Lymphoblastic transformation assay of sheep peripheral blood lymphocytes: a new rapid and easy-to-read technique

Comp Immunol Microbiol Infect Dis. 1985;8(3-4):311-8. doi: 10.1016/0147-9571(85)90010-4.

Abstract

A new micro-method was used to evaluate in vitro sensitivity of ovine peripheral blood lymphocytes (PBL) to different non specific mitogens (pHA, Con A, PWM) and to investigate the interest of a colorimetric assay for measurement of transformed lymphocytes. The results showed that sheep PBL in flat-bottomed microplates responded optimally at a cell density of 8 X 10(6) cells/ml to PHA (2.5 micrograms/ml), Con A (5 micrograms/ml) and PWM (5 micrograms/ml). The colorimetric assay using a tetrazolium salt (MTT), for measuring the transformed lymphocytes, is very well correlated with the classical method of [3H]thymidine incorporation. This new revelation technique of the mitogenic response improve the technical value of the assay, which is more rapid and easy-to-read, without diminishing the biological value.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Division
  • Cell Survival
  • Colorimetry
  • Concanavalin A / pharmacology
  • DNA / biosynthesis
  • Female
  • Lymphocyte Activation*
  • Lymphocytes / metabolism*
  • Lymphocytes, Null
  • Male
  • Mitogens / pharmacology*
  • Phytohemagglutinins / pharmacology
  • Pokeweed Mitogens / pharmacology
  • Sheep
  • Tetrazolium Salts
  • Thiazoles
  • Thymidine / metabolism

Substances

  • Mitogens
  • Phytohemagglutinins
  • Pokeweed Mitogens
  • Tetrazolium Salts
  • Thiazoles
  • Concanavalin A
  • DNA
  • thiazolyl blue
  • Thymidine