Monocyte chemoattractant protein 1-dependent leukocytic infiltrates are responsible for autoimmune disease in MRL-Fas(lpr) mice - PubMed (original) (raw)
Monocyte chemoattractant protein 1-dependent leukocytic infiltrates are responsible for autoimmune disease in MRL-Fas(lpr) mice
G H Tesch et al. J Exp Med. 1999.
Abstract
Infiltrating leukocytes may be responsible for autoimmune disease. We hypothesized that the chemokine monocyte chemoattractant protein (MCP)-1 recruits macrophages and T cells into tissues that, in turn, are required for autoimmune disease. Using the MRL-Fas(lpr) strain with spontaneous, fatal autoimmune disease, we constructed MCP-1-deficient MRL-Fas(lpr) mice. In MCP-1-intact MRL-Fas(lpr) mice, macrophages and T cells accumulate at sites (kidney tubules, glomeruli, pulmonary bronchioli, lymph nodes) in proportion to MCP-1 expression. Deleting MCP-1 dramatically reduces macrophage and T cell recruitment but not proliferation, protects from kidney, lung, skin, and lymph node pathology, reduces proteinuria, and prolongs survival. Notably, serum immunoglobulin (Ig) isotypes and kidney Ig/C3 deposits are not diminished in MCP-1-deficient MRL-Fas(lpr) mice, highlighting the requirement for MCP-1-dependent leukocyte recruitment to initiate autoimmune disease. However, MCP-1-deficient mice are not completely protected from leukocytic invasion. T cells surrounding vessels with meager MCP-1 expression remain. In addition, downstream effector cytokines/chemokines are decreased in MCP-1-deficient mice, perhaps reflecting a reduction of cytokine-expressing leukocytes. Thus, MCP-1 promotes MRL-Fas(lpr) autoimmune disease through macrophage and T cell recruitment, amplified by increasing local cytokines/chemokines. We suggest that MCP-1 is a principal therapeutic target with which to combat autoimmune diseases.
Figures
Figure 1
MCP transcripts increase with advancing disease in MRL-Faslpr kidneys. The renal cortex was isolated from C57BL/6 and MRL-Faslpr kidneys from 2 to 6 mo of age and was analyzed for MCP-1, -3, and -5 by reverse transcriptase (RT)-PCR. MCP-1, -3, and -5 are increased in MRL-Faslpr kidneys with progressive disease. Examples of the amplified PCR products are illustrated in the gel photos. Graph: data = mean ± SD; n = 3; *P < 0.01 vs. MRL-Faslpr (2 mo).
Figure 1
MCP transcripts increase with advancing disease in MRL-Faslpr kidneys. The renal cortex was isolated from C57BL/6 and MRL-Faslpr kidneys from 2 to 6 mo of age and was analyzed for MCP-1, -3, and -5 by reverse transcriptase (RT)-PCR. MCP-1, -3, and -5 are increased in MRL-Faslpr kidneys with progressive disease. Examples of the amplified PCR products are illustrated in the gel photos. Graph: data = mean ± SD; n = 3; *P < 0.01 vs. MRL-Faslpr (2 mo).
Figure 2
MCP-1 increases with advancing kidney disease in MRL-Faslpr mice. MCP-1 was assessed by immunostaining in (a) cortical tubules, (b) glomeruli, and (c) vessels in MRL-Faslpr kidneys. Data = mean ± SD; *P < 0.05; **P < 0.005. (d) The relative proportion of MRL-Faslpr kidney cells expressing MCP-1 was similar at 2, 4, and 6 mo of age. Few infiltrating cells in the interstitium (≤1%) express MCP-1.
Figure 3
MCP-1 expression in MRL-Faslpr kidney, lung, and lymph nodes. Tissues from MCP-1–intact (a–c) and MCP-1–deficient (e and f) MRL-Faslpr mice at 5 mo of age were immunostained for MCP-1. MCP-1 is strongly expressed by TECs and glomerular podocytes (a) but is weak in vessels (inset) in MCP-1–intact MRL-Faslpr kidneys. Bronchiolar epithelial cells are the main source of MCP-1 in MCP-1–intact MRL-Faslpr lungs (b). A large proportion of infiltrating cells surrounding lymphatics express MCP-1 within the enlarged lymph nodes of MCP-1–intact MRL-Faslpr mice (c). MCP-1 is not detected in MCP-1–deficient kidney, lung, and lymph node (d–f). a and d, ×800; b, c, e, and f, ×500.
Figure 4
MCP-1–deficient MRL-Faslpr mice are protected from lethal autoimmune injury. (a) We evaluated survival in MCP-1–intact (+/+, +/−) and –deficient (−/−) MRL-Faslpr mice (<50% male and female per group). The survival of MCP-1–intact compared with –deficient MRL-Faslpr mice is markedly reduced (P < 0.0001). In addition, MCP-1+/− MRL-Faslpr mice survive longer than the MCP-1+/+ strain (P < 0.0001). (b) As the collection of small daily volumes of urine is compromised by evaporation problems, we evaluated fresh urine samples using a spot analysis. However, spot analysis in individual samples has several limitations, including small sample volume and semiquantitative measurement. With these caveats in mind, we now report that MCP-1–deficient MRL-Faslpr mice are protected from proteinuria (2–6 mo) in comparison to the MCP-1–intact MRL-Faslpr strain. The number of surviving MCP-1–intact MRL-Faslpr mice declines rapidly after 6 mo of age; therefore, proteinuria at these ages is limited to a subset of MCP-1–intact MRL-Faslpr mice, which are more resistant to disease (normal = B6/129 wild type). (c) Lymphadenopathy is reduced in MCP-1–deficient MRL-Faslpr mice; however, lymphadenopathy is greater in MCP-1–intact MRL-Faslpr females as compared with males (P < 0.01; females in graph). (d) Inflammatory skin lesions are reduced in MCP-1–deficient compared with –intact strains. Data = mean ± SEM; *P < 0.05; **P < 0.005; and ***P < 0.0001 compared with MCP-1−/−.
Figure 5
Kidney and lung histopathology are reduced in MCP-1–deficient MRL-Faslpr mice. Tubular damage (arrowheads) and glomerular crescents (arrow) are severe in MCP-1–intact MRL-Faslpr kidneys (a; hematoxylin and PAS) and are reduced in the MCP-1–deficient MRL-Faslpr strain (b) at 5 mo of age. Peribronchial and perivascular cell infiltration in MRL-Faslpr lungs are prominent in the MCP-1 (+/+) strain at 5 mo of age (c). By comparison, the peribronchial infiltrate is reduced in the MCP-1 (−/−) MRL-Faslpr strain (d). a and b, ×800; c and d, ×500.
Figure 6
MCP-1–deficient MRL-Faslpr mice are protected from kidney and lung damage during renal disease. MCP-1 intact (+/+, +/−) and deficient (−/−) MRL-Faslpr kidneys were assessed for (a) tubular damage, (b) glomerular damage, and (c) perivascular cell infiltrate at 5 mo of age and compared with age-matched wild-type MRL++ C3H/FeJ strains. MCP-1–intact (+/+, +/−) and deficient (−/−) MRL-Faslpr lungs were analyzed for (d) peribronchiolar and (e) perivascular cell infiltrate at 5 mo of age. Kidney (tubular and glomerular) and lung (bronchiolar) pathology but not vasculitis was reduced in MCP-1–deficient versus MCP-1–intact MRL-Faslpr mice. Data = mean ± SD.
Figure 7
Kidney- and lung-infiltrating leukocytes are reduced in MCP-1–deficient MRL-Faslpr mice. Infiltrating leukocytes were assessed in the kidney (a–c) and lung (d–f) by immunostaining. Macrophage (MØ) accumulation adjacent to kidney parenchymal cells (peritubular, periglomerular, intraglomerular) in MCP-1–intact MRL-Faslpr mice (b) is markedly reduced in MCP-1–deficient MRL-Faslpr mice (c). Similarly, the notable macrophage accumulation adjacent to parenchymal cells in the lung (peribronchiolar) in MCP-1–intact MRL-Faslpr mice (e) is dramatically less in MCP-1–deficient MRL-Faslpr mice (f). Furthermore, T cells, including CD4 and CD8 T cells, are reduced in the peritubular area, whereas CD4 T cells accumulate less surrounding glomeruli in MCP-1–deficient versus MCP-1–intact MRL-Faslpr strains (a). In contrast, perivascular macrophages and T cells are not different in the MCP-1–intact and –deficient MRL-Faslpr lungs (d) and kidneys (not shown). Data = mean ± SD; n = 6; *P < 0.05 and **P < 0.01 compared with MCP-1+/+. F4/80 immunostaining: b and c, ×500; e and f, ×330.
Figure 9
(a) MCP-1 deficiency does not reduce serum Ig isotype levels in MRL-Faslpr mice. Serum from MCP-1–intact and –deficient MRL-Faslpr mice at 5 mo of age was analyzed for Igs by ELISA. (b) MCP-1–deficient MRL-Faslpr mice (5 mo of age) have equivalent amounts of IgG and C3 (not shown) in the kidney despite a reduction in loss of renal function, glomerular and tubular injury, and enhanced survival as compared with the MCP-1–intact strain. Data = mean ± SD; n = 6.
Figure 8
MCP-1 deficiency reduces CSF-1, IFN-γ, and other CCR2 ligands in MRL-Faslpr kidneys. Transcript levels of CSF-1, IFN-γ, MCP-3, and MCP-5 were assessed in comparison to GAPDH in MCP-1–intact and –deficient MRL-Faslpr and B6/129 mouse renal cortices at 5 mo of age by RT-PCR. CSF-1, IFN-γ, MCP-3, and MCP-5 transcripts were reduced in MCP-1–deficient versus MCP-1–intact MRL-Faslpr mice. Data = mean ± SD; n = 6; P values compared with MCP-1–intact MRL-Faslpr mice.
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