OBJECTIVE: Natural killer (NK) cells have long been known to be involved in the recognition and lysis of tumor cells but the mechanisms contributing to deficient NK activity in patients with cancer remains unclear. Manipulation of them is likely essential to the success of cancer immunotherapy protocols although optimal stimulation and maintenance of NK activity remains elusive. Here we studied the stimulatory effects of PHA and K562, on NK activity of human peripheral blood mononuclear cells (PBMCs). MATERIALS AND METHODS: The NK activity of PBMCs against DU-145 was determined with 51Cr-release assay. The PBMCs were stimulated with PHA, either on one occasion or repetitively on three occasions (1-PHA-PBMC or 3-PHA-PBMC, respectively), and were incubated with irradiated K562 (iK562). The expression of CD56, NKG2D and MICA/B were detected on PBMCs and cell lines with flow cytometry. RESULTS: PHA stimulation increased the proportion of CD56+ cells and upregulated NKG2D expression on 1-PHA-PBMC and 3-PHA-PBMC, but co-incubation with iK562 decreased NKG2D expression on 1-PHA-PBMC without change of NKG2D expression on the 3-PHA-PBMC. NK activity of 1-PHA-PBMC appeared to decrease with co-incubation with iK562 compared to a significant increase in activity of 3-PHA-PBMC. A similar increase in interferon-γ (IFN-γ) secretion from 3-PHA-PBMC was demonstrated compared to 1-PHA-PBMC. DISCUSSION AND CONCLUSION: Our results demonstrated that varying the mitogen exposure to PBMCs affect the influence of iK562 on NK activity. This effect appeared to be unrelated to the subsequent expression of NKG2D or IFN-γ secretion. These results may be beneficial in the development of future cancer immunotherapy protocols.
OBJECTIVE: Natural killer (NK) cells have long been known to be involved in the recognition and lysis of tumor cells but the mechanisms contributing to deficient NK activity in patients with cancer remains unclear. Manipulation of them is likely essential to the success of cancer immunotherapy protocols although optimal stimulation and maintenance of NK activity remains elusive. Here we studied the stimulatory effects of PHA and K562, on NK activity of human peripheral blood mononuclear cells (PBMCs). MATERIALS AND METHODS: The NK activity of PBMCs against DU-145 was determined with 51Cr-release assay. The PBMCs were stimulated with PHA, either on one occasion or repetitively on three occasions (1-PHA-PBMC or 3-PHA-PBMC, respectively), and were incubated with irradiated K562 (iK562). The expression of CD56, NKG2D and MICA/B were detected on PBMCs and cell lines with flow cytometry. RESULTS: PHA stimulation increased the proportion of CD56+ cells and upregulated NKG2D expression on 1-PHA-PBMC and 3-PHA-PBMC, but co-incubation with iK562 decreased NKG2D expression on 1-PHA-PBMC without change of NKG2D expression on the 3-PHA-PBMC. NK activity of 1-PHA-PBMC appeared to decrease with co-incubation with iK562 compared to a significant increase in activity of 3-PHA-PBMC. A similar increase in interferon-γ (IFN-γ) secretion from 3-PHA-PBMC was demonstrated compared to 1-PHA-PBMC. DISCUSSION AND CONCLUSION: Our results demonstrated that varying the mitogen exposure to PBMCs affect the influence of iK562 on NK activity. This effect appeared to be unrelated to the subsequent expression of NKG2D or IFN-γ secretion. These results may be beneficial in the development of future cancer immunotherapy protocols.
Authors: Kyo Chul Koo; Doo Hee Shim; Chang Mo Yang; Saet-Byul Lee; Shi Mun Kim; Tae Young Shin; Kwang Hyun Kim; Ho Geun Yoon; Koon Ho Rha; Jae Myun Lee; Sung Joon Hong Journal: PLoS One Date: 2013-11-04 Impact factor: 3.240