Receptor mediated endocytosis by mesangial cells modulates transmigration of macrophages.
BACKGROUND
Accumulation of immune complexes in the mesangium is a common finding. Since migration of macrophages (Mphi) in the mesangium has been demonstrated to be an important event in the development of glomerular lesions, we studied the role of immune complexes and mesangial cell (MC) interaction in the transmigration (Tm) of Mphi.


METHODS
To determine the effect of MC and immune complexes (aggregated IgG, IgGAg) on transmigration of Mphi. MC were incubated with or without IgGAg in the lower compartment of a modified Boyden Chamber. To determine the effects of the secretory products (as a result of endocytosis of IgGAg by mesangial cells), MC-IgAg conditioned media was prepared and placed in the lower compartment of the Boyden chamber. We evaluated the effects of MC alone, MC + IgGAg, or MC-IgGAg conditioned media on the transmigration of macrophages across a filter. To determine the effect of free radicals on MC-IgAg endocytosis-induced Mphi migration we evaluated the effect of free radical scavengers such as dimethyl thiourea (DMTU) and tetramethylthiourea (TMTU) in MC-IgAg endocytosis-induced Mphi migration. To determine the role of chemokines in MC-IgAs endocytosis-induced Mphi migration we evaluated the effect of ani-MCP-1 antibodies on MC-IgAg endocytosis-induced Mphi migration, and also studied the effects of IgAg on MC mRNA expression of MCP-1 and RANTES. In addition, we evaluated the role of Fc receptors and actin cytoskeleton of MC in transmigration of Mphi.


RESULTS
Mesangial cell endocytosis of IgG aggregates (IgGAg) is associated with enhanced (P < 0.001) transmigration of Mphi (control, 11.2 +/- 0.2 vs. MC + IgGAg, 22.1 +/- 0.9 migrated Mphi/field). IgGAg also induced MC mRNA expression for RANTES and MCP-1 on MC. DMTU and TMTU attenuated (P < 0.001) the MC + IgGAg-induced migration of Mphi as well as IgGAg-induced mRNA expression for RANTES and MCP-1. MC and IgGAg interaction products (MC-IgGAg conditioned media) also increased (P < 0.01) transmigration of Mphi (control, 18.3 +/- 1.7 vs. MC-IgGAg conditioned media, 30.7 +/- 0.6 Mphi/field). This effect of MC-IgGAg conditioned media on the migration of macrophages was dose dependent. Anti-MCP-1 antibody partially inhibited MC-IgGAg-induced migration of macrophages. MC and monomeric IgG (MIgG) interaction (MC-MIgG conditioned media) showed a lower (P < 0.05) migration of Mphi, when compared to the MC-IgGAg conditioned media. MC-IgGAg conditioned media prepared from cytochalasin B pretreated MCs also showed a lower (P < 0.001) migration of Mphi when compared with MC-IgGAg conditioned media-induced migration.


CONCLUSIONS
These results indicate that MC-IgGAg conditioned media-induced transmigration of macrophages may be mediated through the generation of RANTES and MCP-1 by MC.
Singhal
P C
PC
Department of Medicine, Long Island Jewish Medical Center, Nev Hyde Park, New York 11040, USA.
Gupta
S
S
Sharma
P
P
Shah
H
H
Shah
N
N
Patel
P
P
D000818:Animals; D000906:Antibodies / Q000494:pharmacology; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects / Q000502:physiology; D018932:Chemokine CCL2 / Q000235:genetics / Q000276:immunology; D018946:Chemokine CCL5 / Q000235:genetics; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003571:Cytochalasin B / Q000494:pharmacology; D004705:Endocytosis / Q000502:physiology*; D016166:Free Radical Scavengers / Q000494:pharmacology; D005920:Glomerular Mesangium / Q000166:cytology / Q000502:physiology* / Q000648:ultrastructure; D007074:Immunoglobulin G / Q000494:pharmacology / Q000502:physiology; D008264:Macrophages / Q000502:physiology*; D051379:Mice; D008854:Microscopy, Electron; D012333:RNA, Messenger / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000502:physiology*; D013890:Thiourea / Q000031:analogs & derivatives* / Q000494:pharmacology
D000906:Antibodies; D018932:Chemokine CCL2; D018946:Chemokine CCL5; D017077:Culture Media, Conditioned; D016166:Free Radical Scavengers; D007074:Immunoglobulin G; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D003571:Cytochalasin B; C038983:1,3-dimethylthiourea; D013890:Thiourea; C019107:tetramethylthiourea