Development of a continuous IL-7-dependent murine pre-B cell line PB-1 suitable for the biological characterisation and assay of human IL-7.
Human interleukin-7 (IL-7) is a cytokine that appears to be critical for early T- and B-cell development and although IL-7 is currently under investigation as a therapeutic agent in a variety of hematolymphopoietic disorders, there have been few instances of the detection or investigation of this cytokine using a biological assay. This has been due, in the main, to the lack of a widely available, stable, easy to maintain and use, IL-7 responsive cell line. We have developed a pre-B-cell line, PB-1, from murine bone marrow, that is dependent on IL-7 for growth and has been maintained continually for up to 1 year without loss of responsiveness. The cells survive freezing and reviving, having been stored for periods of up to 4 years. The IL-7 bioassay is reproducible and sensitive, able to reliably detect 50 pg/ml IL-7. The assay is completely unresponsive to any other stimulatory cytokines tested and is not affected by a wide variety of inhibitory cytokines, with the exception of high levels of interferon alpha. The assay can be made completely specific for human IL-7 by including specific neutralizing antibodies for IL-7 and has been shown to be suitable for the estimation of IL-7 in both plasma and serum samples.
Mire-Sluis
A R
AR
National Institute for Biological Standards and Control, Blanche Lane, South Mimms, Potters Bar, UK. tms@gene.com
Healey
L
L
Griffiths
S
S
Hockley
D
D
Thorpe
R
R
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects*; D001681:Biological Assay / Q000379:methods* / Q000706:statistics & numerical data; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D005069:Evaluation Studies as Topic; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects*; D006801:Humans; D015851:Interleukin-7 / Q000032:analysis* / Q000097:blood / Q000494:pharmacology*; D051379:Mice; D008854:Microscopy, Electron; D012680:Sensitivity and Specificity; D013045:Species Specificity
D015851:Interleukin-7