti-LT beta mAbs in a FACS analysis. The ability of these reagents to discriminate between surface TNF and LT was verified by analysis of surface TNF-positive, LPS-activated murine RAW 264.7 monocytic cells. Primary mouse leukocytes from spleen, thymus, lymph node, and peritoneum were activated in vitro, and CD4+ and CD8+ T cells as well as B cells expressed surface LT ligand but not the LT beta R. Conversely, elicited peritoneal monocytes/macrophages were surface LT negative yet LT beta R positive. This study shows that on mononuclear cells, surface LT complexes and receptor are expressed similarly in mice and man, and the tools described herein form the foundation for study of the functional roles of the LT system in the mouse.
Browning
J L
JL
Department of Immunology, Biogen, Cambridge, MA 02142, USA. Jeff_Browning@biogen.com
Sizing
I D
ID
Lawton
P
P
Bourdon
P R
PR
Rennert
P D
PD
Majeau
G R
GR
Ambrose
C M
CM
Hession
C
C
Miatkowski
K
K
Griffiths
D A
DA
Ngam-ek
A
A
Meier
W
W
Benjamin
C D
CD
Hochman
P S
PS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry; D000918:Antibody Specificity; D001402:B-Lymphocytes / Q000737:chemistry; D002460:Cell Line; D005434:Flow Cytometry; D006801:Humans; D006825:Hybridomas; D007136:Immunoglobulins / Q000235:genetics / Q000378:metabolism; D008214:Lymphocytes / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism*; D016399:Lymphoma, T-Cell; D008233:Lymphotoxin-alpha / Q000737:chemistry* / Q000276:immunology; D053308:Lymphotoxin-beta; D008264:Macrophages; D008565:Membrane Proteins / Q000737:chemistry* / Q000276:immunology; D051379:Mice; D051381:Rats; D018124:Receptors, Tumor Necrosis Factor / Q000235:genetics / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012995:Solubility; D013045:Species Specificity; D013601:T-Lymphocytes / Q000737:chemistry; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism
D000911:Antibodies, Monoclonal; D007136:Immunoglobulins; C505886:LTB protein, human; C505887:Ltb protein, mouse; C505888:Ltb protein, rat; D008233:Lymphotoxin-alpha; D053308:Lymphotoxin-beta; D008565:Membrane Proteins; D018124:Receptors, Tumor Necrosis Factor; D011993:Recombinant Fusion Proteins; D014409:Tumor Necrosis Factor-alpha