Novel method for detection of ex vivo tumor necrosis factor alpha production by monocytes.
Tumor necrosis factor alpha (TNF-alpha) has come into recent focus as a proinflammatory cytokine derived from monocytes/macrophages. We developed a novel system (the SEK-5001 system) for measurement of ex vivo production of TNF-alpha stimulated by lipopolysaccharide (LPS) as a marker of immune function. In the present study, we evaluated the performance of this system as a diagnostic tool. Furthermore, we compared TNF-alpha levels with data from other immune function tests, including lymphocyte blast formation test and differential leukocyte counts. Incubation of whole blood with a stimulation of low-dose LPS (100 EU/mL blood) for 4 hr at 37 degrees C gave acceptable results. After incubation, plasma TNF-alpha levels were determined by enzyme-linked immunosorbent assay (ELISA). The specificity, reproducibility, and recovery of the SEK-5001 system were excellent. No correlation between TNF-alpha levels and total leukocyte counts was found. Lymphocyte blast formation test and monocyte counts, however, were correlated with TNF-alpha levels in blood from patients with hematological malignancy and aplastic anemia before treatment. This assay system may potentially be clinically applicable to assess in vivo immune function.
Suzuki
Kikumi
K
Department of Molecular Physiology, Kyoritsu College of Pharmacy Tokyo, Japan.
Koyama
Takatoshi
T
Kobayashi
Shizuko
S
Kobayashi
Koji
K
Inagaki
Koji
K
Abe
Yoshiko
Y
Kuriyama
Kiyoshi
K
Shiba
Kiyoko
K
D000328:Adult; D000368:Aged; D001772:Blood Cell Count; D004305:Dose-Response Relationship, Drug; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D019337:Hematologic Neoplasms / Q000097:blood / Q000175:diagnosis*; D006801:Humans; D007962:Leukocytes / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D008875:Middle Aged; D009000:Monocytes / Q000737:chemistry / Q000378:metabolism*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis* / Q000378:metabolism
D008070:Lipopolysaccharides; D014409:Tumor Necrosis Factor-alpha