Relationship Between sCD163 and mCD163 and Their Implication in the Detection and Typing of Leprosy

Figures & data

Figure 1 (A) Forward scatter versus side scatter with primary gate on the monocyte region depending on cell size and granularity. (B-D) The monocyte area was selected where double positive for both CD163 PE and CD 14 FITC. The single CD14 FITC positive are in the lower right quadrant, single CD163 PE positive are in the upper left quadrant, double positive are in the upper right quadrant. LL had significantly higher mCD163 (46.68±22.17) than BL (16.80±8.71) and TL (14.72±3.41) (p<0.001), but insignificant difference between BL and TL (p=0.1).

Abbreviations: CD, cluster of differentiation; PE, phycoerythrin; FITC, fluorescein isothiocyanate; mCD, membrane bound cluster of differentiation; LL, lepromatous leprosy; BL, border-line leprosy; TL, tuberculoid leprosy; SSC, side scatter; FSC, forward scatter.

Table 1 Demographic and Clinical Data of Studied Subjects

		Leprosy Group N=70	Control Group N=30	Test of Significance	P-value
Sex N. (%)	Males	42 (60.0%)	19 (63.3%)	χ2=0.098	0.754
	Females	28 (40.0%)	11 (36.7%)
Age (Years)
Mean±SD		55.8±10.1	54.3±9.6	t-test=0.69	0.491
Range		31–77	30–75
Occupation
Working (manual – farmer)		30(42.9%)	20(66.7%)	χ2=4.76	0.029*
Not working		40(57.1%)	10(33.3%)
Residence
Rural		35(50.0%)	14(46.7%)	0.093	0.759
Urban		35(50.0%)	16(53.3%)
Socioeconomic Level
Low		54(77.2%)	10(33.3%)	12.9	0.002*
Middle		11(15.7%)	12(40%)
High		5(7.1%)	8(26.7%)
Family History
Yes		40(57.1%)	–	–	–
No		30(43.9%)
History of Prolonged Contact with Leprosy Patients
Yes		40(57.1%)	–		–
No		30(43.9%)
History of BCG Vaccination
Yes		65(92.9%)	–	–	–
No		5(7.1%)
History of Associated Diseases
No		23(32.9%)	–	–	–
Yes		47(67.1%)

Note: *Significant (p<0.05).

Abbreviations: N, number; χ², Chi- squared test; t-test, Student’s t-test; SD, standard deviation; BCG, Bacillus Chalmette Guerin.

Table 2 Comparison Between Patients and Controls Regarding Complete Blood Count (CBC), Serum sCD163 and Flow Cytometric Assessment

		Leprosy Group N=70	Control Group N=30	Test of Significance	P-value
WBCs *x103/µL		2.5±3.8	8.5±10.7	t-test=13.67	<0.001*
Platelets *x103/µL		202.1±30.2	250.4±38.9	t-test=6.70	<0.001*
Anemia	Yes	36(51.4%)	10 (33.3%)	χ^2=2.76	0.096
Serum sCD163 (ng\mL)
Median		314.65	87.11	MW test =196	0.025*
Range		30.1–11,260	0.282–580.46
Mean±SD		680.90±223.50	27.19±2.09
mCD163 Positive Monocytes (cells/mL)
Median		42.8	4.7	MW test =132	0.042*
Range		(7.3–162)	(1.1–11.2)
Mean±SD		130.94±53.12	9.35±1.15

Note: *Significant (p<0.05).

Abbreviations: CBC, complete blood count; N, number; SD, standard deviation; MW, Mann Whitney test; WBCs, white blood cells; sCD163, soluble cluster of differentiation; mCD163, membrane bound cluster of differentiation.

Table 3 Comparison Between the Studied Leprosy Types Regarding Laboratory Data

	Tuberculoid Leprosy (TL) N=27	Borderline (BB) N=10	Lepromatous Leprosy (LL) N=33	F Test	P-value
Serum sCD163 (ng\mL)
Mean±SD	69.89±8.36	342.04±122.55	1159.63±397.07	48.28	<0.001*
Monocyte Positive
mCD163 (cells/mL) Mean±SD	14.72±0.84	16.80±2.71	46.68±2.17	49.35	<0.001* P^1 <0.001* P^2 <0.001* P^3=0.1
CBC
WBCs x103/µL	3.5±1.2	2.5±2.8	1.5 ±2.2	t= 12.74	0.032*
Platelets x103/µL	240.4±21.5	220.1±21.6	192.1±40.2	t= 4.76	0.024*
Anemia (+ve)	15		29	χ^2= 3.1	0.096

Notes: *Significant (p<0.05), P¹ TL vs LL. P² BB vs LL. P³ BB vs TL.

Abbreviations: SD, standard deviation; F, Kruskal Wallis test; t, t-test; χ², Chi-squared; WBCs, white blood cells; sCD163, soluble cluster of differentiation; mCD163, membrane bound cluster of differentiation; CBC, complete blood count.

Figure 2 Correlation between mCD163 positive monocytes and sCD163 serum levels in leprosy patients (r=0.896, p<0.001).

Abbreviations: m, membrane bound; CD, cluster of differentiation; s, soluble.

Table 4 Baseline Data in Leprosy Patients in Relation to Serum sCD163 and mCD163 Positive Monocytes

Variable	Serum sCD163 (ng\mL)	F test	P-value	mCD163 Positive Monocytes (cells/mL)	t-test	P-value
History of Associated Diseases		1.13	0.276		1.327	0.131
Yes Median Mean±SD Range	905 796 ±2204 0.3–11,260.1			35.2 27.1 ±18.5 7.5–62
No Median Mean±SD Range	440.8 340.8±511.9 0.31–2727.9			30.5 25.2 ±15.6 7.3–61
Family History		11.208	0.029*		49.35	<0.001*
+ve
Median Mean±SD Range	545.23 799.45±277.17 160.3–11,260			53.1 48.3 ±12.7 28–62
-ve
Median Mean±SD Range	329.11 342.04±325.05 65–679.3			17.2 14.87 ±6.5 7.3–23
Socioeconomic Status		48.28	<0.001*		2.705	0.025*
Low
Median Mean±SD Range	623.86 519±355 160.3–11,260			51.6 48.5 ±12.4 7.3–62
Middle
Median Mean±SD Range	243.74 275.33±135 65–679.3			37.2 32.6 ±18.7 8–55
High
Median Mean±SD Range	44.64 52.88±15.2 30.1–189			16.9 14.87 ±6.3 7.3–40
Occupation		11.208	0.002*		2.150	0.029*
Working
Median Mean ±SD Range	692.43 802.45±157.17 260.3–11,260			37.7 33.5 ±22.4 8–62
Not Working
Median Mean±SD Range	397.42 442.04±125.05 56–679.3			26.5 24.3 ±16.1 7.3–62
History of BCG		1.306	0.205		1.541	0.453
No
Median Mean±SD Range	339.28 652.2±435 40–679.8			33.9 27 ±15.5 7.3–23
Yes
Median Mean±SD Range	387.21 741.3±1347.9 0.3–11,260			23.1 22.6 ±4.3 28–62
History of Contact with Leprosy Cases		12.098	0.002*		2.455	0.029*
+ve
Median Mean±SD Range	645.23 699.45±277.17 160.3–11,260			49.7 ±28.9 8–62
-ve
Median Mean±SD Range	489.11 392.04±305.05 65–679.3			33.5 26.8 ±16.4 7.3–62

Note: *Significant (p<0.05).

Abbreviations: SD, standard deviation; +ve, positive; +ve, negative; BCG, Bacillus Chalmette Guerin; sCD163, soluble cluster of differentiation; mCD163, membrane bound cluster of differentiation.

Table 5 Correlation Between WBCs and Platelet Counts with Serum sCD163 and mCD163 Positive Monocytes

	Serum sCD163 Level (pg/mL)		Monocyte Positive mCD163 (cells/mL)
	(r)	P-value
WBCs (x103/mL)	−0.8312	<0.001*	−0.78446	<0.001*
Platelet counts (x103/mL)	−0.845	<0.001*	−0.3774	<0.001*

Notes: r: Spearman correlation. *Significant.

Abbreviations: WBCs, white blood cells; sCD163, soluble cluster of differentiation; mCD163, membrane bound cluster of differentiation.

Figure 3 Receiver operator characteristic (ROC) curve analysis in the detection of leprosy. (A) Serum sCD163: area under the curve was 0.87, best cut off was 100.11 pg/mL, sensitivity of 92.9% and specificity of 90% (p<0.001). (B) Monocytes mCD163: area under the curve was 0.96 and best cut off was 10.95 cells/mL with sensitivity of 91.4% and specificity of 90% (p<0.001).

Abbreviations: m, membrane bound; CD, cluster of differentiation; s, soluble.

Figure 4 Receiver operator characteristic (ROC) curve analysis in the typing of leprosy. (A) Serum sCD163: area under the curve was 0.83 with sensitivity of 84.8% and specificity of 80% for differentiation between border-line and lepromatous leprosy types with best cut off level of 379.13 pg/mL (p =0.002). (B) Monocytes mCD163: area under the curve was 0.93 with sensitivity of 87.9% and specificity of 80% for differentiation between border-line and lepromatous leprosy types with best cut off level of 22.5 cells/mL (p<0.001).

Abbreviations: m, membrane bound; CD, cluster of differentiation; s, soluble.