J Hematol

Journal of Hematology, ISSN 1927-1212 print, 1927-1220 online, Open Access

Article copyright, the authors; Journal compilation copyright, J Hematol and Elmer Press Inc

Journal website https://jh.elmerpub.com

Original Article

Volume 15, Number 4, August 2026, pages 198-213

Could the Immune Modulation of Invariant Natural Killer T Cell and the Expression of CD244 and CD196 on Natural Killer Cell Subtypes Impact the Treatment Outcomes of Acute Myeloid Leukemia Patients?

↓ Figure 1. Flow cytometric detection of lymphocytes, NK cells, NKT cells, and their expression of CD196 and CD244. (a) Forward scatter area and height to define singlets and exclude duplicates. (b) Forward and side scatter dot plot was used to define lymphocyte population. (c) The expression of CD3 on lymphocyte population to detect CD3⁺ and CD3⁻ lymphocytes. (d) The expression of CD16 and CD56 on CD3⁺ lymphocyte population to detect CD3⁺CD16⁺CD56⁺ NKT cells. (e) The expression of CD196 and CD244 on CD3⁺CD16⁺CD56⁺ NKT cells. (f) The expression of CD16 and CD56 on CD3⁻ population to detect different NK cell subtypes according to the positivity or negativity, dimness, or brightness of CD16 and CD56. (g) The expression of CD196 and CD244 on CD3⁻CD16^+brightCD56^+bright NK cells. (h) The expression of CD196 and CD244 on CD3⁻CD16^+brightCD56^+dim NK cells. (i) The expression of CD196 and CD244 on CD3⁻CD16^+dimCD56^+dim NK cells. (j) The expression of CD196 and CD244 on CD3⁻CD16^+dimCD56^+bright NK cells. (k) The expression of CD196 and CD244 on total CD3⁻CD16⁺CD56⁺ NK cells. (l) The expression of CD196 and CD244 on CD3⁻CD16⁺CD56⁻ population. (m) The expression of CD196 and CD244 on CD3⁻CD16⁻CD56⁺ population.

↓ Figure 7. (a) Differential expression of NKT subpopulations according to remission. (b) Differential expression of NK subpopulations. (c) Differential expression of CD244 and CD196 on NK subpopulations. (d) Differential expression of CD16 and CD56 on NK cells according to remission.

↓ **Table 1.** Demographic Data of Study Population
Demographic	Patients (n = 26)	Controls (n = 24)	P-value
Data are expressed as mean ± SD, median, number, and percentages. ^aMann–Whitney test. ^bFisher’s exact test. P ≤ 0.05 is considered significant. SD: standard deviation.
Age^a			0.6
Mean ± SD	45.5 ± 18.1	47.6 ± 6.7
Median	46 years	47 years
Sex^b			0.7
Male (n = 31)	17 (54.8%)	14 (45.2%)
Female (n = 19)	9 (47.4%)	10 (52.6%)

↓ Table 1. Demographic Data of Study Population

Demographic

Patients (n = 26)

Controls (n = 24)

P-value

Data are expressed as mean ± SD, median, number, and percentages. ^aMann–Whitney test. ^bFisher’s exact test. P ≤ 0.05 is considered significant. SD: standard deviation.

Age^a

0.6

Mean ± SD

45.5 ± 18.1

47.6 ± 6.7

Median

46 years

47 years

Sex^b

0.7

Male (n = 31)

17 (54.8%)

14 (45.2%)

Female (n = 19)

9 (47.4%)

10 (52.6%)

↓ **Table 2.** Laboratory Characteristics of AML Patients
Characteristics	Descriptive
Data are expressed as mean ± SD, numbers, and percentages. AML: acute myelocytic leukemia; CRP: C-reactive protein; DIC: disseminated intravascular coagulation; Hb: hemoglobin; LDH: lactate dehydrogenase; SD: standard deviation; WBC: white blood cell.
Hb (mean ± SD)	8.1 ± 1.4
WBCs (mean ± SD)	74.9 ± 77.5
Lymphocytic count (mean ± SD)	15.9 ± 6.1
Neutrophilic count (mean ± SD)	24.3 ± 13.1
Platelet count (mean ± SD)	48.14 ± 38.8
LDH (mean ± SD)	929.3 ± 735.6
Uric acid (mean ± SD)	9.6 ± 4.8
Serum ferritin (mean ± SD)	606.1 ± 643.04
CRP (mean ± SD)	46.4 ± 54.8
Peripheral blasts % (mean ± SD)	52.9 ± 19.5
BM blasts % (mean ± SD)	57.7 ± 19.8
Impaired liver function	10 (38.5%)
Raised renal function	6 (23.1%)
DIC, %	4 (15.4%)
Decreased fibrinogen level, %	6 (23.1%)
Raised D-dimer, %	5 (19.2%)
Impaired coagulation, %	5 (19.2%)

↓ Table 2. Laboratory Characteristics of AML Patients

Characteristics

Descriptive

Data are expressed as mean ± SD, numbers, and percentages. AML: acute myelocytic leukemia; CRP: C-reactive protein; DIC: disseminated intravascular coagulation; Hb: hemoglobin; LDH: lactate dehydrogenase; SD: standard deviation; WBC: white blood cell.

Hb (mean ± SD)

8.1 ± 1.4

WBCs (mean ± SD)

74.9 ± 77.5

Lymphocytic count (mean ± SD)

15.9 ± 6.1

Neutrophilic count (mean ± SD)

24.3 ± 13.1

Platelet count (mean ± SD)

48.14 ± 38.8

LDH (mean ± SD)

929.3 ± 735.6

Uric acid (mean ± SD)

9.6 ± 4.8

Serum ferritin (mean ± SD)

606.1 ± 643.04

CRP (mean ± SD)

46.4 ± 54.8

Peripheral blasts % (mean ± SD)

52.9 ± 19.5

BM blasts % (mean ± SD)

57.7 ± 19.8

Impaired liver function

10 (38.5%)

Raised renal function

6 (23.1%)

DIC, %

4 (15.4%)

Decreased fibrinogen level, %

6 (23.1%)

Raised D-dimer, %

5 (19.2%)

Impaired coagulation, %

5 (19.2%)

↓ **Table 3.** AML Types, Cytogenetics, Molecular Genetics, and Risk Groups
Characteristics	Frequencies
Data are expressed as numbers and percentages. AML: acute myelocytic leukemia; FAB: French-American-British.
AML type
Primary	23 (88.5%)
Secondary	3 (11.5%)
FAB types
M1	2 (7.7%)
M2	1 (3.8%)
M1/M2	4 (15.4%)
M4	2 (7.7%)
M4/M5	14 (53.8%)
M5	3 (11.5%)
Cytogenetics
Favorable	12 (46.2%)
Unfavorable	14 (53.8%)
Molecular genetics
Positive FLT3	9 (34.6%)
Positive CD117	8 (30.8%)
Positive NPM1	8 (30.8%)

↓ Table 3. AML Types, Cytogenetics, Molecular Genetics, and Risk Groups

Characteristics

Frequencies

Data are expressed as numbers and percentages. AML: acute myelocytic leukemia; FAB: French-American-British.

AML type

Primary

23 (88.5%)

Secondary

3 (11.5%)

FAB types

2 (7.7%)

1 (3.8%)

M1/M2

4 (15.4%)

2 (7.7%)

M4/M5

14 (53.8%)

3 (11.5%)

Cytogenetics

Favorable

12 (46.2%)

Unfavorable

14 (53.8%)

Molecular genetics

Positive FLT3

9 (34.6%)

Positive CD117

8 (30.8%)

Positive NPM1

8 (30.8%)

↓ **Table 4.** Differential Expression of CD244 and CD196 on CD3⁻CD16^dimCD56^bright NK Cells Between Patients and Controls
CD3⁻CD16^dimCD56^bright subpopulations	Group	Mean% ± SD	P-value
Data are expressed as mean % ± SD and analyzed by Mann–Whitney test. P ≤ 0.05 is significant.
CD3⁻CD16^dimCD56^bright	Patients	1.1 ± 2.1	0.6
	Controls	0.9 ± 0.4
CD3⁻CD16^dimCD56^brightCD244⁺CD196⁺	Patients	26.7 ± 25.2	0.016
	Controls	12.2 ± 11.3
CD3⁻CD16^dimCD56^brightCD244⁺CD196⁻	Patients	38.4 ± 28.8	< 0.001
	Controls	80.3 ± 8.2
CD3⁻CD16^dimCD56^brightCD244⁻CD196⁻	Patients	0.09 ± 0.4	0.4
	Controls	0.01 ± 0.0
CD3⁻CD16^dimCD56^brightCD244⁻CD196⁺	Patients	0.4 ± 1.3	0.9
	Controls	0.3 ± 1.1

↓ Table 4. Differential Expression of CD244 and CD196 on CD3⁻CD16^dimCD56^bright NK Cells Between Patients and Controls

CD3⁻CD16^dimCD56^bright subpopulations

Group

Mean% ± SD

P-value

Data are expressed as mean % ± SD and analyzed by Mann–Whitney test. P ≤ 0.05 is significant.

CD3⁻CD16^dimCD56^bright

Patients

1.1 ± 2.1

0.6

Controls

0.9 ± 0.4

CD3⁻CD16^dimCD56^brightCD244⁺CD196⁺

Patients

26.7 ± 25.2

0.016

Controls

12.2 ± 11.3

CD3⁻CD16^dimCD56^brightCD244⁺CD196⁻

Patients

38.4 ± 28.8

< 0.001

Controls

80.3 ± 8.2

CD3⁻CD16^dimCD56^brightCD244⁻CD196⁻

Patients

0.09 ± 0.4

0.4

Controls

0.01 ± 0.0

CD3⁻CD16^dimCD56^brightCD244⁻CD196⁺

Patients

0.4 ± 1.3

0.9

Controls

0.3 ± 1.1

↓ **Table 5.** Differential Expression of CD244 and CD196 on CD3⁻CD16^dimCD56^dim NK Cells Between Patients and Controls
CD3⁻CD16^dimCD56^dim subpopulations	Group	Mean % ± SD	P-value
Data are expressed as mean % ± SD and analyzed by Mann–Whitney test. P ≤ 0.05 is significant.
CD3⁻CD16^dimCD56^dim	Patients	22.02 ± 13.8	< 0.001
	Controls	5.7 ± 2.6
CD3⁻CD16^dimCD56^dimCD244⁺CD196⁺	Patients	33.1 ± 11.6	< 0.001
	Controls	18.4 ± 7.2
CD3⁻CD16^dimCD56^dimCD244⁺CD196⁻	Patients	46.6 ± 19.0	0.3
	Controls	54.4 ± 18.2
CD3⁻CD16^dimCD56^dimCD244⁻CD196⁺	Patients	12.6 ± 11.6	0.2
	Controls	16.0 ± 3.7
CD3⁻CD16^dimCD56^dimCD244⁻CD196⁻	Patients	7.7 ± 7.2	< 0.001
	Controls	1.5 ± 1.5

↓ Table 5. Differential Expression of CD244 and CD196 on CD3⁻CD16^dimCD56^dim NK Cells Between Patients and Controls

CD3⁻CD16^dimCD56^dim subpopulations

Group

Mean % ± SD

P-value

Data are expressed as mean % ± SD and analyzed by Mann–Whitney test. P ≤ 0.05 is significant.

CD3⁻CD16^dimCD56^dim

Patients

22.02 ± 13.8

< 0.001

Controls

5.7 ± 2.6

CD3⁻CD16^dimCD56^dimCD244⁺CD196⁺

Patients

33.1 ± 11.6

< 0.001

Controls

18.4 ± 7.2

CD3⁻CD16^dimCD56^dimCD244⁺CD196⁻

Patients

46.6 ± 19.0

0.3

Controls

54.4 ± 18.2

CD3⁻CD16^dimCD56^dimCD244⁻CD196⁺

Patients

12.6 ± 11.6

0.2

Controls

16.0 ± 3.7

CD3⁻CD16^dimCD56^dimCD244⁻CD196⁻

Patients

7.7 ± 7.2

< 0.001

Controls

1.5 ± 1.5

↓ **Table 6.** Type of Chemotherapy Received and Responses
Type	Descriptive
Data are expressed as numbers and percentages. Ara C: cytarabine; ATRA: all-trans retinoic acid.
Chemotherapy
3+7 regimen	16 (61.5%)
Low-dose Ara C	9 (34.6%)
ATRA + low-dose Ara C	1 (3.8%)
Response
Remission	17 (65.4%)
No remission	9 (34.6%)

↓ Table 6. Type of Chemotherapy Received and Responses

Type

Descriptive

Data are expressed as numbers and percentages. Ara C: cytarabine; ATRA: all-trans retinoic acid.

Chemotherapy

3+7 regimen

16 (61.5%)

Low-dose Ara C

9 (34.6%)

ATRA + low-dose Ara C

1 (3.8%)

Response

Remission

17 (65.4%)

No remission

9 (34.6%)

↓ **Table 7.** Differential Expression of Immune Cells According to Risk Groups
Immune cells	Mean ± SD	P-value
Data are expressed as mean ± SD and analyzed by Robust tests. P ≤ 0.05 is significant.
CD3⁺CD16⁺CD56⁺CD244⁻CD196⁻ NKT
Low risk	1.2 ± 0.9	0.05
Intermediate risk	0.42 ± 0.8
High risk	0.42 ± 0.5
CD3⁻ NK
Low risk	32.8 ± 3.5	0.013
Intermediate risk	36.5 ± 9.3
High risk	64.8 ± 25.0
CD3⁻CD16^brightCD56^dim
Low risk	12.4 ± 9.8	0.024
Intermediate risk	10.4 ± 6.2
High risk	4.6 ± 2.7
CD3⁻CD16^brightCD56^bright
Low risk	31.8 ± 11.4	0.019
Intermediate risk	35.1 ± 13.5
High risk	50.2 ± 14.0
CD3⁻CD16^dimCD56^dim
Low risk	30.4 ± 14.3	0.005
Intermediate risk	24.0 ± 12.7
High risk	10.4 ± 3.7
CD3⁻CD16⁺CD56⁻CD244⁺CD196⁻
Low risk	43.9 ± 13.6	0.047
Intermediate risk	33.1 ± 12.3
High risk	40.14 ± 18.6
CD3⁻CD16⁻CD56⁺CD244⁻CD196⁻
Low risk	9.5 ± 5.5	0.046
Intermediate risk	19.8 ± 11.6
High risk	12.5 ± 6.7

↓ Table 7. Differential Expression of Immune Cells According to Risk Groups

Immune cells

Mean ± SD

P-value

Data are expressed as mean ± SD and analyzed by Robust tests. P ≤ 0.05 is significant.

CD3⁺CD16⁺CD56⁺CD244⁻CD196⁻ NKT

Low risk

1.2 ± 0.9

0.05

Intermediate risk

0.42 ± 0.8

High risk

0.42 ± 0.5

CD3⁻ NK

Low risk

32.8 ± 3.5

0.013

Intermediate risk

36.5 ± 9.3

High risk

64.8 ± 25.0

CD3⁻CD16^brightCD56^dim

Low risk

12.4 ± 9.8

0.024

Intermediate risk

10.4 ± 6.2

High risk

4.6 ± 2.7

CD3⁻CD16^brightCD56^bright

Low risk

31.8 ± 11.4

0.019

Intermediate risk

35.1 ± 13.5

High risk

50.2 ± 14.0

CD3⁻CD16^dimCD56^dim

Low risk

30.4 ± 14.3

0.005

Intermediate risk

24.0 ± 12.7

High risk

10.4 ± 3.7

CD3⁻CD16⁺CD56⁻CD244⁺CD196⁻

Low risk

43.9 ± 13.6

0.047

Intermediate risk

33.1 ± 12.3

High risk

40.14 ± 18.6

CD3⁻CD16⁻CD56⁺CD244⁻CD196⁻

Low risk

9.5 ± 5.5

0.046

Intermediate risk

19.8 ± 11.6

High risk

12.5 ± 6.7