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Tumor cell lysate induces the immunosuppression and apoptosis of mouse immunocytes.

Dong B, Dai G, Xu L, Zhang Y, Ling L, Sun L, Lv J - Mol Med Rep (2014)

Bottom Line: Furthermore, the present study found that the immunosuppressive factor, hyaluronan, and the apoptosis inducers, Fas ligand and transforming growth factor-β, are present in TCL.These components may be associated with the emergence of immunosuppressive cells or splenocyte apoptosis.Thus, the present study has enriched our understanding of the composition of TCL and its negative regulatory effect on immunocytes.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Wannan Medical College, Wuhu, Anhui 241002, P.R. China.

ABSTRACT
Although tumor cell lysate (TCL) is a type of immunocyte stimulator, its immunosuppressive function must not be ignored. The present study reported that TCL prepared from a Lewis lung cancer cell was able to induce the development of immunosuppressive macrophages (MΦ) and tolerogenic dendritic cells. In addition, TCL upregulated the expression of CD69 in mouse splenocytes, and cell apoptosis and the percentage of regulatory T cells in mouse splenocytes simultaneously increased. Furthermore, the present study found that the immunosuppressive factor, hyaluronan, and the apoptosis inducers, Fas ligand and transforming growth factor-β, are present in TCL. These components may be associated with the emergence of immunosuppressive cells or splenocyte apoptosis. Thus, the present study has enriched our understanding of the composition of TCL and its negative regulatory effect on immunocytes.

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Fas-L and TGF-β expression in TCL. (A) TCL was prepared from 1×106 Lewis lung cancer cells. Immunoblot analysis was processed to detect the Fas-L in TCL with anti-Fas-L antibody. FasL (Lane 1 of NC membrane) was detected in the TCL, but bovine serum albumin (Lane 2 of NC membrane) and mycobacterial heat shock protein 65 (Lane 3 of NC membrane) control proteins were not detected. (B) TCL was produced from 1×106 or 2×106 Lewis lung cancer cells. ELISA was processed to detect the TGF-β in TCL and the concentration of HA was calculated by comparing the actual OD value of TCL to that of different HA standard samples. The actual OD value of TCL was: TCL OD (TCL-1 or TCL-2 well) - NaCl OD. The actual OD of the standard sample was: HA standard samples OD - Blank well OD. TCL-1 indicates TCL from 2×106 Lewis lung cancer cells. TCL-2 signifies TCL from 1×106 Lewis lung cancer cells. NC, nitrocellulose; Fas-L, Fas ligand; TGF-β, transforming growth factor-β; TCL, tumor cell lysate; HA, hyaluronan; OD, optical density; NaCl, sodium chloride; M, marker.
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f2-mmr-10-06-2827: Fas-L and TGF-β expression in TCL. (A) TCL was prepared from 1×106 Lewis lung cancer cells. Immunoblot analysis was processed to detect the Fas-L in TCL with anti-Fas-L antibody. FasL (Lane 1 of NC membrane) was detected in the TCL, but bovine serum albumin (Lane 2 of NC membrane) and mycobacterial heat shock protein 65 (Lane 3 of NC membrane) control proteins were not detected. (B) TCL was produced from 1×106 or 2×106 Lewis lung cancer cells. ELISA was processed to detect the TGF-β in TCL and the concentration of HA was calculated by comparing the actual OD value of TCL to that of different HA standard samples. The actual OD value of TCL was: TCL OD (TCL-1 or TCL-2 well) - NaCl OD. The actual OD of the standard sample was: HA standard samples OD - Blank well OD. TCL-1 indicates TCL from 2×106 Lewis lung cancer cells. TCL-2 signifies TCL from 1×106 Lewis lung cancer cells. NC, nitrocellulose; Fas-L, Fas ligand; TGF-β, transforming growth factor-β; TCL, tumor cell lysate; HA, hyaluronan; OD, optical density; NaCl, sodium chloride; M, marker.

Mentions: Cancer cells may induce the apoptosis of immune cells by secreting pro-apoptotic factors (Fas-L and TGF-β), which is one of mechanisms underlying the escape of cancer cells from antitumor immunity (14–19). In the present study, western blot analysis and ELISA were performed to detect Fas-L and TGF-β in the TCL from Lewis lung cancer cells (Fig. 2A). Western blot analysis demonstrated that the Fas-L concentration was 200 ng/ml in the TCL from the Lewis cells (1×106) and that the Fas-L detected by the western blot assay had a high specificity and could not react with non-specific proteins (including MHSP65 and bovine serum albumin). ELISA demonstrated that the TGF-β concentration was 239.64 pg/ml in the TCL from the Lewis cells (1×106). However, no TGF-β was identified in the TCL from the Lewis cells in the NaCl group (Fig. 2B).


Tumor cell lysate induces the immunosuppression and apoptosis of mouse immunocytes.

Dong B, Dai G, Xu L, Zhang Y, Ling L, Sun L, Lv J - Mol Med Rep (2014)

Fas-L and TGF-β expression in TCL. (A) TCL was prepared from 1×106 Lewis lung cancer cells. Immunoblot analysis was processed to detect the Fas-L in TCL with anti-Fas-L antibody. FasL (Lane 1 of NC membrane) was detected in the TCL, but bovine serum albumin (Lane 2 of NC membrane) and mycobacterial heat shock protein 65 (Lane 3 of NC membrane) control proteins were not detected. (B) TCL was produced from 1×106 or 2×106 Lewis lung cancer cells. ELISA was processed to detect the TGF-β in TCL and the concentration of HA was calculated by comparing the actual OD value of TCL to that of different HA standard samples. The actual OD value of TCL was: TCL OD (TCL-1 or TCL-2 well) - NaCl OD. The actual OD of the standard sample was: HA standard samples OD - Blank well OD. TCL-1 indicates TCL from 2×106 Lewis lung cancer cells. TCL-2 signifies TCL from 1×106 Lewis lung cancer cells. NC, nitrocellulose; Fas-L, Fas ligand; TGF-β, transforming growth factor-β; TCL, tumor cell lysate; HA, hyaluronan; OD, optical density; NaCl, sodium chloride; M, marker.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4227419&req=5

f2-mmr-10-06-2827: Fas-L and TGF-β expression in TCL. (A) TCL was prepared from 1×106 Lewis lung cancer cells. Immunoblot analysis was processed to detect the Fas-L in TCL with anti-Fas-L antibody. FasL (Lane 1 of NC membrane) was detected in the TCL, but bovine serum albumin (Lane 2 of NC membrane) and mycobacterial heat shock protein 65 (Lane 3 of NC membrane) control proteins were not detected. (B) TCL was produced from 1×106 or 2×106 Lewis lung cancer cells. ELISA was processed to detect the TGF-β in TCL and the concentration of HA was calculated by comparing the actual OD value of TCL to that of different HA standard samples. The actual OD value of TCL was: TCL OD (TCL-1 or TCL-2 well) - NaCl OD. The actual OD of the standard sample was: HA standard samples OD - Blank well OD. TCL-1 indicates TCL from 2×106 Lewis lung cancer cells. TCL-2 signifies TCL from 1×106 Lewis lung cancer cells. NC, nitrocellulose; Fas-L, Fas ligand; TGF-β, transforming growth factor-β; TCL, tumor cell lysate; HA, hyaluronan; OD, optical density; NaCl, sodium chloride; M, marker.
Mentions: Cancer cells may induce the apoptosis of immune cells by secreting pro-apoptotic factors (Fas-L and TGF-β), which is one of mechanisms underlying the escape of cancer cells from antitumor immunity (14–19). In the present study, western blot analysis and ELISA were performed to detect Fas-L and TGF-β in the TCL from Lewis lung cancer cells (Fig. 2A). Western blot analysis demonstrated that the Fas-L concentration was 200 ng/ml in the TCL from the Lewis cells (1×106) and that the Fas-L detected by the western blot assay had a high specificity and could not react with non-specific proteins (including MHSP65 and bovine serum albumin). ELISA demonstrated that the TGF-β concentration was 239.64 pg/ml in the TCL from the Lewis cells (1×106). However, no TGF-β was identified in the TCL from the Lewis cells in the NaCl group (Fig. 2B).

Bottom Line: Furthermore, the present study found that the immunosuppressive factor, hyaluronan, and the apoptosis inducers, Fas ligand and transforming growth factor-β, are present in TCL.These components may be associated with the emergence of immunosuppressive cells or splenocyte apoptosis.Thus, the present study has enriched our understanding of the composition of TCL and its negative regulatory effect on immunocytes.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry, Wannan Medical College, Wuhu, Anhui 241002, P.R. China.

ABSTRACT
Although tumor cell lysate (TCL) is a type of immunocyte stimulator, its immunosuppressive function must not be ignored. The present study reported that TCL prepared from a Lewis lung cancer cell was able to induce the development of immunosuppressive macrophages (MΦ) and tolerogenic dendritic cells. In addition, TCL upregulated the expression of CD69 in mouse splenocytes, and cell apoptosis and the percentage of regulatory T cells in mouse splenocytes simultaneously increased. Furthermore, the present study found that the immunosuppressive factor, hyaluronan, and the apoptosis inducers, Fas ligand and transforming growth factor-β, are present in TCL. These components may be associated with the emergence of immunosuppressive cells or splenocyte apoptosis. Thus, the present study has enriched our understanding of the composition of TCL and its negative regulatory effect on immunocytes.

Show MeSH
Related in: MedlinePlus