INDEX [link.springer.com]978-1-62703-493-7/1.pdf · detection of complement-fixing HLA antibodies...

405

A

Adaptive immunity ......................................... 71, 75, 76, 359 Adhesion molecules

β 2 integrins ....................................................................7 7 CD62L (L-selectin) .....................................................80 ICAM-1 .................................................................77, 78

Allelic ambiguities ............................................................1 97 Alloantibody ................................... 9 , 50, 52, 53, 61, 77, 131,

258, 270, 286 Allograft rejection

antibody mediated .................... 4 2, 49, 51, 53, 56, 57, 60, 79, 128, 133

cell mediated ..........................................7 2, 75, 76, 78, 79 Alloreactive T cells

frequencies of naïve and memory T cells ................1 0–11 polyspecificity .........................................................1 1–15

Alloreactivity MHC plasticity ................................................ 1 4, 22, 27 MHC restriction .............................3 , 4, 8, 10, 15–18, 28 molecular basis .................................................. 1 1–19, 30 structural basis ................................................................4 T cell receptor frequencies ............................ 4 , 10–12, 16 T cell receptor repertoire ............................. 8 , 10–12, 14,

15, 18, 24, 27 T cell receptor specificity ..............................................1 6

Allorecognition direct ...................................... 8 , 9, 11, 19–30, 72, 79, 360 indirect .....................................................8 , 9, 72, 73, 360 semi-direct ..............................................................7 2, 73

Amplification refractory mutation system (ARMS) ..............................................147

Angiotensin II-type 1 receptor (AT1R) antibody assay AT1R antibodies and obliterative vasculopathy in

autoimmune diseases ......................................3 31 AT1R antibodies and role in heart and

kidney antibody mediated rejection ................3 31 calculation of results ...........................................3 32–333 ELISA procedure ...............................................3 31, 332 sensitivity and specificity compared to cardiomyocyte

bioassay ...........................................................3 32 standard curve .....................................................3 32, 333

Antibody binding capacity ............................................................4 7

donor specific antibody (DSA) .................. 41–45, 47, 55, 57, 59, 108, 118–123, 129, 130, 135, 179, 258–260, 277, 278, 285, 297, 298, 306, 4953

endothelial cell reactive (AECA) .......... 59, 319, 320, 328 HLA specific antibody ...............................................2 98 non-HLA antibody ........................................ 4 1–62, 259

Antibody induced leukocyte recruitment ......................5 6–57 infiltration in antibody-mediated rejection ...................5 6

Antibody mediated rejection (AMR) complement activation ............................................5 1, 53 Fc receptor dependent ..................................................7 9 in vivo models .........................................................4 8–49 mechanisms ...........................................4 6, 49, 52, 57, 79

Antibody monitoring ................................................1 23, 140 Antibody screening ............................1 28–131, 133, 134, 297 Antigen presenting cells (APC) ................................ 4, 9, 58,

72–75, 77, 91, 93, 108, 111, 222, 314, 316, 321, 323, 324, 359–367

tolerogenic ..........................................................3 59–367 Apoptosis ................................... 8 , 53, 58, 61, 62, 79, 93, 354 ATP synthesis and measure of immune function .............3 43 Autoantibody

angiotensin II-type 1-receptor ............................3 31–333 collagen, I,II,IV,V ...............................................3 35–340 K-alpha-1 tubulin .......................................................1 33 MICA/MICB ........................................................5 9, 60 myosin ................................................................3 35–340 tubulin ................................................................3 35–340 vimentin ..............................................................3 35–340

B

Biomarker analysis ..............................................................8 7 B lymphocytes

activation ................................................................7 5–76 antigen presenting cells .................................................5 8 intragraft B cells ...........................................................7 5

C

Calcineurin inhibitors cyclosporine A ............................................................1 04 tacrolimus ...........................................................1 04, 127

Calculated panel reactive antibody (CPRA) ......................41, 118, 128, 132, 133

INDEX

Andrea A. Zachary and Mary S. Leffell (eds.), Transplantation Immunology: Methods and Protocols, Second Edition, Methods in Molecular Biology, vol. 1034, DOI 10.1007/978-1-62703-493-7, © Springer Science+Business Media, LLC 2013

Carboxyfluorescein succinyl ester (CFSE) assay .............4 8, 361, 362, 365, 366

C4d assay anti-mouse IgG-phycoerythrin (PE)

conjugate .................................................2 99, 301 complement binding antibodies ..................................2 98 MFI values ..................................................................3 02 modification for Luminex® beads .......................3 01, 302 mouse anti-human C4d ......................................2 99, 300 normal human plasma as

complement source .................................2 98–301 sensitivity ....................................................................3 03 single antigen kits ...............................................2 99, 302 two stage process for detection of complement

deposition .......................................................2 98 CDC crossmatch ...................................... 1 31, 258–260, 279 C4d deposition ........................ 5 1, 59, 87, 108, 130, 297–303 Cellular proliferation .............................4 8, 53–55, 57, 61, 62 Chemokines

CXCL1-8 .....................................................................77 MIG/CXCL9 ...............................................................76 RANTES/CCL5 ..........................................................76

Chimerism lineage specific ....................................................2 22, 224 macrochimerism ...........................................................9 6 microchimerism .............................9 6, 139, 140, 222, 236 split chimerism ...........................................................2 22

Chimerism testing engraftment monitoring ............................. 2 21, 222, 224 informative markers ............................ 2 22–226, 229–236 insertion and deletion markers (Indel) ........................223 sensitivity of quantitative PCR ...................................2 23 STR-PCR, short tandem repeat or

microsatellite PCR ....................................9 6, 222 Chimerism testing by qPCR with Indel markers

cell separation .....................................................2 24–226 data analysis ........................................................2 24–226 DNA isolation ....................................................2 24–226 quantitation of recipient cells post- transplant .............2 24 quantitation test ..........................................................2 33 screening for informative markers ..................... 2 22–226,

229–233, 235, 236 screening test ...................................... 2 26–229, 234, 235

Chronic rejection HLA class II antibodies ............................. 1 29, 288, 321 mechanisms ................................................ 4 9, 52–56, 61

Circulating T cells and tolerance CD4 + /CD25 + ................................................................91 CD4 + /CD25+hi/CD62L + hi .........................................9 1 gamma-delta T cells ...............................................9 0, 91

Clonal amplification .........................................................1 98 Clonal pyrosequencing .....................................................1 98 Complement

activation .............................................. 9 , 50–53, 62, 306

C4d deposition ...........................................................3 01 components ....................... 4 9, 77, 78, 294, 301, 305, 306 C1q binding ........................................................3 05–311 proteolytic cascade ........................................................7 7

Complement dependent cytotoxicity (CDC) AHG-enhanced CDC ............................... 2 59, 269–270 AHG evaluation .................................................2 73–275 antihuman globulin (AHG) .......259, 260, 264, 269–270,

272–275, 277–281 B cell positive control ................................. 2 61, 264, 272 carboxyfluorescein diacetate (CFDA) ......... 261, 268–270 cell counting and viability determination ....................2 67 complement evaluation ....................... 2 71–274, 280, 281 deoxyribonuclease (DNase) ........................ 261, 263, 267 dithiothreitol (DTT) ...........259, 261, 264–265, 275, 276 DTT serum treatment ........................ 2 64–265, 275, 276 ethidium bromide ............................... 2 61, 262, 269, 270 ethylenediaminetetraacetic acid (EDTA) ...........261, 262 immunomagnetic isolation of T and B cells .......2 67–269 interpretation of CDC results .....................................2 79 modifications ......................................................2 58, 277 preparation of donor cells ...................................2 65–267 rabbit serum complement ...................................2 61, 262 scoring assay results ............................................2 70–271 standard CDC crossmatch .................................2 70–271 T cell positive control .........................................2 61, 273 trypan blue .................................................. 2 63, 267, 277

Composite tissue transplantation acute rejection .....................................................1 06–111 antibody mediated rejection ........................................1 08 C4d deposition ...........................................................1 08 chronic rejection .........................................................1 06 functional outcomes .................................... 1 04, 105, 111 history of development .......................................1 03–104 immunosuppression ............................ 1 04–106, 108–113 mechanisms of skin rejection ..............................1 07–108 novel concepts for immune modulation ..............1 11–113 peripheral nerve regeneration .....................................1 04 protocols for prophylaxis and treatment

of skin rejection .......................................1 08–109 skin rejection, histopathologic

characterization .......................................1 06, 107 unique biologic and immunologic

features of VCA ......................................1 09–111 vascularized composite

allotransplantation (VCA) ..............................103 Costimulation

B7/CD28 ......................................................................73 CD40/CD154 ..............................................................73 costimulatory molecules ..........................................7 3, 74 signal 1, TCR-CD3 complex ........................................7 3

C1q binding assay analysis and interpretation ..........................................3 09 biotin-labeled human C1q ..........................................3 06

406 TRANSPLANTATION IMMUNOLOGY: METHODS AND PROTOCOLS

Index

detection of complement-fixing HLA antibodies ................................................3 05–311

sensitivity compared to CDC .....................................2 59 serum preparation by absorption ........................3 07–308

Crossmatch assays complement dependent cytotoxicity

(CDC) ....................................................257–281 endothelial cell precursor crossmatch

(ECXM) .................................................319–328 flow cytometric (FCXM) ....................................319–328 virtual ..................................................................1 31, 135

Cylex® ImmunoKnow™ cell function assay ATP release and measurement ............................3 45–346 bioactivity of immunosuppressive drugs .....................3 43 calculation of results ...........................................3 47–348 calibration ...................................................................3 47 CD4 T cell activation .................................................3 43 cell selection ........................................................3 45–346 cell stimulation ...........................................................3 45 interpretation of results ...............................................3 48 intracellular ATP synthesis .........................................3 43 levels of response with infection .........................3 43–344 luciferin/luciferase ......................................................3 44 phytohemagglutinin (PHA) stimulation ....................3 43 quality control .....................................................3 48–349

Cytotoxicity antibody dependent cell-mediated

(ADCC) ....................................50, 52, 60, 77, 79 complement dependent cytotoxicity

(CDC) ..................................43–45, 52, 128, 129, 131–133, 135, 257–281, 285, 297, 298, 301, 302, 305, 306

Cytotoxic T cells (CTLs) Fas ligand (FasL) ..........................................................79 granzyme B ...................................................................7 9 perforin .........................................................................7 9

D

Dendritic cells immature (iDC) ............................................ 93, 364, 365 mature .........................................................................3 60 plasmacytoid ...............................................................3 61 tolerogenic ..........................................................3 59–369

Desensitization antibody characterization ............................................1 20 antibody monitoring ...................................................1 40 donor specific antibody (DSA) specificity

and strength ....................................................1 18 high dose intravenous immunoglobulin

(IVIg) ......................................................118–121 kidney paired donation (KPD) combined with

desensitization ................................ 1 17, 120–121 patient evaluation ...............................................1 19–120 plasmapheresis and low dose IVIg ..............................1 18

post-transplant monitoring .........................................1 21 risk assessment ............................................................1 23 sensitized patients ...............................................1 17–121 solid phase immunoassays ...........................................1 18

Detection of antibodies to self- antigens by ELISA antibodies to collagens in liver

transplantation ................................................3 36 antibodies to myosin and cardiac allograft

vasculopathy ....................................................3 36 antibodies to tubulin and/or collagen in

bronchiolitis obliterans syndrome ...................3 36 antibodies to vimentin and coronary

atherosclerosis .................................................3 36 blocking and loading of serum samples ..............3 37, 338 coating plates with antigen solutions ..........................3 38 collagen I,II,IV,V ................................................3 35–340 data analysis and sample calculation ...................3 38–340 K-alpha 1 tubulin ...............................................3 35–340 myosin ................................................................3 35–340 role of auto-antibodies in allograft rejection ...............3 31 secondary antibody incubation and detection .............3 31 vimentin ..............................................................3 35–340

Detection of HLA specific B cells allophycocyanin conjugated mouse

anti-human CD19 ..........................................314 B cell enrichment ................................................3 15–316 conformational epitopes .............................................3 13 flow cytometric analysis ......................................3 16–317 fluorescein conjugated mouse

anti- human CD45 ..........................................314 HLA sensitization ..............................................3 13–318 HLA tetramers conjugated with

phycoerythrin ..................................................3 14 isolation of peripheral blood

mononuclear cells ...................................3 14–315 surface immunoglobulin receptors ..............................3 13 tetramer staining .................................................3 13–318

Detection of intracellular cytokines activation of cells with phorbol

myristate acetate (PMA) .................................354 brefeldin A .................................................. 3 53, 355, 356 cell permeabilization ...........................................3 54, 355 controls ...............................................................3 54, 357 data analysis ................................................................3 57 flow cytometer calibration ..................................3 55, 357 protein transport inhibitor ..........................................3 53 sample collection ........................................................3 55 staining of cell surface markers ...........................3 53, 357

Direct TCR allorecognition, structural insights 2C system ...............................................................1 9–23 LC13 system ...........................................................2 3–27 YAe62 system .........................................................2 7–30

Discovery of antibody targets by protein arrays conventional ELISA method ..............................3 73–383 data analysis with ProtoArray® Prospector .................3 75

TRANSPLANTATION IMMUNOLOGY: METHODS AND PROTOCOLS

407

Index

Discovery of antibody targets by protein arrays (cont.)data analysis with significant analysis

of microarrays (SAM) .............................379–380 data generation with GenePix® Pro data

acquisition software ........................................3 79 detection antibody ..............................................3 78, 381 high density protein arrays ..........................................3 74 MSD ELISA .............................................. 3 74, 381, 383 MSD platform ............................................................3 75 non-HLA antigens .....................................................3 74 protein arrays, Protoarrays® ........................ 374–375, 379 protoarray blocking ..................................... 3 74, 376–378 sample preparation ......................................................3 76 scanning of slides ................................................3 78–379 validation of antibody targets

by indirect ELISA ..................................3 73–383 DNA sequencing for HLA typing

allele group-specific amplification of HLA class I loci .................. 1 68–169, 181–182

allele group-specific amplification of HLA-DRB loci .......................... 1 69–170, 182

ambiguous allele assignments, alternative genotypes ........................................................1 86

antigen recognition site (ARS) ...........................162, 164 co-amplification of both alleles at

an HLA class I locus ............... 1 66–168, 175–181 DNA preparation ...............................................1 66, 174 HLA allele isolation by cloning .................. 1 73, 186–187 HLA alleles .................................1 62, 164, 173, 186–187 HLA loci ............................................................1 62, 165 locus specific PCR amplification ................................1 66 PCR amplification of DQB1 ...................... 170, 182–183 polymorphism ..............................1 63, 181, 185, 188, 193 purification of PCR amplicons ...........................1 71, 183 Sanger sequencing ......................................................1 62 sequence analysis ................................ 1 73, 181, 184–186 sequencing primers .............................1 62, 163, 171, 172,

177, 178, 180, 181, 186, 188–190, 193, 194 use in clinical practice .................................................1 65

Donor-specific antibody (DSA) clinical relevance .........................................................1 08 frequency .......................................................... 4 1–46, 59

E

Emulsion PCR (emPCR) ................................ 198–200, 202, 204–205, 210, 212–215, 218

Endothelial cell activation ..................................................5 6 anti-endothelial cell antibodies

(AECA) .............................................. 59, 60, 319 Endothelial cell precursor crossmatch

(ECXM) acquisition and analysis ....................3 24 agrin ............................................................................3 19 angiotensin II type 1 receptor (AT1R) .......................319 anti-endothelial cell antibodies

(AECAs) ........................................ 319, 320, 328

appropriate reactivity of controls ................................3 24 differentiation of EPCs from lymphocytes

by forward and side scatter .....................3 24, 325 endothelial cell precursor (ECP)

isolation ..................................................3 20–325 fluorochrome-conjugated antibody

incubation .......................................................3 24 major histocompatibility class I chain related

gene A (MICA) ..............................................319 serum preparation and incubation ......................3 23–324 XM-ONE® ................................................................320

Enzyme-linked immunosorbent assay (ELISA) ......................................... 44, 45, 94–96, 129, 331, 332, 335–340, 373–383

Experimental techniques to measure antibody effects analysis of intracellular signaling ..................................4 7 determination of cell growth .........................................4 8 determination of cytoskeletal changes and

cell migration ....................................................4 8 in vitro techniques ..................................................4 7–48 in vivo models of antibody-mediated

rejection ......................................................4 8–49 measurement of HLA antibody binding

capacity .............................................................4 7 measurement of leukocyte adherence ...........................4 8 siRNA and pharmacological inhibitors ........................4 8

F

Flow cytometric crossmatch (FCXM) addition of anti-human IgG and monoclonal

antibodies ........................................................2 90 anti-human CD3 ........................................................288 anti-human CD19 ......................................................314 anti-human IgG ................................................ 3 20, 323,

337, 339 data acquisition and analysis ....................... 2 90–294, 338 fluorescein-conjugated goat,

anti- human IgG ..............................................2 88 median channel shift (MCS) ..............................129, 130 molecular equivalent of soluble fluorochrome

(MESF) .......................................... 291, 292, 294 molecular equivalent of standardized

fluorescence (MESF) ......................................129 pronase treatment .......................................................2 87 sensitivity ............................................................2 94, 295 serum/cell incubation ..........................................2 89–290 specimen requirements ...............................................2 87 standardized fluorescence intensity (SFI) ...........129, 130 three color fluorescence ..............................................2 86

G

Gene arrays B cell signature .............................................................8 9 granulysin RNA and acute rejection .............................8 7 tribbles-1 expression and chronic rejection ...................8 7


Index

Global gene expression profiling ...........................3 85, 387, 388, 393–403

Growth factors insulin-like growth factor-1 (IGF-1) ......................56, 76 platelet-derived growth factor (PDGF) ..................56, 76 transforming growth factor beta (TGF-β) ........ 76, 93–95

H

HLA class I and II sequencing .........................................1 98 HLA genotype ambiguity reduction ........................2 01, 202 HLA specific antibody ..................................... 2 97–311, 321 HLA specific B cells ......................................... 1 20, 313–318

I

Immune monitoring Allomap® ....................................................................140 ImmunKnow® assay .....................................................8 9

Indoleamine 2,3-dioxygenase (IDO) ............................95–96 Inhibitory receptors

IL10, IFN-α, IFN-β upregulation ..............................3 60 ILT3/ILT4 ..................................................................360 induction by allospecific T suppressor cells (Ts) .........360 induction by 1, 25-dihydroxy vitamin D3 ...................360

Innate immunity damage associated molecular

patterns (DAMPS) ...........................................76 inflammasome ..............................................................7 6 innate immune receptors ..............................................7 6 pattern recognition receptors (PRRs) .....................71, 76

Interleukin 10 (IL-10) ........................................................94 Intragraft Foxp3 ...........................................................91, 92 Ischemia reperfusion injury ..........................................7 4, 76

K

Kidney paired donation combined with desensitization ...........................1 17, 119 patient evaluation .......................................................1 21 post-transplant monitoring .........................................1 21

Killer-cell immunoglobulin-like receptors (KIR) absence of class I ligands and reduced

long-term renal graft survival ..........................2 41 association with disease risk ........................................2 52 expression

CD8 + T cells .........................................................2 39 NK cells ................................................................2 39

genotyping classification of KIR genotypes by centromeric and

telomeric gene-clusters ...........................2 51–252 data analysis, basic rules ................................2 48–252 diversity ................................................................2 52 framework genes ...................................................2 48 gel electrophoresis ......................... 2 42–243, 246–247 genes .............................................................2 48–252 group A haplotypes .......................................2 48, 250

group B haplotypes ............................... 2 48, 250, 251 interpretation of gel results ...........................2 47–248 KIR genotyping ........................................... 2 41, 243,

245, 247–252 PCR amplification for SSP ...................................2 41 verification of raw data .................................2 48–250

group B haplotypes and improved relapse-free survival with AML ......................2 41

leukocyte receptor complex (LRC) .............................239 KIR. See Killer-cell immunoglobulin-like receptors (KIR)

L

Leukocyte adherence ..........................................................4 8 Leukocyte receptor complex (LRC) .................................239 Leukocyte recruitment

P-selectin ................................................................5 6, 57 von Willebrand factor (vWF) .......................................56 Weibel-Palade bodies .............................................5 6, 78

LRC. See Leukocyte receptor complex (LRC)

M

Major histocompatibility complex (MHC) allotypes .......................................................... 4 , 9, 11, 16 class I chain-related gene A (MICA) ...........................49 class I molecules ...........................................................7 7 class II molecules ........................................................1 08 crystal structure ............................................... 5 , 8, 12, 16 peptide binding groove ...................................................5 restriction .........................................3 , 4, 8, 10, 15–18, 28

Mean/median fluorescence intensity (MFI) ................................ 53, 121–123, 129, 135, 301, 302, 306, 309–311

Mechanisms of antibody-mediated injury antibody-dependent cell-mediated

cytotoxicity ..................................................5 0, 52 Fc dependent effects of antibodies ..........................5 0–52 target cell signaling .................................................5 3–57

MHC. See Major histocompatibility complex (MHC) miH. See Minor histocompatibility antigens (miH) Minor histocompatibility

antigens (miH) ................................. 8, 72, 75, 79 Mixed lymphocyte reaction

(MLR) .........................................9, 19, 24, 87, 92 MLR. See Mixed lymphocyte reaction (MLR) MMF. See Mycophenolate mofetil (MMF) Multiplexed bead assays

FlowPRA® .................................................................129 Luminex® ...................................................................129 phenotype panels ........................................................1 23 single antigen assays ...................................................1 29

Mycophenolate mofetil (MMF) ....................... 104, 105, 361 Myeloid-derived suppressor cells (MDSC)

arginase (Arg-1) .....................................................92, 93 inducible nitric oxidase synthase (iNOS) ................92, 93


409

Index

N

Natural killer cells (NK) KIR .......................................................................7 7, 239 NKG2A/CD94 ............................................................77 receptors .................................................................4 9, 77

Next-generation sequencing (NGS) amplicon purification ..........................................2 08–209 choosing the amplicons for emulsion PCR .................2 10 clonal pyrosequencing .................................................1 98 emulsion PCR ............................ 2 04–205, 210, 213–214 genomic PCR .....................................................2 02–203 genotype guidelines ............................................2 13–214 HLA class I and II sequencing ...................................1 98 HLA genotype ambiguity reduction ...................2 01, 202 454 Life Sciences

GS FLX 454 system ....................... 1 97, 199, 206 454 Life Sciences

GS FLX Titanium chemistry .........................1 98 multiplex identifiers (MIDs) ......................................199 sample preparation ......................................................2 02

O

Operational tolerance ..................................... 7 5, 88, 89, 112 B cell specific gene expression ......................................8 9

P

Polymerase chain reaction (PCR) .............................. 89, 136, 162, 166–167, 169–170, 175, 182, 187

Proteomics analysis of protein-protein interaction ........................3 73 analysis of protein-small molecule interactions ..........3 73 enzyme-substrate profiling .........................................3 73 high density protein arrays ..........................................3 74 immune monitoring ......................................................3 0

Q

Quantitative PCR .................................9 6, 97, 221–236, 396 Quantitative reverse transcription polymerase

chain reaction (RT-PCR) .......................... 89, 90, 92, 95, 229, 366

R

RNA expression analysis Affimetrix GeneTitan ........................................3 86, 387 Ambion WT expression kit ........................................3 93 deep RNA sequencing ........................................3 85–403 fragmentation and labeling of cDNA .........................3 94 high quality RNA purification ....................................3 86 high-throughput microarray technology .....................3 97 microarrays .........................................................3 85–403 NuGEN Encore Complete DR protocol ...................3 98 NuGEN Ovation Pico WTA system V2 ....................395

PAXGene RNA preparation protocol .........................3 86 preparation of sequencing library ................................3 97 RNA deep sequencing ........................................3 85–403 RNA extraction from cells and

tissues ...................................... 3 88–393, 402–403 RNAseq library ...................................................3 86, 398 target hybridization and processing ............................3 87 Trizol hybrid protocol .........................................3 89–390

S

Sensitization antibody breadth .........................................................1 17 antibody strength ........................................................1 19 factors affecting route to transplantation ............1 17–119 probability of a compatible donor ...............................1 19

Sequence-specific primers for HLA typing (PCR-SSP)

design of typing system for unambiguous results ..............................................................1 48

interpretation ..............................................................1 49 multiplexed PCR reactions .........................................1 48 primer design ......................................................1 48, 154 reaction components ...................................................1 52 reaction design ............................................................1 56 resolution levels ..........................................................1 62 template DNA requirements ..............................1 52–153 troubleshooting ...........................................................1 58 typical setup ................................................................1 53

Signaling induced by HLA I antibodies cell proliferation ......................................................5 4–55 survival and accommodation .........................................5 3

Small interfering RNA (siRNA) ........................................48 Solid phase immunoassays

correlation with crossmatch results .............................1 21 C1q binding assay ...............................................3 05–311 elimination of interference ..........................................1 19 FlowPRA® .................................................................129 Luminex® ...................................................................129 phenotype panels ........................................................1 23 sensitivity ............................................................1 18, 121 single antigen assays ...................................................1 33 variability ............................................................1 18, 121

Suppression ...................................9 2–94, 137, 360, 361, 365

T

T cell differentiation .....................................................7 4–75 T cell receptor (TCR)

complementarity determining regions (CDR) ................7 intracellular signaling ....................................................5 5 MHC restriction ................................................ 4 , 17, 18 molecular mimicry ........................................................2 7 polyspecificity .........................................................1 3, 14 receptor cross-reactivity

(heterologous immunity) ..................................80


Index

recognition of peptide-major histocompatibility complex .............................4 –8

repertoire ..................................................................8 , 24 thymic selection ................................................ 1 1, 15, 17

Thoracic organ transplantation AlloMap .............................................................1 36–137 antibody screening and characterization .............1 28–130 biomarkers ..........................................................1 38–139 calculated panel reactive antibodies (CPRA) ......128, 132 complement fixing antibodies .............................1 34–135 desensitization ............................................................1 33 FlowPRA® .................................................................129 immune monitoring ............................................1 37–138 multiplex bead assay on Luminex® platform ..............1 29 non-HLA antibodies ..........................................1 33–134 post-transplant monitoring .................................1 35–136 virtual crossmatch ...............................................1 30–132

T lymphocytes (T cells) activation ...................................4 , 72–74, 77, 93, 96, 343 CD4 + class II restricted .................................................7 4 CD8 + class I restricted ..................................................7 4 clonal expansion ..............................................................4 costimulation .................................................... 7 3–74, 81 gamma-delta T cells ...............................................9 0, 91 homeostatic proliferation ..............................................8 0 Th subsets .....................................................................7 4

Tolerance barriers ..........................................................................8 6 central .....................................................................8 5, 91 definition ................................................................8 5–86 operational ...........................................7 5, 88, 89, 92, 112 peripheral .......................................................... 3 0, 85, 86

Tolerogenic dendritic cells cDNA preparation ......................................................3 69 cell sorting using magnetic beads ................................3 62 induction by immunosuppressive agents .....................3 43 induction by vitamin D receptor ligands .....................3 61 induction of tolerogenic phenotypic

markers on APC .............................................3 61 measurement of serum ILT3/ILT4

inducing capacity ....................................3 66–367 monitoring for decreasing

immunosuppression ........................................3 61 phenotype ...........................................................3 62–365 proliferation assay ...............................................3 65–366 real time PCR .............................................................3 69 RNA extraction ..........................................................3 68

Transcription factors Foxp3 ....................................................................75, 139 Tbet ..............................................................................7 4

Transplantation of the sensitized patient antibody characterization ............................................1 28 antibody monitoring ...................................................1 23 CDC crossmatch ........................................................2 69 desensitization ....................................................1 19–120 flow cytometric crossmatch ........................................1 20 histocompatibility testing ...................................1 17–124 identifying the most likely route

to transplantation ....................................1 17–119 kidney paired donation (KPD) ...........................120–121 KPD combined with desensitization ..........................1 19 risk assessment ............................................ 1 20, 121, 123 solid phase immunoassays ...........................................1 18 virtual crossmatch .......................................................1 21

Transplant quiescence ...............................................1 12, 138 Transplant vasculopathy .....................................................5 4 T regulatory cells (Treg)

inducible Treg (iTreg) ...................................................91 natural Treg (nTreg) .....................................................91

T suppressor cells CD8 + CD28 - T cells .............................................3 61, 363 CD8 + Ts ...............................................................3 60, 361

U

Urinary biomarkers chemokines CXCL9, CXCL10,

CXCL11, CCL2 ...............................................88 Foxp3 mRNA ...............................................................8 8 granzyme A mRNA .....................................................8 8

V

Vascular injury ..................................................................1 38 Vascularized composite allotransplantation (VCA)

infectious complications .............................................1 10 unique biologic and immunologic

features ....................................................1 09–111 Virtual crossmatch .............................1 21, 130–132, 135, 140

W

Wound healing assays cell migration ................................................................4 8

cytoskeletal changes ......................................................4 8


411

Index

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