Brief Communication to Cell Transplantation
A Preliminary Gene Expression Profile of Acute Graft-Versus-Host Disease Running head: Gene expression profile of acute GVHD
Matthew P. Buzzeo1, Jie Yang2, George Casella2 and Vijay Reddy1 1
Department of Medicine, Division of Hematology/Oncology, University of Florida,
Gainesville, Florida, 32610, USA; 2
Department of Statistics, University of Florida, Gainesville, Florida, 32610, USA
Correspondence:
Vijay Reddy, M.D. Ph.D, University of Florida, 1600 SW Archer Road,
ARB R4-204, P.O. Box 100277, Gainesville, FL 32610-0277, Telephone: 352-846-1749, FAX: 352-392-2323, E-mail:
[email protected]
Submitted: June 6, 2007 Revised: August 22, 2007 Accepted: August 27, 2007
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Abstract Allogeneic hematopoietic stem cell transplantation (HSCT) is an effective treatment for high-risk hematological malignancies, yet a major complication associated with this therapy is acute graftversus-host disease (GVHD). Despite a well-defined pathophysiological mechanism, there are no definitive markers for predicting acute GVHD development or progression to advanced stages. In the current study, we enrolled 4 acute GVHD and 4 acute GVHD-free recipients of allogeneic HSCT and collected peripheral blood just prior to onset of clinical acute GVHD for analysis on Affymetrix GeneChip® Human Genome U133 Plus 2.0 microarrays. We noted significant differences in expression of 1,658 genes between control and acute GVHD patients, based on an analysis of covariance (ANCOVA) by type of transplant, a pooled error estimate and a false discovery rate (FDR) of 10%. In conclusion, we offer the first report of a preliminary molecular signature of acute GVHD in allogeneic HSCT patients.
Keywords: GVHD, HSCT, microarray, gene expression
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Brief Communication The major complication associated with allogeneic hematopoietic stem cell transplantation (HSCT) is graft-versus-host disease (GVHD). Acute GVHD develops within 100 days posttransplant and is clinically manifest predominantly in the skin, liver and gastrointestinal tract. A poor clinical prognosis is imminent particularly in cases where tissue damage is extensive, disease progression is rapid, and patients are not responsive to immunosuppressive therapies (9). The classical model of acute GVHD pathophysiology is divided into three phases: (I) conditioning regimen, (II) donor T cell activation and (III) cytolytic effector responses (10). In Phase I, pre-transplant conditioning renders damage to mucosal tissue and initiates a “cytokine storm” marked by increased levels of inflammatory cytokines such as tumor necrosis factor- (TNF-), interleukin-1 (IL-1), IL-6 and IL-12. These stress signals activate host dendritic cells (DC) to up-regulate MHC, co-stimulatory molecules and adhesion proteins that facilitate alloantigen presentation to naïve donor T cells. In Phase II, alloreactive T cell proliferation is increased in response to interferon- (IFN-) and subsequently, naïve T cells become polarized toward T helper-type 1 (Th1). In Phase III, acute GVHD becomes clinically evident as damage to host tissues by mature natural killer (NK) cells and cytotoxic T lymphocytes (13,18). Despite this knowledge, no clinical disease markers exist for increased acute GVHD risk or acute GVHD progression to potentially fatal stages. In the past decade, it has become more feasible to uncover such molecular markers through the use of high-throughput technologies, such as DNA microarray analysis. Recently, we have demonstrated the applicability of Affymetrix microarray analysis in analyzing allograft composition (4). Prior studies have established the use of gene expression profiling in differentiating allograft tolerance and rejection (2).
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In the current study we analyzed the peripheral blood leukocyte gene expression profiles of 8 allogeneic HSCT recipients (Table 1) using Affymetrix Human Genome U133 Plus 2.0 GeneChip® microarrays. For each patient, 4mL of venous blood was collected at engraftment. We employed a leukocyte RNA isolation technique shown previously to have higher reliability than a whole blood RNA collection system when using the Affymetrix platform(5). In brief, immediately after sample collection leukocytes were concentrated by centrifugation, and Buffer EL (Qiagen, Valencia, CA) was added to lyse erythrocytes. Cells were washed again in Buffer EL to lyse residual erythrocytes and thus eliminate the abundance of globin species from the mRNA pool. Total RNA was purified with RNeasy mini kits (Qiagen) and 2.5 g total RNA was used to generate cRNA for microarray hybridization according to standard Affymetrix protocol. Concurrently, white blood counts were assessed and flow cytometry was used to measure lymphocyte (T cells, CD3+, CD4+, CD8+; NK, CD56+CD16+) and DC (plasmacytoid DC, Lin- HLA-DR+ CD123+; myeloid DC, Lin- HLA-DR+ CD11c+) subsets as previously described (11). Four patients developed clinical acute GVHD within 5 to 10 days after blood sample collection. The remaining 4 patients were GVHD-free after 3 months of follow-up, and were designated as study controls. This study received approval by the University of Florida Institutional Review Board and all patients provided informed consent prior to enrollment. We proposed that the period of engraftment would be most opportune for measuring gene expression changes occurring in acute GVHD for three reasons. First, we have previously shown that changes in immune cell counts as early as engraftment are associated with later onset of GVHD (11). Second, acute GVHD onset invariably occurs in the post-engraftment period. Third, since patients are routinely administered immunosuppressive drugs at the onset of acute GVHD, conducting a gene expression analysis prior to acute GVHD onset would eliminate
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concerns that changes in immune gene responses were masked by effects of immunosuppressive intervention. We analyzed 54,613 probe sets on each microarray. The probe signal intensities were first log transformed and then an analysis of covariance (ANCOVA) adjusting for type of transplant (e.g., unrelated vs. related donor) was carried out to assess if the expression level significantly differed among the acute GVHD and control groups. A global pooled error estimate was employed to gain more power for hypothesis testing by assuming equal variability in each gene's expression. Differential gene expression among the two groups [(-)acute GVHD vs. (+)acute GVHD] was assessed by controlling the false discovery rate (FDR) at 10% (12). The data discussed in this publication have been deposited in NCBIs Gene Expression Omnibus (GEO, http://www.ncbi.nlm.nih.gov/geo/) and are accessible through GEO Series accession number GSE75101. All probe set annotations were generated using the NetAffx tool available through the Affymetrix website. No significant differences in lymphocyte or DC counts were noted between acute GVHD and control groups at engraftment (Table 1). Yet, of the 54,613 probe sets measured on each microarray, 1,658 showed significant changes in expression based on a 10% FDR (Figure 1). A representative list of genes was compiled, with particular consideration for immune-related genes and those genes showing the greatest magnitude fold change (FC) in expression difference (Table 2). To summarize, down-regulated genes in acute GVHD patients included: the B lymphocyte-specific activation-induced cytidine deaminase (AICDA) and terminal deoxynucleotidyltransferase (DNTT) genes, pro-inflammatory genes such as toll-interleukin-1 1
(note: data will be made available upon publication of this work)
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domain containing adaptor protein (TIRAP), interferon- 16 (IFNA16), interleukin-27 (IL27), chemokine-ligand 1 (CCL1) advanced glycosylation end product-specific receptor (AGER) and components of the IL-1 receptor/NF-kappaB pathway (IRAK1BP1, NFKB1). Additionally, genes involved in immune cell activation such as the cysteinyl leukotriene receptor 2 (CYSLTR2), MHC I (HLA-C), inducible T-cell co-stimulator ligand (ICOSLG) and CD244, were down-regulated. Significantly up-regulated were many pro-inflammatory cytokine, cytokine receptor and cytokine regulatory genes such as IL-2 (IL2), IL-2R (IL2RA), IL-22R2 (IL22RA2), tumor necrosis factor (TNF), lymphotoxin beta (LTB), type 1 tumor necrosis factor receptor shedding aminopeptidase regulator (ARTS1) and CCAAT/enhancer binding protein A (CEBPA). Prior, investigators have measured changes in gene expression occurring in cutaneous or hepatic acute GVHD using mouse models of bone marrow transplantation (7,17,21). The consensus gained from these studies was that gene expression changes due to acute GVHD are dominated by IFN--related processes, leukocyte cell adhesion and leukocyte trafficking. More recently, it has been suggested that the gene expression profile of purified cell populations from allogeneic donors can predict development of GVHD in the recipient (1). In the current study, we observed a predominantly pro-inflammatory gene expression profile in acute GVHD patients consistent with knowledge of the early pathophysiological events surrounding acute GVHD. Significantly, we noted a strong up-regulation of IL-2 and its receptor, IL-2r. Given the role of IL-2 is as a growth factor for T cells, this likely reflects phase II of the acute GVHD pathophysiological process, which is marked by donor T cell expansion. Increased autoimmune regulator (AIRE) expression further suggests enhanced T cell activation, as it has been sown that AIRE is up-regulated in activated IL-2-responsive CD4+ T cells (9). Up-
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regulation of the complement component 3a receptor (C3AR1)—an important component of cell migration and marker for DC and lymphocyte activation (6,19), similarly agrees with early acute GVHD pathophysiology. Up-regulation of TNF, a critical pro-inflammatory cytokine that enhances the proliferation and maturation of donor immune cells and potentiates graft-versushost and graft-versus-malignancy effects, agrees with the “cytokine storm” described during early acute GVHD (14). Of important note is down-regulation of the IL-27 gene in acute GVHD patients. IL-27 is a novel IL-12 family cytokine with an apparently dichotomous role in regulating immune responses. It has been reported that IL-27 exhibits pro-inflammatory properties, activates Th1 cells and enhances immunological responses to tumor cells (15). Alternatively, it has been shown that IL-27 exerts anti-inflammatory effects through antagonism of IL-17-producing Th17 cells (16). Th17 cells produce the pro-inflammatory cytokine IL-22 which has been implicated in autoimmune disorders such as Crohn’s disease and dermal acanthosis (3,20). Notably, the receptor for IL-22 (IL-22RA2) was up-regulated in acute GVHD patients. Thus, perhaps the most valuable knowledge gained from this study is in uncovering the possible role for IL-27, IL22 and Th17 cellular responses in the development of acute GVHD. In conclusion, our work represents a pilot study investigating the genome-wide transcriptional response occurring in engraftment blood of allogeneic HSCT patients just before acute GVHD onset. Although measuring engraftment cell counts by flow cytometry did not have predictive potential in this relatively limited sample size, a microarray analysis yielded several candidate transcriptional markers for the development of acute GVHD. It should be noted that in our previous studies, not all of the patients with low DC counts developed GVHD
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suggesting that there are other mechanisms driving the pathophysiology. This observation provided the rationale for employing a hypothesis-generating microarray analysis. Although we analyzed engraftment blood based on our previous studies showing important immunologic changes occurring at this point, future studies may desire to address gene expression profiling of acute GVHD immediately after noting clinical symptoms, yet before administration of immunosuppressive drugs. Given the applicability of microarray analysis in predicting patient transplant outcomes, future investigations into molecular prognostic and diagnostic tests for GVHD are warranted. We have now demonstrated the feasibility of using peripheral blood for such analyses, thereby circumventing the need to obtain biopsies of patient tissue if otherwise not available to the investigator.
Acknowledgements We thank Christina Cline for data management support and the University of Florida Institute for Biotechnology Research Gene Expression Core for technical assistance. This study was funded in part by the University of Florida Shands Cancer Center.
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References 1. Baron C, Somogyi R, Greller LD et al. Prediction of graft-versus-host disease in humans by donor gene-expression profiling. PLoS Med. 4:e23; 2007. 2. Berg T.; Wu T.; Levay-Young B.; Heuss N.; Pan Y.; Kirchhof N.; Sutherland D.E.; Hering B.J.; Guo Z. Comparison of tolerated and rejected islet grafts: a gene expression study. Cell Transplant. 13:619-629; 2004. 3. Brand S.; Beigel F.; Olszak T.; Zitzmann K.; Eichhorst S.T.; Otte J.M.; Diepolder H.; Marquardt A.; Jagla W.; Popp A.; Leclair S.; Herrmann K.; Seiderer J.; Ochsenkühn T.; Göke B.; Auernhammer C.J.; Dambacher J. IL-22 is increased in active Crohn's disease and promotes proinflammatory gene expression and intestinal epithelial cell migration. Am J Physiol Gastrointest Liver Physiol. 290:827-838; 2006. 4. Buzzeo M.P; Yang J.; Casella G.; Reddy V. Hematopoietic stem cell mobilization with GCSF induces innate inflammation yet suppresses adaptive immune gene expression as revealed by microarray analysis. Exp Hematol. In Press. 5. Feezor R.J; Baker H.V.; Mindrinos M.; Hayden D.; Tannahill C.L.; Brownstein B.H.; Fay A.; MacMillan S.; Laramie J.; Xiao W.; Moldawer L.L.; Cobb J.P.; Laudanski K.; MillerGraziano C.L.;Maier R.V.; Schoenfeld D.; Davis R.W; Tompkins R.G.; and the Inflammation and Host Response to Injury, Large-Scale Collaborative Research Program. Whole blood and leukocyte RNA isolation for gene expression analyses. Physiol Genomics. 19:247-254; 2004. 6. Gutzmer R.; Lisewski M.; Zwirner J.; Mommert S.; Diesel C.; Wittmann M.; Kapp A.; Werfel T. Human monocyte-derived dendritic cells are chemoattracted to C3a after upregulation of the C3a receptor with interferons. Immunology. 111:435-443; 2004.
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7. Ichiba T.; Teshima T.; Kuick R.; Misek D.E.; Liu C.; Takada Y.; Maeda Y.; Reddy P.; Williams D.L.; Hanash S.M.; Ferrara J.L. Early changes in gene expression profiles of hepatic GVHD uncovered by oligonucleotide microarrays. Blood. 102:763-771; 2003. 8. Iwasaki T. Recent Advances in the Treatment of Graft-Versus-Host Disease. Clin Med Res. 2:243-252; 2004. 9. Nagafuchi S.; Katsuta H.; Koyanagi-Katsuta R.; Yamasaki S.; Inoue Y.; Shimoda K.; Ikeda Y.; Shindo M.; Yoshida E.; Matsuo T.; Ohno Y.; Kogawa K.; Anzai K.; Kurisaki H.; Kudoh J.; Harada M.; Shimizu N. Autoimmune Regulator (AIRE) Gene Is Expressed in Human Activated CD4(+) T-Cells and Regulated by Mitogen-Activated Protein Kinase Pathway. Microbiol Immunol. 50:979-987; 2006. 10. Reddy P., Ferrara J.L. Immunobiology of acute graft-versus-host disease. Blood Rev. 17:187-194; 2003. 11. Reddy V.; Iturraspe J.A.; Tzolas A.C.; Meier-Kriesche H.U.; Schold J.; Wingard J.R. Low dendritic cell count after allogeneic hematopoietic stem cell transplantation predicts relapse, death, and acute graft-versus-host disease. Blood. 103:4330-4335; 2004. 12. Reiner A.; Yekutieli D.; Benjamini Y. Identifying differentially expressed genes using false discovery rate controlling procedures. Bioinformatics. 19:368-375; 2003. 13. Schmaltz C.; Alpdogan O.; Horndasch K.J.; Muriglan S.J.; Kappel B.J.; Teshima T.; Ferrara J.L.; Burakoff S.J.; van den Brink M.R. Differential use of Fas ligand and perforin cytotoxic pathways by donor T cells in graft-versus-host disease and graft-versusleukemia effect. Blood. 97:2886-2895; 2001. 14. Schmaltz C.; Alpdogan O.; Muriglan S.J.; Kappel B.J.; Rotolo J.A.; Ricchetti E.T.; Greenberg A.S.; Willis L.M.; Murphy G.F.; Crawford J.M.; van den Brink M.R. Donor T
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cell-derived TNF is required for graft-versus-host disease and graft-versus-tumor activity after bone marrow transplantation. Blood. 101:2440-2445; 2003. 15. Shimizu M.; Shimamura M.; Owaki T.; Asakawa M.; Fujita K.; Kudo M.; Iwakura Y.; Takeda Y.; Luster A.D.; Mizuguchi J.; Yoshimoto T. Antiangiogenic and antitumor activities of IL-27. J Immunol. 176:7317-7324; 2006. 16. Stumhofer J.S.; Laurence A.; Wilson E.H.; Huang E.; Tato C.M.; Johnson L.M.; Villarino A.V.; Huang Q.; Yoshimura A.; Sehy D.; Saris C.J.; O'Shea J.J.; Hennighausen L.; Ernst M.; Hunter C.A. Interleukin 27 negatively regulates the development of interleukin 17producing T helper cells during chronic inflammation of the central nervous system. Nat Immunol. 7: 937-945; 2006. 17. Sugerman P.B.; Faber S.B.; Willis L.M.; Petrovic A.; Murphy G.F.; Pappo J.; Silberstein D.; van den Brink M.R. Kinetics of gene expression in murine cutaneous graft-versus-host disease. Am J Pathol. 164:2189-2202; 2004. 18. van den Brink M.R.; Burakoff S.J. Cytolytic pathways in haematopoietic stem-cell transplantation. Nature Rev Immunol. 2:273-281; 2002. 19. Werfel T.; Kirchhoff K.; Wittmann M.; Begemann G.; Kapp A.; Heidenreich F.; Götze O.; Zwirner J. Activated human T lymphocytes express a functional C3a receptor. J Immunol. 165:6599-6605; 2000. 20. Zheng Y.; Danilenko D.M.; Valdez P.; Kasman I.; Eastham-Anderson J.; Wu J.; Ouyang W. Interleukin-22, a T(H)17 cytokine, mediates IL-23-induced dermal inflammation and acanthosis. Nature. 445:648-651; 2007.
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21. Zhou L.; Askew D.; Wu C.; Gilliam A.C. Cutaneous Gene Expression by DNA Microarray in Murine Sclerodermatous Graft-Versus-Host Disease, a Model for Human Scleroderma. J Invest Dermatol. 127:281-292; 2007.
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Figure Legends Figure 1. Differential gene expression between acute GVHD and acute GVHD-free patients. Shown is a dot plot representing engraftment blood gene expression changes for 1,658 genes significantly differentially expressed between patients who developed acute GVHD (n=4), or remained GVHD-free (n=4) after allogeneic hematopoietic stem cell transplantation. Each point on the plot represents an Affymetrix microarray probe set. X and Y coordinates were calculated as the log of the average signal intensity for (-)acute GVHD and (+)acute GVHD patient groups, respectively. All points above the line (y=x) are up-regulated genes (N=704); all points below are down-regulated genes (N=954).
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Table 1. Patient Characteristics and Engraftment Cell Numbers (-)acute GVHD (+)acute GVHD 1
2
3
4
5
6
7
8
Age/Sex
41/m
42/m
52/m
52/m
58/f
58/m
60/m
60/m
Disease
AML
AML
AML
AML
AML
AML
ALL
AML
Stem cell source
PBSC
PBSC
PBSC
PBSC
PBSC
PBSC
PBSC
PBSC
Graft CD34+ (x 106/kg)
5.20
10.15
7.01
7.02
5.00
6.00
5.36
5.29
Graft CD3+ (x 108/kg)
0.97
1.92
4.44
6.86
1.45
1.47
1.93
4.5
Pa
WBC (x10^3/uL)
6.00
6.70
2.80
2.60
5.60
3.30
4.20
3.70
0.79
#b CD3+
4.8
107.87
155.12
422.24
110.32
33.99
92.4
334.48
0.78
# CD4+
3.0
46.9
34.72
124.28
85.68
2.31
75.6
237.54
0.40
# CD8+
6.0
37.52
120.96
281.58
15.68
24.75
23.1
122.47
0.40
# CD16+CD56+
21.6
38.86
5.32
67.86
10.64
5.28
189
20.35
0.60
# Lin- HLA-DR+ CD123+
0
2.01
5.88
2.08
0
6.27
2.1
5.55
0.63
# Lin- HLA-DR+ CD11c+
0.6
8.71
8.96
1.56
0.56
9.57
8.82
7.03
0.63
a
T-test (two-tailed) comparing (+)acute GVHD vs. (-)acute GVHD mean cell count. Cell counts reported as absolute number of cells per mm3. Abbreviations: AML, acute myeloid leukemia; ALL, acute lymphocytic leukemia; PBSC, peripheral blood stem cells; WBC, white blood count b
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Table 2. Representative Gene Expression Changes Prior to acute GVHD Onset Probe ID Gene Symbol Gene Title
FCa
223629_at
PCDHB5
protocadherin beta 5
5.81
237493_at
IL22RA2
interleukin 22 receptor, alpha 2
3.82
224399_at
PDCD1LG2
programmed cell death 1 ligand 2
3.75
207849_at
IL2
interleukin 2
3.22
241090_at
PKD1
Polycystic kidney disease 1 (autosomal dominant)
3.00
212013_at
PXDN
peroxidasin homolog (Drosophila)
2.81
216851_at
IGL@
Immunoglobulin lambda locus
2.78
208090_s_at
AIRE
autoimmune regulator
2.71
206341_at
IL2RA
interleukin 2 receptor, alpha
2.51
207113_s_at
TNF
tumor necrosis factor (TNF superfamily, member 2)
2.35
202585_s_at
NFX1
nuclear transcription factor, X-box binding 1
2.12
1559754_at
LTB
Lymphotoxin beta (TNF superfamily, member 3)
2.04
209906_at
C3AR1
complement component 3a receptor 1
1.94
212580_at
ARTS-1
Type 1 tumor necrosis factor receptor shedding aminopeptidase
1.89
regulator 214398_s_at
IKBKE
inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase
1.74
epsilon 216852_x_at
IGL@
Immunoglobulin lambda locus
1.63
225527_at
CEBPG
CCAAT/enhancer binding protein (C/EBP), gamma
1.52
222233_s_at
DCLRE1C
DNA cross-link repair 1C (PSO2 homolog, S. cerevisiae)
1.44
204212_at
ACOT8
acyl-CoA thioesterase 8
1.39
214574_x_at
LST1
leukocyte specific transcript 1
1.36
200759_x_at
NFE2L1
nuclear factor (erythroid-derived 2)-like 1
1.35
208771_s_at
LTA4H
leukotriene A4 hydrolase
1.33
200758_s_at
NFE2L1
nuclear factor (erythroid-derived 2)-like 1
1.30
204039_at
CEBPA
CCAAT/enhancer binding protein (C/EBP), alpha
1.28
15
Table 2. Continued 212684_at
ZNF3
zinc finger protein 3
1.27
200778_s_at
SEPT2
septin 2
1.27
208097_s_at
TXNDC
thioredoxin domain containing
1.26
205603_s_at
DIAPH2
diaphanous homolog 2 (Drosophila)
1.22
214459_x_at
HLA-C
major histocompatibility complex, class I, C
-1.16
218520_at
TBK1
TANK-binding kinase 1
-1.22
213749_at
MASP1
mannan-binding lectin serine peptidase 1
-1.28
206584_at
LY96
lymphocyte antigen 96
-1.35
220813_at
CYSLTR2
cysteinyl leukotriene receptor 2
-1.35
228891_at
C9orf164
chromosome 9 open reading frame 164
-1.54
244811_at
IRAK1BP1
Interleukin-1 receptor-associated kinase 1 binding protein 1
-1.56
239876_at
NFKB1
Nuclear factor of kappa light polypeptide gene enhancer in B-cells 1
-1.67
(p105) 242903_at
IFNGR1
Interferon gamma receptor 1
-1.75
207037_at
TNFRSF11A
tumor necrosis factor receptor superfamily, member 11a, NFKB
-1.75
activator 209079_x_at
PCDHG
protocadherin gamma subfamily A,B,C
-1.85
1555086_at
STAT5B
signal transducer and activator of transcription 5B
-1.85
208448_x_at
IFNA16
interferon, alpha 16
-1.92
210000_s_at
SOCS1
suppressor of cytokine signaling 1
-1.96
210081_at
AGER
advanced glycosylation end product-specific receptor
-2.17
216846_at
IGL@
Immunoglobulin lambda locus
-2.38
211199_s_at
ICOSLG
inducible T-cell co-stimulator ligand
-2.50
1552804_a_at
TIRAP
toll-interleukin 1 receptor (TIR) domain containing adaptor protein
-2.63
234320_at
CD244
CD244 molecule, natural killer cell receptor 2B4
-2.63
234340_at
PROCR
Protein C receptor, endothelial (EPCR)
-3.03
211129_x_at
EDA
ectodysplasin A
-3.13
203954_x_at
CLDN3
claudin 3
-3.70
16
Table 2. Continued 210487_at
DNTT
deoxynucleotidyltransferase, terminal
-3.85
1554091_a_at
TIRAP
toll-interleukin 1 receptor (TIR) domain containing adaptor protein
-4.35
1560861_at
SCAP1
Src family associated phosphoprotein 1
-4.55
228895_s_at
ASB1
Ankyrin repeat and SOCS box-containing 1
-4.55
243706_at
CDO1
Cysteine dioxygenase, type I
-4.76
224499_s_at
AICDA
activation-induced cytidine deaminase
-5.88
228704_s_at
CLDN23
Claudin 23
-5.88
203854_at
CFI
complement factor I
-6.25
207533_at
CCL1
chemokine (C-C motif) ligand 1
-6.67
222285_at
IGHD
immunoglobulin heavy constant delta
-7.14
1552995_at
IL27
interleukin 27
-8.33
232099_at
PCDHB16
protocadherin beta 16
-9.09
Log2 fold change (FC) in signal intensity calculated as [2log2 [avg (+)acute GVHD – avg (-)acute GVHD]]. FC>0, up-regulated; FC
