Decreased levels of CXC-chemokines in serum of benzene-exposed workers identified by array-based proteomics.

Benzene is an important industrial chemical and environmental contaminant that causes leukemia. To obtain mechanistic insight into benzene's mechanism of action, we examined the impact of benzene on the human serum proteome in a study of exposed healthy shoe-factory workers and unexposed controls. Two sequential studies were performed, each using sera from 10 workers exposed to benzene (overall mean benzene air level >30 ppm) and 10 controls. Serum samples were subjected to anion-exchange fractionation and bound to three types of ProteinChip arrays (Ciphergen Biosystems, Fremont, CA) [hydrophobic (H50), metal affinity (IMAC3-Cu), and cation exchange (WCX2)]. Protein-expression patterns were detected by surface-enhanced laser desorption/ionization (SELDI)-TOF MS. Three proteins (4.1, 7.7, and 9.3 kDa) were consistently down-regulated in exposed compared with control subjects in both studies. All proteins were highly inversely correlated with individual estimates of benzene exposure (r > 0.75). The 7.7- and 9.3-kDa proteins were subsequently identified as platelet factor (PF)4 and connective tissue activating peptide (CTAP)-III. Initial proteomic results for PF4 and CTAP-III were subsequently confirmed in a single experiment using a ProteinChip-array-based immunoassay(Ciphergen Biosystems). The altered expression of the platelet-derived CXC-chemokines (40% and 63% for PF4 and CTAP-III, respectively) could not be explained by changes in absolute platelet counts. Thus, SELDI-TOF analysis of a limited number of exposed and unexposed subjects revealed that lowered expression of PF4 and CTAP-III proteins is a potential biomarker of benzene's early biologic effects and may play a role in the immunosuppressive effects of benzene.

[1]  N. Rothman,et al.  Discovery of Novel Biomarkers by Microarray Analysis of Peripheral Blood Mononuclear Cell Gene Expression in Benzene-Exposed Workers , 2005, Environmental health perspectives.

[2]  D. Savitz,et al.  Review of epidemiologic evidence on benzene and lymphatic and hematopoietic cancers. , 1997, American journal of industrial medicine.

[3]  F. Boehlen,et al.  Platelet chemokines and their receptors: what is their relevance to platelet storage and transfusion practice? , 2001, Transfusion medicine.

[4]  G. Rosenthal,et al.  Modulation of the immune response to Listeria monocytogenes by benzene inhalation. , 1985, Toxicology and applied pharmacology.

[5]  M Dosemeci,et al.  An expanded cohort study of cancer among benzene-exposed workers in China. Benzene Study Group. , 1996, Environmental health perspectives.

[6]  M. Deeg,et al.  Serum protein profiling by SELDI mass spectrometry: detection of multiple variants of serum amyloid alpha in renal cancer patients , 2004, Laboratory Investigation.

[7]  H Austin,et al.  Benzene and leukemia. A review of the literature and a risk assessment. , 1988, American journal of epidemiology.

[8]  J. Krijgsveld,et al.  Thrombocidins, Microbicidal Proteins from Human Blood Platelets, Are C-terminal Deletion Products of CXC Chemokines* , 2000, The Journal of Biological Chemistry.

[9]  H. Flad,et al.  The β‐thromboglobulins and platelet factor 4: blood platelet‐derived CXC chemokines with divergent roles in early neutrophil regulation , 2000, Journal of leukocyte biology.

[10]  R. Hayes,et al.  Detailed exposure assessment for a molecular epidemiology study of benzene in two shoe factories in China. , 2004, The Annals of occupational hygiene.

[11]  M. Aksoy,et al.  Malignancies due to occupational exposure to benzene. , 1980, American journal of industrial medicine.

[12]  A. Koch,et al.  Chemokines and angiogenesis , 2001, Current opinion in rheumatology.

[13]  R. Shore,et al.  Hematological changes among Chinese workers with a broad range of benzene exposures. , 2002, American journal of industrial medicine.

[14]  M Dosemeci,et al.  Benzene and the dose-related incidence of hematologic neoplasms in China. Chinese Academy of Preventive Medicine--National Cancer Institute Benzene Study Group. , 1997, Journal of the National Cancer Institute.

[15]  B. Adam,et al.  Identification of patients with head and neck cancer using serum protein profiles. , 2004, Archives of otolaryngology--head & neck surgery.

[16]  M Dosemeci,et al.  Hematotoxicity among Chinese workers heavily exposed to benzene. , 1996, American journal of industrial medicine.

[17]  H. Flad,et al.  Platelet‐derived CXC chemokines: old players in new games , 2000, Immunological reviews.

[18]  J. Roboz,et al.  Putative protein markers in the sera of men with prostatic neoplasms , 2003, BJU international.

[19]  Nathaniel Rothman,et al.  Hematotoxicity in Workers Exposed to Low Levels of Benzene , 2004, Science.

[20]  Z. Han,et al.  Platelet factor 4 and other CXC chemokines support the survival of normal hematopoietic cells and reduce the chemosensitivity of cells to cytotoxic agents. , 1997, Blood.

[21]  Down-regulation of neutrophil functions by the ELR(+) CXC chemokine platelet basic protein. , 2000 .

[22]  H. Hansen,et al.  Lung cancer. , 1990, Cancer chemotherapy and biological response modifiers.

[23]  A. Koch,et al.  Chemokines and chemokine receptors in rheumatoid arthritis. , 2003, Seminars in immunology.

[24]  Y. Benjamini,et al.  More powerful procedures for multiple significance testing. , 1990, Statistics in medicine.

[25]  Richard B. Hayes,et al.  An expanded cohort study of cancer among benzene-exposed workers in China. Benzene Study Group. , 1996 .

[26]  P. Sperryn,et al.  Blood. , 1989, British journal of sports medicine.

[27]  T. Zhukov,et al.  Discovery of distinct protein profiles specific for lung tumors and pre-malignant lung lesions by SELDI mass spectrometry. , 2003, Lung cancer.

[28]  R. Strieter,et al.  Chemokines: angiogenesis and metastases in lung cancer. , 2004, Novartis Foundation symposium.

[29]  R. Hayes,et al.  Mortality among benzene-exposed workers in China. , 1996, Environmental health perspectives.

[30]  M. Ernst,et al.  Platelet Factor 4 Inhibits Proliferation and Cytokine Release of Activated Human T Cells1 , 2002, The Journal of Immunology.