Table 3B:
Effect of cryopreservation alone and combined effect of delayed cell processing and cryopreservation on immunophenotyping of T cell subsets using blood collected in CPT™ tubes
| Reference (WB-D0) |
Cryopreser vation+ (PBMC-D0) |
Delayed cell processing and cryopreservation+ | P for trend | |||
|---|---|---|---|---|---|---|
| Cell subsets# | 0hr Mean (95% CI) |
0hr Mean (95% CI) |
24hr Mean (95% CI) |
48hr Mean (95% CI) |
72hr Mean (95% CI) |
|
| Cell Viability | 93.0 (91.9, 94.1) |
94.5 (93.5, 95.4) |
91.3 (89.0, 93.5) |
73.4*** (67.5, 79.3) |
58.9*** (54.1, 63.7) |
<0.0001 |
| T cells | 73.1 (69.6, 76.7) | 73.3 (68.3,78.3) |
67.4*** (63.3, 71.5) |
71.0 (66.9, 75.2) |
85.3*** (82.2, 88.3) |
<0.0001 |
| Helper T cells | 63.1 (56.2, 69.9) |
58.1 (51.5, 64.7) |
58.7 (52.4, 64.9) |
62.1 (55.4, 68.7) |
69.3 (64.4, 74.2) |
0.008 |
| Helper T cells: CM | 41.5 (36.2, 46.8) |
33.7***
(27.5, 39.9) |
34.9 (29.5, 40.3) |
38.0 (30.7, 45.2) |
40.5 (33.6, 47.5) |
0.98 |
| Helper T cells: EFF | 0.4 (0.0, 0.8) |
1.0 (0.2, 1.9) |
1.0 (0.2, 1.7) |
0.5 (0.2, 0.8) |
0.5 (0.1, 0.9) |
0.74 |
| Helper T cells: EM | 18.6 (13.8, 23.3) |
23.0 (16.9, 29.1) |
22.4 (16.9, 28.0) |
16.6 (11.7, 21.5) |
21.5 (15.5, 27.6) |
0.66 |
| Helper T cells: Naïive | 36.3 (28.8, 43.7) |
38.3 (31.1, 45.5) |
37.7 (30.2, 45.1) |
41.0 (32.1, 50.0) |
33.0 (24.2, 42.0) |
0.47 |
| Cytotoxic T cells | 28.3 (23.7, 33.0) |
32.2 (26.7, 37.7) |
32.0 (26.4, 37.7) |
27.2 (22.4, 32.0) |
23.3*** (19.3, 27.3) |
0.004 |
| Cytotoxic T cells: CM | 9.5 (5.7, 13.3) |
7.3 (2.6, 11.9) |
7.4 (2.5, 12.2) |
9.4 (5.5, 13.3) |
10.4 (6.0, 14.7) |
0.26 |
| Cytotoxic T cells: EFF | 21.1 (9.9, 32.3) |
35.8***
(22.0, 49.6) |
36.1***
(22.9, 49.3) |
23.7 (12.8, 34.6) |
22.1 (12.0, 32.1) |
0.49 |
| Cytotoxic T cells: pE | 49.5 (38.1, 70.0) |
53.1 (41.8, 64.4) |
55.6 (44.7, 66.5) |
54.0 (42.4, 65.5) |
48.9 (37.3, 60.4) |
0.70 |
| Cytotoxic T cells:pE1 | 16.9 (10.1, 23.6) |
15.3 (9.4, 21.3) |
13.7 (8.6, 18.7) |
15.2 (8.9, 21.5) |
21.4 (13.0, 29.7) |
0.08 |
| Cytotoxic T cells:pE2 | 31.3 (22.3, 40.4) |
30.0 (21.7, 38.4) |
29.1 (20.5, 37.8) |
28.8 (20.3, 37.2) |
27.9 (19.4, 36.4) |
0.10 |
| Cytotoxic T cells: EM | 39.2 (29.9, 48.5) |
32.3 (22.3, 42.2) |
31.7 (21.4, 42.0) |
30.8 (22.0, 39.7) |
34.8 (24.9, 44.7) |
0.19 |
| Cytotoxic T cells:EM1 | 62.7 (54.0, 71.4) |
62.1 (54.6, 69.7) |
59.1 (52.2, 66.0) |
58.1 (52.4, 63.9) |
65.2 (58.1, 72.3) |
0.48 |
| Cytotoxic T cells: EM2 | 12.2 (9.7, 14.6) |
12.4 (9.1, 15.6) |
12.9 (9.3, 16.6) |
12.6 (8.9, 16.3) |
10.7 (6.7, 14.8) |
0.23 |
| Cytotoxic T cells: EM3 | 18.8 (10.6, 27.0) |
17.8 (10.7, 24.8) |
17.8 (11.1, 24.5) |
17.4 (11.8, 23.0) |
14.7 (9.1, 20.3) |
0.08 |
| Cytotoxic T cells: EM4 | 6.3 (4.6, 8.1) |
7.8 (5.2, 10.3) |
10.1 (7.1, 13.2) |
11.5*** (9.3, 13.8) |
9.4*** (7.8, 11.0) |
0.01 |
| Cytotoxic T cells: Naïive | 20.9 (13.8, 28.0) |
18.8 (11.9, 25.6) |
18.7 (13.1, 24.3) |
28.2*** (19.2, 37.1) |
25.3 (17.2, 33.5) |
0.003 |
Indicates statistically significant pairwise comparisons (p<0.007) between WB-DO (reference) and the individual time points
Blood collected in EDTA tube and processed at time o (WB-D0) was used as the reference for all comparisons
CD4+ and CD8+ subsets of T-cells were expressed as percentage of T-cells. Subsets of CD4+ and CD8+ cells (effector, effector memory, central memory and naïve cells) were expressed as percentage of CD4+ and CD8+ cells respectively. The cytotoxic pE, pE1 and pE2 subsets were expressed as a percentage of cytotoxic effector cells. Cytotoxic EM1, EM2, EM3 and EM4 subsets were expressed as a percentage of cytotoxic effector memory cells.