2. Cluster of Differentiation

3. C ONTENTS What Is Cluster of Differentiation 1 CD Nomenclature 2 Immunophenotyping 3 CD Markers in Solid Tumors 4 List of Human Cluster of Differentiation 5

4. 1 What Is Cluster of Differentiation

5. 1 Cluster of differentiation (CD) is a surface marker that identifies a particular differentiation lineage recognized by a group of monoclonal antibodies. 2 CD antigens are molecules originally defined as being present on the cell surface of leucocytes and recognized by specific antibody molecules, but now including some intracellular molecules and molecules present on cells other than leucocytes. 3 Physiologically, CD antigens do not belong in any particular class of molecules. What Is Cluster of Differentiation X

6. 1 Initially, CD designation was intended to describe a cluster of mAbs that display the same cellular reactivity and identify the same molecular species. Later, the CD designations were used to describe the recognized molecules, but had to be clarified by attaching the term antigen or molecule to the designation. Currently, CD2 is generally used to designate the molecule, and CD2 antibody is used to designate the antibody. What Is Cluster of Differentiation X

7. Characterization and formal designation of more than 400 molecules. Tools for the treatment of several malignancies and autoimmune diseases. CD molecules are commonly used as cell markers. mAbs against these molecules are essential for biomedical research, diagnosis, and biotechnology. What Is Cluster of Differentiation X

8. 2 CD Nomenclature

9. X CD# a/b/ A lowercase letter following the CD number CD1a CD1b Several molecules that share a common chain CD66a CD66b CD66c Name different members of the same gene family CD15s CD60b Modification of the same carbohydrate sequence CD60b = 9-O- acetylated ganglioside GD3 CDw1 86 A lowercase “w” stands for “workshop”, if the molecule has not been well characterized, or has only one mAb hi, mid or low CD34+ CD31−

10. Initiate multi-laboratory studies on important differentiation markers, irrespective of their cellular location Allocate CD numbers to new antibodies and new molecules where adequate data was submitted Clarify and bring up to date earlier provisional (CDw) designations Fill in some of the CD numbers that had previously been reserved for members of molecule families such as chemokine receptors. CD Nomenclature X

11. 6 The level of the cut-points were selected such that the "distances" between mAb within a cluster were minimized while the "distances" between mAb in separate clusters were maximized. 5 A hierarchical tree was established, linking together in decreasing order of similarity all mAb tested. 4 The calculated "distance" for all mAb pairs were then analyzed using a hierarchical clustering algorithm. 3 In order to identify mAb detecting similar antigens, all mAb were compared pair-wise and their similarities or differences designated as the "distance" between them. 2 Other participating researchers perform additional tests, such as immunohistochemistry on tissue sections or the biochemical characterization of the target. 1 Participating laboratories perform specific blind studies with the mAbs included in the panel. Process CD Nomenclature X

12. 3 Immunophenotyping

13. 1 A process that uses antibodies to identify cells based on the types of antigens or markers on the surface of the cells. This process is used to diagnose specific types of leukemia lymphoma and other cells of the immune system. Clusters of Differentiation (CD) markers are widely used for immunophenotyping. Immunophenotyping X

14. X Type of cellCD markers Stem cellsCD34+, CD31-, CD117 all leukocyte groupsCD45+ GranulocyteCD45+, CD11b, CD15+, CD24+, CD114+, CD182+ MonocyteCD4, CD45+, CD14+, CD114+, CD11a, CD11b, CD91+, CD16+ T lymphocyteCD45+, CD3+ T helper cellCD45+, CD3+, CD4+ T regulatory cellCD4, CD25, FOXP3 (a transcription factor) Cytotoxic T cellCD45+, CD3+, CD8+ B lymphocyteCD45+, CD19+, CD20+, CD24+, CD38, CD22 ThrombocyteCD45+, CD61+ Natural killer cellCD16+, CD56+, CD3-, CD31, CD30, CD38 CD markers for immunophenotyping

15. Immunophenotyping X T cell CD antigenB cell CD antigen

16. Immunophenotyping X Thymopoiesis T Cell CD3/TCR

17. Function CD3/TCR 1 Recognition of antigen leads to signal transduction mediated by the invariant chains and subsequently T cell activation. Consequences of binding by TCR depend on antigen density and affinity of TCR for antigen and may result in unresponsiveness. Signal transduction involves tyrosine kinase and phospholipase C activation followed by phosphoinositide turnover and activation of several second messenger pathways. Immunophenotyping X CD3/TCR

18. CD4 is expressed on most thymocytes and approximately two-thirds of peripheral blood T cells, which constitute the CD8 - cells. In human and rat but not in mouse, CD4 is expressed on monocytes and macrophages. Immunophenotyping X CD4

19. ） Immunophenotyping X CD4

20. CD8 is expressed on most thymocytes and approximately one-third of peripheral blood T cells, which constitute the CD4 - cells. CD8αβ heterodimers are expressed only on TCRαβ cells whereas CD8α homodimers can be expressed on αβ and γδ T cells and some NK cells. Distribution 1 The IgSF domain of CD8α binds to the α3 domain of MHC Class I. Like CD4, a CXCP motif in the cytoplasmic domain of CD8α mediates binding to the tyrosine kinase Lck. Ligands 3 CD8 is expressed as a heterodimer of CD8α and CD8β or as a CD8α homodimer. CD8α is required for expression of CD8β. Structure 2 CD8 acts as a co-receptor with MHC Class I- restricted TCRs in antigen recognition. Analysis of mice lacking CD8α or CD8β show that the co-receptor function of CD8 is important for selection of MHC Class I-restricted CD8 + T cells during development. Function 4 Immunophenotyping X CD8

21. 4 CD Markers in Solid Tumors

22. Effect of epigenetic changes on the expression of CD markers CD Markers in Solid Tumors X miRNAExpressionDiseaseCD markerMechanismsPrognosis miR-199-5bDownMedullobla stomaCD15 Methylation in the CpG islands at the upstream promoter Binding Hes1 factor to promotoer miT-199-5b binding site in the 3'-UTR of CD15 Poor miR-199aDownOvarian cancerCD44Binding site in the 3'-UTRPoor miR-193aDownOvarian cancerCD117 Binding of E2F6 to polycombcomplex of miR-193a Binding site in the 3'-UTR of CD117 Poor miR-370DownLung cancerCD105 Binding TRAF4 in the 3'-UTR of miR-370 Binding site in the 3'-UTR of CD105 Poor miR-142-3pDown Hepatic and colon cancer CD133 Binding OCT4 to promoter of miR-142-3p Binding site in the 3'-UTR of CD133 Poor miR-34aDownProstate cancerCD44 BExpression of miR-34a is repressed by p53 Binding site in the 3'-UTR of CD44 Poor miR-224 miR-486 Down pancreatic ductal adenocarcin omas CD40Binding site in tpa 3'-UTR of CD40Poor miR-200DownLung cancerCD274Binding site in the 3'-UTR of CD274Poor

23. CD Markers in Solid Tumors X The expression of CD makers in cancers and their diagnostic and prognostic value Type of Cancer Commonly indentified CD markers Prognostic CD markers Diagnostic CD markers Type of sample Type of diagostic technique Liver CD133, CD44, CD90, CD105, CD34, CD151, CD206, CD68 CD151, CD68, CD206 ------ Tissue, Cell line IHC, Flow cytometery, RT- PCR Colerctal CD66, CD110, CD133, CD44, CD2, CD89, CD200 CD200 CD110, CD133, CD44, CD200 CD66 Tissue, Cell line, Peripheral blood IHC, Flow cytometery, RT- PCR Lung CD117, CD176, CD166, CD88, CD103CD66 CD88, CD103, CD66 Tissue, Cell line, Peripheral blood IHC, Flow cytometery, RT- PCR Breast CD44, CD24, CD133, CD14, CD200, CD4 CD8, CD4 CD25 CD44, CD133, CD14 ------ Tissue, Cell line, Peripheral blood IHC, Flow cytometery, ELISA

24. CD Markers in Solid Tumors X Expression of CD makers and their response to treatment Cancerpredictive CD markersFunctionType of treatmentResponse Liver CD133Activation of ERK pathwaySorafenibIneffective CD279Phosphorylation of ERK Sorafenib +anti-PD-1 Good CD152Suppression of T-cellsTremelimumab+TACEGood Colorectal CD66ApoptosisChemotherapy-5-FUGood CD47 Mediated phagocytosis by macrophage mAbGood CD147 Apoptosis and arrested cell cycle 5-FUGood Lung CD133 Expression ABCG2 and CXCR4 CisplatinIneffective CD317Induced ADCC and CDCmAbGood CD66Sensitive to anoikis8F5m Ab+PaclitaxelGood Breast CD166----- Taxane-free adjuvant chemotherapy Good HER2 with CD3Stimulate T-cellSpecific AbGood CD24 Reduce expression drug resistance factor Chemotherapy-5-FUGood

25. 5 List of Human Cluster of Differentiation

26. List of Human Cluster of Differentiation X list of human clusters of differentiation 1-50 CD1(a-c 1A 1D 1E) CD2 CD3(γ δ ε) CD4 CD5 CD6 CD7 CD8 a CD9 CD10 CD11(a b c d) CD13 CD14 CD15 CD16 A B CD18 CD19 CD20 CD21 CD22 CD23 CD24 CD25 CD26 CD27 CD28 CD29 CD30 CD31 CD32 A B CD33 CD34 CD35 CD36 CD37 CD38 CD39 CD40 CD41 CD42(a b c d) CD43 CD44 CD45 CD46 CD47 CD48 CD49(a b c d e f) CD50 51-100 CD51 CD52 CD53 CD54 CD55 CD56 CD57 CD58 CD59 CD61 CD62(E L P) CD63 CD64 A B C CD66(a b c d e f) CD68 CD69 CD70 CD71 CD72 CD73 CD74 CD78 CD79a/b CD80 CD81 CD82 CD83 CD84 CD85(a d e h j k) CD86 CD87 CD88 CD89 CD90 CD91 - CD92 CD93 CD94 CD95 CD96 CD97 CD98 CD99 CD100CD59 101-150 CD101 CD102 CD103 CD104 CD105 CD106 CD107a/b CD108 CD109 CD110 CD111 CD112 CD113 CD114 CD115 CD116 CD117 CD118 CD119 CD120a/b CD121a/b CD122 CD123 CD124 CD125 CD126 CD127 CD129 CD130 CD131 CD132 CD133 CD134 CD135 CD136 CD137 CD138 CD140b CD141 CD142 CD143 CD144 CD146 CD147 CD148 CD150 151-200 CD151 CD152 CD153 CD154 CD155 CD156(a b c) CD157 CD158 (a d e i k) CD159a/c CD160 CD161 CD162 CD163 CD164 CD166 CD167 a b CD168 CD169 CD170 CD171 CD172 a b g CD174 CD177 CD178 CD179 a b CD180 CD181 CD182 CD183 CD184 CD185 CD186 CD191 CD192 CD193 CD194 CD195 CD196 CD197 CDw198 CDw199 CD200

27. 201-250 CD201 CD202b CD204 CD205 CD206 CD207 CD208 CD209 CDw210a/b CD212 CD213a 1 2 CD217 CD218a/b CD220 CD221 CD222 CD223 CD224 CD225 CD226 CD227 CD228 CD229 CD230 CD233 CD234 CD235a/b CD236 CD238 CD239 CD240CE CD240D CD241 CD243 CD244 CD246 CD247 - CD248 CD249 251-300 CD252 CD253 CD254 CD256 CD257 CD258 CD261 CD262 CD263 CD264 CD265 CD266 CD267 CD268 CD269 CD271 CD272 CD273 CD274 CD275 CD276 CD278 CD279 CD280 CD281 CD282 CD283 CD284 CD286 CD288 CD289 CD290 CD292 CDw293 CD294 CD295 CD297 CD298 CD299 301-350 CD300A CD301 CD302 CD303 CD304 CD305 CD306 CD307 CD309 CD312 CD314 CD315 CD316 CD317 CD318 CD320 CD321 CD322 CD324 CD325 CD326 CD328 CD329 CD331 CD332 CD333 CD334 CD335 CD336 CD337 CD338 CD339 CD340 CD344 CD349 CD350 Surface antigens Major histocompatibility complex/Human leukocyte antigen MHC class I HLA-A HLA-B HLA-C HLA-E HLA-F HLA-G MHC class II HLA-DM α β HLA-DO α β HLA-DP α1 β1 HLA-DQ α1 α2 β1 β2 β3 HLA-DR α β1 β3 β4 β5 Minor histocompatibility antigen Other Arrestin Calgranulin Blood group antigens Cell adhesion molecules Differentiation antigens List of Human Cluster of Differentiation X

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