J Vet Sci.  2016 Dec;17(4):453-458. 10.4142/jvs.2016.17.4.453.

Paclitaxel inhibits the hyper-activation of spleen cells by lipopolysaccharide and induces cell death

• Kim HJ
• Joo HG

• KMID: 2412600
• DOI: http://doi.org/10.4142/jvs.2016.17.4.453

Abstract

Paclitaxel was isolated from the bark of the Pacific yew, Taxus brevifolia, and used as an anticancer agent. Paclitaxel prevents cancer cell division by inhibiting spindle fiber function, inducing cell death. A recent study demonstrated that paclitaxel binds to myeloid differentiation protein-2 of Toll-like receptor 4 and prevents the signal transduction of lipopolysaccharide (LPS). Paclitaxel converts immune cells hypo-responsive to LPS. In this study, we investigated whether paclitaxel can inhibit the phenotype and function of immune cells. To accomplish this, we used spleen cells, a major type of immune cell, LPS, a representative inflammatory agent and a mitogen for B lymphocytes. LPS profoundly increased the activation and cytokine production of spleen cells. However, paclitaxel significantly inhibited LPS-induced hyper-activation of spleen cells. Furthermore, we found that paclitaxel induced cell death of LPS-treated spleen cells. These results suggest that paclitaxel can inhibit the hyper-immune response of LPS in spleen cells via a variety of mechanisms. These findings suggest that paclitaxel can be used as a modulating agent for diseases induced by hyper-activation of B lymphocytes. Taken together, these results demonstrate that paclitaxel inhibits the function of spleen cells activated by LPS, and further induces cell death.

Keyword

cell death; hyper-activation; lipopolysaccharide; paclitaxel; spleen cells

MeSH Terms

Animals
Antineoplastic Agents, Phytogenic/*pharmacology
Apoptosis/*drug effects
Cell Proliferation/*drug effects
Cytokines/*metabolism
Lipopolysaccharides/toxicity
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL
Paclitaxel/*pharmacology
Spleen/*drug effects
Antineoplastic Agents, Phytogenic
Cytokines
Lipopolysaccharides
Paclitaxel

Figure

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  Fig. 1 Paclitaxel decreases the activity and proliferation of lipopolysaccharide (LPS)-treated spleen cells. The cells were seeded at a concentration of 2 × 106 cells/mL in 96-well culture plates, then treated with 5-fold serial dilutions of LPS and 5.0 µg/mL paclitaxel for 3 days. MTT assay was performed as described in the Materials and Methods. The optical density (O.D.) of samples was measured at 570 nm using a microplate reader and expressed as mean ± standard deviation (SD).

• 

  Fig. 2 Paclitaxel alters the cell size and number of LPS-activated spleen cells. Cells were seeded at a concentration of 2 × 106 cells/mL in 6-well culture plates, then treated with a control (medium alone), paclitaxel (5.0 µg/mL), LPS (1.0 µg/mL), or LPS + paclitaxel. After 3 days of treatment, (A) the cells were analyzed using flow cytometry. The circles in the dot plots indicate the population of activated and big cells (forward scattered light [FSC]high cells). (B) The cell morphology was observed by an inverted optical microscope with a digital camera. The circle in the picture indicates a cluster of cells proliferating and the arrow indicates swollen cells.

• 

  Fig. 3 Paclitaxel decreases the action potential of mitochondria and induces death of LPS-activated spleen cells. The cells were setup and treated as described in Fig. 2. (A) Cells were stained with Rhodamine 123 solution. (B) Cells were stained with propidium iodide (PI) solution. Cells in the area of FSClow and PIhigh indicate dead cells.

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  Fig. 4 Paclitaxel decreases the expression of cluster of differentiation (CD)25, an activation marker on LPS-activated spleen cells. The cells were cultured and treated as described in Fig. 2. The cells were stained for mouse CD25 or CD69 molecules, then analyzed by flow cytometry. The number of histograms indicates geometric mean fluorescence intensity.

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  Fig. 5 Paclitaxel decreases the production of tumor necrosis factor-alpha (TNF-α) and IL-10 of LPS-activated spleen cells. The cells were treated with 5-fold serial dilutions of LPS and 5.0 µg/mL paclitaxel for 3 days in 96-well culture plates, after which the culture supernatants were used for TNF-α and IL-10 enzyme-linked immunosorbent assay. Data are presented as the means ± SD.

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