Figure 4.
Figure 4. Culture assay of the human PBMCs in vitro. (A) Culture assay of human PBMCs exposed to Elo and NK cells. The frequency of fibrocytes significantly increased in the NK cell group (n = 6) on days 8, 9, and 10 compared with the control group (n = 6). In contrast, the frequency of fibrocytes significantly decreased in the Elo group (n = 6) and Elo plus NK cell group (n = 6). In addition, the frequency of fibrocytes significantly decreased in the Elo plus NK cell group compared with the Elo group on days 8, 9 and 10. (B) The long-term culture assay of human PBMCs exposed to Elo. The frequency of fibrocytes significantly decreased in the Elo 50 μg/mL group (n = 6) and Elo 100 μg/mL group (n = 6) compared with the control group (n = 6) throughout the observation period (from days 8 to 31). No significant difference was observed in the frequency of fibrocytes between the Elo 50 μg/mL and Elo 100 μg/mL groups. (C) Culture assay of human PBMCs exposed to Elo and other agents. Compared with the Rom group (n = 6), the frequency of fibrocytes significantly decreased in the Rom plus Elo group (n = 6), Rom plus Elo plus Rux group (n = 6), and Rom plus IFN-α2 group (n = 6) throughout the observation period (from days 8 to 10). In contrast, there was no significant difference between the Rom plus Elo and Rom plus Elo plus Rux groups in terms of fibrocyte differentiation. Rom plus IFN-α2 had the strongest suppressive effect on fibrocyte differentiation; however, approximately 80% to 90% of the cultured cells were dead and floated in the Rom plus IFN-α2 group, and we considered that IFN-α2 might be too cytotoxic in this setting. (D) Culture assay of PBMCs of from MF patients. Elo significantly decreased the frequency of fibrocytes derived from MPN patients harboring a CALR mutation, similar to fibrocytes derived from MPN patients with a JAK2V617F mutation. Results were reported as the mean ± standard deviation values for data obtained from 3 individual experiments with 6 HCs or 6 patients in each group. Statistical significance of the data for each day was calculated by 1-way ANOVA with post hoc Tukey’s honestly significant difference test. ns: not significant. *P < .05; **P < .01.

Culture assay of the human PBMCs in vitro. (A) Culture assay of human PBMCs exposed to Elo and NK cells. The frequency of fibrocytes significantly increased in the NK cell group (n = 6) on days 8, 9, and 10 compared with the control group (n = 6). In contrast, the frequency of fibrocytes significantly decreased in the Elo group (n = 6) and Elo plus NK cell group (n = 6). In addition, the frequency of fibrocytes significantly decreased in the Elo plus NK cell group compared with the Elo group on days 8, 9 and 10. (B) The long-term culture assay of human PBMCs exposed to Elo. The frequency of fibrocytes significantly decreased in the Elo 50 μg/mL group (n = 6) and Elo 100 μg/mL group (n = 6) compared with the control group (n = 6) throughout the observation period (from days 8 to 31). No significant difference was observed in the frequency of fibrocytes between the Elo 50 μg/mL and Elo 100 μg/mL groups. (C) Culture assay of human PBMCs exposed to Elo and other agents. Compared with the Rom group (n = 6), the frequency of fibrocytes significantly decreased in the Rom plus Elo group (n = 6), Rom plus Elo plus Rux group (n = 6), and Rom plus IFN-α2 group (n = 6) throughout the observation period (from days 8 to 10). In contrast, there was no significant difference between the Rom plus Elo and Rom plus Elo plus Rux groups in terms of fibrocyte differentiation. Rom plus IFN-α2 had the strongest suppressive effect on fibrocyte differentiation; however, approximately 80% to 90% of the cultured cells were dead and floated in the Rom plus IFN-α2 group, and we considered that IFN-α2 might be too cytotoxic in this setting. (D) Culture assay of PBMCs of from MF patients. Elo significantly decreased the frequency of fibrocytes derived from MPN patients harboring a CALR mutation, similar to fibrocytes derived from MPN patients with a JAK2V617F mutation. Results were reported as the mean ± standard deviation values for data obtained from 3 individual experiments with 6 HCs or 6 patients in each group. Statistical significance of the data for each day was calculated by 1-way ANOVA with post hoc Tukey’s honestly significant difference test. ns: not significant. *P < .05; **P < .01.

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