Cytokine Release Patterns in Mixed Lymphocyte Culture (MLC) of T-Cells with Dendritic Cells (DC) Generated from AML Blasts Contribute to Predict anti-Leukaemic T-Cell Reactions and Patients’ Response to Immunotherapy

Figures & data

Table 1. Characteristics of AML-/MDS-pts (age, gender, FAB-type, stages of the disease and response to immunotherapy) and samples used as stimulator cells (blast containing MNC or DC_leu containing DC) or as effector cells (T-cells: allogeneic, autologous, after SCT) in mixed lymphocyte cultures.

Patient No.	Age at dgn	Gender	FAB-type	Stage	Blast phenotype [CD]	IC Bla%	Source ofT-cells	Response to SCT/DLI	GVH	Infection
P 326	58	Male	AML M2	Relapse after SCT	34, 13, 33	57	Allogeneic	No CR after SCT	GVH II	No infection
P 328	59	Male	AML M2	Relapse before SCT	34, 33, 13, 117	49	Allogeneic	CR after SCT	GVH III-IV	No infection
P 362*	53	Female	MDS RAEB	Diagnosis	13, 33, 34, 117	40	Autologous	Early death after SCT	GVH III	No data available
P 419*	68	Female	AML M4	Relapse after SCT	15, 33, 34, 117	94	Allogeneic	No CR after SCT	GVH II	Unknown infection
P 428	59	Female	AML M4	Relapse after SCT	13, 33,117	47	After SCT	No CR after SCT	No GVH	Liver aspergillosis
P 454	52	Male	MDS CMML	Diagnosis	14, 16, 33, 65	37	Autologous	No SCT	–	No data available
P 460	36	Male	AML M0	Relapse after SCT	7, 15, 13, 33, 56, 65	75	After SCT	No response to SCT	No GVH	No infection
P 466*	62	Female	AML M6a	Persistence after SCT	13, 33, 34, 65, 117	20	Allogeneic	CR after SCT	GVH III	No infection
P 481*	66	Female	AML M4/M5	Relapse before SCT	13, 15, 33, 34, 56, 117	93	Allogeneic	} CR after SCT	No GVH	No infection
P 481	66	Female	AML M4/M5	Relapse before SCT	13, 15, 33, 34, 56, 117	93	Autologous
P 483	34	Male	AML M5	Relapse after SCT	13, 33, 34, 117, 7, 65, 19, 15	29	After SCT	CR after 2nd SCT	GVH I	No infection
P 502	41	Male	AML M4 eo	Persisting disease	13, 15, 33, 34, 117	92	Allogeneic	} CR after SCT	GVH I	No infection
P 502	41	Male	AML M4 eo	Persisting disease	13, 15, 33, 34, 117	92	Autologous
P 518	55	Male	AML M2	Diagnosis	13, 33, 117	81	Autologous	No SCT	–	No infection
P 520*	46	Female	AML M1/M2	Diagnosis	65, 14, 15, 19, 33, 34	70	Autologous	No SCT	–	No data available
P 538	12	Male	AML M0	Relapse after SCT	34, 33, 65	93	Allogeneic	No response to DLI	No GVH	No infection
P 545	44	Female	AML M1	Relapse before SCT	33, 34, 117	83	Autologous	Early death after SCT	–	No data available
P 546	39	Male	AML M2	Diagnosis	15, 33, 34, 65, 117	76	Autologous	No SCT	–	No infection
P 560	53	Male	AML	Diagnosis	34, 117, 33	53	Allogeneic	CR after SCT	GVH II	No infection
P 561	43	Male	AML M2	Diagnosis	13, 15, 33, 65, 117	65	Autologous	CR after SCT	GVH III	Pneumonia
P 565	68	Female	AML M5	Relapse after SCT	13, 15, 33, 34, 65	88	After SCT	No CR after SCT	GVH III	Pneumonia
P 584	52	Female	AML M4	Relapse before SCT	13, 33, 117	95	Allogeneic	CR after SCT	GVH I	No infection
P 618	66	Male	MDS RAEBt	Persisting disease	33, 117	42	Allogeneic	CR after SCT	GVH I	No infection
P 652	66	Male	AML M1	Persisting disease	13, 15, 33, 117	68	Autologous	No response to SCT	GVH II	No infection
P 655	66	Female	AML M2	Persisting disease	33, 34, 117	95	Allogeneic	CR after SCT	No GVH	No infection

* No CTX, only MLC performed; dgn: first diagnosis; IC-Bla: immunocytologically determined blasts; autologous: patients’ T-cells; allogeneic: allogeneic stem cell donors’ T-cells; after SCT: T-cells from pts at relapse after SCT; CR: complete remission; GVH: graft-versus-host-disease.

Figure 1. Kinetic of cytokines in serum of AML-/MDS-patients and in MLC-supernatants of T-cells after ‘DC’-stimulation (results with autologous, allogeneic T-cells or T-cells after SCT pooled). Median cytokine concentrations in serum and during different phases of MLC after stimulation with ‘DC’ are given as box-and-whisker plot. The ends of the whiskers represent the minimum and maximum of all of the data. The ordinate is presented as logarithmic scale to the base 10. Z1: Serum (n = 16), Z2: first restimulation of MLC (n = 19), Z3: second restimulation of MLC (n = 19), Z4: cell harvest of MLC (n = 11).

Figure 2. Correlation of anti-leukaemic T-cell activity with cytokine release patterns after stimulation with ‘DC’ (results with autologous, allogeneic T-cells or T-cells after SCT pooled). Median cytokine concentrations are given with cases subdivided in cases with or without gain of anti-leukaemic lytic activity after ‘DC’-stimulation as box-and-whisker plot. The ends of the whiskers represent the minimum and maximum of all of the data. The ordinate is presented as logarithmic scale to the base 10. (a) At second restimulation of MLC: cases with lysis (n = 7), cases with no lysis (n = 9), (b) At cell harvest of MLC: cases with lysis (n = 5), cases with no lysis (n = 3).

Figure 3. Cytokine release patterns in MLC-supernatants in correlation with clinical response vs. non response to immunotherapy (results with allogeneic T-cells or T-cells after SCT pooled). Median cytokine concentrations in MLC-supernatants of T-cells after ‘DC’-stimulation in cases subdivided in cases with or without clinical response to immunotherapy are given as box-and-whisker plot. The ends of the whiskers represent the minimum and maximum of all of the data. The ordinate is presented as logarithmic scale to the base 10. (a) At second restimulation of MLC: responders to immunotherapy (resp. n = 8), non responders to immunotherapy (no resp. n = 5), (b) At cell harvest of MLC: responders to immunotherapy (resp. n = 4), non responders to immunotherapy (no resp. n = 2)

Figure 4. Predictive values of cytokine concentrations for antileukaemic lytic capability of T-cells after stimulation with ‘DC’: cut-off analyses. Correlations of cytokine concentrations with antileukaemic lytic activities of T-cells are given at second restimulation (R) or at cell harvest (H) of MLC with ‘DC’, as available (results with autologous, allogeneic T-cells or T-cells after SCT pooled). All cases were separated in two prognostic groups evaluating a cut-off value for the best differentiation of cases into those with or without antileukaemic activity.