Impact of CD105 Flow-Cytometric Expression on Childhood B-Acute Lymphoblastic Leukemia

Figures & data

Figure 1 Flow cytometry analysis of CD105 positive case. (A) Dot plot showed gating using CD45/SSC strategy, blast cells (R1), normal lymphocytes (R2) and granulocytes (R3). (B) Scatter dot plot showing blast cells positive for CD105 expression in combination with normal lymphocytes and granulocytes negative for CD105 expression. (C) Dot plot showing the mouse IgG1 isotypic negative control. (D) Dot plot showing positive CD105 expression on gated blast cells. (E) Histogram showing positive CD105 (solid violet curve) versus negative control (green colored curve).

Figure 2 Flow cytometry of analysis of CD105 negative case. (A) Dot plot showed gating using CD45/SSC strategy, blast cells (R1), normal lymphocytes (R2) and granulocytes (R3). (B) Scatter dot plot showing blast cells, normal lymphocytes and granulocytes all negative for CD105 expression. (C) Dot plot showing the mouse IgG1 isotypic negative control. (D) Dot plot showing negative CD105 expression on gated blast cells. (E) Histogram showing negative CD105 (solid violet curve) versus negative control (green colored curve).

Table 1 The Pattern and Range of CD105 Expression in B-ALL Cases as Regards Risk Stratification

	Standard Risk (54 Patients)	High Risk (17 Patients)	Very-High (9 Patients)	P-value	Post Hoc Test
CD105 SFIs
Median (Min. – Max.)	1.2 (0.5–6)	1.3 (0.5–6)	6 (1–8)	0.035*	P1:0.239, P2:0.013*, P3:0.168
CD105%
Median (Min. – Max.)	4 (0.5–60)	4 (2–60)	50 (1–90)	0.045*	P1:0.169, P2:0.022*, P3:0.282

Notes: *Significant at P ≤ 0.05. P₁: p-value for comparing between standard and high risk. P₂: p-value for the association between standard and very high. P₃: p-value for the association between high risk and very high.

Abbreviation: SFIs, specific fluorescence indices.

Table 2 The Impact of CD105 Expression Pattern on B-ALL Patient’s Characteristics

Parameters	CD105 Negative Group (47 Patients)	CD105 Positive Group (33 Patients)	P-value
Age (years)			0.109
Median (Min. – Max.)	6.5 (1.5–14)	8.5 (1–15)
Hemoglobin (gm/dl)			0.394
Mean ± SD.	8.1 ± 1.6	7.7 ± 2
Total leucocytic count (×10^3/cmm)			0.052
Median (Min. – Max.)	18 (1–110)	30 (1.7–96)
Platelets count (×10^3/cmm)			0.384
Median (Min. – Max.)	57 (10–260)	70 (15–185)
Peripheral blood blasts (%)			0.324
Mean ± SD.	36.2 ± 20.8	40.6 ± 26.7
Bone marrow blasts (%)
Median (Min. – Max.)	80 (50–98)	90 (59–98)	0.634
Lactate dehydrogenase (IU/L)			0.095
Median (Min. – Max.)	787 (340–2300)	900 (444–3200)
Erythrocyte sedimentation rate (m/h)			0.667
Mean ± SD.	82.6 ± 15.4	81.1 ± 15.3
Sex			^FEp= 0.218
Male	32 (68.1%)	18 (54.5%)
Female	15 (31.9%)	15 (45.5%)
Central nervous system infiltration			^FEp= 0.644
Absent	45 (95.7%)	30 (90.9%)
Present	2 (4.3%)	3 (9.1%)
Philadelphia chromosome			^FEp= 0.077
Negative	46 (97.9%)	28 (84.8%)
Positive	1 (2.1%)	5 (15.2%)
Testicular infiltration			^FEp= 0. 224
Present	2 (4.3%)	4 (12.1%)
Absent	45 (95.7%)	29 (87.9%)

Abbreviation: FE, Fisher exact.

Table 3 Correlation Between CD105 SFIs and CD105% with Different Parameters

	CD105 SFIs		CD105%
	rs	P-value	rs	P-value
Age (years)	0.298	0.007*	0.293	0.008*
Hemoglobin (gm/dl)	−0.151	0.181	−0.105	0.354
Total leucocytic count (×10^3/cmm)	0.261	0.019*	0.230	0.040*
Platelets count (×10^3/cmm)	−0.039	0.731	−0.025	0.824
Peripheral blood blasts (%)	0.059	0.606	0.081	0.473
Bone marrow blasts (%)	0.113	0.320	0.189	0.092
Lactate dehydrogenase (IU/L)	0.236	0.035*	0.199	0.077
Erythrocyte sedimentation rate (m/h)	0.018	0.871	−0.036	0.754
CD19 (%)	0.041	0.721	0.018	0.874
CD20 (%)	−0.232	0.038*	−0.179	0.113

Notes: *Significant at P ≤ 0.05. r_s: Spearman coefficient.

Abbreviation: SFIs, specific fluorescence indices.

Table 4 Univariate and Multivariate Analysis for the Parameters Affecting Response to Induction Therapy

	Univariate		Multivariate
	P-value	OR (95% C.I)	P-value	OR (95% C.I)
Age (>8 versus. ≤8 years)	0.232	2.325 (0.582–9.283)
Sex (female versus. male)	0.213	2.400 (0.605–9.522)
Hemoglobin (>7.8 gm/dl versus. ≤7.8 gm/dl)	0.195	0.381 (0.088–1.640)
Total leucocytic count (>20×10^3/cmm versus. ≤20×10^3/cmm)	0.280	2.143 (0.538–8.540)
Platelets count (>65×10^3/cmm versus ≤65×10^3/cmm)	0.865	1.125 (0.290–4.366)
Peripheral blood blasts (>40% versus. ≤40%)	0.385	1.833 (0.468–7.187)
Bone marrow blasts (>89% versus. ≤89%)	0.161	2.841 (0.659–12.239)
Lactate dehydrogenase (>856 IU/L versus. ≤856 IU/L)	0.998	–
Erythrocyte sedimentation rate (>85 m/h versus. ≤85 m/h)	0.683	0.750 (0.189–2.980)
Central nervous system infiltration (present versus. absent)	0.635	1.778 (0.166–19.065)
Philadelphia chromosome (positive versus. negative)	0.049*	12.500 (1.012–154.397)	0.110	14.820 (0.54–405.82)
CD105 (positive versus negative)	0.005*	21.600 (2.511–185.80)	0.007*	22.807 (2.34–221.84)

Notes: *Significant at P ≤ 0.05. The number between brackets in columns 1 represent cutoff used was the median of the cases included.

Abbreviations: OR, odds ratio; C.I, confidence interval.

Table 5 Performance (AUC, Sensitivity, Specificity) for CD105 SFIs and CD 105% to Discriminate Between Poor Responder (n=10) and Good Responder (n=51)

	AUC	P-value	95% C.I	Cutoff	Sensitivity	Specificity	PPV	NPV
CD105 SFIs	0.928	<0.001*	0.800–1.057	>2.5	90.0	92.16	69.2	97.9
CD105%	0.948	<0.001*	0.867–1.029	>35	90.0	94.12	75.0	98.0

Note: *Significant at P ≤0.05.

Abbreviations: AUC, area under the curve; C.I, confidence interval; NPV, negative predictive value; PPV, positive predictive value.

Figure 3 ROC curve for CD105 SFIs and CD105%to discriminate between poor and good responder.