TABLE 1.
Collagenase cleavage of surface molecules
| Marker | Subset tested | Blend F (gMFI) |
Type IV (gMFI) |
Trypsin (gMFI) |
Statisticsa | Best enzyme | |||
|---|---|---|---|---|---|---|---|---|---|
| Means | sd | Means | sd | Means | sd | ||||
| Population markers | |||||||||
| HLA-DR | LC | 6094 | 2128 | 4975 | 2947 | 3259 | 894 | NS | NA |
| CD45 | LC | 7544 | 4211 | 12,974 | 7724 | 4799 | 2629 | NS | NA |
| CD1a | LC | 41,052 | 11,087 | 38,823 | 10,006 | 5638 | 1395 | NS | NA |
| CD1c | CD1a | 2193 | 108 | 8771 | 5543 | 1063 | 213 | ** | Type IV |
| CD11c | CD1a | 2118 | 41 | 3094 | 218 | 106 | 21 | NS | NA |
| CD14 | CD14 | 2254 | 796 | 3262 | 2291 | 548 | 24 | NS | NA |
| CD141 | CD141 | 3534 | 2213 | 2403 | 1239 | 245 | 83 | NS | NA |
| Langerin | LC | 4753 | 2915 | 20262 | 18157 | 7516 | 5349 | * | Type IV |
| Epcam | LC | 21,227 | 15,762 | 8448 | 9146 | 10,768 | 1249 | NS | NA |
| Maturation markers | |||||||||
| CD80 | ALLb | — | — | — | — | — | NA | ||
| CD83 | CD1a | — | — | 522 | 150 | ** | Type IV | ||
| CD86 | CD1a | 180 | 90 | 406 | 105 | * | Type IV | ||
| HIV entry receptors | |||||||||
| CD4 | CD1a | 322 | 125 | 843 | 257 | 427 | 49 | * | Type IV |
| CXCR4 | CD14 | — | — | 156 | 51 | 189 | 97 | ** | Type IV |
| CCR5 | CD1a | 454 | 323 | 39 | 53 | 203 | 38 | ** | Blend F |
| CLRs | |||||||||
| CLEC4A (DCIR) | CD1a | 13770 | 4813 | 631 | 142 | 250 | 88 | ** | Blend F |
| CLEC4D | CD14 | 575 | 271 | 136 | 123 | ** | Blend F | ||
| CLEC4E | CD14 | 7021 | 3008 | 4890 | 1840 | NS | NA | ||
| CLEC4G | ALL | 207 | 31 | 74 | 40 | ** | Blend F | ||
| CLEC4L (DC-SIGN) | Mac | 1264 | 450 | 152 | 205 | 0 | — | ** | Blend F |
| CLEC4M | CD14 | 2077 | 621 | 165 | 135 | ** | Blend F | ||
| CLEC5A | CD1a | 18,772 | 7665 | 16,767 | 4976 | 3698 | 488 | NS | NA |
| CLEC5B | ALLb | — | — | — | — | — | NA | ||
| CLEC5C | ALLb | — | — | — | — | — | NA | ||
| CLEC6A | ALLb | — | — | — | — | — | NA | ||
| CLEC7A | CD1a | 508 | 139 | 702 | 137 | NS | NA | ||
| CLEC8A | CD14 | 263 | 99 | 90 | 118 | * | Blend F | ||
| CLEC9A | CD141 | 186 | 66 | 188 | 116 | NS | NA | ||
| CLEC10A | CD14 | 561 | 334 | 44 | 43 | * | Blend F | ||
| CLEC12A | CD14 | 5042 | 869 | 1439 | 1211 | * | Blend F | ||
| CLEC13B (DEC-205) | CD1a | 518 | 321 | 428 | 276 | 2823 | 488 | NS | NA |
| CLEC13D (MR) | CD14 | 6571 | 712 | 2219 | 582 | 0 | — | ** | Blend F |
| CLEC14A | CD141 | 110 | 105 | 119 | 118 | NS | NA | ||
| Siglecs | |||||||||
| Siglec-1 | CD14 | 1168 | 578 | 316 | 306 | * | Blend F | ||
| Siglec-3 | CD1a | 12,616 | 12,452 | 5644 | 4117 | 1938 | 190 | NS | Blend F |
| Siglec-5 | CD1a | 668 | 126 | 298 | 350 | NS | Blend F | ||
| Siglec-6 | ALLb | — | — | — | — | — | NA | ||
| Siglec-9 | CD1a | 3757 | 2304 | 4827 | 3767 | NS | NA | ||
| Siglec-16 | CD1a | 181 | 175 | 144 | 99 | NS | NA | ||
| Other | |||||||||
| CD69 | CD1a | 3123 | 895 | 2876 | 660 | NS | NA | ||
| CD91 | CD14 | 212 | 43 | 0 | 0 | ** | Blend F | ||
Human skin mononuclear phagocytes were isolated from human skin with either blend F or type IV collagenase, and the surface expression was determined for a range of markers on a specific cell subset by flow cytometry with the gating strategy shown in Fig. 3. For each marker, the cell subset was analyzed, and the MFI of surface marker expression after extraction with each enzyme blend (n = 5) was determined. Epcam, epithelial cell adhesion molecule; FC, fold change; gMFI, geometric mean fluorescence intensity; NA, not applicable; NS, not significant. *P ≤ 0.05; **P ≤ 0.01.
a
Mann-Whitney U test.
b
No expression on any subset, confirmed by RNAseq.