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An Ectopic Chemokine Expression Model for Testing Macrophage Recruitment In Vivo
Published on: September 25, 2019
Chemokine expression in experimental tubulointerstitial nephritis
1Department of Pathology, Amgen Inc., Thousand Oaks, CA 91320, USA.
Insights
Monocyte chemotactic protein-1 (MCP-1) and interferon-inducible protein-10 (IP-10) chemokines are upregulated in puromycin aminonucleoside (PAN) nephrosis, driving leukocyte infiltration. Neutralizing MCP-1 significantly reduced macrophage infiltration in tubulointerstitial nephritis.
Area of Science:
- Nephrology
- Immunology
- Molecular Biology
Background:
- Tubulointerstitial nephritis is a key feature of glomerular disease.
- Chemokines mediate leukocyte infiltration in inflammatory kidney diseases.
Purpose of the Study:
- To investigate the role of specific chemokines in puromycin aminonucleoside (PAN) nephrosis-induced tubulointerstitial nephritis.
- To determine the cellular source and temporal expression of chemokines during PAN nephrosis.
Main Methods:
- Quantitative analysis of renal cortical chemokine mRNA expression (IP-10, MCP-1, RANTES, CINC, MIP-2, MIP-1alpha) at various time points post-PAN induction.
- Localization of chemokine mRNA production using in situ hybridization.
- Measurement of MCP-1 protein levels via ELISA.
- Assessment of tubulointerstitial leukocyte infiltration (T lymphocytes, macrophages).
- In vivo blockade of MCP-1 using a neutralizing antibody.
Main Results:
- A significant, transient increase in renal cortical IP-10 and MCP-1 mRNA expression was observed 6-8 days after PAN administration.
- IP-10 and MCP-1 mRNA were produced by intrinsic tubulointerstitial cells.
- MCP-1 protein levels correlated with increased T lymphocyte and macrophage infiltration, peaking around day 10.
- Neutralization of MCP-1 led to a 45% reduction in tubulointerstitial macrophage infiltration on day 6.
Conclusions:
- MCP-1 is a critical chemokine involved in monocyte/macrophage recruitment in PAN nephrosis.
- IP-10 may also play a role in the inflammatory process.
- Targeting MCP-1 could be a therapeutic strategy for tubulointerstitial nephritis associated with glomerular disease.
Abstract:
Chemokines may be important in the pathogenesis of leukocyte infiltration in tubulointerstitial nephritis associated with glomerular disease. We studied the renal cortical expression of the C-C (macrophage inflammatory protein-1alpha (MIP-1alpha)), monocyte chemotactic protein-1 (MCP-1), and RANTES) and C-X-C (interferon-inducible protein-10 (IP-10), MIP-2, and cytokine-induced neutrophil chemoattractant (CINC)) chemokines 4, 6, 8, 10, 14, and 21 days after the induction of puromycin aminonucleoside (PAN) nephrosis. There was a 7- to 10-fold increase in the steady state mRNA expression of IP-10 and MCP-1 in the renal cortex of rats 6 to 8 days after the administration of PAN that declines thereafter reaching control values by day 21. The site of IP-10 and MCP-1 mRNA production was localized to intrinsic tubulointerstitial cells and not to infiltrating monocytes or macrophages. By comparison, there was a low basal expression of RANTES mRNA in the renal cortex of nephrotic rats that did not differ from those of control rats. In contrast, CINC, MIP-2, and MIP-1alpha mRNAs were not detected. Translation of MCP-1 mRNA into protein was confirmed with an ELISA. These changes in chemokine gene expression were associated with a tubulointerstitial T lymphocyte and macrophage infiltration beginning on day 6 that peaked on day 10. Administration of a neutralizing Ab to rat MCP-1 (n = 5) beginning on day 4 resulted in a 45% decline in tubulointerstitial macrophage infiltration from 8.4 +/- 1.3% to 4.6 +/- 0.4% (p < 0.001) on day 6. These data provide evidence that MCP-1, and possibly IP-10, are important in the pathogenesis of monocyte/macrophage infiltration in the tubulointerstitial nephritis associated with PAN nephrosis.
