A number of genes active in granulocytes have been intensively studied as to the function of their products and their expression controls. However, the intensities and relative order of these gene activities have not been studied. This report describes an expression profile of 748 different species of active genes in human peripheral granulocytes obtained by analyzing a 3′-directed cDNA library that faithfully represents the mRNA population in the source cells. A significant fraction (20.3% of the total) of the expressed genes in granulocytes consisted of nuclear proteins such as DNA binding proteins, of secretory proteins such as cytokines, and of membrane proteins such as major histocompatibility complex (MHC) proteins and receptors. By comparing this expression profile with 11 profiles similarly obtained with unrelated human cells/tissues, we discovered 10 novel genes that are likely to act specifically in granulocytes. Comparison of this expression profile with that obtained with granulocytoids widely used as a granulocyte model by inducing a cultured promyelocytic leukemia cell line HL60 showed similarities and dissimilarities of gene expressions.

© 1998 by The American Society of Hematology.

Topics:
genes, granulocytes, rna, messenger, dna, complementary

GRANULOCYTES ARE short-lived cells for body defense, with a half-life of 6 to 7 hours in blood. Because of this short life span, their protein synthetic apparatus is poorly developed,1,2 but they maintain active genes that are responsible for their unique activities. These genes have been the focus of intensive study.

We have initiated a systematic survey of active genes, as well as the relative abundance of mRNA expression, in granulocytes using an expression profiling method that is based on quantitative analysis of mRNA populations.3 This is performed by using 3′-directed cDNA libraries that faithfully represent the mRNA population and by obtaining short base sequences just upstream of polyA, called gene signatures (GSs), by single-pass sequencing of randomly selected clones from such libraries.3-5 Active genes are identified by sequences and the gene activities are identified by their recurrences. The resulting list showing the expressed gene species and the abundance of their transcript is called an expression profile, which illuminates the gene-product–based cellular phenotype.

In the profile are represented several known genes as well as novel genes. These genes can be categorized as those that are commonly expressed in different types of cells (candidate genes for housekeeping functions) and those that are expressed uniquely in granulocytes (candidate genes for granulocyte-specific functions). We used 11 expression profiles obtained with different human cells/tissues for the gene categorization and discovered some genes that are likely to be granulocyte-specific, even though we do not yet know their functions.

A promyelocyte cell line, HL60,6 is converted into granulocytoid cells by treatment with dimethylsulfoxide (DMSO). The cells stopped proliferating, the nucleus became pyknotic and polymorphic, and the cells expressed several genes unique to granulocytes. Thus, because they are morphologically and functionally similar to granulocytes, the cells are referred to as granulocytoids and are widely used as a model system for studies of granulocytes.7-10 We monitored the expression profile of these cells. Comparison of this profile with that of the peripheral granulocytes showed advantages and disadvantages in using the granulocytoids as a model system.

Preparation of human granulocytes.

Freshly obtained venous blood was diluted with the same volume of phosphate-buffered saline (PBS), and the suspension was centrifuged on a ficoll step gradient (upper density, 1.077; lower density, 1.119) for 20 minutes at 3,000g.11,12 Granulocytes collected from the interface were washed twice with PBS and resuspended in the same buffer. The purity of the preparation was examined under a microscope upon Giemsa staining. At least 99% of total cells were mature granulocytes.

Induction of granulocytoid cells.

HL60 cells (Japanese Cancer Research Resource Bank, Osaka, Japan) were grown in RPMI 1640 medium (Nissui, Tokyo, Japan) supplemented with 10% (vol/vol) bovine fetal serum (Hyclone, Logan, UT) and harvested in the logarithmic phase (106 cells/mL). Granulocytoid cells were prepared by seeding HL60 in a plate at a concentration of 2 × 105cells/mL and incubating in the presence of 1.3% (vol/vol) DMSO (Sigma, St Louis, MO) for 72 hours.

cDNA library construction and sequencing.

RNAs were prepared from the cytoplasmic fraction of DMSO-induced HL60 cells as described3 and from the total cell lysate13 of granulocytes. Purification of polyA RNA was not attempted, because of the presence of RNase. Construction of the 3′-directed cDNA libraries and transformation intoEscherichia coli were performed by synthesizing cDNA using pUC19-based vector primer, digesting with dam-sensitive four-base cutter Mbo I, followed by circularization and transformation into E coli.4 The transformant colonies were randomly selected and cultured in 96-well plates, and the inserted cDNAs were amplified with flanking primers and subjected to cycle sequencing.

Data analysis.

The polyA tail was removed from the sequencing data after checking the electropherograms, leaving 3 As as a marker. From the resulting sequence data, those having inserts shorter than 20 bp or those having more than 5 ambiguous bases (N) within the initial 100 bases were discarded. The sequences of the remaining clones were truncated where the N content exceeded 5%, and the final N was replaced by an X to mark the point of truncation. The resulting sequences are referred to as GSs.

The GSs were compared using the FastA program.14 Identical signatures were lumped together, and a representative sequence that had the lowest content of ambiguous bases was selected to represent the group and deposited in the DDBJ, wherein the locus name corresponds to the GS number (such as HUMGS01234 to GS01234). All the representative GSs were searched against GenBank (Re95) using the FastA program,14 and those that had greater than 90% similarity to the 3′ end of the mRNA entries or to the reported terminal exon of genes were regarded as representing the corresponding genes.

Expression profiles of active genes in granulocytes.

From the 3′-directed cDNA library constructed from human peripheral granulocytes, we randomly selected 1,142 independent clones and sequenced them. Among the resulting short sequences called GSs, representing just upstream of the polyA, sequences that were considered essentially identical were lumped together to represent the same gene species. After this treatment, 748 independent GS species resulted. Among them, 216 (28.9%) represented by 493 clones were identified in GenBank (Re95), and the remaining 532 (71.1%), represented by 649 clones, were from novel genes. Table 1A shows an expression profile of active genes as represented by their GSs and their activities with their relative abundance. We listed here only those 64 GSs that appeared 3 times or more in descending order of appearance. Those genes that appeared twice or less can be seen in www bodymapper server (http://www.imcb.osaka-u.ac.jp/bodymap). We believe that this is the first publication describing relative activities of genes in granulocytes that are expressed abundantly. The profile reflects several unique features of the granulocytes physiology. First, it includes several genes that have been well known in peripheral neutrophils, such as genes for β2-microglobulin,15granulocyte colony-stimulating factor receptor,16-18 major histocompatibility complex (MHC) class I, and so on. Genes whose activity has been detected in granulocytes15-36 are marked with asterisks. High activities of genes for spermidine/spermine N1-acetyltransferase, pre-B–cell enhancing factor (PBEF), and B94 protein are also noted. The purity of the source material guarantees that this result reflects the relative activities of those genes.

Table 1.

Expression Profile of Active Genes in Human Peripheral Granulocytes (A) and Granulocytoids by Inducing HL60 With DMSO (B)

(A)
GS lib PM GR HL MOHE FL IL AL AP VF LG CM KC CORE Gene Name
00565  ▪  24  3  4  5   1  1  1   1  4  2   1  β 2-microglobulin* 
00402  ▪  19    1  1 2     1  1   1  2  Spermidine/spermine N1-acetyltransferase  
00254  ▪ 18    2  1    1  1  1  2   3   MHC class I HLA-Cw1  
01823   18   2             Pre-B cell enhancing factor (PBEF)  
02821   15         1  2  2   1   CL 100 protein tyrosine phosphatase  
00937  ▪  14  4 1        5  11   1    HLA-E heavy chain (3′ untranslated region)  
08362  □  13               Granulocyte colony-stimulating factor receptor* 
03341   11        1  2       B94 protein  
01476   10  3   2     1  1  
08339  □  9               IL-8* 
01267   8  1              1-8D 
01811   8    1         2   1  H3.3 histone  
08347  □  8  
01367 ▪  7  3   1      1  1  3    1  hnRNP-E1  
00155  ▪  6  3  8  1   3   2   2  4  1  4  Thymosin β-4  
02490  ▪  6       1  1  1  2    2  1 
04345  □  6               B4-2 protein  
05209  □  6  
01724   6   2            Pleckstrin (P47) 
00816   5  5  1  1        1     Lymphocyte-specific protein 1 (LSP1)  
00994 ▪  5  2   1      1     1  1  Glutamine synthetase  
01809  5    3         6   1  Lysosomal membrane glycoprotein CD63  
04290  □ 5  
02015   5    11            LD78 α/macrophage inflammatory protein (GOS19-1)* 
07795   5         1       B-cell lymphoma 3-encoded protein (bcl-3)  
08383  □  5               Zinc-finger protein (bc1-6)  
00244  ▪  4  14   1  2     1   3  4  1  1  β-actin 
01371  □  4  1  
02161   4     1           AMP deaminase isoform L (AMPD2)  
05014  □  4               Histone H3.3  
05242  □  4  
03554  □  4  
08325 □  4  
08395  □  4  
08438  □  4               Nramp 
01087   3  6        2  1      Brain-expressed HHCPA78 homolog  
00685  3  5  2  4     1        L-plastin  
00273  ▪  3 4  6  11  9  2  11  2  12  2  1  5 3  3  5  Translationally controlled tumor protein 
01228   3  4       1    1     Anonymous mRNA  
00304  ▪ 3  2  1  2  2  1   1  3  4  1  3  3   Ubiquitin  
01473   3  2  2        1  
01187   3  2  1        1      Monocyte activation antigen (Mo3)  
01334   3  2   1 
01406   3  2   1  
01585   3  2  3             Tumor necrosis factor receptor* 
01325  ▪  3  1        1   1   2  1 sui1iso1  
00239   3  1    1  
01304  3  1   2       1      Transmembrane glycoprotein (CD53) 
01594  □  3  1              Leukocyte common antigen T200 (CD45)* 
01919  ▪  3    5   1   5  6  3  2  5  19  Calcyclin  
03575   3           1  2    Histone H3.3  
01877   3    1       1   1    Paxillin 
00729   3   2          1    CLP mRNA 
06701   3            1 
04074   3            1  1   Transcription factor ETR101  
04103   3            1    HLA-B27 antigen  
01826   3   1  
03216   3         1       Long-chain acyl-CoA synthetase 
08336  □  3               Secretory granule proteoglycan peptide  
05103  □  3               ICAM-3* 
08389  □  3               E4BP4 gene  
08375  □  3               MAD-3 
08424  □  3  
08548  □  3  
Total   370 74  17  75  17  6  16  8  32  30  33 37  21  43  12   
(B)  
GS  lib  GR  PM  HL  MO  HE FL  IL  AL  AP  VF  LG  CM  KC  CO  RE Gene Name  
00565  ▪  3  24  4  5    1 1  1   1  4  2   1 β-2-microglobulin  
00937  ▪  4  14  1       5  11   1    HLA-E heavy chain (3′ untranslated region)  
01476   3  10  2      1  1  
01367  ▪  3  7  1      1  1  3     1 hnRNP-E1  
00155  ▪  3  6   8  1  3   2   2  4  1  4   Thymosin β-4  
00816   5  5  1  1       1      Lymphocyte-specific protein 1 (LSP1)  
00244  ▪  14  4   1  2     1   3  4  1  1  β-actin 
01087   6  3        2  1      Brain-expressed HHCPA78 homolog  
00685  5  3  2  4     1        L-plastin  
01228   4 3       1     1    Anonymous mRNA  
00273  ▪  4  3  6 11  9  2  11  2  12  2  1  5  3  3 5  Translationally controlled tumor protein  
01394 □  4  2             1  
01106   4  2   3            Lysozyme  
01321 ▪  3  2   2   1  2   1   1  1  2  2  
01007  ▪  3  2       1   1  2  2   Stimulatory GTP-binding protein alpha subunit 
01609   4  1       1    2 1     γ-interferon-inducible protein (IP-30) 
01603  □  4  1  
01224   4  1         1      KIAAO184 protein  
01075   4  1     1   1  
00543  ▪  4  1  9  5    2 1  2  2   2  2  2  3  23-kD highly basic protein  
01365  □  3  1  
00995  ▪  3 1   1   1   1  1   1  2   1 1  Eukaryotic initiation factor 4AII  
00783   3  1  1  
00383  ▪  3  1  1  3  3  1   2    1  2  
total  □  6  2 2  3  31  14  25  16  56  30  42  49 34  37  29  
00583  ▪  5   2  3    1  1  2  3  2  1   4  Ribosomal protein L3  
00249   5     1    1  1    1    Liver glycogen phosphorylase type IV  
00114  ▪  5   1   1 1     1  3  1     Cytoskeletal gamma-actin  
01572  □  4  
01345  □  4               Bactericidal permeability increasing protein (BPI)  
01445   4        1    1  
01024   4   2             Leukocyte adhesion protein/LFA-1/Mac-1  
00096  ▪  4   5 2  6    1  6  2  2  3  3   1 α NAC  
01478   3             2  hnRNP-E2  
01372   3         1     2   Antigen migration inhibitory factor-related protein 14  
01244 ▪  3    1      2  2   2  1 1  2  Mitochondrial ubiquinone-binding protein  
01141  3      1    2  1    7  
01091   3        1    1    Transcriptional coactivator PC4  
00997  □ 3  
00875  ▪  3   2  1   1  1      1  
00839   3   1          1  1   Cercopithecus aethiops UV-damaged DNA-binding protein  
00732  ▪  3  6  4    1  4  1   1  1   2 1  Ribosomal protein L38  
00716  ▪  3   5 3      1  3  2  1  2  1  1 
00697   3   2  2            Plasminogen activator inhibitor-2 (PAI-2)  
00687   3   1  5            Glycoprotein 
00335  ▪  3   15  3  1  1  1   7 2  3  6  1  5  1  Ribosomal protein L7a 
00290  ▪  3   9  8  5  4  2  1  1  1  1  5  2  4  Ribosomal protein L37a 
00162  ▪  3   4  2  1  1   1  5 2  2  4  1  1   Acidic ribosomal phosphoprotein P2  
00071   3     1            
(A)
GS lib PM GR HL MOHE FL IL AL AP VF LG CM KC CORE Gene Name
00565  ▪  24  3  4  5   1  1  1   1  4  2   1  β 2-microglobulin* 
00402  ▪  19    1  1 2     1  1   1  2  Spermidine/spermine N1-acetyltransferase  
00254  ▪ 18    2  1    1  1  1  2   3   MHC class I HLA-Cw1  
01823   18   2             Pre-B cell enhancing factor (PBEF)  
02821   15         1  2  2   1   CL 100 protein tyrosine phosphatase  
00937  ▪  14  4 1        5  11   1    HLA-E heavy chain (3′ untranslated region)  
08362  □  13               Granulocyte colony-stimulating factor receptor* 
03341   11        1  2       B94 protein  
01476   10  3   2     1  1  
08339  □  9               IL-8* 
01267   8  1              1-8D 
01811   8    1         2   1  H3.3 histone  
08347  □  8  
01367 ▪  7  3   1      1  1  3    1  hnRNP-E1  
00155  ▪  6  3  8  1   3   2   2  4  1  4  Thymosin β-4  
02490  ▪  6       1  1  1  2    2  1 
04345  □  6               B4-2 protein  
05209  □  6  
01724   6   2            Pleckstrin (P47) 
00816   5  5  1  1        1     Lymphocyte-specific protein 1 (LSP1)  
00994 ▪  5  2   1      1     1  1  Glutamine synthetase  
01809  5    3         6   1  Lysosomal membrane glycoprotein CD63  
04290  □ 5  
02015   5    11            LD78 α/macrophage inflammatory protein (GOS19-1)* 
07795   5         1       B-cell lymphoma 3-encoded protein (bcl-3)  
08383  □  5               Zinc-finger protein (bc1-6)  
00244  ▪  4  14   1  2     1   3  4  1  1  β-actin 
01371  □  4  1  
02161   4     1           AMP deaminase isoform L (AMPD2)  
05014  □  4               Histone H3.3  
05242  □  4  
03554  □  4  
08325 □  4  
08395  □  4  
08438  □  4               Nramp 
01087   3  6        2  1      Brain-expressed HHCPA78 homolog  
00685  3  5  2  4     1        L-plastin  
00273  ▪  3 4  6  11  9  2  11  2  12  2  1  5 3  3  5  Translationally controlled tumor protein 
01228   3  4       1    1     Anonymous mRNA  
00304  ▪ 3  2  1  2  2  1   1  3  4  1  3  3   Ubiquitin  
01473   3  2  2        1  
01187   3  2  1        1      Monocyte activation antigen (Mo3)  
01334   3  2   1 
01406   3  2   1  
01585   3  2  3             Tumor necrosis factor receptor* 
01325  ▪  3  1        1   1   2  1 sui1iso1  
00239   3  1    1  
01304  3  1   2       1      Transmembrane glycoprotein (CD53) 
01594  □  3  1              Leukocyte common antigen T200 (CD45)* 
01919  ▪  3    5   1   5  6  3  2  5  19  Calcyclin  
03575   3           1  2    Histone H3.3  
01877   3    1       1   1    Paxillin 
00729   3   2          1    CLP mRNA 
06701   3            1 
04074   3            1  1   Transcription factor ETR101  
04103   3            1    HLA-B27 antigen  
01826   3   1  
03216   3         1       Long-chain acyl-CoA synthetase 
08336  □  3               Secretory granule proteoglycan peptide  
05103  □  3               ICAM-3* 
08389  □  3               E4BP4 gene  
08375  □  3               MAD-3 
08424  □  3  
08548  □  3  
Total   370 74  17  75  17  6  16  8  32  30  33 37  21  43  12   
(B)  
GS  lib  GR  PM  HL  MO  HE FL  IL  AL  AP  VF  LG  CM  KC  CO  RE Gene Name  
00565  ▪  3  24  4  5    1 1  1   1  4  2   1 β-2-microglobulin  
00937  ▪  4  14  1       5  11   1    HLA-E heavy chain (3′ untranslated region)  
01476   3  10  2      1  1  
01367  ▪  3  7  1      1  1  3     1 hnRNP-E1  
00155  ▪  3  6   8  1  3   2   2  4  1  4   Thymosin β-4  
00816   5  5  1  1       1      Lymphocyte-specific protein 1 (LSP1)  
00244  ▪  14  4   1  2     1   3  4  1  1  β-actin 
01087   6  3        2  1      Brain-expressed HHCPA78 homolog  
00685  5  3  2  4     1        L-plastin  
01228   4 3       1     1    Anonymous mRNA  
00273  ▪  4  3  6 11  9  2  11  2  12  2  1  5  3  3 5  Translationally controlled tumor protein  
01394 □  4  2             1  
01106   4  2   3            Lysozyme  
01321 ▪  3  2   2   1  2   1   1  1  2  2  
01007  ▪  3  2       1   1  2  2   Stimulatory GTP-binding protein alpha subunit 
01609   4  1       1    2 1     γ-interferon-inducible protein (IP-30) 
01603  □  4  1  
01224   4  1         1      KIAAO184 protein  
01075   4  1     1   1  
00543  ▪  4  1  9  5    2 1  2  2   2  2  2  3  23-kD highly basic protein  
01365  □  3  1  
00995  ▪  3 1   1   1   1  1   1  2   1 1  Eukaryotic initiation factor 4AII  
00783   3  1  1  
00383  ▪  3  1  1  3  3  1   2    1  2  
total  □  6  2 2  3  31  14  25  16  56  30  42  49 34  37  29  
00583  ▪  5   2  3    1  1  2  3  2  1   4  Ribosomal protein L3  
00249   5     1    1  1    1    Liver glycogen phosphorylase type IV  
00114  ▪  5   1   1 1     1  3  1     Cytoskeletal gamma-actin  
01572  □  4  
01345  □  4               Bactericidal permeability increasing protein (BPI)  
01445   4        1    1  
01024   4   2             Leukocyte adhesion protein/LFA-1/Mac-1  
00096  ▪  4   5 2  6    1  6  2  2  3  3   1 α NAC  
01478   3             2  hnRNP-E2  
01372   3         1     2   Antigen migration inhibitory factor-related protein 14  
01244 ▪  3    1      2  2   2  1 1  2  Mitochondrial ubiquinone-binding protein  
01141  3      1    2  1    7  
01091   3        1    1    Transcriptional coactivator PC4  
00997  □ 3  
00875  ▪  3   2  1   1  1      1  
00839   3   1          1  1   Cercopithecus aethiops UV-damaged DNA-binding protein  
00732  ▪  3  6  4    1  4  1   1  1   2 1  Ribosomal protein L38  
00716  ▪  3   5 3      1  3  2  1  2  1  1 
00697   3   2  2            Plasminogen activator inhibitor-2 (PAI-2)  
00687   3   1  5            Glycoprotein 
00335  ▪  3   15  3  1  1  1   7 2  3  6  1  5  1  Ribosomal protein L7a 
00290  ▪  3   9  8  5  4  2  1  1  1  1  5  2  4  Ribosomal protein L37a 
00162  ▪  3   4  2  1  1   1  5 2  2  4  1  1   Acidic ribosomal phosphoprotein P2  
00071   3     1            

Genes are registered according to their order of abundance. Numbers represent the frequency among 1142 cDNA clones analyzed in (A) and 1042 cDNA analyzed in (B). For simplicity’s sake, only those genes that appeared three times or more are listed. The expression profile of low abundant genes can be accessed electronically in www bodymapper server (http://www.imcb.osaka-u.ac.jp/bodymap). Abundances of the GSs that appeared in other libraries are also shown for comparison’s sake. Numbers represent abundance among 1142 cDNA clones. Number of libraries in which a given GS has been detected is shown in the column ‘lib’ by the shading pattern: (solid area) the GS has been detected in 6 libraries or more representing ubiquitous expression; (open area) only in the PM and GR libraries, possibly representing granulocyte-specific expression; (hatched area) in 2 through 5 libraries, possibly representing common expression. Genes that have so far been reported in GenBank are listed in the column “Gene Name.”

Abbreviations: PM, peripheral granulocytes; GR, granulocytoid cells induced by DMSO; HL, HL60 cells; MO, monocytoid cells obtained by inducing HL60 with tetradecanoylphorbol-13 acetate (TPA); HE, HepG2 cells; FL, fetal liver; IL, infant liver; AL, adult liver; AP, subcutaneous fat; VF, visceral fat; LG, lung; CM, colonic mucosa; KC, kerationcyte; CO, cornea; RE, retina.

*Active in granulocytes (only in A).

View Large

We categorized the 493 known genes active in the granulocytes into subgroups according to their function and subcellular localization. The results are collectively shown in Table2. The most prominent feature of granulocytes is the high activity with genes for cell surface membrane components. Thirty GS species represented by 114 clones (Table 2J), amounting to 10% of the mRNA population, were of this category. Genes for nuclear DNA binding protein, components for secretory protein, and components for signal transduction were also noticeably active. Genes for energy production, lysosomal proteins, protein synthesis machinery, and cytoskeleton are not so active in the granulocytes, as had been expected.

Table 2.

Identified Genes Detected in Peripheral Granulocytes

GS PM GR HL MO HE FL ILAL AP VF LG CM KC CO RE Gene Name
A. Energy production  
00943 1  1  3             Phosphoglycerate kinase 1  
02200  1      1        1   Transaldolase 
Total  2  1  3    1        1    
B. Miscellaneous  
00402  19   1  1  2     1  1   1  2  Spermidine/spermine N1-acetyltransferase  
02161  4     1           AMP deaminase isoform L (AMPD2)  
00304  3  2  1  2 2  1   1  3  4  1  3   3  Ubiquitin  
03216  3        1       Long-chain acyl-CoA synthetase 
01202  2  1              Neutrophil oxidase factor (NCF2)/p67-phox  
00483  2     1           IMP dehydrogenase type 1  
03596  2               Spermidine/spermine N1-acetyltransferase  
07377  2         1       Inosine monophosphate dehydrogenase type II  
08337  2               Grancalcin 
08513  2               Proteasome subunit p40 (Mov34)  
01263  1  2              Vacuolar H+-ATPase Mr 56,000 subunit (HO57)  
00253  1  2  1  2   1      2     Importin beta subunit  
01328  1  1              Calpastatin=glycoprotein BS-17 component  
02147  1      1    2       Protein disulfide isomerase related protein (ERp72)  
00154  1     1         1   Glutaredoxin 
00533  1   1       2     3 6  1  Ferritin H chain  
01766  1    6  15  2  4   1  2  5  10   1 Metallothionein from cadmium-treated cells  
07546  1               Type 1 inositol 1,4,5-trisphosphate receptor  
08483  1               Ubiquitin conjugating enzyme (UBC4)  
08328  1               Ribonuclease A  
08418  1               Erythrocyte-type AMP deaminase  
08504  1               Lysyl hydroxylase (PLOD)  
08643  1               Histidase 
08690  1               Ferritin H chain  
Total  55  8  3 9  7  20  3  5  6  9  4  10  14  12 2   
C. Lysosomal 
01809  5    3         6  1   Lysosomal membrane glycoprotein CD63  
01106 2  4   3            Lysozyme  
01533 1  1   2       2  3  1   1 1  Lysosomal membrane glycoprotein-1  
08663  1               Arylsulphatase A  
08679  1               26S protease (S4) regulatory subunit  
08396  1               Giantin/golgi antigen gcp372  
08464  1               Iduronate sulphate sulphatase (IDS)  
08664  1               Iduronate sulphate sulphatase (IDS)  
Total 13  5   8       2  3  7   2 1   
D. Signal transduction  
02821  15         1  2  2   1   CL 100 protein tyrosine phosphatase  
01724  6    2            Pleckstrin (P47) 
00816  5  5  1  1        1     Lymphocyte-specific protein 1 (LSP1)  
01919 3    5    1   5  6  3  2  5 19   Calcyclin  
00651  2  1  4          1    Rho GDP-dissociation Inhibitor 2  
01089  2  1              Tyrosine kinase (HCK)  
07614  2               Cyclin G2  
01524  1  1     1          TSE1=protein kinase A regulatory subunit  
00124  1   1   2     1     1   Ras-related protein (Krev-1)  
00286  1     2           ERK activator kinase (MEK1)  
00637  1   1             56K autoantigen annexin XI  
01687  1    1            c-raf-1 proto-oncogene  
02144  1      1   1        Protein kinase C ζ  
03705 1               α-subunit of Gi2  
04134  1           1    Rapamycin- and FK506-binding protein  
06319 1              1  Protein kinase  
08680  1               5-lipoxygenase 
Total  45  8  7  9  4 2  1  1  5  8  6  5  6  22    
E. Ribosomal component and translation factor  
01325  3  1         1  1   2  1  sui1iso1  
00314  2  1  3 3  1   1  1  1  4  5    3  2 Ribosomal protein S11  
00919  2  1  1    1          Splicing factor (CC1.4)  
00305  2   6  2  1  2  3  1 1  2  2  2  4  1   Ribosomal protein L28 
00995  1  3   1   1   1  1   1 2   1  1  Eukaryotic initiation factor 4AII 
00650  1  2  14  5    2   1  2  1 6  1  2  2  Ribosomal protein S18  
00418  1 1  22  10  5  1  5  1  10  1    7  9  Ribosomal protein L9  
00795  1 1  1  2    1  2  5  3     6 3  Ribosomal protein L30  
00818  1  1  4  1   2  1     2    2  Ribosomal protein L5  
00889  1   1    1  1  1   1   2    EF-1 δ  
00356  1  16  2  1  2  1  2   1    1   L23 putative ribosomal protein  
03424  1            1   Translational initiation factor 2 β subunit 
Total  17  11  68  26  8  7  17  10  19 14  10  13  9  22  20   
GS  PM  GR  HL  MO  HE  FL  IL AL  AP  VF  LG  CM  KC  CO  RE  Gene Name 
F. Cytoskeltal  
00155  6 3   8  1   3   2   2  4  1  4  Thymosin β-4  
00244  4  14   1  2     1   3  4  1  1  β-actin 
00685  3  5  2  4     1        L-plastin  
01877  3   1        1   1   Paxillin  
04080  2           1  1   Cofilin  
01030  1  2   1     2   1      Myosin regulatory light chain  
01507  1  1              α-actinin 
00258  1     1    1   1   1 1    2.5-kb cytoskeletal tropomyosin TM30 (nm) 
01702  1    1            Alkali myosin light chain  
01726  1    2       1      Striated muscle contraction regulatory protein  
02833  1         1  1   1    Nonmuscle myosin heavy chain (NMHC)  
Total  24  25  2 18  4   3  2  5  3  5  9  9  5  1  
G. Mitochondrial 
00994  5  2   1      1     1  1  Glutamine synthetase  
00313 2  1    1  1      1     SOD-2 manganese superoxide dismutase  
00360 1  1  2   2     1   1    1  Mitochondrial phosphate carrier protein  
00934  1 1  3  1    1  3  1  5      2 coxVIb  
08345  1               ATP synthase c subunit  
08434  1               Manganese superoxide dismutase  
08667  1               Glutamine synthetase  
01995  1    3    1   1 1   1   4   Glutathione peroxidase  
08536 1                SOD-2 gene for manganese superoxide dismutase  
04986  1               Catalase 
Total  15  5  5  5  3 1  2  3  4  6  2  1   6  3   
H. Nuclear  
01811  8   1         2    1  H3.3 histone  
01367  7  3   1      1 1  3     1  hnRNP-E1  
07795  5         1       B-cell lymphoma 3-encoded protein (bc1-3)  
08383  5               Zinc-finger protein (bc1-6)  
05014  4               H3.3 histone  
03575  3           1  2    H3.3 histone  
04074  3           1   1  Transcription factor ETR101  
08375  3               MAD-3 
08389  3               E4BP4 gene  
01431  2  1         3      jun-D  
01719  2    1            Myeloid cell nuclear differentiation antigen  
06259  2               spi-1 proto-oncogene  
08044  2             1   Zinc finger transcriptional regulator  
01566  1  1     1    1     1   fus 
00827  1  1  1     2        2  dUTP pyrophosphatase (hdut) 
08621  1               hnRNP A2/hnRNP B1  
04003  1           1     Receptor of retinoic acid  
01772  1    1            Interferon-γ induced protein (IFI 16)  
01912  1    1     1        CDEI binding protein/amyloid protein homologue  
02730  1          2    2   fos proto-oncogene  
00487  1     1       1     Transformation upregulated nuclear protein  
00648  1   1          2   1  Proliferating cell nucleolar protein P120  
01803  1    1            BAT1 nuclear RNA helicase (DEAD family)  
01832  1    1      1  2  1  1    jun-B 
03413  1               tpr 
06299  1               Arginine-rich nuclear protein  
08013  1             1  Replication protein A 70 kD subunit  
08366  1               Pre-mRNA splicing factor SR p75  
08411  1               Transcription factor SUPT4H  
08454  1               Nuclear factor erythroid 2 isoform f  
08546  1               Transcription factor  
08647  1               Putative RNA helicase HRH1  
Total  68 6  2  7  1   3  1  1  7  7  7  5 6  5   
I. Secretory and granular  
01823  18    2            Pre-B cell enhancing factor (PBEF)  
08339  9               IL-8 
02015  5    11            LD78 α/macrophage inflammatory protein  
08336  3               Secretory granule proteoglycan peptide  
08638  2               N-formylpeptide receptor (fMLP-R26)  
01820  2    1            Antisecretory factor-1  
00592  1  1  1  1    1       1   ATL-derived factor/thioredoxin  
01616  1  1   3   1         1  Cystatin B  
08342 1               Neutrophil peptide/defensin1/HP-1  
08553  1               Cathepsin S  
08318  1               N-formylpeptide receptor (fMLP-R98)  
00111  1    1  7  9 13  6         α 1-antitrypsin 
02007  1    1      1     1  1  Globin  
08501  1               Metalloproteinase-2 inhibitor (TIMP-2)  
08530  1               Platelet-derived endothelial cell growth factor  
08644  1               Platelet-derived endothelial cell growth factor  
08674  1               Monocyte/neutrophil elastase inhibitor  
Total  50  2  1  20  7 10  14  6  1      2  2   
GS  PM  GR  HL  MO  HE  FL  IL AL  AP  VF  LG  CM  KC  CO  RE  Gene Name 
J. Surface membrane  
00565 24  3  4  5    1  1  1   1  4 2   1  β-2-microglobulin  
00254  18   2  1    1  1  1  2    3  MHC class I HLA-Cw1  
00937  14  4  1       5  11   1    HLA-E heavy chain  
08362  13               Granulocyte colony-stimulating factor receptor  
08438  4               Nramp 
01187  3  2   1        1     Monocyte activation antigen (Mo3)  
01585 3  2   3            Tumor necrosis factor receptor  
01304  3  1   2       1      Transmembrane glycoprotein (CD53) 
01594  3  1              Leukocyte common antigen T200 (CD45)  
04103  3           1     HLA-B27 antigen  
05103  3               ICAM-3 
01624  2  2              Aminopeptidase N/CD13 encoding aminopeptidase N  
00303  2   2  2    1      1    (chromosome 3p25) membrane protein  
08355  2               C5a anaphylatoxin receptor  
08594  2               TAP1 
01048  1  1              MHC antigen (HLA-B)  
01395  1  1   1   2      1     Transmembrane receptor protein  
01229  1  1              Migration inhibitory factor-related protein 8  
00474  1    1           Transmembrane carcinoembryonic antigen BGPb  
02079  1     1          Palmitoylated erythrocyte membrane protein (MPP1)  
02156  1      1     1      CD14 myelid cell-specific leucine-rich glycoprotein  
06689  1           1     Leukocyte antigen CD97  
08349  1               Leukocyte adhesion molecule-1 (LAM-1)  
08524  1               Interferon-γ receptor  
08563  1               Low-affinity IgG Fc receptor  
08564  1               Complement receptor type 1  
08592  1               Common acute lymphoblastic leukemia antigen (CALLA)  
08661  1               Putative voltage-gated potassium channel (KVLQT1)  
08678  1               MHC class I-related protein  
08686  1               IL-8 receptor β (IL8RB)  
Total 114  18  7  14  4  4  1  3  2  7  16 7  4  3  1   
K. Unknown Function  
03341 11         1  2      B94 protein  
01267  8  1              1-8D 
04345  6               B4-2 protein  
01087  3  6        2  1      Brain-expressed HHCPA78 homolog  
00273 3  4  6  11  9  2  11  2  12  2  1 5  3  3  5  Translationally controlled tumor protein  
01228  3  4       1    1     Anonymous mRNA  
00729  3  2           1   CLP  
01288  2  1              Bcl-2 related (Bfl-1) mRNA  
00080  2     1           Myeloid cell differentiation protein (MCL1)  
00600  2   2        1     1  CIRP 
01926  2    2       1  1   1  1  ras-related C3 botulinum toxin substrate (rac) mRNA  
08056  2             1  Helix-loop-helix basic phosphoprotein (GOS8)  
08235 2              1  ERF-1  
08444  2               GLUDP2 
00543  1  4  9  5    2  1  2  2  2  2  2  3  23-kD highly basic protein 
01224  1  4          1     KIAA0184 protein  
01609  1  4      1    2  1    γ-interferon-inducible protein (IP-30)  
00185 1  2  1  1  2        1  2   Putative p64 CLCP protein  
01032  1  2    1     1    4  2   polyA binding protein  
01608  1  2       1  2    1   5 APPH-amyloid precursor protein homolog  
01665  1  2              KIAAO168 protein  
00751  1  1  1             (clone SAA7C) sequence  
00936  1  1  2        2 1      MLN51  
01000  1  1              EVI2B3P 
00129  1    1  1      1  1   2   fau  
00241  1    2  1    1     1  1   Acute myeloid leukemia associated protein (AML1/EAP)  
00363  1    1  1   1   1  1  2  2  1 1  ORF (complete cds) and HepG2 identical sequence 
00434  1    2  1  1       1 1  1   MLN50  
02011  1    1      1       SH3 domain-containing proline-rich kinase (sprk)  
02135  1      1      1     KIAA0183 protein 
02443  1        1        HVLCAD gene  
02662  1          1      Anonymous mRNA  
02820  1         1  3  1  1    1-8U  
02835  1          1   1   Transformer-2 α (htra-2 α) mRNA  
03864  1           1     (clone 05) liver expressed protein fragment  
04100  1           1     Anonymous mRNA  
05241  1          1    1   ZFM1 protein  
06150  1         1       OS-9 precursor mRNA  
07841  1         2      Brain-expressed HHCPA78 homolog  
08011 1              1  Phorbolin I  
08353  1               H sapiens (clone pS6) Alu repeat  
08372  1               DNA sequence from cosmid U61B11  
08393  1               fus-chop fusion protein  
08400  1               MG44 
08410  1               Putative 32-kD heart protein PHP32 mRNA 
GS  PM  GR  HL  MO  HE  FL  IL AL  AP  VF  LG  CM  KC  CO  RE  Gene Name 
08472  1               Retinoic acid-inducible E3 protein  
08492  1               Fetal brain cDNA 5′-end  
08529  1               Bcl-2 related (Bfl-1) mRNA  
08558  1               Anonymous mRNA  
08600  1               ftp-3 
08627  1               Fragile × mental retardation protein (FMR-1)  
08696  1               Transcript ch138  
12544  1               Sp17 gene  
Total  90  39  23  25  16  6  13 7  20  23  12  16  19  17  16  
GS PM GR HL MO HE FL ILAL AP VF LG CM KC CO RE Gene Name
A. Energy production  
00943 1  1  3             Phosphoglycerate kinase 1  
02200  1      1        1   Transaldolase 
Total  2  1  3    1        1    
B. Miscellaneous  
00402  19   1  1  2     1  1   1  2  Spermidine/spermine N1-acetyltransferase  
02161  4     1           AMP deaminase isoform L (AMPD2)  
00304  3  2  1  2 2  1   1  3  4  1  3   3  Ubiquitin  
03216  3        1       Long-chain acyl-CoA synthetase 
01202  2  1              Neutrophil oxidase factor (NCF2)/p67-phox  
00483  2     1           IMP dehydrogenase type 1  
03596  2               Spermidine/spermine N1-acetyltransferase  
07377  2         1       Inosine monophosphate dehydrogenase type II  
08337  2               Grancalcin 
08513  2               Proteasome subunit p40 (Mov34)  
01263  1  2              Vacuolar H+-ATPase Mr 56,000 subunit (HO57)  
00253  1  2  1  2   1      2     Importin beta subunit  
01328  1  1              Calpastatin=glycoprotein BS-17 component  
02147  1      1    2       Protein disulfide isomerase related protein (ERp72)  
00154  1     1         1   Glutaredoxin 
00533  1   1       2     3 6  1  Ferritin H chain  
01766  1    6  15  2  4   1  2  5  10   1 Metallothionein from cadmium-treated cells  
07546  1               Type 1 inositol 1,4,5-trisphosphate receptor  
08483  1               Ubiquitin conjugating enzyme (UBC4)  
08328  1               Ribonuclease A  
08418  1               Erythrocyte-type AMP deaminase  
08504  1               Lysyl hydroxylase (PLOD)  
08643  1               Histidase 
08690  1               Ferritin H chain  
Total  55  8  3 9  7  20  3  5  6  9  4  10  14  12 2   
C. Lysosomal 
01809  5    3         6  1   Lysosomal membrane glycoprotein CD63  
01106 2  4   3            Lysozyme  
01533 1  1   2       2  3  1   1 1  Lysosomal membrane glycoprotein-1  
08663  1               Arylsulphatase A  
08679  1               26S protease (S4) regulatory subunit  
08396  1               Giantin/golgi antigen gcp372  
08464  1               Iduronate sulphate sulphatase (IDS)  
08664  1               Iduronate sulphate sulphatase (IDS)  
Total 13  5   8       2  3  7   2 1   
D. Signal transduction  
02821  15         1  2  2   1   CL 100 protein tyrosine phosphatase  
01724  6    2            Pleckstrin (P47) 
00816  5  5  1  1        1     Lymphocyte-specific protein 1 (LSP1)  
01919 3    5    1   5  6  3  2  5 19   Calcyclin  
00651  2  1  4          1    Rho GDP-dissociation Inhibitor 2  
01089  2  1              Tyrosine kinase (HCK)  
07614  2               Cyclin G2  
01524  1  1     1          TSE1=protein kinase A regulatory subunit  
00124  1   1   2     1     1   Ras-related protein (Krev-1)  
00286  1     2           ERK activator kinase (MEK1)  
00637  1   1             56K autoantigen annexin XI  
01687  1    1            c-raf-1 proto-oncogene  
02144  1      1   1        Protein kinase C ζ  
03705 1               α-subunit of Gi2  
04134  1           1    Rapamycin- and FK506-binding protein  
06319 1              1  Protein kinase  
08680  1               5-lipoxygenase 
Total  45  8  7  9  4 2  1  1  5  8  6  5  6  22    
E. Ribosomal component and translation factor  
01325  3  1         1  1   2  1  sui1iso1  
00314  2  1  3 3  1   1  1  1  4  5    3  2 Ribosomal protein S11  
00919  2  1  1    1          Splicing factor (CC1.4)  
00305  2   6  2  1  2  3  1 1  2  2  2  4  1   Ribosomal protein L28 
00995  1  3   1   1   1  1   1 2   1  1  Eukaryotic initiation factor 4AII 
00650  1  2  14  5    2   1  2  1 6  1  2  2  Ribosomal protein S18  
00418  1 1  22  10  5  1  5  1  10  1    7  9  Ribosomal protein L9  
00795  1 1  1  2    1  2  5  3     6 3  Ribosomal protein L30  
00818  1  1  4  1   2  1     2    2  Ribosomal protein L5  
00889  1   1    1  1  1   1   2    EF-1 δ  
00356  1  16  2  1  2  1  2   1    1   L23 putative ribosomal protein  
03424  1            1   Translational initiation factor 2 β subunit 
Total  17  11  68  26  8  7  17  10  19 14  10  13  9  22  20   
GS  PM  GR  HL  MO  HE  FL  IL AL  AP  VF  LG  CM  KC  CO  RE  Gene Name 
F. Cytoskeltal  
00155  6 3   8  1   3   2   2  4  1  4  Thymosin β-4  
00244  4  14   1  2     1   3  4  1  1  β-actin 
00685  3  5  2  4     1        L-plastin  
01877  3   1        1   1   Paxillin  
04080  2           1  1   Cofilin  
01030  1  2   1     2   1      Myosin regulatory light chain  
01507  1  1              α-actinin 
00258  1     1    1   1   1 1    2.5-kb cytoskeletal tropomyosin TM30 (nm) 
01702  1    1            Alkali myosin light chain  
01726  1    2       1      Striated muscle contraction regulatory protein  
02833  1         1  1   1    Nonmuscle myosin heavy chain (NMHC)  
Total  24  25  2 18  4   3  2  5  3  5  9  9  5  1  
G. Mitochondrial 
00994  5  2   1      1     1  1  Glutamine synthetase  
00313 2  1    1  1      1     SOD-2 manganese superoxide dismutase  
00360 1  1  2   2     1   1    1  Mitochondrial phosphate carrier protein  
00934  1 1  3  1    1  3  1  5      2 coxVIb  
08345  1               ATP synthase c subunit  
08434  1               Manganese superoxide dismutase  
08667  1               Glutamine synthetase  
01995  1    3    1   1 1   1   4   Glutathione peroxidase  
08536 1                SOD-2 gene for manganese superoxide dismutase  
04986  1               Catalase 
Total  15  5  5  5  3 1  2  3  4  6  2  1   6  3   
H. Nuclear  
01811  8   1         2    1  H3.3 histone  
01367  7  3   1      1 1  3     1  hnRNP-E1  
07795  5         1       B-cell lymphoma 3-encoded protein (bc1-3)  
08383  5               Zinc-finger protein (bc1-6)  
05014  4               H3.3 histone  
03575  3           1  2    H3.3 histone  
04074  3           1   1  Transcription factor ETR101  
08375  3               MAD-3 
08389  3               E4BP4 gene  
01431  2  1         3      jun-D  
01719  2    1            Myeloid cell nuclear differentiation antigen  
06259  2               spi-1 proto-oncogene  
08044  2             1   Zinc finger transcriptional regulator  
01566  1  1     1    1     1   fus 
00827  1  1  1     2        2  dUTP pyrophosphatase (hdut) 
08621  1               hnRNP A2/hnRNP B1  
04003  1           1     Receptor of retinoic acid  
01772  1    1            Interferon-γ induced protein (IFI 16)  
01912  1    1     1        CDEI binding protein/amyloid protein homologue  
02730  1          2    2   fos proto-oncogene  
00487  1     1       1     Transformation upregulated nuclear protein  
00648  1   1          2   1  Proliferating cell nucleolar protein P120  
01803  1    1            BAT1 nuclear RNA helicase (DEAD family)  
01832  1    1      1  2  1  1    jun-B 
03413  1               tpr 
06299  1               Arginine-rich nuclear protein  
08013  1             1  Replication protein A 70 kD subunit  
08366  1               Pre-mRNA splicing factor SR p75  
08411  1               Transcription factor SUPT4H  
08454  1               Nuclear factor erythroid 2 isoform f  
08546  1               Transcription factor  
08647  1               Putative RNA helicase HRH1  
Total  68 6  2  7  1   3  1  1  7  7  7  5 6  5   
I. Secretory and granular  
01823  18    2            Pre-B cell enhancing factor (PBEF)  
08339  9               IL-8 
02015  5    11            LD78 α/macrophage inflammatory protein  
08336  3               Secretory granule proteoglycan peptide  
08638  2               N-formylpeptide receptor (fMLP-R26)  
01820  2    1            Antisecretory factor-1  
00592  1  1  1  1    1       1   ATL-derived factor/thioredoxin  
01616  1  1   3   1         1  Cystatin B  
08342 1               Neutrophil peptide/defensin1/HP-1  
08553  1               Cathepsin S  
08318  1               N-formylpeptide receptor (fMLP-R98)  
00111  1    1  7  9 13  6         α 1-antitrypsin 
02007  1    1      1     1  1  Globin  
08501  1               Metalloproteinase-2 inhibitor (TIMP-2)  
08530  1               Platelet-derived endothelial cell growth factor  
08644  1               Platelet-derived endothelial cell growth factor  
08674  1               Monocyte/neutrophil elastase inhibitor  
Total  50  2  1  20  7 10  14  6  1      2  2   
GS  PM  GR  HL  MO  HE  FL  IL AL  AP  VF  LG  CM  KC  CO  RE  Gene Name 
J. Surface membrane  
00565 24  3  4  5    1  1  1   1  4 2   1  β-2-microglobulin  
00254  18   2  1    1  1  1  2    3  MHC class I HLA-Cw1  
00937  14  4  1       5  11   1    HLA-E heavy chain  
08362  13               Granulocyte colony-stimulating factor receptor  
08438  4               Nramp 
01187  3  2   1        1     Monocyte activation antigen (Mo3)  
01585 3  2   3            Tumor necrosis factor receptor  
01304  3  1   2       1      Transmembrane glycoprotein (CD53) 
01594  3  1              Leukocyte common antigen T200 (CD45)  
04103  3           1     HLA-B27 antigen  
05103  3               ICAM-3 
01624  2  2              Aminopeptidase N/CD13 encoding aminopeptidase N  
00303  2   2  2    1      1    (chromosome 3p25) membrane protein  
08355  2               C5a anaphylatoxin receptor  
08594  2               TAP1 
01048  1  1              MHC antigen (HLA-B)  
01395  1  1   1   2      1     Transmembrane receptor protein  
01229  1  1              Migration inhibitory factor-related protein 8  
00474  1    1           Transmembrane carcinoembryonic antigen BGPb  
02079  1     1          Palmitoylated erythrocyte membrane protein (MPP1)  
02156  1      1     1      CD14 myelid cell-specific leucine-rich glycoprotein  
06689  1           1     Leukocyte antigen CD97  
08349  1               Leukocyte adhesion molecule-1 (LAM-1)  
08524  1               Interferon-γ receptor  
08563  1               Low-affinity IgG Fc receptor  
08564  1               Complement receptor type 1  
08592  1               Common acute lymphoblastic leukemia antigen (CALLA)  
08661  1               Putative voltage-gated potassium channel (KVLQT1)  
08678  1               MHC class I-related protein  
08686  1               IL-8 receptor β (IL8RB)  
Total 114  18  7  14  4  4  1  3  2  7  16 7  4  3  1   
K. Unknown Function  
03341 11         1  2      B94 protein  
01267  8  1              1-8D 
04345  6               B4-2 protein  
01087  3  6        2  1      Brain-expressed HHCPA78 homolog  
00273 3  4  6  11  9  2  11  2  12  2  1 5  3  3  5  Translationally controlled tumor protein  
01228  3  4       1    1     Anonymous mRNA  
00729  3  2           1   CLP  
01288  2  1              Bcl-2 related (Bfl-1) mRNA  
00080  2     1           Myeloid cell differentiation protein (MCL1)  
00600  2   2        1     1  CIRP 
01926  2    2       1  1   1  1  ras-related C3 botulinum toxin substrate (rac) mRNA  
08056  2             1  Helix-loop-helix basic phosphoprotein (GOS8)  
08235 2              1  ERF-1  
08444  2               GLUDP2 
00543  1  4  9  5    2  1  2  2  2  2  2  3  23-kD highly basic protein 
01224  1  4          1     KIAA0184 protein  
01609  1  4      1    2  1    γ-interferon-inducible protein (IP-30)  
00185 1  2  1  1  2        1  2   Putative p64 CLCP protein  
01032  1  2    1     1    4  2   polyA binding protein  
01608  1  2       1  2    1   5 APPH-amyloid precursor protein homolog  
01665  1  2              KIAAO168 protein  
00751  1  1  1             (clone SAA7C) sequence  
00936  1  1  2        2 1      MLN51  
01000  1  1              EVI2B3P 
00129  1    1  1      1  1   2   fau  
00241  1    2  1    1     1  1   Acute myeloid leukemia associated protein (AML1/EAP)  
00363  1    1  1   1   1  1  2  2  1 1  ORF (complete cds) and HepG2 identical sequence 
00434  1    2  1  1       1 1  1   MLN50  
02011  1    1      1       SH3 domain-containing proline-rich kinase (sprk)  
02135  1      1      1     KIAA0183 protein 
02443  1        1        HVLCAD gene  
02662  1          1      Anonymous mRNA  
02820  1         1  3  1  1    1-8U  
02835  1          1   1   Transformer-2 α (htra-2 α) mRNA  
03864  1           1     (clone 05) liver expressed protein fragment  
04100  1           1     Anonymous mRNA  
05241  1          1    1   ZFM1 protein  
06150  1         1       OS-9 precursor mRNA  
07841  1         2      Brain-expressed HHCPA78 homolog  
08011 1              1  Phorbolin I  
08353  1               H sapiens (clone pS6) Alu repeat  
08372  1               DNA sequence from cosmid U61B11  
08393  1               fus-chop fusion protein  
08400  1               MG44 
08410  1               Putative 32-kD heart protein PHP32 mRNA 
GS  PM  GR  HL  MO  HE  FL  IL AL  AP  VF  LG  CM  KC  CO  RE  Gene Name 
08472  1               Retinoic acid-inducible E3 protein  
08492  1               Fetal brain cDNA 5′-end  
08529  1               Bcl-2 related (Bfl-1) mRNA  
08558  1               Anonymous mRNA  
08600  1               ftp-3 
08627  1               Fragile × mental retardation protein (FMR-1)  
08696  1               Transcript ch138  
12544  1               Sp17 gene  
Total  90  39  23  25  16  6  13 7  20  23  12  16  19  17  16  

Two hundred sixteen gene species matched to known genes were categorized according to their functions and subcellular localizations. Numbers represent frequency of appearance of the gene transcripts among 1,000 cDNA analyzed. Abundances of the GSs that appeared in other libraries are also shown for comparison’s sake. For some cells, see legend to Table 1. The bottom line (total) shows the total count of the collection. Numbers of the expressed genes shown in columns PM through RE are from about 1,000 mRNA analyzed in each library.

Abbreviation: PM, peripheral granulocytes.

View Large
Comparison of gene activities in granulocytes and DMSO-induced granulocytoids.

An expression profile of the granulocytoid cells is represented in Table 1B (column GR). In the same table are collectively displayed the relevant gene activities with HL60 cells (HL) and the monocytoids derived from HL60 by tetradecanoylphorbol-13 acetate (TPA)10 treatment (MO). Comparison of Table 1B with Table1A or column PM versus GR in Table 1A with column GR versus PM in Table1B shows that about 50% (24/48) of the abundantly expressed genes in the DMSO-induced HL60 are also present in peripheral granulocytes, although the abundances differ. Scarcity of highly abundant transcripts is characteristic of the mRNA population in granulocytoid cells. Generally, genes for cytoskeleton and protein synthesis machinery are moderately active, unlike genes for cell surface membrane components in granulocytoid cells. Further discussions will be presented in the Discussion.

Identification of granulocyte-specific genes.

We have prepared expression profiles of active genes in 11 other human cells/tissues.3,10,37 Genes listed in Table 1A and B were extracted from each of these profiles, and their activities (abundance of the transcripts among 1,000 mRNA molecules) were compiled. The resulting Table 1A, although incomplete, allows us to categorize genes into those whose expression is peripheral granulocyte-specific, limited to certain types of cells, or ubiquitous. When the genes have been detected in 6 libraries or more, we categorized them as ubiquitous (solid area in column “lib”). Genes known to perform house-keeping functions, such as ubiquitin or ribosomal proteins, are seen in this category. A gene expressed only in granulocytes or in granulocytes and/or granulocytoid cells may be categorized as unique to this type of cell (open area). The rest were categorized as common or intermediate (hatched area). We categorized 22 GSs as unique, among which 12 were identified in GenBank and 10 represent novel genes. Among the 12 known genes are granulocyte colony-stimulating factor receptor, interleukin-8 (IL-8), leukocyte common antigen T200 (CD45), and ICAM-3, which have been known to act mainly in granulocytes. No data have been reported so far as to the cell type specificity of the remaining 8 genes. Thus, at least one third of the genes in the unique category were indeed those that represent specific functions of the granulocytes. We argue that we can extrapolate this finding to the novel genes.

dbEST and the granulocyte GS.

As a result of a rapid expansion in the collection of expressed sequence tags (ESTs), more than 400,000 fragmentary human cDNA sequences from more than 20 tissues have been collected in dbEST. This database can be readily compared with the expression profiles as described here, because the quality of the source cells for this library construction has not been biased and because the cDNA libraries that have been subjected to normalizing protocol do not reflect the composition of the mRNA population in the original source cells/tissues. Nevertheless, they can provide some information as to what RNA species are present in tissues so far examined. We queried the 22 GS sequences that were categorized as unique with dbEST. The results, shown in Table 3, demonstrate that 5 of them, GS08339 (IL-8), GS05242, GS01594 (leukocyte common antigen T200), GS08389 (E4BP4 gene), and GS08424 have not been registered in dbEST. Considering that no granulocytes or related tissues were used for the EST collection, it is not surprising that these sequences failed to appear in dbEST. From an opposite point of view, the absence in dbEST strengthens the idea that such genes are unique to the granulocytes. Five GSs, GS08362 (granulocyte colony-stimulating factor receptor), GS08347, GS08438 (Nramp), GS08336 (secretory granule proteoglycan peptide), and GS08375 (MAD-3) matched ESTs from fetal liver spleen, placenta, lung, and other tissues. This observation shows the limit of the application of dbEST for the categorization under discussion: it may simply show that tissues used in the dbEST data construction contained some granulocytes. As with novel genes, one GS (GS08347) is highly likely to represent a gene that is unique to granulocytes. The other 9 genes were subjected to further examination, because they were found recurrently in tissues not related to granulocytes in the dbEST.

Table 3.

Representation of Peripheral Granulocyte GS in dbEST

GS PM GR EST Lib Fetal Liver SpleenFetal Spleen Ovary Breast Placenta LiverLung Infant Brain Adult Brain Retina OthersGene Name
08339  9               IL-8 
05242  4  
01594  3  1              Leukocyte common antigen T200 (CD45)  
08389  3               E4BP4 gene  
08424  3  
08362  13   3  1     3        Granulocyte colony-stimulating factor receptor  
08347  8   2  1 2  
08438  4   1  1      1       Nramp  
08336  3   1  1           1  Secretory granule proteoglycan peptide  
08375  3   2  1       2      MAD-3  
01267 8  1  31  13  1  2  1  5  10  5  1  1  1  4  1-8D  
04345  6   7  6 1  2   1  1      1  1  B4-2 protein  
05209  6   14  7   1   3  2   5  2   1  
08383  5   5  3  1       3    1  Zinc-finger protein (bcl-6)  
04290  5   9  3      4      5  
01371  4  1  3  2    1     2  
03554  4   2  2        1  1  
08325  4   6  4 2    2     1    1  
08395  4  10  6  3     2   1     4 
05014  4   11  7  4  1    2   2  1   1  Histone H3.3  
05103  3  2  2  1    1        ICAM-3  
08548  3   6 4  1   1      1    3  
GS PM GR EST Lib Fetal Liver SpleenFetal Spleen Ovary Breast Placenta LiverLung Infant Brain Adult Brain Retina OthersGene Name
08339  9               IL-8 
05242  4  
01594  3  1              Leukocyte common antigen T200 (CD45)  
08389  3               E4BP4 gene  
08424  3  
08362  13   3  1     3        Granulocyte colony-stimulating factor receptor  
08347  8   2  1 2  
08438  4   1  1      1       Nramp  
08336  3   1  1           1  Secretory granule proteoglycan peptide  
08375  3   2  1       2      MAD-3  
01267 8  1  31  13  1  2  1  5  10  5  1  1  1  4  1-8D  
04345  6   7  6 1  2   1  1      1  1  B4-2 protein  
05209  6   14  7   1   3  2   5  2   1  
08383  5   5  3  1       3    1  Zinc-finger protein (bcl-6)  
04290  5   9  3      4      5  
01371  4  1  3  2    1     2  
03554  4   2  2        1  1  
08325  4   6  4 2    2     1    1  
08395  4  10  6  3     2   1     4 
05014  4   11  7  4  1    2   2  1   1  Histone H3.3  
05103  3  2  2  1    1        ICAM-3  
08548  3   6 4  1   1      1    3  

The 22 GSs categorized as unique (see text) are regrouped according to their presence or absence in dbEST. BLAST N program was used for this purpose. Abundance values for our libraries are listed in the columns PM and GR. ESTs matched to these GSs are listed with their frequency of appearance and the source tissues. The tissue names show the source organ from which ESTs were collected. Notice that recurrence of EST reflects an abundance of mRNA, but incompleteness of the cDNA library normalization.

Abbreviations: PM, peripheral granulocytes; GR, the granulocytoids; EST, dbEST; Lib, libraries from dbEST.

View Large
The cell physiology reflected in the expression profile of active genes.

Granulocytes are a major player in the defense of the body against foreign materials. About 90% to 95% of granulocytes are neutrophils, with the remainder being eosinophils and basophils in circulating human blood. Hence, peripheral granulocytes represent the activities of neutrophils.1 The cytoplasm of these cells has highly developed cytomatrixes, as well as granules that contain microbicidal proteins and digestive enzymes. The plasma membranes carry a number of receptors and other structures needed for recognition and disposal of invading pathogens.2 

Although gene activities are not necessarily reflected by the abundance of mRNA, other methods being not available (except for quantitizing two-dimensionally separated protein bands), gene expression profiling3 leads to the best approximation. Table 1A shows several genes well studied in conjunction with the functions in granulocytes. The quantitative ratios should help us understand the regulatory systems acting in the granulocytes. An abundant expression of genes for cell surface membrane proteins drew our attention; eg, genes for β2-microglobulin, MHC class I HLA-Cw, and HLA-E heavy chain, which are components of cell surface receptors. That a lot of genes for cell surface membrane proteins are active in granulocytes supports the notion that granulocyte responses can be evoked by a variety of stimuli caused by particulate and soluble materials. On the other hand, most of the genes for cytoskeleton were not so active, except those for thymosin β4 and β-actin, in accord with the notion that these cells do not maintain a rigid shape.

The list has shown several genes not known to be active in granulocytes. This study points out the importance of elucidating the role of gene products in granulocytes. B94 protein and B4-2 protein are good examples. Expected changes in the expression of genes in association with inflammation or changes in adhesive properties during chemotaxis and phagocytosis remain to be examined. Fibronectins, β2-integrins, and the L-selectins, which are notably associated with adhesiveness as mediators,2 were not detected in our expression profile. Actins, which play important roles in production of pseudopodium for locomotion, were not expressed strongly. Activation of these genes is yet another feature of the activation of granulocytes worth investigating. Genes for chemotactic factor receptors were moderately expressed, including tumor necrosis factor receptor, N-formylpeptide receptor, C5a anaphylatoxin receptor, and IL-8 receptor. On the other hand, expression of receptors for C3b and C3bi were not detected, in line with the fact that our granulocytes were in a resting stage.38 Here again, examination of the profile in induced cells, including the time course of activation and their relative order of activities, is of utmost interest.

Genes for secretory proteins, such as cytokines, are not particularly active in circulating granulocytes and, indeed, only 2 genes, for pre-B–cell colony-enhancing factor and IL-8, were detected. Thus, the relative activities of these important granulocyte-specific genes in resting cells have been determined. As with secretable bactericidal components, there were cathepsin S, neutrophil oxidase factor (NCF2)/p67-phox, proteasome subunit p40, and defensin, in addition to lysosomal proteins. Thus, these proteins are constitutively produced at a level of 4.0% or more of total protein synthesis. Bactericidal/permeability-increasing protein (BPI) was found in granulocytoid cells, but not in the peripheral granulocytes, probably because its expression level is just at the border of the level of detection.

Whereas active expressions of genes for cell surface membrane proteins, including receptors for chemotactic factors as well as genes for bactericidal proteins such as lysosomes, are characteristic to our granulocytes, so is poor expression of components for protein synthesis machinery, as it is for cells in the resting stage.

Among the mRNAs in granulocytes identified in GenBank (Table 1A) are genes for granulocyte colony-stimulating factor receptor, tumor necrosis factor receptor, and T200. These gene products are related to neutrophilic granulopoiesis and their maturation. Thus, these findings strongly suggest that granulocytes in circulating blood wait for stimuli exposing granulopoietic receptors. The list also included IL-8, a neutrophilic chemoattractant and activator. In contrast to IL-8 receptor (Table 2J), IL-8 is highly and specifically expressed. This mRNA has been known to be induced in neutrophils in response to granulocyte/macrophage colony-stimulating factor.31,39Although granulocyte colony-stimulating factor regulates the expression of IL-8 receptor,40 whether granulocyte colony-stimulating factor can induce the expression of IL-8 is not clear. Our results indicate that granulocyte colony-stimulating factor induces the expression of IL-8 mRNA.

The uniquely active genes in granulocytes.

Among the 22 genes categorized as unique in Table 1A, 10 were novel genes. It is of utmost importance to characterize these genes, although such categorization can be performed only with abundantly expressed genes, and yet some misleading categorizations are unavoidable due to the limited number of GS collections.

Comparison of expression profiles between granulocytes and granulocytoid cells induced from HL60.

β2-microglobulin and HLA-E heavy chain were commonly expressed in granulocytes and granulocytoid cells. Proteins in lysosomes, BPI, and leukocyte adhesion protein (Mac-1), which are known to be expressed in neutrophils, were also expressed in granulocytoid cells. In total, 20% or more of the mRNAs are commonly expressed in both types of cells (data not shown).

However, their relative proportions in granulocytoids differ from those in granulocytes. In general terms, expression profiles of genes in granulocytes and granulocytoid cells differ from each other qualitatively and quantitatively. In Table 1A, 24 gene species represented by 74 clones were commonly expressed in granulocytes and granulocytoid cells. These observations demonstrate the capacity and limitations of this model system.

Address reprint requests to Kenichi Matsubara, PhD, Nara Institute of Science and Technology 8916-5, Takayama, Ikoma, Nara 630-01, Japan; e-mail: kenichi@bs.asist-nara.ac.jp.

The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" is accordance with 18 U.S.C. section 1734 solely to indicate this fact.

1
Fawcett DW: A Text Book of Histology (ed 12). New York, NY, Chapman & Hall, 1994
2
Beutler E, Lichtman MA, Coller BS, Kipps TJ: Hematology (ed 5). New York, NY, McGraw-Hill, 1995
3
Okubo
K
Hori
N
Matoba
R
Niyama
T
Fukushim
A
Kojima
Y
Matsubara
K
Large scale cDNA sequencing for analysis of quantitative and qualitative aspects of gene expression.
Nat Genet
2
1992
173
Google Scholar
Crossref
Search ADS
PubMed
 
4
Okubo
K
Hori
N
Matoba
R
Niyama
T
Matsubara
K
A novel system for large-scale sequencing of cDNA by PCR amplification.
DNA Seq
2
1991
137
Google Scholar
Crossref
Search ADS
PubMed
 
5
Matsubara
K
Okubo
K
cDNA analysis in the human genome project.
Gene
135
1993
265
Google Scholar
Crossref
Search ADS
PubMed
 
6
Collins
SJ
Gallo
RC
Gallagher
RE
Continuous growth and differentiation of human myeloid leukaemic cells in suspension culture.
Nature
270
1977
347
Google Scholar
Crossref
Search ADS
PubMed
 
7
Collins
SJ
Ruscetti
FW
Gallagher
RE
Gallo
RC
Terminal differentiation of human promyelocytic leukemia cells induced by dimethyl sulfoxide and other polar compounds.
Proc Natl Acad Sci USA
75
1978
2458
Google Scholar
Crossref
Search ADS
PubMed
 
8
Collins
SJ
Ruscetti
FW
Gallagher
RE
Gallo
RC
Normal functional characteristics of cultured human promyelocytic leukemia cells (HL60) after induction of differentiation by dimethylsulfoxide.
J Exp Med
149
1979
969
Google Scholar
Crossref
Search ADS
PubMed
 
9
Harris
P
Ralph
P
Human leukemic models of myelomonocytic development: A review of the HL-60 and U937 cell lines.
J Leukoc Biol
37
1985
407
Google Scholar
Crossref
Search ADS
PubMed
 
10
Okubo
K
Itoh
K
Fukushima
A
Yoshii
J
Matsubara
K
Monitoring cell physiology by expression profiles and discovering cell type-specific genes by compiled expression profiles.
Genomics
30
1995
178
Google Scholar
Crossref
Search ADS
PubMed
 
11
Ficoll-Paque and Ficoll-Paque ET. Uppsala, Sweden, Pharmacia Biotech, 1994
12
Boyum
A
Isolation of mononuclear cells and granulocytes from blood.
Scand J Clin Lab Invest Suppl
97
1968
77
Google Scholar
PubMed
 
13
Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor, NY, Cold Spring Harbor Laboratory, 1989
14
Pearson
WR
Lipman
DJ
Improved tools for biological sequence comparison.
Proc Natl Acad Sci USA
85
1988
2444
Google Scholar
Crossref
Search ADS
PubMed
 
15
Bjerrum
OW
Bjerrum
OJ
Borregaard
N
Beta 2-microglobulin in neutrophils: An intragranular protein.
J Immunol
138
1987
3913
Google Scholar
PubMed
 
16
Fukunaga
R
Seto
Y
Mizushima
S
Nagata
S
Three different mRNAs encoding human granulocyte colony-stimulating factor receptor.
Proc Natl Acad Sci USA
87
1990
8702
Google Scholar
Crossref
Search ADS
PubMed
 
17
Fukunaga
R
Ishizaka-Ikeda
E
Nagata
S
Growth and differentiation signals mediated by different regions in the cytoplasmic domain of granulocyte colony-stimulating factor receptor.
Cell
74
1993
1079
Google Scholar
Crossref
Search ADS
PubMed
 
18
Jubinsky
PT
Laurie
AS
Nathan
DG
Yetz-Aldepe
J
Sieff
CA
Expression and function of the human granulocyte-macrophage colony-stimulating factor receptor alpha subunit.
Blood
84
1994
4174
Google Scholar
Crossref
Search ADS
PubMed
 
19
Ito
Y
Seto
Y
Brannan
CI
Copeland
NG
Jenkins
NA
Fukunaga
R
Nagata
S
Structural analysis of the functional gene and pseudogene encoding the murine granulocyte colony-stimulating-factor receptor.
Eur J Biochem
220
1994
881
Google Scholar
Crossref
Search ADS
PubMed
 
20
Shieh
JH
Gordon
M
Jakubowski
A
Peterson
RH
Gabrilove
JL
Moore
MA
Interleukin-1 modulation of cytokine receptors on human neutrophils: In vitro and in vivo studies.
Blood
81
1993
1745
Google Scholar
Crossref
Search ADS
PubMed
 
21
Kuijpers
TW
Tool
AT
van-der-Schoot
CE
Ginsel
LA
Onderwater
JJ
Roos
D
Verhoeven
AJ
Membrane surface antigen expression on neutrophils: A reappraisal of the use of surface markers for neutrophil activation.
Blood
78
1991
1105
Google Scholar
Crossref
Search ADS
PubMed
 
22
Parolini
I
Sargiacomo
M
Lisanti
MP
Peschle
C
Signal transduction and glycophosphatidylinositol-linked proteins (lyn, lck, CD4, CD45, G proteins, and CD55) selectively localize in Triton-insoluble plasma membrane domains of human leukemic cell lines and normal granulocytes.
Blood
87
1996
3783
Google Scholar
Crossref
Search ADS
PubMed
 
23
Diaz-Gonzalez
F
Gonzalez-Alvaro
I
Campanero
MR
Mollinedo
F
del Pozo
MA
Munoz
C
Pivel
JP
Sanchez-Madrid
F
Prevention of in vitro neutrophil-endothelial attachment through shedding of L-selectin by nonsteroidal antiinflammatory drugs.
J Clin Invest
95
1995
1756
Google Scholar
Crossref
Search ADS
PubMed
 
24
de Fougerolles
AR
Diamond
MS
Springer
TA
Heterogenous glycosylation of ICAM-3 and lack of interaction with Mac-1 and p150,95.
Eur J Immunol
25
1995
1008
Google Scholar
Crossref
Search ADS
PubMed
 
25
Briggs
RC
Briggs
JA
Ozer
J
Sealy
L
Dworkin
LL
Kingsmore
SF
Seldin
MF
Kaur
GP
Athwal
RS
Dessypris
EN
The human myeloid cell nuclear differentiation antigen gene is one of at least two related interferon-inducible genes located on chromosome 1q that are expressed specifically in hematopoietic cells.
Blood
83
1994
2153
Google Scholar
Crossref
Search ADS
PubMed
 
26
Buhl
AM
Osawa
S
Johnson
GL
Mitogen-activated protein kinase activation requires two signal inputs from the human anaphylatoxin C5a receptor.
J Biol Chem
270
1995
19828
Google Scholar
Crossref
Search ADS
PubMed
 
27
Durstin
M
Gao
JL
Tiffany
HL
McDermott
D
Murphy
PM
Differential expression of members of the N-formylpeptide receptor gene cluster in human phagocytes.
Biochem Biophys Res Commun
201
1994
174
Google Scholar
Crossref
Search ADS
PubMed
 
28
Furie
MB
Burns
MJ
Tancinco
MC
Benjamin
CD
Lobb
RR
E-selectin (endothelial-leukocyte adhesion molecule-1) is not required for the migration of neutrophils across IL-1-stimulated endothelium in vitro.
J Immunol
148
1992
2395
Google Scholar
PubMed
 
29
Prado
GN
Suzuki
H
Wilkinson
N
Cousins
B
Navarro
J
Role of the C terminus of the interleukin 8 receptor in signal transduction and internalization.
J Biol Chem
271
1996
19186
Google Scholar
Crossref
Search ADS
PubMed
 
30
Mollinedo
F
Vaquerizo
MJ
Naranjo
JR
Expression of c-jun, jun B and jun D proto-oncogenes in human peripheral-blood granulocytes.
Biochem J
273
1991
477
Google Scholar
Crossref
Search ADS
PubMed
 
31
Takahashi
GW
Andrews
DF
3d
Lilly
MB
Singer
JW
Alderson
MR
Effect of granulocyte-macrophage colony-stimulating factor and interleukin-3 on interleukin-8 production by human neutrophils and monocytes.
Blood
81
1993
357
Google Scholar
PubMed
 
32
Avalos
BR
Bartynski
KJ
Elder
PJ
Kotur
MS
Burton
WG
Wilkie
NM
The active monomeric form of macrophage inflammatory protein-1 alpha interacts with high- and low-affinity classes of receptors on human hematopoietic cells.
Blood
84
1994
1790
Google Scholar
Crossref
Search ADS
PubMed
 
33
Remold-O’Donnell
E
Chin
J
Alberts
M
Sequence and molecular characterization of human monocyte/neutrophil elastase inhibitor.
Proc Natl Acad Sci USA
89
1992
5635
Google Scholar
Crossref
Search ADS
PubMed
 
34
Ziegler
SF
Marth
JD
Lewis
DB
Perlmutter
RM
Novel protein-tyrosine kinase gene (hck) preferentially expressed in cells of hematopoietic origin.
Mol Cell Biol
7
1987
2276
Google Scholar
Crossref
Search ADS
PubMed
 
35
Grinstein
S
Butler
JR
Furuya
W
L’Allemain
G
Downey
GP
Chemotactic peptides induce phosphorylation and activation of MEK-1 in human neutrophils.
J Biol Chem
269
1994
19313
Google Scholar
PubMed
 
36
Sariban
E
Mitchell
T
Kufe
D
Expression of the c-raf protooncogene in human hematopoietic cells and cell lines.
Blood
69
1987
1437
Google Scholar
Crossref
Search ADS
PubMed
 
37
Nishida
K
Adachi
W
Shimizu-Matsumoto
A
Kinoshita
S
Mizuno
K
Matsubara
K
Okubo
K
A gene expression profile of human corneal epithelium and the isolation of human keratin 12 cDNA.
Invest Ophthalmol Vis Sci
37
1996
1800
Google Scholar
PubMed
 
38
Berger
M
O’Shea
J
Cross
AS
Folks
TM
Chused
TM
Brown
EJ
Frank
MM
Human neutrophils increase expression of C3bi as well as C3b receptors upon activation.
J Clin Invest
74
1984
1566
Google Scholar
Crossref
Search ADS
PubMed
 
39
van Pelt
LJ
Huisman
MV
Weening
RS
von dem Borne
AE
Roos
D
van Oers
RH
A single dose of granulocyte-macrophage colony-stimulating factor induces systemic interleukin-8 release and neutrophil activation in healthy volunteers.
Blood
87
1996
5305
Google Scholar
Crossref
Search ADS
PubMed
 
40
Lloyd
AR
Biragyn
A
Johnston
JA
Taub
DD
Xu
L
Michiel
D
Sprenger
H
Oppenheim
JJ
Kelvin
DJ
Granulocyte-colony stimulating factor and lipopolysaccharide regulate the expression of interleukin 8 receptors on polymorphonuclear leukocytes.
J Biol Chem
270
1995
28188
Google Scholar
Crossref
Search ADS
PubMed
 
Copyright © 1998 American Society of Hematology
1998
Sign in via your Institution