New monoclonal antibodies (mAbs) of the 9th HLDA Workshop: an immunohistochemical analysis of the FACS and IHC panel mAbs on normal and activated skin and tonsil
Cross reactivity and multiple reactivities are very frequently detected in tissue
studies; this study summarize the pattern of tissue staining of normal and pathological skin (affected from cutaneous lymphomas) and normal tonsil observed by using FACS and IHC panels of the 9th HLDA Workshop.
FACS panel: cryostat sections of frozen tissues were dried for 12 hours, fixed 10" in
aceton and stained by an avidin-biotin immunoalkaline phosphatase method (DAKO). IHC panel: formalin fixed paraffin-embedded sections were dewaxed, heated in EDTA buffer (PH-8) for 15 min in a pressure cooker and immunostained by an avidin-biotin immunoalkaline-phosphatase method (DAKO). A) Skin reactivity results: N\ub0 7 (FcRLB) and N\ub08 (CD20L) labelled epithelial cells (EP) and
polymophonuclear leucocytes (PMN); N\ub0 16 (XBP-1s) smooth muscle (SM), EP, dendritic cells (DR) and melanocytes (MEL); N\ub0 18 DC, SM and EP; N\ub0 20/21 (FCRL1-2) EP, collagen fibers (COLL) and nerves (N); N\ub0 24 EP, COLL fibers, macrophages (MO) and fibroblasts (Fb); N\ub0 26 (SC3) hair and eccrine gland (ADN); N\ub0 30(CD210) EP; N\ub0 32(CDW329) DR, MO, EP; N\ub0 33 DC, EP and ADN; N\ub0 34 MO, EP and AND; N\ub0 35 MO; N\ub0 39 (CD124) EP; N\ub0 40(CD200) vessels (Vs) and ADN; N\ub0 41(CD130) Vs and EP; N\ub0 48(CD196) EP and ADN; 49(CD267) ADN; N\ub0 52(CD191) EP and AND; 53(Int.beta5) and 54 (CD320) Vs, EP, AND, Fb; N\ub0 55(DR4), N\ub0 56(CMKLR1) EP and AND; N\ub0 57(DR5) MO, Fb, EP, and AND; N\ub0 58(Notch-1) Vs, EP and AND; N\ub0 59 (4IgB7-H3) EP, Vs, striated muscle; N\ub0 74(Int.beta6) EP and AND; N\ub0 75(Dectin-1) Vs, EP and AND; N\ub0 76-101(TREM-1) Vs, EP, MO and PMN; N\ub0 77(Siglec-5/14) PMN and MO; N\ub0 78(AMICA) Epand AND;
N\ub0 80(Siglec-9) MO, DR and PMN; N\ub0 82(int.alfaVbeta5) MO, DR, EP, COLL, Fb, Vs; N\ub0 84(DCIR) PMN; N\ub0 85/99(GITR) PMN, EP and AND; N\ub0 86(CD229) MO; N\ub0 87(CD319) MO and plasmocytes (PC); N\ub0 88(CD48) MO, DR and EP; N\ub0 84(CD84) MO, DR and basal membrane (BM); N\ub0 93(NTBA) MO, DR and PC; N\ub0 94(HVEM)
MO, Vs and EP; N\ub0 95(TSLPR) nerves (Nv), EP and ADN; N\ub0 98 (Galectin-3/Mac-2)
MO, PMN, TBM, VS,COLL and EP; N\ub0 100(Lymphotoxin betaR) EP; N\ub0 106(CD150)
MO-DR; N\ub0 115(BLK 10B/2) Vs; N\ub0 116(Bclx/8H) SM, EP, ADN, COLL, Fb; N\ub0 128(CD38) EP; N\ub0 223(AID) EP, ADN and MEL; N\ub0 263(EVI2) MO and PC.
B) Tonsil reactivity with mononuclear cells and resident cells were detected in all
mAbs, also if some reagents showed in our method high background (n\ub0 6, 16, 17, 19, 24, 26, 27, 59, 69, 72, 81, 115, 116) or weak staining (n\ub0 7, 8, 9, 10, 29, 32, 62,
67, 71, 78, 90, 91, 92). Results: N\ub0 11(TRAIL-R2/DR5), 223 (AID), 263 (EVI2b) and
268 (LSP1) showed a pan leukocyte reaction; interestingly BLIMP1 specific N\ub0 6/19 showed a cytoplasmic wide reactivity with FOL, PC, MO and T cell areas, whereas N\ub0103/105 showed a strong nuclear staining of the PC and of the EP cells; N\ub013/107(BTLA) and 48(CD196-CCR6) stained PC and MG/MT zone of the FOL; N\ub0 12/37/66 (PD1) and CD152 intrafollicular activated cells (IAC), whereas N\ub050(CXCR5) stained IAC and MG and MT zone of the FOL; N\ub0 14(GCET-1),
17(LMO2), 18 (KLHL-6), N\ub070 (B7H4) stained GC; N\ub0 29 GC and DR; N\ub0 20/108(FCRL1), 21/110(FCRL2) MG/MT/GC and PC; N\ub0 94 (HVEM), 114 (BLK154), 235 (BAFF-R) MG/MT zone; N\ub0 97 (TCL1) nucleus of the FOL/MT zone; N\ub0 27
(weak), 128-CD38 PC. We hope that our efforts may be useful to better understanding the tissue distribution of the analyzed reagents
