Table 2. Status of NF-κB and STAT3... | American Society of Hematology

Table 2.

Status of NF-κB and STAT3 activation in CD138⁺ cells from bone marrow aspirates of multiple myeloma patients

							Nuclear^*
Patient no.	% plasma cells in BM differential	MNC count, × 10⁶	CD138⁺ MNCs, %	Enriched CD138⁺, %	Viability, %	CD138⁺ count, × 10⁶	NF-κB	STAT3
1	NA	NA	NA	NA	NA	NA	+++	+
2	8	14	NA	93	90	1.1	+++	++
3	47	26	NA	NA	70	NA	+	+
4	40	36	NA	98	95	5.6	+++	+
5	35	24	NA	NA	95	0.8	++	+++
6	96	40	NA	97	99	0.5	+++	++
7	45	21	NA	95	99	0.5	+++	+
8	18	12	NA	NA	95	0.2	+++	+
9	30	21	NA	92	98	2	+++	+++
10	25	18	NA	95	95	1	++	++
11	66	10	NA	NA	95	0.3	++	+++
12	63	38	NA	98	98	22	++	+++
13	14	6.6	NA	80	98	1	+++	+++
14	44	9	3.59	60	98	0.6	+++	+++
15	50	29	3.64	52	60	1.2	++	+++
16	22	6	7.68	87	90	0.6	+++	+++
17	3	18	5.29	NA	98	0.3	+	−
18	22	21	NA	72	50	1.1	+++	+++
19	25	12	NA	98	95	1.5	+++	+++
20	49	17	24.7	97	95	1.7	+++	+++
21	46	10.2	NA	98	98	1.2	+++	+++
22	14	5.4	NA	85	98	1.2	+	++

							Nuclear^*
Patient no.	% plasma cells in BM differential	MNC count, × 10⁶	CD138⁺ MNCs, %	Enriched CD138⁺, %	Viability, %	CD138⁺ count, × 10⁶	NF-κB	STAT3
1	NA	NA	NA	NA	NA	NA	+++	+
2	8	14	NA	93	90	1.1	+++	++
3	47	26	NA	NA	70	NA	+	+
4	40	36	NA	98	95	5.6	+++	+
5	35	24	NA	NA	95	0.8	++	+++
6	96	40	NA	97	99	0.5	+++	++
7	45	21	NA	95	99	0.5	+++	+
8	18	12	NA	NA	95	0.2	+++	+
9	30	21	NA	92	98	2	+++	+++
10	25	18	NA	95	95	1	++	++
11	66	10	NA	NA	95	0.3	++	+++
12	63	38	NA	98	98	22	++	+++
13	14	6.6	NA	80	98	1	+++	+++
14	44	9	3.59	60	98	0.6	+++	+++
15	50	29	3.64	52	60	1.2	++	+++
16	22	6	7.68	87	90	0.6	+++	+++
17	3	18	5.29	NA	98	0.3	+	−
18	22	21	NA	72	50	1.1	+++	+++
19	25	12	NA	98	95	1.5	+++	+++
20	49	17	24.7	97	95	1.7	+++	+++
21	46	10.2	NA	98	98	1.2	+++	+++
22	14	5.4	NA	85	98	1.2	+	++

A total of 2 to 10 mL bone marrow was aspirated from the upper iliac crest or sternum and was layered over Ficoll-Paque to isolate MNCs. The cell suspension was then incubated with anti-CD138 microbeads. The cell suspension was centrifuged, and the cell pellet was loaded onto the magnetic column of the AutoMACS system. Anti-CD138⁺ cells were isolated by positive selection. The purity of the isolated CD138⁺ plasma cell population was determined by FACS analysis using anti-CD 138–PE antibody. NF-κB and STAT3 activation status was determined by fixing these cells on slides by cytospin followed by immunocytochemistry for NF-κB (p65) and STAT3 as described in “Patients, materials, and methods.”

MNC indicates mononuclear cells; STAT3, signal transducer and activator of transcription 3; NA, not available.

Grading: — indicates 0% cells with nuclear positivity; +, less than 10% cells with nuclear positivity; ++, 10% to 50% cells with nuclear positivity; +++, more than 51% cells with nuclear positivity.

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