Lymphocyte subpopulations: a potential predictor of a response in patients with immune thrombocytopenia

Figures & data

Table 1. Patient characteristics.

Characteristics
Patients (N)	35
Median age / years (range)	54 (21–98)
Sex (F/M)	20/15
Median duration / years (range)	5.5 (0–39)
Median platelet count / x 10^9 per L (range)*	20 (0–72)
Newly diagnosed (N)	4
Median number of prior treatment lines (range)**	2.5 (1–7)
Previous splenectomy (N)	3 (9%)
Outpatient visits per year (range)	13 (4–40)

* At the time of enrolment.

** In previously treated patients.

Figure 1. Legend: Gating strategy for T cell analysis: (A) helper CD4+ T cells as CD3+/CD4+ T cells; (B) IFNγ+, IL-4 + and IFNγ+/IL-4+ CD4+ T cells within CD4+ T cells; (C) IL-17a + CD4+ T cells within CD4+ T cells; (D) cytotoxic CD8+ T cells as CD3+/CD8+ T cells; (E) IFNγ+, IL-4 + and IFNγ+/IL-4+ CD8+ T cells within CD8+ T cells

Table 2. Treatment discontinuation during follow-up: after 6 and 12 months, respectively.

Patients (N)	Therapy	6 months	12 months
Responders	none	12	14
	ongoing	9	7
Nonresponders	none	4	4
	ongoing	10	10
Total		35	35

Figure 2. Legend: The results of CD8+ T lymphocytes (%) stratified with respect to platelet count achieved at the 6-month time point, divided into two groups – PLT ≥ 100 and PLT < 100 × 10⁹ /L.

Figure 3. Legend: The results of IRI stratified with respect to platelet count achieved at the 6-month time point, divided into two groups – PLT ≥ 100 and PLT < 100 × 10⁹ /L.

Figure 4. Legend: The results of antiplatelet autoantibodies (%) stratified with respect to platelet count achieved at the 6-month time point, divided into two groups – PLT ≥ 100 and PLT < 100 × 10⁹ /L.

Figure 5. The results of CD8+ T lymphocytes (%) stratified with respect to platelet count achieved at the 12-month time point, divided into two groups – PLT ≥ 100 and PLT < 100 × 10⁹ /L.

Figure 6. The results of CD8+/IL-4+/IFN-γ+ T lymphocytes (%) stratified with respect to platelet count achieved at the 12-month time point, divided into two groups – PLT ≥ 100 and PLT < 100 × 10⁹ /L.

Figure 7. The results of IRI stratified with respect to platelet count achieved at the 12-month time point, divided into two groups – PLT ≥ 100 and PLT < 100 × 10⁹ /L.

Figure 8. The results of antiplatelet autoantibodies (%) stratified with respect to platelet count achieved at the 12-month time point, divided into two groups – PLT ≥ 100 and PLT < 100 × 10⁹ /L.

Table 3. Summary of statistical analysis of our data: Comparison with use of the T – test or Mann – Whitney test according to normality of the analyzed data comparing patients divided into two groups, responders (PLT ≥ 100 × 10⁹ /L) and nonresponders (PLT < 100 × 10⁹/L).

Parameter		6 months			12 months
		P value	median R (IQR)	median NR (IQR)	P value	median R (IQR)	median NR (IQR)
CD4+ T cells	relative	0.40	44.35 (38.10–54.18)	42.30 (30.61–48.83)	0.59	43.5 (37.50–54.72)	43.0 (37.08–48.93)
	absolute	0.95	9.23 (4.64–12.80)	9.67 (2.94–13.03)	0.58	8.64 (3.55–12.80)	9.67 (5.14–13.03)
CD8+ T cells	relative	0.02	20.40 (18.84–26.78)	28.46 (23.82–42.94)	0.02	20.40 (18.84–26.65)	27.99 (23.86–36.69)
	absolute	0.20	4.86 (2.49–6.28)	5.93 (4.25–8.73)	0.07	4.46 (2.17–6.28)	6.02 (5.09–8.73)
CD4+/INF-γ+ T cells	relative	0.51	21.81 (18.98–30.57)	23.62 (20.71–43.37)	0.80	24.13 (18.98–32.73)	23.05 (20.65–29.21)
	absolute	0.65	2.34 (1.44–3.58)	2.57 (1.27–3.03)	0.98	2.34 (1.11–3.58)	2.57 (1.55–3.06)
CD4+/IL-4+ T cells	relative	0.36	1.99 (0.60–2.53)	1.60 (0.64–1.93)	0.52	1.99 (0.60–2.53)	1.70 (0.71–1.87)
	absolute	0.52	0.18 (0.04–0.30)	0.12 (0.04–0.26)	0.89	0.15 (0.03–0.30)	0.13 (0.08–0.24)
CD4+/IL-4+/IFN-γ+ T cells	relative	0.23	0.74 (0.42–1.57)	1.51 (0.42–5.06)	0.28	0.69 (0.42–1.83)	1.30 (0.62–3.01)
	absolute	0.18	0.09 (0.02–0.13)	0.19 (0.06–0.30)	0.10	0.09 (0.02–0.20)	0.18 (0.10–0.33)
CD4+/IL-17+ T cells	relative	0.71	1.36 (1.01–2.26)	1.42 (0.66–1.80)	0.11	1.56 (1.01–2.50)	1.17 (0.66–1.48)
	absolute	0.99	0.11 (0.06–0.30)	0.15 (0.05–0.25)	0.78	0.12 (0.04–0.30)	0.10 (0.07–0.27)
CD8+/IFN-γ+ T cells	relative	0.08	60.6 (35.59–76.47)	75.2 (59.13–87.02)	0.17	60.6 (35.59–82.13)	68.8 (59.13–82.66)
	absolute	0.21	2.32 (1.06–3.94)	4.39 (1.59–6.87)	0.06	2.32 (0.72–3.93)	4.68 (2.81–7.19)
CD8+/IL-4+ T cells	relative	0.61	0.70 (0.25–2.47)	0.49 (0.09–2.49)	0.64	0.36 (0.19–1.64)	1.07 (0.16–2.88)
	absolute	0.90	0.03 (0.008–0.14)	0.04 (0.008–0.14)	0.15	0.03 (0.01–0.09)	0.11 (0.03–0.21)
CD8+/IL-4+/IFN-γ+ T cells	relative	0.27	0.86 (0.22–1.57)	1.16 (0.74–3.03)	0.04	0.68 (0.22–1.48)	1.43 (0.97–3.50)
	absolute	0.13	0.03 (0.005–0.09)	0.08 (0.03–0.34)	0.010	0.03 (0.005–0.07)	0.09 (0.08–0.42)
IRI (CD4 + to CD8+ T cells ratio)		0.04	2.1 (1.60–3.03)	1.5 (0.85–2.16)	0.06	2.20 (1.43–3.03)	1.54 (1.08–2.02)
Antiplatelet autoantibodies	%	0.01	57.65 (24.30–81.85)	20.30 (7.15–44.10)	0.02	54.55 (24.30–81.40)	20.30 (7.15–44.10)
WBC	x 10^9 /L	0.84	9.7 (6.51–13.34)	10.3 (6.23–12.87)	0.68	10.39 (6.51–14.23)	9.54 (6.23–12.12)
MCV	fL	0.73	86.2 (83.35–89.40)	85.4 (82.33–88.18)	0.46	86.9 (83.70–90.20)	85.4 (82.63–93.05)
Hb	g/L	0.38	135 (126.5–149.5)	143 (136.0–155.3)	0.55	135 (124–144)	150.5 (134–156.3)
Neutrophiles	%	0.75	69.9 (59.15–79.35)	64.6 (60.90–85.90)	0.67	70.0 (59.15–83.05)	64.6 (60.90–79.90)
Monocytes	%	0.42	6.4 (3.80–8.10)	7.3 (5.60–9.60)	0.10	6.4 (3.35–8.0)	7.8 (5.8–9.6)
Lymphocytes	%	0.54	21.6 (11.55–27.0)	23.2 (9.0–26.40)	0.98	21.1 (9.25–27.0)	23.2 (12.20–26.40)
Plt (0M)	x 10^9 /L	0.80	24.00 (7–28.5)	20.00 (2.75–35.25)	0.15	16.00 (4.0–26.0)	25.50 (14.5–35.1)
IPF	%	0.89	23.8 (9.70–33.75)	20.5 (13.20–34.95)	0.68	23.8 (9.70–34.80)	20.5 (13.20–34.40)
Age	years	0.27	50.0 (37.5–66.0)	55.5 (42.0–77.0)	0.91	50.00 (37–68)	53.50 (42–63.75)
Treatment line		0.07	2.00 (1.5–3.5)	2.50 (2.0–6.25)	0.92	2.00 (2.0–4.0)	2.00 (2.0–4.75)
Disease duration	years	0.14	0.34 (0.2–9.50)	5.50 (0.50–9.25)	0.79	3.50 (8.0–18.25)	5.00 (0.42–7.0)

Legend: IQR – interquartil range, WBC – white blood cells, MCV – mean cell corpuscular volume, Hb – hemoglobin.

Figure 9. Receiver operating characteristic (ROC) curve comparing responders (PLT ≥ 100 × 10⁹/L) and nonresponders (PLT < 100 × 10⁹/L) at 12 months, comparing IRI (P = 0.06).

Figure 10. Receiver operating characteristic (ROC) curve comparing responders (PLT ≥ 100 × 10⁹/L) and nonresponders (PLT < 100 × 10⁹/L) at 12 months, comparing antiplatelet autoantibodies (%, P = 0.02).

Data availability

Data supporting the findings of this study are available from the corresponding author on a reasonable request.