The activity level of follicular helper T cells in the peripheral blood of osteosarcoma patients is associated with poor prognosis

Figures & data

Figure 1. Flow diagram of study design.

OS: osteosarcoma; BT: benign tumor; Healthy: healthy controls; PBMC: peripheral blood mononulcear cell; Tfh: follicular helper T cell.

Table 1. Comparison of baseline information

Item	OS group (N = 148)	BT group (N = 102)	Healthy group (N = 133)	P value
Age (years), n				0.679
≦20	94	68	91
>20	54	34	42
Gender, n				0.878
Male	97	70	89
Female	51	32	44
Tumor location, n
Femur	62
Tibia	53
Others	33
Tumor size (cm), n
<5	102
≧5	46
Tumor stage, n
IIA	71
IIB	47
III	30
Tumor differentiation, n
Poorly differentiated	114
Well-differentiated	34
Metastasis, n
Yes	118
No	30
Tfh cell activity, n
Low activity	74
High activity	74

OS, Osteosarcoma; BT, benign bone tumor; Tfh, follicular helper T cells.

Figure 2. Expression of CD4⁺CXCR5⁺ Tfh cells in peripheral blood mononuclear cells (a) Flow cytometry-based measurement of expression of CD4⁺CXCR5⁺ Tfh cells in peripheral blood mononuclear cell (PBMCs); (b)Proportion of CD4⁺CXCR5⁺ Tfh cells in CD4⁺ T cells. **P < 0.01; ***P < 0.001.

Figure 3. Expression of CD4⁺CXCR5⁺CD69⁺ cells in the peripheral blood (a-c) Flow cytometry-based measurement of expression of CD4⁺CXCR5⁺CD69⁺ cells in peripheral blood mononuclear cell (PBMCs) in the OS group (a), BT group (b), and Healthy group (c); (d) Proportion of CD4⁺CXCR5⁺CD69⁺ cells in CD4⁺ T cells. ***P < 0.001.

Figure 4. Correlation analysis between the activity level of Tfh cells and clinicopathological characteristics of osteosarcoma. (a) Tumor location; (b) Tumor size; (c) Tumor stage; (d) Tumor differentiation; (e) Presence or absence of distant metastasis. ***P < 0.001; ns, no significant.

Figure 5. Kaplan-Meier survival curve. Overall survival of osteosarcoma patients with high versus low activation levels of Tfh cells in peripheral blood.

Table 2. Univariate and multivariate Cox analysis of overall survival in osteosarcoma patients

Item	Type	Univariate		Multivariate
		HR (95%CI)	P	HR (95%CI)	P
Tumor location	Femur/Tibia/ Others	0.998 (0.655–1.521)	0.994	1.036 (0.778–1.379)	0.809
Tumor size	≦5 cm/>5 cm	5.245 (2.227–12.352)	<0.001	0.708 (0.349–1.435)	0.338
Tumor stage	IIA/IIB/III	6.636 (3.436–12.814)	<0.001	4.034 (2.282–7.132)	<0.001
Tumor differentiation	Poorly differentiated/ well-differentiated	10.897 (3.150–37.699)	<0.001	2.071 (1.066–4.023)	0.032
Tumor metastasis	With/without metastasis	8.847 (2.543–30.777)	0.001	0.236 (0.085–0.655)	0.006
Tfh activity level	Low/high activation	9.739 (3.554–26.691)	<0.001	2.280 (1.202–4.325)	0.012

OS, Osteosarcoma; BT, benign bone tumor; Tfh, follicular helper T cells.