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Neelapu and Kwak Figure 3. Tumor-specific cellular immune response induced following membrane proteoliposomevaccination in a representative patient with follicular lymphoma. Cryopreserved prevaccine and postvaccine peripheral bloodmononuclear cells were cultured in either medium alone, or with soluble CD40 ligand–activated autologous tumor cells or solubleCD40 ligand–activated autologous normal B cells and cytokine production was detected by IFN-γ enzyme-linked immunopsot(ELISPOT) assay. Representative wells of ELISPOT demonstrating IFN-γ spots are shown. Significantly increased number of IFN-γspots were detected in response to autologous tumor cells but not in response to autologous normal B cells by the postvaccine Tcells.

Neelapu and Kwak Figure 2. Schema for manufacture of membrane proteoliposomal vaccine. Following an excisional biopsyof at least a 2 cm lymph node, the tumor sample is processed into a single cell suspension and proteins from whole cell membranesare directly extracted from 2 × 109 lymph node biopsy cells with detergent. The membrane proteins are incorporated into liposomesalong with interleukin (IL)-2 to produce membrane-patched proteoliposome vaccine. The liposomes are initially in the form of smallunilamellar vesicles (SUV) when synthesized from dimyristoylphosphatidylcholine (DMPC). These SUVs coalesce followingincubation with IL-2 to form large multilamellar vesicles with the incorporation of IL-2 and various membrane proteins within the lipidbilayers.Abbreviations: Id, idiotype; MHC Ag, major histocompatibility antigen complex; B7, T-cell costimulatory molecule.

Appendix: Color Figures

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Barker Figures 1A and 1B. Umbilical cord blood (UCB) unit procurements and UCB transplants facilitated by the NationalMarrow Donor Program (NMDP) by year (1A); NMDP facilitated transplants by year according to hematopoietic stem cell(HSC) source (1B). (Slides courtesy of Dr Dennis Confer, NMDP).

Pedersen-Bjergaard et al Figure 1. Alternative genetic pathways in therapy-relatedmyelodysplasia (t-MDS) and therapy-related acute myeloid leukemia (t-AML).

indicates association with transformation from MDS to AML

From Leukemia 2006;20:1943-1949 with permission.

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High Figure 3. (A) The phage φC31 uses anintegrase-driven system to insert its geneticcontent into a host bacterium. The system requiresφC31 integrase and a pair of complementary sequencesin the phage (attP) and the bacterial host genome (attB).(B) Two plasmids, one containing φC31 integrase, theother containing the attP site and the therapeutictransgene, are used to co-transfect mammalian cells.The φC31 integrase effects transgene insertion into themammalian genome at pseudo-attB sites, recognizedby the attP site in the therapeutic gene plasmid.

High Figure 5. Translational bypass therapy using a small-molecule drug. (A) In normal translation, the ribosometranslates mRNA to produce the polypeptide chain. (B) Anonsense mutation introduces a premature stop codon,resulting in synthesis of a truncated (non-functional) protein. (C)A small molecule drug permits readthrough of the prematurestop codon, restoring normal translation.

HIgh Figure 4. Genome editing by zinc finger nucleases.(A) A zinc finger nuclease consisting of DNA recognitiondomains and an endonuclease are designed to recognize aspecific site near a mutation. (B) The nuclease domainsintroduce a double-strand break into the DNA. (C) A wild-typesequence in introduced into the cells and used as the templatefor a cellular repair process, homologous recombination. (D) Themutant sequence is corrected. Adapted from High, Nature2005;435:577-579.

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Natkunam Figure 1. Schematic representation ofnormal B-cell ontogeny in the germinal center.Naïve B-cells encounter antigen and undergoproliferation, clonal expansion, somatic hypermutationand class-switch recombination in the germinal centermicroenvironment. The resulting B-cell clones with highaffinity antibodies are positively selected todifferentiate into plasma cells and secrete largequantities of specific antibodies or become memory B-cells that recognize and respond to repeat antigenexposure. The remaining B-cell clones are eliminatedby apoptosis.

Natkunam Figure 2. The t(14;18) translocation andexpression of the BCL2 protein. Schematicrepresentations show the germline configuration of theBCL2 gene as well as the chromosomal breakpointsinvolved in the t(14;18)(q21;q32) translocation. TheBCL2 protein is absent in normal germinal center B-cells whereas it is expressed in the neoplastic folliclesof follicular lymphoma, as shown by immunohistologicstaining for the BCL2 protein.

Shah Figure 1. Schematic of binding modes of clinically activeBCR-ABL kinase inhibitors. “Active” refers to the open conformationof BCR-ABL that is compatible with ATP binding and hydrolysis.“Inactive” refers to the closed conformation that is not catalytically active.The ability to bind to various BCR-ABL kinase domain conformationswas determined by co-crystal studies, except when marked with anasterisk (*), which reflects results of computer modeling–based studiesof binding. “Other targets” lists notable kinases that are reported to beinhibited at concentrations required to inhibit BCR-ABL activity.

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Natkunam Figure 3. Histologic andimmunohistologic images ofnormal germinal centers.(A) Hematoxylin and eosin-stainedsections of a normal tonsil shows agerminal center with polarized darkand light zones. The dark zone showsrapidly proliferating cells as evidencedby increased mitotic activity andtingible body macrophages. The lightzone is pale stained and shows a lowproliferative activity; (B) An immuno-histologic stain for Ki-67 (MIB1)highlights increased proliferation inone pole of the secondary folliclecorresponding to the dark zone; (C)An immunohistologic stain for CD21highlights the follicular dendritic cellmeshwork within the germinal center;(D) An immunohistologic stain for IgMhighlights immune complexescaptured on follicular dendritic cellprocesses that are involved in antigenpresentation to germinal center B-cells.

Ballas Figure 1. Molecular mechanisms of pain.Abbreviations: DH, dorsal horn; T, thalamus; LS, limbic system;RF, reticular formation.Reprinted with permission from Semin Hematol. 2001;38(4):309.

Natkunam Figure 4. Schematic representation oft(14;18)+ B-cell clones and the genesis offollicular lymphoma. Defective VDJ recombinationleads to the t(14;18) translocation and aberrant BCL2protein expression. Recent work by Roulland andcolleagues52 suggests that antigen stimulation allowsthe t(14;18)+ B-cell in normal healthy individuals toenter the germinal center and undergo the germinalcenter reaction. Ectopic expression of BCL2 in thegerminal center favors extended B-cell survival suchthat additional genetic changes are acquired by this Bcell and may ultimately result in transformation tofollicular lymphoma. In addition, N-glycosylation sitesin Ig V regions by directly binding oligomannosesugars could modulate B-cell receptor-mediatedsignaling and contribute to follicular lymphomapathogenesis.

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O’Connor Figure 3. A theoretical model for howproteasome inhibitors could affect the balance betweenpro- and antiapoptotic forces in MCL. Proteasome inhibitionleads to increased NOXA and MCL-1, the latter of which couldtheoretically mitigate some of the therapeutic merit of targetingthe proteaseome. However, the selective increase in NOXAresults in liberation of the pro-apoptotic BAX, which thenmediates induction of apoptosis.

O’Connor Figure 1. Schematic representation of cell cycleregulatory influences, depiciting the role of cyclin D, thecell cycle–dependent kinases, and the cell cycle–dependent kinase inhibitors.

O’Connor Figure 2. The interplay of established cell cycleevents in the pathogenesis in MCL. Loss of cell cycle–dependent kinase inhibitor influence (i.e., p16, p21 or p27)synergizes with the driving force of cyclin D1 overexpression.P53 is more commonly mutated in cases of blastic mantle celllymphoma

Kelly and Rickinson Figure 1. Characteristic features ofBurkitt lymphoma (BL) cells. Top: characteristic ‘starry sky’histology associated with BL tumors brought about by thepresence of round monomorphic B cells interspersed withmacrophages. Middle: BL cells grow as round single cells inculture. Bottom: BL cells express markers characteristic ofgerminal center B cells such as CD10, CD38 and BCL6.

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Figure 1. Conceptual models of CML progression. The upper panel shows theconcept of stepwise progression of CML, with distinct clinical and pathological features.However, the data suggest the lower panel concept of a continual evolution of chronicphase through an advanced phase (accelerated and blast phases), with progressionstarting at the acquisition of the Philadelphia chromosome.

Radich Figure 2A. CML progression. Stemcells acquire the Philadelphia chromosome.This begins a process of instability, andincreased proliferation yielding multipleclones of progenitors. With unchecked Bcr-Abl activity, instability causes geneticalterations, including point mutations in Abl(and elsewhere), and gross cytogeneticchanges. This causes a block indifferentiation, increased proliferation ofprimitive cells, and a block of apoptosis. Inthis model, progression from chronic to blastphase begins from the outset of disease. Theability of TKI to arrest the progressiondepends on how long Bcr-Abl activity wenton unimpeded.

Radich Figure 2B. Progression in CML.

Shipp Figure 3. DLBCL, gray zone lymphomas, andHodgkin lymphomas. MLBCL and T-cell/histocyte-rich large B-cell lymphoma (T/HRBCL) are considered “gray zonelymphomas,” which share certain characteristics of large B-celllymphomas and Hodgkin lymphomas (cHL and NLPHL,respectively), including increased host inflammatory response.The similarities among these entities point to a group of tumorsthat are partially defined and possibly driven by their interactionwith the host microenvironment. Reprinted with permission fromAbramson and Shipp.2

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Kuypers Figure 1. The red cell membrane bilayer. Phospholipid molecular species move rapidly in the plane as well as across thebilayer in a dynamic but highly organized fashion. Areas are enriched in certain lipids essential for proper protein function, andphospholipids are asymmetrically distributed across the lipid bilayer, with the amino phospholipids (PS and PE) on the inside andcholine containing phospholipids (PC and SM) on the outside.

Kuypers Figure 2. Maintenance of phospholipid organization in red blood cells. Reactive oxygen species (ROS) will modifythe unsaturated fatty acyl side chains in phospholipids (PL). The oxidized lipids (oxPL) will have a different orientation in the bilayerand are recognized by phospholipases (PLA2) that remove the oxidized fatty acid to generate lysophospholipid (LPL). Fatty acid(FA) taken up from plasma is activated to fatty acyl CoA (FA-CoA) by acylCoA synthetase (ACSL6) using adenosine triphosphate(ATP) and Coenzyme A (CoA). FA-CoA pools are modulated by FA-CoA–binding protein (FA-CoABP). Lysophospholipid acylCoAacyltransferases (LPLAT) use FA-CoA and LPL to generate PL, completing the de-acylation/re-acylation repair cycle. Theasymmetric distribution of phospholipids across the bilayer is maintained by the Magnesium ATP-dependentaminophospholipidtranslocase or flippase, which transfers PS and PE from outer to inner monolayer at the expense of ATP. Loss ofphospholipid asymmetry and the exposure of PS is triggered by a phospholipid scrambling activity.

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Sandlund Figure 3. Histologic types of non-Hodgkinlymphoma by age group, SEER, 2001-2004, ICDO-3(source - SEER 17).

Sandlund Figure 1. Survival rates for non-Hodgkinlymphoma, SEER, 1975-19982

Sandlund Figure 2. Histologic types of non-Hodgkin lymphoma inchildren and adults, SEER, 2001-2004, ICDO-3 (source – SEER 17)

Richardson et al Figure 1. Schematic representation ofmain caspase-mediated pathways for the direct anti–multiple myeloma (MM) effect of proteasome inhibitors(bortezomib), immunomodulatory thalidomide derivatives(IMIDs) and dexamethasone (Dex).

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Skoda Figure 2. Model of the clonal progression in myeloproliferative disorder (MPD). The onset of MPD is preceded by asomatic mutation (sporadic MPD) or germ-line mutation (familial MPD) in an as yet unknown gene(s). The onset of MPD coincideswith the acquisition of a mutation in JAK2 or other genes (gray box). Acute leukemia can arise de novo or from the MPD clone.Leukemic transformation of a “pre-JAK2” progenitor or stem cell could explain the absence of JAK2-V617F in leukemic blasts fromMPD patients previously positive for JAK2-V617F. Alternatively, JAK2-V617F could be lost during mitotic recombination of a cellheterozygous for JAK2-V617F (dashed line).

Skoda Figure 1. Effect of THPO gene mutations on the composition of uORFs in THPO mRNA. The main THPO mRNAtranscribed from promoter 2 is shown. Boxes numbered in italic represent exons. The uORFs are drawn as thick red lines and areplaced into one of the three reading frames (+1, 0, and –1). The THPO coding region is shown as a thick blue arrow. Numbersindicate the order in which the uAUGs appear in the full length THPO mRNA (uAUGs 1-4 are not shown). The eighth AUG is thephysiological initiation codon. (A) Translation of normal THPO mRNA is physiologically almost completely inhibited by the presence ofuORFs in the 5′′′′′-UTR. In particular, the uORF 7 is a potent inhibitor of translation, most likely because of its extension beyond thephysiological start site. (B) A splice donor mutation in the Dutch HT family causes exon 3 skipping (DE3) that deletes uORF7 andshifts the THPO coding sequence into reading frame +1. Tpo translation now initiates from the fifth and sixth AUG. (C) The Japanesemutation I consists of a single G nucleotide deletion (DG) that shifts the THPO coding sequence into reading frame -1. Tpo translationnow initiates from the seventh AUG. Note that both the Dutch and the Japanese mutation I create altered THPO signal peptides, butdo not alter the sequence of the mature Tpo protein. Both signal peptides remain functionally active and promote secretion of abiologically active Tpo protein. (D) The Japanese mutation II creates a premature stop codon in uORF7. This allows re-initiation oftranslation at the physiological start site (the eighth AUG).

Figure reprinted from Cazzola and Skoda, Blood. 2000;95:3280-3288 © the American Society of Hematology

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Dunbar Figure 1. Typical hematopoietic stem cell (HSC) gene therapy trial design. Marrow or mobilized peripheral blood cellsare collected from patients, enriched for hematopoietic stem and progenitor cells via CD34+ selection, and exposed to retrovirusvector particles during in vitro culture. Transduced cells are then reinfused into the patient, either with or without priormyelosuppressive or ablative conditioning radiochemotherapy. Patients are then monitored for the presence of vector and correctivetransgene expression in hematopoietic progeny cells.

Dunbar Figure 2. Insertional mutagenesis by integrating vectors. Anintegrated provirus with strong enhancers in the vector LTRs can activateadjacent proto-oncogenes or inactivate tumor repressor genes.

Döhner Figure 1. Model of leukemogenesis withtwo cooperating classes of mutations.

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Alter Figure 1A. Schematic representationof protein interactions in the FA pathway.DNA lesions arising during normal S-phase orafter DNA damage activate ATR-dependentphosphorylation and FA core complex–dependent monoubiquitination of both FANCD2and FANCI. The ID complex consisting ofmonoubiquitinated FANCD2 and FANCI formsnuclear foci in chromatin. USP1 deubiquitinatesboth proteins.Reprinted from Developmental Cell, vol. 23.Grompe M, van de Vrogt H. The Fanconi familyadds a fraternal twin; pp 661-662, 2007, withpermission from Elsevier.37

Alter Figure 1B. Telomere maintenancepathway. Telomeres contain hexanucleotiderepeats (TTAGGG)n and associated proteinsinvolved in the addition of these base pairsduring mitosis. The major components of thetelomerase pathway are the RNA template andthe telomerase enzyme, but multiple proteinsare involved, only some of which are shown inthe figure.Reprinted by permission from MacmillanPublishers Ltd: Nature Reviews Immunology.2002;2:699-706.38

Ataga and Key Figure 1. Schematic representation of pathophysiological mechanisms (described or postulated) leadingto activation of coagulation in sickle cell disease. Based on Virchow’s triad, the illustrated pathways contribute to activation ofcoagulation (and possibly eventual thrombosis) by one of three broad mechanisms: (1) changes in the vessel wall; (2) changes inblood flow; and/or (3) changes in the composition of blood components (‘hypercoagulability’). The numbers refer to the specificreference describing the particular pathway (represented by a joining arrow); references are listed at the end of the text.Abbreviations: NO, nitric oxide; PS, phosphatidylserine; RBC, red blood cell; MPs, microparticles; TF, tissue factor; SCD, sickle celldisease