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Chemokine receptor expression in peripheral blood monocytes from patients with neovascular age-related macular
Michelle Grunin1, Tal Burstyn-Cohen, Shira Hagbi-Levi
1Department of Ophthalmology, Hadassah–Hebrew University Medical Center, Jerusalem, Israel.
Purpose:
Chemokine signaling and monocytes/macrophages were implicated in the pathogenesis of AMD. We tested the association between chemokines involved in monocyte recruitment and AMD.
Methods:
Immunophenotyping for white blood cell (WBC) populations including CD14++CD16- and CD14+CD16+ monocytes, CD19+, CD3+, and CD16+ lymphocytes, and chemokine receptors CCR1, CCR2, CCR5, CX(3)CR1, and CXCR4 was performed on peripheral blood from treatment-naïve neovascular AMD (NV-AMD) patients and controls. The mRNA level of chemokine receptors in monocytes was measured with quantitative-PCR. Systemic levels of major chemokine ligands CCL2, CCL5, CCL3, and CXCL10 were evaluated by ELISA. Genotyping was performed for risk SNPs for AMD in the CFH, C3, and HTRA1 genes.
Results:
The percentage of WBC subpopulations tested was similar between NV-AMD patients (n = 18) and controls (n = 20). CD14+CD16+ monocyte subpopulation showed a 3.5-fold increased expression of CCR1 (P = 0.039; t-test) and a 2.2-fold increased expression of CCR2 (P = 0.027) in patients compared with controls. Increased CCR1 and CCR2 expression was correlated with each other in patients (R(2) = 0.64, P < 0.0001), but not controls (R(2) = 0.02, P = 0.57). Increased mRNA levels of CCR1 (1.6-fold, P = 0.037) and CCR2 (1.6-fold, P = 0.007) were found in monocytes from NV-AMD patients. Chemokine receptor expression was not correlated with the presence of risk SNPs, and was not associated with blood chemokine levels.
Conclusions:
CCR1 and CCR2 are coupregulated on the CD14+CD16+ monocyte population in NV-AMD patients. These data implicate CD14+CD16+ monocytes and chemokine signaling in AMD. Additional investigation is needed to elucidate the role of these monocytes and their potential as a biomarker or therapeutic target for AMD.
Insights
Chemokine signaling is linked to age-related macular degeneration (AMD). Increased CCR1 and CCR2 expression on CD14+CD16+ monocytes in AMD patients suggests their role in disease pathogenesis.
Area of Science:
- Ophthalmology
- Immunology
- Genetics
Background:
- Chemokine signaling and monocyte/macrophage involvement are implicated in age-related macular degeneration (AMD) pathogenesis.
- Understanding the specific roles of chemokines in monocyte recruitment is crucial for AMD research.
Purpose of the Study:
- To investigate the association between chemokines involved in monocyte recruitment and neovascular AMD (NV-AMD).
- To analyze chemokine receptor expression on monocyte subpopulations in NV-AMD patients.
Main Methods:
- Immunophenotyping of peripheral blood white blood cells (WBCs), including monocyte subsets (CD14++CD16-, CD14+CD16+), and lymphocyte populations.
- Quantitative PCR to measure mRNA levels of chemokine receptors (CCR1, CCR2, CCR5, CX(3)CR1, CXCR4) in monocytes.
- ELISA to assess systemic levels of chemokine ligands (CCL2, CCL5, CCL3, CXCL10).
- Genotyping for AMD risk single nucleotide polymorphisms (SNPs) in CFH, C3, and HTRA1 genes.
Main Results:
- No significant difference in overall WBC subpopulations between NV-AMD patients (n=18) and controls (n=20).
- CD14+CD16+ monocytes showed significantly increased expression of CCR1 (3.5-fold) and CCR2 (2.2-fold) in NV-AMD patients compared to controls.
- Increased CCR1 and CCR2 expression in patients was correlated (R²=0.64, P<0.0001) and correlated mRNA levels were observed.
- Chemokine receptor expression did not correlate with AMD risk SNPs or systemic chemokine levels.
Conclusions:
- CCR1 and CCR2 are co-upregulated on the CD14+CD16+ monocyte population in NV-AMD patients.
- These findings implicate CD14+CD16+ monocytes and chemokine signaling in the pathogenesis of AMD.
- Further research is warranted to explore the role of these monocytes as potential biomarkers or therapeutic targets for AMD.
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