Galectin-3 regulates the innate immune response of human monocytes - PubMed (original) (raw)

. 2013 Mar 15;207(6):947-56.

doi: 10.1093/infdis/jis920. Epub 2012 Dec 18.

Peter A Sieling, Mirjam Schenk, Rosane M B Teles, Stephan R Krutzik, Daniel K Hsu, Fu-Tong Liu, Euzenir N Sarno, Thomas H Rea, Steffen Stenger, Robert L Modlin, Delphine J Lee

Affiliations

Galectin-3 regulates the innate immune response of human monocytes

Andrew W Chung et al. J Infect Dis. 2013.

Abstract

Galectin-3 is a β-galactoside-binding lectin widely expressed on epithelial and hematopoietic cells, and its expression is frequently associated with a poor prognosis in cancer. Because it has not been well-studied in human infectious disease, we examined galectin-3 expression in mycobacterial infection by studying leprosy, an intracellular infection caused by Mycobacterium leprae. Galectin-3 was highly expressed on macrophages in lesions of patients with the clinically progressive lepromatous form of leprosy; in contrast, galectin-3 was almost undetectable in self-limited tuberculoid lesions. We investigated the potential function of galectin-3 in cell-mediated immunity using peripheral blood monocytes. Galectin-3 enhanced monocyte interleukin 10 production to a TLR2/1 ligand, whereas interleukin 12p40 secretion was unaffected. Furthermore, galectin-3 diminished monocyte to dendritic cell differentiation and T-cell antigen presentation. These data demonstrate an association of galectin-3 with unfavorable host response in leprosy and a potential mechanism for impaired host defense in humans.

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Figures

Figure 1.

Figure 1.

Galectin-3 expression in leprosy lesions. Galectin-3 protein expression in the skin lesions of lepromatous leprosy (L-lep; n = 3) and tuberculoid leprosy (T-lep; n = 3) patients was investigated by immunohistochemical staining with monoclonal mouse anti-human galectin-3 (B2C10). Four-micrometer tissue sections of leprosy skin biopsy specimens were incubated with anti–galectin-3 and labeled with an immunoperoxidase method followed by hematoxylin for visualization of histology. Tissue sections labeled with isotype control antibodies were negative. Photographs were taken using ×10 and ×40 objective lenses. Scale bars = 50 μm.

Figure 2.

Figure 2.

Coexpression of macrophage markers with galectin-3 (Gal-3) expression in lepromatous leprosy (L-lep) skin lesions. Skin lesions from L-lep patients were sectioned and immunolabeled with monoclonal antibodies as indicated and visualized by confocal laser microscopy. Images were photographed using a ×63 objective. Scale bars = 30 μm. CD14 and CD68 were visualized as green fluorescence, and Gal-3 (9C4) was visualized as red fluorescence. Nuclei were labeled with DAPI. Coexpression (yellow) revealed the relatively close proximity of Gal-3 to macrophage markers CD14 and CD68.

Figure 3.

Figure 3.

Effect of galectin-3 (Gal-3) on TLR2/1-induced cytokine profiles. Cytokine expression of (A) interleukin 10 (IL-10; n = 9 donors) and (B) interleukin 12p40 (IL-12p40; n = 7 donors) from monocytes treated with medium or 19-kD (a mycobacterial TLR2/1 ligand) in the presence of Gal-3 or control buffer (2 days) as measured by enzyme-linked immunosorbent assay. Values are expressed as the mean ± SEM of triplicates. Monocytes treated with medium alone secreted no IL-10, and IL-12p40 within a range from 0 to 132 pg/mL. Mixed-models statistical analysis was performed to compare Gal-3 with control buffer. In (A) compound symmetry variance–covariance structure was assumed, and in (B) unstructured variance–covariance structure was assumed. C, IL-10 (n = 3 donors) secretion from monocytes treated as in A and B, with the addition of lactose or sucrose (1 mM each). Each individual line represents 1 donor and the data points are the mean ± SEM of the triplicates for each experiment. Statistical analysis was performed using general linear model, and Tukey method was used for multiple comparison adjustment; *P = .001; **P < .0001. D, Dose titration of sugar inhibition of the effect of Gal-3 on IL-10 secretion. Results shown are from the same 3 donors as in (C). Values are reported as the percentage of decrease of 19-kD stimulated IL-10 secretion: [(pg/mL in the presence of Gal-3 – pg/mL in the presence of Gal-3 and sugar)/(pg/mL in the presence of Gal-3 – pg/mL in the absence of Gal-3)] × 100%. Statistical analysis was performed using mixed models assuming compound symmetry covariance matrix among repeated measures over several concentration levels.

Figure 4.

Figure 4.

Effect of galectin-3 (Gal-3) on granulocyte macrophage colony-stimulating factor (GM-CSF)–treated monocytes. CD1b and HLA-DR expression on monocytes treated with medium or GM-CSF in the presence of Gal-3 or control buffer for 2 days as measured by flow cytometry is shown. Cell surface protein expression levels are expressed as the percentage positive for CD1b from 14 donors (A) and HLA-DR from 3 donors (B). Monocytes treated with medium alone expressed CD1b within a range of 0.3% to 14.5%, and HLA-DR within a range of 81.5% to 98.3%. Statistical analysis was performed using paired t test (A) and sign test (B), which is a nonparametric version of the paired t test. C, CD1b expression on monocytes treated as in (A) from 3 donors, with the addition of lactose or sucrose (1 mM each). Statistical analysis was performed with general linear model using Tukey method for multiple comparison adjustment. D, Dose titration of sugar inhibition of the effect of Gal-3 on CD1b expression. Results shown are from the same 3 donors as in (C). Values are reported as the percentage of decrease in the percentage of CD1b-positive cells induced by GM-CSF: [|(percent in the presence of Gal-3 minus percent in the presence of Gal-3 and sugar)|/|(percent in the presence of Gal-3 minus percent in the absence of Gal-3)|] ×100%. Statistical analysis was performed using mixed models assuming compound symmetry covariance matrix among repeated measures over several concentrations.

Figure 5.

Figure 5.

Effect of galectin-3 (Gal-3) on antigen-presenting cells (APCs) on the ability to present antigen to T cells from leprosy patients. T-cell proliferation using APCs previously treated with granulocyte macrophage colony-stimulating factor in the presence of Gal-3 or control buffer as in Figure 4 are shown. In triplicate, CD1b-restricted T cells and APCs (1:1) were cultured in the presence or absence of mycobacterial antigen (M. tuberculosis sonicate or mycobacterial lipomannan) for 3 days, and proliferation was measured by 3H thymidine incorporation. Proliferative response was reported as average of counts per minute (cpm) of wells in the presence of antigen minus the average of cpm of corresponding wells in medium alone (Δ cpm). T cells cultured with medium alone–derived APCs proliferated at a mean of 65 Δ cpm (range, 2–207 cpm). Shown are results from 5 experiments, 4 donors. Statistical analysis was performed with paired t test.

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