Introduction
Materials and methods
Subjects
RRMS patients | Control groups | |||||
---|---|---|---|---|---|---|
Median age | Female% | Median EDSS | Duration | Mean age | Female% | |
Whole blood studies
| 34.0 (28.0 to 39.0) | 59% | 2.0 (1.0 to 3.0) | 5.0 (2.0 to 7.0) | 32.0 (29.0 to 38.0) | 58% |
Cell subset studies
| 38.0 (30.5 to 47.0) | 75% | 1.5 (1.0 to 3.1) | 5.0 (1.3 to 12.5) | 33.5 (24.5 to 36.5) | 75% |
CSF cell studies
| 37.5 (31.3 to 42.8) | 59% | 3.5 (3.0 to 6.4) | 6.0 (1.5 to 9.75) | 51.5 (45.0 to 59.0) | 70% |
Gene expression studies
Gene Symbol | Assay number | GenBank | Whole blood | PBMC populations | CSF cells |
---|---|---|---|---|---|
GAPDH
| Hs99999905_m1 | NM_002046 | · | · | · |
IFNG
| Hs99999041_m1 | NM_000619 | · | · | · |
IL1B
| Hs00174097_m1 | NM_000576 | · | · | · |
IL1RN
| Hs00277299_m1 | NM_000577 | · | ||
IL7
| Hs00174202_m1 | NM_000880 | · | · | · |
IL10
| Hs00174086_m1 | NM_000572 | · | · | |
IL12A
| Hs00168405_m1 | NM_000882 | · | · | · |
IL15
| Hs00542571_m1 | NM_172175 | · | · | · |
IL18
| Hs00155517_m1 | NM_001562 | · | ||
IL23
| Hs00372324_m1 | NM_016584 | · | · | |
IL27
| Hs00377366_m1 | NM_145659 | · | · | · |
EBI3
| Hs00194957_m1 | NM_005755 | · | ||
LTA
| Hs00236874_m1 | NM_000595 | · | · | · |
LTB
| Hs00242739_m1 | NM_009588 | · | · | · |
TGFB1
| Hs99999918_m1 | NM_000660 | · | ||
TNF
| Hs00174128_m1 | NM_000594 | · | ||
PRF1
| Hs00169473_m1 | NM_005041 | · | ||
CD3d
| Hs00174158_m1 | NM_000732 | · | · | |
CD14
| Hs00169122_g1 | NM_000591 | · | · | |
CD19
| Hs01047407_m1 | NM_001770 | · | · | |
CD56
| Hs00941824_m1 | NM_181351 | · | · |
Statistical analysis
Results
Gene expression in whole blood from RRMS patients in remission
Gene expression in PBMC cell subpopulations from RRMS patients in remission
Gene expression in PBMCs and CSF-cells from RRMS relapse patients
RRMS relapse | NIND | RRMS relapse vs NIND | |||||||
---|---|---|---|---|---|---|---|---|---|
Mean CT value | N | Median (IQR) | CSF/PBMC p-value | N | Median (IQR) | CSF/PBMC p-value | p-value | q-value | |
IFNG
PBMC NR
| 32.3 | 15 | 4.10 (2.74 to 10.21) | 10 | 5.25 (2.65 to 9.12) | 0.78 | 0.37 | ||
IFNG
CSF NR
| 34.9 | 15 | 7.22 (0.00 to 16.42) | 10 | 0.00 (0.00 to 0.00) |
0.004
|
0.02
| ||
IFNG
CSF/PBMC Ratio
| 15 | 0.71 (0.00 to 2.31) | 0.87 | 10 | 0.00 (0.00 to 0.00) |
0.004
|
0.02
| ||
IL1B
PBMC NR
| 32.4 | 16 | 1.13 (0.88 to 2.14) | 10 | 1.81 (1.05 to 2.70) | 0.19 | 0.15 | ||
IL1B
CSF NR
| 36.7 | 14 | 0.21 (0.00 to 1.09) | 10 | 2.70 (0.00 to 6.94) | 0.36 | 0.23 | ||
IL1B
CSF/PBMC Ratio
| 13 | 0.18 (0.00 to 0.92) |
0.02
| 10 | 1.96 (0.00 to 6.40) | 0.29 | 0.42 | 0.24 | |
IL7
PBMC NR
| 34.0 | 16 | 0.41 (0.21 to 1.17) | 10 | 0.31 (0.18 to 0.46) | 0.40 | 0.24 | ||
IL7
CSF NR
| 36.1 | 13 | 0.00 (0.00 to 0.86) | 10 | 0.00 (0.00 to 0.00) | 0.06 | 0.09 | ||
IL7
CSF/PBMC Ratio
| 12 | 0.00 (0.00 to 2.51) | 8 | 0.00 (0.00-0.00) | 0.08 | 0.09 | |||
IL10
PBMC NR
| 34.3 | 16 | 2.75 (2.02 to 3.90) | 10 | 2.95 (2.38 to 4.26) | 0.54 | 0.27 | ||
IL10
CSF NR
| 35.3 | 17 | 23.51 (3.22 to 46.39) | 10 | 116.78 (72.22 to 156.28) |
0.001
|
0.01
| ||
IL10
CSF/PBMC Ratio
| 16 | 8.57 (0.79 to 12.71) |
0.003
| 10 | 42.21 (24.04 to 66.18) |
0.01
|
0.002
|
0.01
| |
IL12A
PBMC NR
| 37.5 | 16 | 0.49 (0.30 to 0.66) | 10 | 0.54 (0.42 to 0.82) | 0.53 | 0.27 | ||
IL12A
CSF NR
| 38.3 | 15 | 0.00 (0.00 to 0.00) | 10 | 0.00 (0.00 to 0.00) | 0.24 | 0.17 | ||
IL12A
CSF/PBMC Ratio
| 12 | 0.00 (0.00 to 0.00) | 9 | 0.00 (0.00 to 0.00) | 0.21 | 0.16 | |||
IL15
PBMC NR
| 36.3 | 16 | 1.19 (0.91 to 1.43) | 10 | 1.10 (0.57 to 1.60) | 0.79 | 0.37 | ||
IL15
CSF NR
| 37.3 | 14 | 0.00 (0.00 to 1.10) | 10 | 0.00 (0.00 to 0.00) | 0.07 | 0.09 | ||
IL15
CSF/PBMC Ratio
| 13 | 0.00 (0.00 to 0.97) | 10 | 0.00 (0.00 to 0.00) | 0.06 | 0.09 | |||
IL23
PBMC NR
| 34.6 | 16 | 0.80 (0.65 to 1.18) | 10 | 0.80 (0.62 to 1.15) | 1.00 | 0.44 | ||
IL23
CSF NR
| 36.8 | 14 | 1.12 (0.00 to 1.98) | 10 | 0.00 (0.00 to 0.48) | 0.10 | 0.10 | ||
IL23
CSF/PBMC Ratio
| 13 | 1.54 (0.00 to 3.17) | 0.28 | 10 | 0.00 (0.00 to 1.50) | 0.09 | 0.10 | ||
IL27
PBMC NR
| 35.1 | 16 | 4.74 (2.49 to 7.60) | 10 | 8.36 (6.17 to 12.70) |
0.045
| 0.09 | ||
IL27
CSF NR
| ND | 14 | 0.00 (0.00 to 0.00) | 10 | 0.00 (0.00 to 0.00) | 1.00 | 0.44 | ||
IL27
CSF/PBMC Ratio
| 12 | 0.00 (0.00 to 0.00) | 10 | 0.00 (0.00 to 0.00) | 1.00 | 0.44 | |||
LTA
PBMC NR
| 34.3 | 16 | 1.74 (1.43 to 2.30) | 10 | 2.11 (1.59 to 2.73) | 0.37 | 0.23 | ||
LTA
CSF NR
| 36.7 | 14 | 1.41 (0.00 to 3.20) | 10 | 0.00 (0.00 to 0.00) | 0.05 | 0.09 | ||
LTA
CSF/PBMC Ratio
| 13 | 1.08 (0.00 to 1.82) | 0.92 | 10 | 0.00 (0.00 to 0.00) |
0.04
| 0.09 | ||
LTB
PBMC NR
| 28.2 | 16 | 4.09 (2.44 to 4.99) | 10 | 3.71 (3.06 to 5.54) | 0.79 | 0.37 | ||
LTB
CSF NR
| 33.5 | 13 | 8.05 (6.62 to 13.37) | 10 | 8.82 (3.30 to 12.27) | 0.42 | 0.23 | ||
LTB
CSF/PBMC Ratio
| 12 | 2.37 (1.76 to 3.12) |
0.002
| 10 | 1.70 (0.97 to 3.12) |
0.04
| 0.13 | 0.11 |
CSF-cell subsets and cytokine gene expression in RRMS relapse patients
RRMS relapse | NIND | RRMS relapse vs NIND | |||||||
---|---|---|---|---|---|---|---|---|---|
Mean CT | N | Median (IQR) | CSF/PBMC p-value | N | Median (IQR) | CSF/PBMC p-value | p-value | q-value | |
CD3d
PBMC NR
| 31.1 | 15 | 0.06 (0.04 to 0.08) | 10 | 0.05 (0.04 to 0.07) | 0.47 | 0.26 | ||
CD3d
CSF NR
| 35.6 | 13 | 0.10 (0.08 to 0.23) | 10 | 0.06 (0.02 to 0.15) | 0.07 | 0.09 | ||
CD3d
CSF/PBMC Ratio
| 11 | 2.37 (1.30 to 3.67) |
0.01
| 10 | 1.12 (0.42 to 3.07) | 0.39 | 0.23 | 0.17 | |
CD14
PBMC NR
| 30.1 | 15 | 0.33 (0.27 to 0.38) | 10 | 0.33 (0.29 to 0.49) | 0.38 | 0.23 | ||
CD14
CSF NR
| 34.9 | 13 | 0.38 (0.07 to 0.52) | 10 | 1.52 (0.94 to 2.40) |
0.0004
|
0.01
| ||
CD14
CSF/PBMC Ratio
| 11 | 1.39 (0.30 to 2.15) | 0.42 | 10 | 4.01 (3.54 to 5.16) |
0.01
|
0.003
|
0.02
| |
CD19
PBMC NR
| 36.1 | 15 | 0.14 (0.09 to 0.23) | 10 | 0.08 (0.05 to 0.16) | 0.05 | 0.09 | ||
CD19
CSF NR
| 37.1 | 13 | 0.00 (0.00 to 0.35) | 10 | 0.00 (0.00 to 0.00) |
0.03
| 0.09 | ||
CD19
CSF/PBMC Ratio
| 11 | 0.00 (0.00 to 0.89) | 10 | 0.00 (0.00 to 0.00) |
0.04
| 0.09 | |||
CD56
PBMC NR
| 34.9 | 15 | 0.15 (0.09 to 0.23) | 10 | 0.16 (0.07 to 0.32) | 0.96 | 0.43 | ||
CD56
CSF NR
| 37.8 | 13 | 0.00 (0.00 to 0.03) | 10 | 0.00 (0.00 to 0.00) | 0.11 | 0.10 | ||
CD56
CSF/PBMC Ratio
| 11 | 0.00 (0.00 to 0.00) | 10 | 0.00 (0.00 to 0.00) | 0.17 | 0.14 |
RRMS relapse and NIND | RRMS relapse | |||||||||
---|---|---|---|---|---|---|---|---|---|---|
CSF NR | N | R | p-value | q-value | N | R | p-value | q-value | ||
IFNG
| vs |
CD3d
| 22 | 0.383 | 0.08 | 0.06 | 12 | 0.441 | 0.151 | 0.39 |
IFNG
| vs |
CD14
| 22 | −0.606 |
0.003
|
0.008
| 12 | 0.018 | 0.956 | 0.90 |
IFNG
| vs |
CD19
| 22 | 0.538 |
0.010
|
0.02
| 12 | 0.111 | 0.731 | 1.04 |
IFNG
| vs |
CD56
| 22 | 0.372 | 0.09 | 0.06 | 12 | 0.019 | 0.954 | 0.93 |
IL1B
| vs |
CD3d
| 23 | −0.453 |
0.03
|
0.03
| 13 | 0.064 | 0.837 | 0.89 |
IL1B
| vs |
CD14
| 23 | 0.126 | 0.57 | 0.27 | 13 | 0.067 | 0.829 | 0.91 |
IL1B
| vs |
CD19
| 23 | 0.168 | 0.44 | 0.21 | 13 | 0.435 | 0.137 | 0.39 |
IL1B
| vs |
CD56
| 23 | 0.110 | 0.62 | 0.28 | 13 | 0.285 | 0.345 | 0.73 |
IL7
| vs |
CD3d
| 23 | 0.090 | 0.68 | 0.30 | 13 | −0.121 | 0.694 | 1.03 |
IL7
| vs |
CD14
| 23 | −0.169 | 0.44 | 0.22 | 13 | 0.087 | 0.777 | 0.98 |
IL7
| vs |
CD19
| 23 | 0.600 |
0.002
|
0.009
| 13 | 0.513 | 0.073 | 0.36 |
IL7
| vs |
CD56
| 23 | 0.412 | 0.05 |
0.04
| 13 | 0.241 | 0.429 | 0.86 |
IL10
| vs |
CD3d
| 23 | −0.506 |
0.01
|
0.02
| 13 |
−0.589
|
0.034
| 0.23 |
IL10
| vs |
CD14
| 23 | 0.773 |
0.00002
|
0.00011
| 13 | 0.541 | 0.056 | 0.32 |
IL10
| vs |
CD19
| 23 | −0.438 |
0.04
|
0.04
| 13 | −0.151 | 0.623 | 1.01 |
IL10
| vs |
CD56
| 23 | −0.242 | 0.27 | 0.15 | 13 | 0.086 | 0.781 | 0.95 |
IL12A
| vs |
CD3d
| 23 | 0.075 | 0.73 | 0.31 | 13 | −0.096 | 0.755 | 1.03 |
IL12A
| vs |
CD14
| 23 | −0.327 | 0.13 | 0.08 | 13 | −0.236 | 0.437 | 0.78 |
IL12A
| vs |
CD19
| 23 |
0.577
|
0.004
|
0.008
| 13 | 0.508 | 0.076 | 0.32 |
IL12A
| vs |
CD56
| 23 |
0.756
|
0.00003
|
0.00014
| 13 | 0.697 |
0.008
| 0.09 |
IL15
| vs |
CD3d
| 23 | 0.226 | 0.30 | 0.16 | 13 | 0.067 | 0.828 | 0.94 |
IL15
| vs |
CD14
| 23 |
−0.519
|
0.01
|
0.02
| 13 | −0.450 | 0.123 | 0.42 |
IL15
| vs |
CD19
| 23 |
0.847
|
0.0000003
|
0.0000048
| 13 | 0.758 |
0.003
| 0.09 |
IL15
| vs |
CD56
| 23 |
0.469
|
0.02
|
0.03
| 13 | 0.349 | 0.242 | 0.59 |
IL23
| vs |
CD3d
| 23 | 0.004 | 0.98 | 0.39 | 13 | 0.034 | 0.912 | 0.91 |
IL23
| vs |
CD14
| 23 | −0.396 | 0.06 | 0.05 | 13 | −0.040 | 0.898 | 0.92 |
IL23
| vs |
CD19
| 23 | 0.340 | 0.11 | 0.07 | 13 | 0.177 | 0.562 | 0.96 |
IL23
| vs |
CD56
| 23 | 0.234 | 0.28 | 0.15 | 13 | 0.092 | 0.766 | 1.00 |
LTA
| vs |
CD3d
| 23 | 0.002 | 0.99 | 0.39 | 13 | −0.121 | 0.693 | 1.07 |
LTA
| vs |
CD14
| 23 | −0.420 | 0.05 | 0.04 | 13 | −0.078 | 0.800 | 0.94 |
LTA
| vs |
CD19
| 23 |
0.563
|
0.005
|
0.009
| 13 | 0.461 | 0.113 | 0.43 |
LTA
| vs |
CD56
| 23 | 0.384 | 0.07 | 0.05 | 13 | 0.238 | 0.433 | 0.82 |
LTB
| vs |
CD3d
| 23 |
0.579
|
0.004
|
0.009
| 13 | 0.440 | 0.133 | 0.41 |
LTB
| vs |
CD14
| 23 | −0.057 | 0.80 | 0.33 | 13 | 0.305 | 0.310 | 0.70 |
LTB
| vs |
CD19
| 23 | −0.337 | 0.12 | 0.07 | 13 | −0.702 |
0.008
| 0.13 |
LTB
| vs |
CD56
| 23 | −0.304 | 0.16 | 0.09 | 13 | −0.598 |
0.031
| 0.26 |