Directs effects of granulocyte-macrophage colony stimulating factor (GM-CSF) on adaptive immunogenesis

Figures & data

Table 1. Percentages of CD14⁺, CD16⁺, CD119⁺, CD124⁺, CD197⁺ cells in non-activated Mc/Mph cell cultures

CD marker	GM-CSF (ng/ml)
	0.0	0.01	0.1	1.0	10.0
CD14	25.05 (23.07–32.16)	32.30 (11.94–38.25)	22.76 (11.43–34.53)	23.82 (9.55–33.49)	19.72 (11.69–41.79)
CD16	11.06 (4.87–16.35)	5.95 (3.82–11.60)	4.59 (3.52–17.39)	8.7 (3.32–17.09)	8.35 (3.32–18.32)
CD119	83.16 (78.07–88.75)	74.58 (58.49–82.18)	80.79 (69.83–84.08)	81.32 (70.01–88.82)	82.65 (78.62–89.37)
CD124	47.01 (14.76–57.57)	39.60 (11.55–54.93)	47.43 (11.22–55.10)	44.45 (11.41–55.48)	47.05 (9.64–55.13)
CD197	8.31 (6.65–66.05)	3.68* (3.58–7.73)	3.23* (3.08–5.67)	6.29* (2.51–41.62)	5.4* (2.69–47.58)

Notes: Data are presented as Median with the first and third quartiles (Q1-Q3); *p < 0.05 – as compared to Mc/Mph cells cultured in the absence of GM-CSF.

Figure 1. Histograms of staining for CD197⁺ Mc/Mph

1 – Mc/Mphs in the absence of GM-CSF;2 – Mc/Mphs in the presence of GM-CSF.

Table 2. Percentages of CD14⁺, CD16⁺, CD119⁺, CD124⁺, CD197⁺ cells in LPS-activated Mc/Mph cell cultures

CD marker	GM-CSF (ng/ml)
	0.0	0.01	0.1	1.0	10.0
CD14	25.01 (11.37–33.46)	18.42* (7.56–30.70)	20.99* (7.98–28.95)	20.91* (7.99–31.94)	20.35* (23.07–32.16)
CD16	12.05 (4.94–24.10)	9.11* (3.27–14.29)	8.13* (3.60–16.96)	7.75* (3.55–13.47)	8.13* (3.77–12.70)
CD119	84.55 (65.02–86.62)	77.92 (74.54–79.66)	77.15* (65.45–80.28)	78.31* (68.12–80.31)	79.29* (67.94–79.68)
CD124	45.12 (15.91–56.74)	45.70 (14.31–53.22)	43.79 (16.83–54.3)	42.62 (16.92–53.26)	44.92 (16.11–53.05)
CD197	20.38 (7.88–50.75)	12.34* (6.59–46.22)	16.55* (5.44–39.62)	11.60* (5.55–38.97)	11.52* (5.46–41.86)

Notes: Data are presented as Median with the first and third quartiles (Q1-Q3); *p < 0.05 – as compared to Mc/Mph cells activated by LPS in the absence of GM-CSF.

Figure 2. Histograms of staining for CD14⁺, CD16⁺, CD119⁺, and CD197⁺ Mc/Mphs

1 – Mc/Mphs activated by LPS in the absence of GM-CSF;2 – Mc/Mphs activated by LPS in the presence of GM-CSF.

Table 3. Concentrations of cytokines (pg/ml) in Mc/Mph cell culture supernatants

Cytokine	Without activation	Activation + GM-CSF (ng/ml)
		0.0	0.01	0.1	1.0	10.0
TNF-α	320.8 (74.2–421.6)	1198.2 (745.1–1553.6)	1284 (533.8–1638)	1166 (496.3–1516)	1282.2 (551.1–1599.8)	1418.6* (1002.4–1849.9)
IL-1β	5.7 (0–157.6)	459.0 (265.8–507.6)	300.6 (118.2–621.6)	372.6 (183.6–683.4)	369.0 (337.8–754.8)	399 (288–757.8)
IL-6	124.0 (36.0–733.5)	8339 (4734–14577)	10881 (6328–13633)	9336 (5711–13269)	9192 (5103–13315)	12216* (8921–17225)
IL-10	0.8 (0–31.6)	118.98 (95.87–127.54)	124.95 (86.25–161.03)	139.2 (92.93–165.3)	93.6 (78.75–140.1)	89.85 (75.68–139.65)

Notes: Data are presented as Median with the first and third quartiles (Q1-Q3); * p < 0.05 – as compared to LPS-activated Mc/Mph cells cultured in the absence of GM-CSF.

Table 4. Percentages of CD38⁺ T cells in T cell subpopulations studied

T cell subpopulation		Without activation	Activation + GM-CSF (ng/ml)
			0	0.01	0.10	1.00	10.00
CD4^+	Naïve cells	23.4 (15.5–30.5)	47.1 (29.1 − 62.7)	49.5 (39.8–66.4)	51.0 (36.0–67.5)	50.8* (41.5–63.3)	49.9 (35.7–61.6)
	Central memory cells	9.1 (7.1–12.8)	28.6 (24.3–29.9)	29.1 (24.6–32.6)	31.3* (27.6–32.3)	29.1 (25.8–33.4)	28.9* (25.7–30.0)
	Effector memory cells	3.7 (2.2–5.3)	14.2 (10.3–19.0)	15.5 (9.8–20.0)	15.7 (9.7–21.1)	16.8 (11.0–20.9)	15.1 (11.1–19.5)
	Terminally-differentiated effector cells	8.3 (5.7–15.2)	40.3 (29.8–51.0)	45.7 (37.6–54.6)	47.1 (38.6–55.5)	44.2 (38.1–53.4)	42.3 (36.9–54.6)
CD4^−	Naïve cells	9.5 (5.3–24.4)	25.7 (19.2–37.6)	28.6 (22.6–38.1)	29.3* (22.5–40.6)	26.5 (21.0–38.8)	24.6 (21.8–37.5)
	Central memory cells	12.6 (9.6–18.4)	25.4 (17.7–33.3)	26.0 (19.8–34.3)	26.9* (21.7–34.0)	26.3 (19.1–33.9)	23.8 (19.6–29.2)
	Effector memory cells	2.8 (1.7–4.9)	8.0 (4.9–10.2)	6.3 (5.4–10.0)	8.1 (5.5–11.8)	8.0 (5.0–10.0)	7.3 (4.6–10.6)
	Terminally-differentiated effector cells	6.7 (2.4–8.9)	15.4 (11.7–18.9)	15.5 (12.7–22.3)	16.8 (14.9 − 22.3)	17.1 (15.1–19.5)	15.4 (13.8–21.5)

Notes: T cell activation was performed in the presence of MACSiBead particles conjugated with Abs against human CD2, CD3, and CD28. Data are presented as Median with the first and third quartiles (Q1-Q3); * p < 0.05 – as compared to T cells activated in the absence of GM-CSF.

Table 5. Cytokine concentrations in T cell culture supernatants (pg/ml)

Cytokine	Without activation	Activation + GM-CSF (ng/ml)
		0	0.01	0.10	1.00	10.00
IL-2	<10	590.0 (121.0–869.5)	580.0 (96.0–987.5)	540.0 (106.0–1034.5)	468.0* (244.0–1120.0)	614.0* (272.0–1142.0)
INF-γ	<10	5094.0 (4039.8–9688.3)	2652.0* (2290.0–5530.0)	3312.0 (2524.0–7669.5)	5764.0 (3941.5–9071.5)	6276.0 (3716.0–9346.0)
IL-4	<2	11.6 (5.4–29.5)	7.1 (0.5–30.6)	7.4 (2.4–31.5)	8.8 (4.6–31.6)	13.9* (10.8–30.8)
IL-10	<5	620.6 (484.8–870.9)	528.9 (452.7–668.4)	509.4 (430.2–732.3)	512.1 (366.9–934.8)	452.4* (322.2–769.5)

Notes: T cell activation was performed in the presence of MACSiBead particles conjugated with Abs against human CD2, CD3, and CD28. Data are presented as Median with the first and third quartiles (Q1-Q3); * p < 0.05 – as compared to cell culture supernatants derived from activated T cells cultured in the absence of GM-CSF.

Table

Ab	antibody
DC	dendritic cell
GM-CSF	granulocyte-macrophage colony stimulating factor
IFN	interferon
IL	interleukin
GM-CSFR	receptor for GM-CSF
Мс	monocyte
Mph	macrophage
PBMC	peripheral blood mononuclear cell
TNF	tumor-necrosis factor.