Presentazione workshop sies 2015 muccio

SUITABLE SURFACE ANTIGENS FOR

MYELOMA AND NORMAL PLASMA

CELLS IDENTIFICATION AFTER

MONOCLONAL ANTIBODIES

THERAPY.

Vittorio Emanuele Muccio

Divisione Universitaria di Ematologia

Prof. M. Boccadoro

A.O.U Città della Salute e della Scienza di

Torino


MULTIPLE MYELOMA

SURVIVAL


Surface markers to identify PCs

Surface markers to identify

MALIGNANT Cs

PLASMA CELLS AND FLOW CYTOMETRY


CD38 why?

CD138 why?

CD40 B lymphocytes and Antigen-Presenting Cells

CD31 Platelets, monocytes, neutrophils and T

lymphocytes

Surface markers to identify PCs


CD117

CD13

CD14 CD15

CD20

CD10

CD27 CD19

CD9 CD11b CD40

CD31

CD41

CD56 CD45

CD28

CD23

CD22

CD33 CD24

CD25

CD37

CD39

CD71

HLA-DR

CD44 CD4

CD8

CD81

CD38

CD200

Surface markers to identify malignant PCs


EuroFlow panel for MM MRD study

Tube 1

CD138 CD27 CD38 CD56 CD45 CD19 CD117 CD81

Tube 2

CD138 CD27 CD38 CD56 CD45 CD19 cyIgKappa cyIgLambda


Immunotherapies in MM

Anti-CD38

Anti-CD138

Anti-CD56

Anti-CD319 Anti-CD40


EuroFlow panel for MM MRD study

Tube 1: CD138 CD27 CD38 CD56 CD45 CD19 CD117 CD81

Tube 2: CD138 CD27 CD38 CD56 CD45 CD19 cyIgKappa cyIgLambda


9th International Conference on

Human Leukocyte Differentiation Antigens (HLDA9)

82 Surface molecules

SIALOADESINES

SLAM

INTEGRINS TNF/TNFR

IRTA CITOCHINES-R

CHEMOKINES-R CD28/B7 TLR/C-TYPE LEC R


# mAb Specificity Conjugated fluorochrome CD Isotype Ab Species % of positive MM samples

20 FCRL1 PE CD307a IgG1k Mouse 0 21 FCRL2 PE CD307b IgG2Ak Mouse 0 22 FCRL3 PE CD307c IgG2Bk Mouse 0 23 FCRL4 PE CD307d IgG2Ak Mouse 0 28 Siglec-7 PE CD328 IgG1k Mouse 0 29 B7-2 PE CD86 IgG1k Mouse 64* 30 IL-10R PE CD210 IgG2ak Rat 0 31 CCR7 PE CD197 IgG2ak Rat 0 32 Siglec-9 PE CDw329 IgG1k Mouse 0 33 HB15 PE CD83 IgG1k Mouse 0 34 CTLA-4 PE CD152 IgG2ak Mouse 0 35 B7-1 PE CD80 IgG1k Mouse 0 36 IL6-Rα PE CD126 IgG1k Mouse 45 37 PD1 PE CD279 IgG1k Mouse 0 38 B7-DC PE CD273 IgG1k Mouse 0 39 IL-4Rα PE CD124 IgG1k Mouse 0 40 OX2 PE CD200 IgG1k Mouse 70* 41 gp130 PE CD130 IgG1k Mouse 45 42 IL-12R PE CD212 IgG1k Mouse 0 43 Siglec PE CD328 IgG1k Mouse 0 44 CCR2 A647 CD192 IgG2b Mouse 18 45 ICOS PE CD278 IgG1 Mouse 0 46 CXCR4 PE CD184 IgG2ak Mouse 64* 47 BTLA PE CD272 IgG1k Mouse 64* 48 CCR6 PE CD196 IgG1k Mouse 18 49 TACI PE CD267 IgG2ak Rat 0 50 CXCR5 A647 CD185 IgG2bk Rat 45 51 Integrin beta 7 chain PE NONE IgG2ak Rat 0 52 CCR1 A647 CD191 IgG2bk Mouse 27 53 Integrin beta 5 PE NONE IgG1 k Mouse 0 54 B7-H4 PE NONE IgG1 k Mouse 0 55 DR4 PE CD271 IgG1 k Mouse 0 56 CMKLR1 PE NONE IgG2a k Rat 0 57 DR5 PE CD262 IgG1 k Mouse 18 74 Integrin beta 6 PE NONE IgG2b Mouse 0 75 Dectin-1/CLEC7A PE NONE IgG2b Mouse 0 76 TREM-1 PE NONE IgG1 Mouse 10 77 Siglec-5/Siglec-14 APC CD170 IgG1 Mouse 0 78 AMICA PE NONE IgG2a Mouse 0 79 BLT1/LTB4R1 PE NONE IgG1 Mouse 10 80 Siglec-9 APC CD329 IgG2a Mouse 0 81 IL-23-R PE NONE IgG2b Mouse 9 82 Integrin alfaVbeta5 PE NONE IgG1 Mouse 0 83 CRTAM PE CD355 IgG2b Mouse 0 84 DCIR/CLEC4A PE NONE IgG1 Mouse 0 85 GITR PE NONE IgG1k Mouse 0 86 Ly9 PE CD229 IgG1k Mouse 100* 87 CRACC PE CD319 IgG2b k Mouse 100* 88 BLAST-1 PE CD48 IgG1k Mouse 100* 89 SLAMF5 PE CD84 IgG2a k Mouse 17 90 Toll-like Receptor 3 PE CD283 IgG2a k Mouse 0 91 DcR3 PE NONE IgG1k Mouse 9 92 DR3 (TRAMP) PE NONE IgG1k Mouse 0 93 NTBA PE NONE IgG1k Mouse 100* 94 HVEM PE CD270 IgG1k Mouse 45 95 TSLPR PE NONE IgG1k Mouse 0 96 TWEAK PE CD255 IgG3 k Mouse 0 97 TCL1 PE NONE IgG2b k Mouse 0 98 Galectin-3 (Mac-2) PE NONE IgG1k Mouse 0 99 GITR-L PE NONE IgG1k Mouse 0 100 Lymphotoxin beta receptor PE NONE IgG2b k Mouse 0 101 TREM-1 PE NONE IgG1k Mouse 0 106 SLAM PE CD150 IgG1k Mouse 67* 109 S1PR1 PE CD363 IgG2B Mouse 0 234 IL-15Ralfa PE NONE IgG2b k Mouse 0 235 BAFF-R PE CD268 IgG1 k Mouse 20 236 LIGHT PE CD258 IgG2b k Mouse 20 237 BAFF PE CD257 IgG1 k Mouse 40 238 CD274 PE CD274 IgG2b k Mouse 45 239 B7-H2 PE CD275 IgG2b k Mouse 20 240 CD267 PE CD267 IgG2a k Rat 20 241 NKG2D PE CD314 IgG1 Mouse 0 242 LAIR1 PE CD305 IgG2b Mouse 0 243 DEP-1 PE CD148 IgG1 Mouse 20 244 Syndecan-2 PE NONE IgG2b Rat 20 245 CXCR7/RDC-1 PE NONE IgG2a Mouse 0 246 IL21R PE NONE IgG1 Mouse 0 247 Toll-like Receptor 9 PE CD289 IgG2a k Rat 0 249 TACI PE CD267 IgG2a Mouse 40 250 ABCG2 PE CDw338 IgG2b k Mouse 0 251 BR3 PE CD268 IgG2a k Mouse 0 252 BLyS PE CD257 IgG1 k Mouse 0

In the right column the percentage of positive samples at diagnosis of

MM is reported. Only antigens expressed in >50% of patients were selected for the second

phase of the study,

# mAb

Specificity Conjugated fluorochrome

CD Isotype Ab Species % of positive MM samples

29 B7-2 PE CD86 IgG1k Mouse 64* 40 OX2 PE CD200 IgG1k Mouse 70* 46 CXCR4 PE CD184 IgG2ak Mouse 64* 47 BTLA PE CD272 IgG1k Mouse 64* 86 Ly9 PE CD229 IgG1k Mouse 100* 87 CRACC PE CD319 IgG2b k Mouse 100* 88 BLAST-1 PE CD48 IgG1k Mouse 100* 93 NTBA PE NONE IgG1k Mouse 100* 106 SLAM PE CD150 IgG1k Mouse 67*

TESTED ANTIBODIES


35 NEWLY DIAGNOSED MM 15 HEALTHY SUBJECTS

CD86 CD200 CD272 CD184

CD229 CD319 CD352 CD150

CD48

SECOND PHASE OF THE STUDY


PERCENTAGES OF POSITIVE PCs

*

* *

* *

*

*

*


MFI OF POSITIVE PCs

* * *

*


CONCLUSIONS

CD150, CD86, CD200 and CD352

allow the differentiation between healthy and tumor PCs

Useful in the MRD study after

therapy with Lorvotuzumab

(anti-CD56), Dacetuzumab or

Lucatumumab (anti-CD40)

CD352


Therapeutic targets?

CD272, CD319, CD229 e CD48 are highly expressed by PCs

CONCLUSIONS

Lymphocytes Monocytes Granulocytes

CD150 + + +- - - - - - -

CD352 ++ ++ ++ - - - - - -

CD86 - - - + + + - - -

CD319 -+ -+ -+ - - - - - -

CD272 + + + - - - - - -

CD184 + + + - - - - - -

CD48 + + + + + + - - -

CD229 - - - - - - - - -

CD200 - - - - - - - - -


Useful to carachterize PCs in the MRD study after

therapy with BT062 (anti-CD138) or Daratumumab

(anti-CD38)

CONCLUSIONS


A.O.U Città della Salute e della Scienza di

Torino

Divisione Universitaria di Ematologia

Prof. M. Boccadoro

Dott.ssa Paola Omedè

Gilestro Milena Saraci Elona

Astolfi Monica Oddolo Daniela

Caltagirone Simona Ruggeri Marina

Marzanati Eleonora Aschero Simona

Gattei Valter Zucchetto Antonella


Presentazione workshop sies 2015 muccio

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