A Potential Role of B7-H4 Expression in Predicting the Recurrence of Chronic Rhinosinusitis with Nasal Polyps

Figures & data

Table 1 The Demographic and Clinical Characteristics Among Three Groups

Variables	HC (n=32)	CRSsNP (n=27)	CRSwNP (n=80)	P-value
Age, years	40.9±13.2	37.9±14.6	43.5±13.2	0.167
Gender (male/female)	19/13	14/13	42/38	0.781
BMI, kg/m^2	23.0±3.1	23.5±3.1	24.4±3.5	0.085
Allergic rhinitis (yes/no)	0/32	8/19	26/54	0.001
Asthma (yes/no)	0/32	2/25	12/68	0.051
Peripheral eosinophil count, 10^9/L	0.05±0.03	0.09±0.06	0.27±0.11	<0.001
Peripheral eosinophil percentage, %	0.6±0.3	0.1±0.0	5.1±2.2	<0.001
VAS score	–	4.9±1.6	6.8±1.8	<0.001
Lund-Mackay score	–	12.0±2.5	17.4±3.0	<0.001

Abbreviations: HC, healthy control; CRSsNP, chronic rhinosinusitis without nasal polyps; CRSwNP, chronic rhinosinusitis with nasal polyps; BMI, body mass index; VAS, visual analogue scale. Bold font indicates statistical difference.

Table 2 The Demographic and Clinical Parameters Between pCRSwNP Group and rCRSwNP

Variables	pCRSwNP Group (n=35)	r CRSwNP Group (n=34)	P-value
Age, years	43.7±12.5	43.3±14.0	0.907
Gender (male/female)	25/15	22/18	0.051
BMI, kg/m2	25.2±3.5	23.7±3.3	0.597
Allergic rhinitis (yes/no)	4/36	14/26	0.014
Asthma (yes/no)	3/37	7/33	0.311
Peripheral eosinophil count, 10^9/L	0.3±0.1	0.3±0.1	0.379
Peripheral eosinophil percentage, %	4.7±1.8	5.4±2.6	0.187
Tissue eosinophil count, 10^9/L	18.8±12.5	32.4±24.2	0.003
Tissue eosinophil percentage, %	11.6±7.7	16.8±10.9	0.016
VAS score	6.6±1.7	7.1±1.9	0.209
Lund-Mackay score	17.9±3.2	16.9±2.8	0.141
Serum B7-H4 level	3.6±0.8	4.7±0.8	< 0.001
Tissue B7-H4 mRNA level	1.0±0.5	1.7±0.6	< 0.001

Abbreviations: pCRSwNP, primary chronic rhinosinusitis with nasal polyps; rCRSwNP, recurrent chronic rhinosinusitis with nasal polyps; BMI, body mass index; VAS, visual analogue scale. Bold font indicates statistical difference.

Table 3 Logistic Regression Analysis of Factors Associated with the Rate of Recurrence in CRSwNP Patients

Variables	OR	95% CI	P-value
Allergic rhinitis	0.393	0.149–1.038	0.059
Peripheral eosinophil count, 10^9/L	0.155	0.003–9.427	0.374
Peripheral eosinophil percentage, %	1.148	0.935–1.409	0.187
Tissue eosinophil count, 10^9/L	1.048	1.014–1.083	0.005
Tissue eosinophil percentage, %	1.064	1.009–1.121	0.021
Serum B7-H4 level	6.837	2.808–16.649	< 0.001
Tissue B7-H4 mRNA level	7.674	2.794–21.078	< 0.001

Abbreviations: CRSwNP, chronic rhinosinusitis with nasal polyps; Bold font indicates statistical difference.

Figure 1 Comparison of tissue B7-H4 mRNA levels among CRSwNP, CRS and HC groups. (A) Tissue B7-H4 mRNA levels were increased in the CRSwNP group than the CRS and HC groups. (B) The expression levels of B7-H4 in the CRSwNP with AR group and CRSwNP without AR group. (C) Compared to CRSwNP without AS group, B7-H4 levels were also no significantly difference in the CRSwNP with AS group. *P<0.05, **P<0.01.

Abbreviations: CRSwNP, chronic rhinosinusitis with nasal polyps; CRSsNP, chronic rhinosinusitis without nasal polyps; HC, healthy control; AR, allergic rhinitis; AS, asthma; ns, no significance.

Figure 2 The associations between tissue B7-H4 levels and clinical variables (A-H) in CRSwNP patients. Elevated B4-H4 mRNA levels exhibited positively correlation with tissue eosinophil count and percentage (G and H).

Abbreviations: CRSwNP, chronic rhinosinusitis with nasal polyps; BMI, body mass index; VAS, visual analog scale.

Figure 3 Comparison of tissue B7-H4 mRNA levels between two. (A) Tissue B7-H4 expression levels were higher in the rCRSwNP group than the pCRSwNP group. (B) ROC curve analysis of tissue B7-H4 in predicting CRSwNP recurrence. ****P < 0.0001.

Abbreviations: pCRSwNP, primary chronic rhinosinusitis with nasal polyps; rCRSwNP, recurrent chronic rhinosinusitis with nasal polyps; ROC, Receiver operating characteristic; AUC, area under the curve.

Figure 4 Immunofluorescence and B7-H4 protein expression in the tissue among three groups. Representative immunofluorescence images of HC, CRSsNP and CRSwNP (magnification x400).

Abbreviations: HC, healthy control; CRSsNP, chronic rhinosinusitis without nasal polyps; CRSwNP, chronic rhinosinusitis with nasal polyps, DAPI, 4’,6-diamidino-2-phenylindole.

Figure 5 Immunofluorescence and B7-H4 protein expression in primary and recurrent CRSwNP patients. Representative immunofluorescence images of pCRSwNP and rCRSwNP patients (magnification, x400).

Abbreviations: CRSwNP, chronic rhinosinusitis with nasal polyps; pCRSwNP, primary chronic rhinosinusitis with nasal polyps; rCRSwNP, recurrent chronic rhinosinusitis with nasal polyps; DAPI, 4’,6-diamidino-2-phenylindole.

Figure 6 Quantification of B7-H4+ cells in immunofluorescence staining. (A) Compared to, the increased the number of B7-H4+ cell in the CRSwNP group was higher the CRSsNP and HC groups. (B) The B7-H4+ cell number were significantly higher in the rCRSwNP group than the pCRSwNP group. **P<0.01; ***P<0.001; ****P<0.0001.

Abbreviations: HC, healthy control; CRSsNP, chronic rhinosinusitis without nasal polyps; CRSwNP, chronic rhinosinusitis with nasal polyps; pCRSwNP, primary chronic rhinosinusitis with nasal polyps; rCRSwNP, recurrent chronic rhinosinusitis with nasal polyps; HPF, high power field; ns, no significance.

Figure 7 Differential expressions of serum B7-H4 concentrations in different groups. (A) Serum B7-H4 concentrations were increased in the CRSwNP group than the CRSsNP and HC groups. (B) Serum B7-H4 concentrations in the rCRSwNP group were higher than the pCRSwNP group. (C) ROC curve analysis of serum B7-H4 in predicting CRSwNP recurrence. **P<0.01; ***P<0.001; ****P<0.0001.

Abbreviations: HC, healthy control; CRSsNP, chronic rhinosinusitis without nasal polyps; CRSwNP, chronic rhinosinusitis with nasal polyps; pCRSwNP, primary chronic rhinosinusitis with nasal polyps; rCRSwNP, recurrent chronic rhinosinusitis with nasal polyps; ROC, receiver operating characteristic; AUC, area under the curve; ns, no significance.