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Galectin Hco-gal-m from Haemonchus contortus modulates goat monocytes and T cell function in different patterns.

Wang W, Wang S, Zhang H, Yuan C, Yan R, Song X, Xu L, Li X - Parasit Vectors (2014)

Bottom Line: Different members of the galectin family show multiple and distinct regulatory effects on different cell types.Previous studies have demonstrated that the galectin from Haemonchus contortus (Hco-gal-m) performed immunomodulatory effects on goat PBMC, however, which subpopulation of PBMC is the primary target of Hco-gal-m and whether the immune modulations share the same mechanism remain unclear.Consequently, T cell proliferations were potently inhibited by rHco-gal-m.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China. lixiangrui@njau.edu.cn.

ABSTRACT

Background: Monocytes and T cells are two major subpopulations of peripheral blood mononuclear cells (PBMC) and play an essential role in the innate and adaptive immune systems. Different members of the galectin family show multiple and distinct regulatory effects on different cell types. Previous studies have demonstrated that the galectin from Haemonchus contortus (Hco-gal-m) performed immunomodulatory effects on goat PBMC, however, which subpopulation of PBMC is the primary target of Hco-gal-m and whether the immune modulations share the same mechanism remain unclear.

Methods: In this study, the developmental expression of Hco-gal-m was analyzed by RT-PCR and Western blot analysis. The distribution of Hco-gal-m in adult worm was detected by an immunohistochemical test. The binding activity of the recombinant Hco-gal-m (rHco-gal-m) on goat monocytes and T cells were assessed by flow cytometry. The immunomodulatory effects of Hco-gal-m on cytokine secretion, cell activation and apoptosis were observed by co-incubation of rHco-gal-m with goat monocytes and T cells.

Results: Hco-gal-m was expressed in L4 as well as adult worms and predominantly localized at the internal surface of the worm guts. rHco-gal-m could bind to both monocytes and T cells. The engagement of rHco-gal-m decreased the production of IL-6, IL-10 and TNF-α in T cells, however, it significantly increased the secretion of IL-10 in monocytes. After rHco-gal-m exposure, the expression of MHC-II on monocytes and that of CD25 on T cells were restricted. Consequently, T cell proliferations were potently inhibited by rHco-gal-m. In addition, rHco-gal-m induced apoptosis in T cells, but not significantly in monocytes.

Conclusions: Our results indicated that rHco-gal-m modulated goat monocytes and T cell function in different patterns.

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Related in: MedlinePlus

Inhibitory effect of rHco-gal-m on T cell activation and proliferation. (A) T cells were activated with ConA and incubated at the same time with serial concentrations of rHco-gal-m at 37°C and 5% CO2. The proliferation was measured by CCK-8 incorporation after 72 h. Cell proliferation index was calculated considering the OD450 values in controls as 100%. (B) T cells were simultaneously treated with ConA and rHco-gal-m at 37°C and 5% CO2 for 24 h. The expression of CD25, CCNA1, CCNB1 and CCND1 was analyzed by real-time PCR. All experiments were set up in triplicate. The data are representative of three independent experiments (*p < 0.05 and **p < 0.01).
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Fig6: Inhibitory effect of rHco-gal-m on T cell activation and proliferation. (A) T cells were activated with ConA and incubated at the same time with serial concentrations of rHco-gal-m at 37°C and 5% CO2. The proliferation was measured by CCK-8 incorporation after 72 h. Cell proliferation index was calculated considering the OD450 values in controls as 100%. (B) T cells were simultaneously treated with ConA and rHco-gal-m at 37°C and 5% CO2 for 24 h. The expression of CD25, CCNA1, CCNB1 and CCND1 was analyzed by real-time PCR. All experiments were set up in triplicate. The data are representative of three independent experiments (*p < 0.05 and **p < 0.01).

Mentions: As demonstrated by incorporation of CCK-8, rHco-gal-m significantly inhibited the proliferation of T cells in vitro (Figure 6A). This inhibitory effect was further supported by the expression of some marker genes. Real-time PCR analyses proved that, rHco-gal-m significantly inhibited the transcription of CD25, a sensitive marker for T cell activation, and the cell cycle genes, CCNA1 (cyclin A1) and CCND1 (cyclin D1) (Figure 6B).Figure 6


Galectin Hco-gal-m from Haemonchus contortus modulates goat monocytes and T cell function in different patterns.

Wang W, Wang S, Zhang H, Yuan C, Yan R, Song X, Xu L, Li X - Parasit Vectors (2014)

Inhibitory effect of rHco-gal-m on T cell activation and proliferation. (A) T cells were activated with ConA and incubated at the same time with serial concentrations of rHco-gal-m at 37°C and 5% CO2. The proliferation was measured by CCK-8 incorporation after 72 h. Cell proliferation index was calculated considering the OD450 values in controls as 100%. (B) T cells were simultaneously treated with ConA and rHco-gal-m at 37°C and 5% CO2 for 24 h. The expression of CD25, CCNA1, CCNB1 and CCND1 was analyzed by real-time PCR. All experiments were set up in triplicate. The data are representative of three independent experiments (*p < 0.05 and **p < 0.01).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4117971&req=5

Fig6: Inhibitory effect of rHco-gal-m on T cell activation and proliferation. (A) T cells were activated with ConA and incubated at the same time with serial concentrations of rHco-gal-m at 37°C and 5% CO2. The proliferation was measured by CCK-8 incorporation after 72 h. Cell proliferation index was calculated considering the OD450 values in controls as 100%. (B) T cells were simultaneously treated with ConA and rHco-gal-m at 37°C and 5% CO2 for 24 h. The expression of CD25, CCNA1, CCNB1 and CCND1 was analyzed by real-time PCR. All experiments were set up in triplicate. The data are representative of three independent experiments (*p < 0.05 and **p < 0.01).
Mentions: As demonstrated by incorporation of CCK-8, rHco-gal-m significantly inhibited the proliferation of T cells in vitro (Figure 6A). This inhibitory effect was further supported by the expression of some marker genes. Real-time PCR analyses proved that, rHco-gal-m significantly inhibited the transcription of CD25, a sensitive marker for T cell activation, and the cell cycle genes, CCNA1 (cyclin A1) and CCND1 (cyclin D1) (Figure 6B).Figure 6

Bottom Line: Different members of the galectin family show multiple and distinct regulatory effects on different cell types.Previous studies have demonstrated that the galectin from Haemonchus contortus (Hco-gal-m) performed immunomodulatory effects on goat PBMC, however, which subpopulation of PBMC is the primary target of Hco-gal-m and whether the immune modulations share the same mechanism remain unclear.Consequently, T cell proliferations were potently inhibited by rHco-gal-m.

View Article: PubMed Central - PubMed

Affiliation: College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China. lixiangrui@njau.edu.cn.

ABSTRACT

Background: Monocytes and T cells are two major subpopulations of peripheral blood mononuclear cells (PBMC) and play an essential role in the innate and adaptive immune systems. Different members of the galectin family show multiple and distinct regulatory effects on different cell types. Previous studies have demonstrated that the galectin from Haemonchus contortus (Hco-gal-m) performed immunomodulatory effects on goat PBMC, however, which subpopulation of PBMC is the primary target of Hco-gal-m and whether the immune modulations share the same mechanism remain unclear.

Methods: In this study, the developmental expression of Hco-gal-m was analyzed by RT-PCR and Western blot analysis. The distribution of Hco-gal-m in adult worm was detected by an immunohistochemical test. The binding activity of the recombinant Hco-gal-m (rHco-gal-m) on goat monocytes and T cells were assessed by flow cytometry. The immunomodulatory effects of Hco-gal-m on cytokine secretion, cell activation and apoptosis were observed by co-incubation of rHco-gal-m with goat monocytes and T cells.

Results: Hco-gal-m was expressed in L4 as well as adult worms and predominantly localized at the internal surface of the worm guts. rHco-gal-m could bind to both monocytes and T cells. The engagement of rHco-gal-m decreased the production of IL-6, IL-10 and TNF-α in T cells, however, it significantly increased the secretion of IL-10 in monocytes. After rHco-gal-m exposure, the expression of MHC-II on monocytes and that of CD25 on T cells were restricted. Consequently, T cell proliferations were potently inhibited by rHco-gal-m. In addition, rHco-gal-m induced apoptosis in T cells, but not significantly in monocytes.

Conclusions: Our results indicated that rHco-gal-m modulated goat monocytes and T cell function in different patterns.

Show MeSH
Related in: MedlinePlus