|
|
|
Specificity/antibody Mono/Poly Species Working dilution Sourcea PGP 9.5 (RA95101) Poly Rabbit 1 : 100 UltraClone Substance P Poly Rabbit prediluted Zymed CGRP Poly Rabbit 1 : 10000 Affiniti VIP Poly Rabbit prediluted Zymed Somatostatin Poly Rabbit 1 : 300 Dako Neuropeptide Y (B48-1) Poly Rabbit 1 : 600 Euro-Diagnostica PMN (neutrophil elastase) + + Mono Mouse 1 : 150 Dako Langerhans cells (CD1a) Mono Mouse 1 : 100 Dako Macrophages (CD68) Mono Mouse 1 : 100 Dako Ki-67 Mono Mouse 1 : 50 Dako NGF Mono Mouse 1 : 200 Austral Biologicals NGF-receptor (Trk A) Mono Mouse 1 : 10 Boehringer Mannheim NGF-receptor (p75NTR) Mono Mouse 1 : 50 Dako BDNF Mono Mouse 1 : 1000 Santa Cruz NT-3 Poly Rabbit 1 : 200 Santa Cruz Neutral endopeptidase (CD10) Mono Mouse 1 : 80 Novocastra Angiotensin-converting enzyme Mono Mouse 1 : 5000 Chemicon IFN-γ Poly Mouse 1 : 100 Dako TNF-α Poly Mouse 1 : 100 Dako IL-1α Poly Mouse 1 : 500 Dako IL-1β Poly Mouse 1 : 100 Dako IL-2 Poly Mouse 1 : 500 Dako IL-4 Poly Mouse 1 : 500 Dako IL-5 Poly Mouse 1 : 30 Genzyme-Techne IL-6 Poly Mouse 1 : 100 Dako IL-8 Poly Mouse 1 : 500 Dako IL-10 Poly Mouse 1 : 200 Biogenesis IL-12 Poly Mouse 1 : 100 Dako PECAM-1 (CD31) Mono Mouse 1 : 40 Dako ICAM-1 (CD54) Mono Mouse 1 : 500 Dako ELAM-1 (CD62E) Mono Mouse 1 : 200 Chemicon VCAM-1 (CD106) Mono Mouse 1 : 50 Dako BDNF, brain-derived neurotrophic factor; CGRP, calcitonin gene-related peptide; ELAM, endothelial leucocyte adhesion molecule; ICAM, intercellular cell adhesion molecule; IFN, interferon; IL, interleukin; NGF, nerve growth factor; NT-3, neurotrophin-3; PECAM, platelet endothelial cell adhesion molecule; PGP, protein gene product; PMN, polymorphonuclear leucocyte; TNF, tumour necrosis factor; VCAM, vascular cell adhesion molecule; VIP, vasoactive intestinal peptide.
aAffiniti Research Products Ltd, Exeter, U.K.; Austral Biologicals, San Ramon, CA, U.S.A.; Biogenesis Inc., Kingston, NH, U.S.A.; Boehringer Mannheim Biochemica, Mannheim, Germany; Chemicon International Inc, Temecula, CA, U.S.A.; Dako Japan Co. Ltd, Kyoto, Japan; Euro-Diagnostica, Westbury, NY, U.S.A.; Genzyme-Techne, Wako Pure Chemical Industries Ltd, Osaka, Japan; Novocastra Laboratories Ltd, Newcastle upon Tyne, U.K.; Santa Cruz Biotechnology Inc., Santa Cruz, CA, U.S.A.; UltraClone Limited, Isle of Wight, U.K.; Zymed Laboratories Inc., San Francisco, CA, U.S.A.
Pruritus (-)
(n = 15)Pruritus (+)
(n = 23)P-valuea PASI (0-73) 20·65 ± 8·31 18·73 ± 10·11 NS Lymphocytes (D) (cells mm-2) 99·66 ± 12·50 100·63 ± 16·21 NS Neutrophils (E) (cells mm-2) 74·60 ± 22·04 65·55 ± 18·24 NS Neutrophil elastase (E) (0-5) 2·41 ± 0·55 2·24 ± 0·32 NS Eosinophils (E) (cells mm-2) 19·17 ± 6·54 18·90 ± 4·76 NS Mast cells (D) (cells mm-2) 36·44 ± 10·22 61·75 ± 8·02 P < 0·01 Activated mast cells (D) (0-4) 0·83 ± 0·24 2·18 ± 0·45 P < 0·001 CD1a (+) cells (E) (cells mm-2) 29·25 ± 7·10 35·69 ± 13·63 NS CD68 (+) cells (D) (cells mm-2) 20·82 ± 3·05 17·01 ± 6·23 NS Ki-67 (+) cells (E) (per mm epidermis) 122·80 ± 36·13 108·57 ± 24·61 NS D, dermis; E, epidermis, NS, not significant.
aAll P-values were determined using the unpaired Student's t-test.
Pruritus (-) (n = 15) Pruritus (+) (n = 23) P-valuea Neurotrophins NGF (E) (0-5)b 1·25 ± 0·74 3·85 ± 1·55 P < 0·001 NGF content (pg g-1 tissue) 96·84 ± 25·7 154·33 ± 34·1 P < 0·01 BDNF (E) (0-5) 1·45 ± 0·30 1·24 ± 0·41 NS NT-3 (E) (0-5) 0·84 ± 0·31 0·88 ± 0·27 NS Nerves and neuropeptides PGP 9.5 (E) (No. per mm2)b 4·45 ± 1·75 9·01 ± 2·32 P < 0·05 (D) (No. per mm2) 9.54 ± 1·46 14·46 ± 2·41 P < 0·01 SP (D) (No. per mm2) 6·96 ± 1·71 11·50 ± 2·85 P < 0·05 CGRP (D) (No. per mm2) 2·32 ± 0·50 2·50 ± 0·38 NS VIP (D) (No. per mm2) 5·21 ± 1·21 5·2 ± 1·19 NS NPY (D) (No. per mm2) 1·68 ± 0·30 1·23 ± 0·22 NS SOM (D) (No. per mm2) 2·08 ± 0·29 1·68 ± 0·41 NS NGF receptor NGF-R (Trk A) (E) (0-5)b 0·69 ± 0·14 2·31 ± 0·40 P < 0·005 NGF-R (p75NTR) (E) (0-5) 0·43 ± 0·09 0·62 ± 0·17 NS NGF-R (Trk A) (D) (0-5) 2·04 ± 0·53 3·41 ± 1·08 P < 0·01 NGF-R (p75NTR) (D) (0-5) 0·95 ± 0·38 0·74 ± 0·26 NS Neuropeptide-degrading enzymes NEP (E) (0-5) 2·65 ± 0·55 1·07 ± 0·38 P < 0·005 (V) (0-5) 2·10 ± 0·65 0·75 ± 0·29 P < 0·001 ACE (V) (0-5) 1·86 ± 0·44 1·52 ± 0·69 NS ACE, angiotensin-converting enzyme; BDNF, brain-derived neurotrophic factor; CGRP, calcitonin gene-related peptide; NEP, neutral endopeptidase; NGF, nerve growth factor; NPY, neuropeptide Y; NT-3, neurotrophin-3; PGP, protein gene product; SOM, somatostatin; SP, substance P; VIP, vasoactive intestinal peptide; D, dermis; E, epidermis; V, vessel; NS, not significant.
aAll P-values were determined using the unpaired Student's t-test.
bA significant positive correlation with the severity of pruritus expressed by visual analogue scale and by the verbal pruritic scale using Spearmann rho test (see text for detail).
Pruritus (-)
(n = 15)Pruritus (+)
(n = 23)P-valuea IFN-γ (D) (0-5) 1·45 ± 0·12 1·36 ± 0·35 NS TNF-α (E) (0-5) 2·84 ± 0·43 2·50 ± 0·22 NS (D) (0-5) 1·77 ± 0·25 1·82 ± 0·40 NS IL-1α (E) (0-5) 1·21 ± 0·26 1·59 ± 0·38 NS (D) (0-5) 1·38 ± 0·04 1·27 ± 0·09 NS IL-1β (D) (0-5) 1·15 ± 0·23 1·12 ± 0·30 NS IL-2 (D) (0-5) 1·44 ± 0·31 2·79 ± 0·54 P < 0·005 IL-4 (D) (0-5) 0·87 ± 0·15 0·63 ± 0·17 NS IL-5 (D) (0-5) 1·10 ± 0·33 1·32 ± 0·27 NS IL-6 (E) (0-5) 2·50 ± 0·69 2·74 ± 0·83 NS (D) (0-5) 1·54 ± 0·44 1·17 ± 0·36 NS IL-8 (E) (0-5) 2·26 ± 0·55 2·68 ± 0·29 NS (D) (0-5) 1·79 ± 0·47 1·38 ± 0·35 NS IL-10 (D) (0-5) 1·07 ± 0·18 1·33 ± 0·25 NS IL-12 (D) (0-5) 1·17 ± 0·41 1·30 ± 0·25 NS IFN, interferon; IL, interleukin; TNF, tumour necrosis factor; D, dermis; E, epidermis; NS, not significant.
aAll P-values were determined using the unpaired Student's t-test.
Pruritus (-)
(n = 15)Pruritus (+)
(n = 23)P-valuea CD31 (PECAM-1) (V) (0-4) 3·25 ± 0·51 3·11 ± 0·66 NS ICAM-1 (E) (0-5) 1·02 ± 0·17 1·19 ± 0·34 NS (V) (0-5) 2·34 ± 0·37 2·19 ± 0·54 NS ELAM-1 (V) (0-5)b 1·98 ± 0·25 4·06 ± 0·80 P < 0·001 VCAM-1 (V) (0-5) 2·10 ± 0·64 2·55 ± 0·75 NS ELAM, endothelial leucocyte adhesion molecule; ICAM, intercellular cell adhesion molecule; PECAM, platelet endothelial cell adhesion molecule; VCAM, vascular cell adhesion molecule; E, epidermis; V, vessel; NS, not significant.
aAll P-values were determined using the unpaired Student's t-test.
bA significant positive correlation with the severity of pruritus expressed by VAS and by the verbal pruritic scale using Spearmann rho test (see text for detail).