SPECIFIC INVITRO LYMPHOCYTE-RESPONSE OF CHICKENS INJECTED WITH KILLED MYCOBACTERIUM-TUBERCULOSIS

  • 1 January 1977
    • journal article
    • research article
    • Vol. 38  (3) , 395-397
Abstract
An in vitro microassay for lymphocyte transformation, using 3H-thymidine incorporated into avian peripheral blood leukocytes, is described. The transformation responses of 1 .times. 106 leukocytes stimulated by the nonspecific mitogen, phytohemmaglutinin-M (PHA-M), were equivalent with 68- and 92-h incubation durations. PHA-M at a concentration of 50 .mu.l/ml produced greater stimulation than PHA-M used at a concentration of 25 or 100 .mu.l/ml. The transformation response to PHA-M was significantly affected by the lot of bovine fetal serum used to supplement the RPMI 1640 culture medium. The specific antigen, purified protein derivative (PPD) of tuberculin, stimulated significant transformation in cultures of 1 .times. 106 leukocytes from M. tuberculosis-sensitized fowl. PPD at concentration of 50 .mu.g/ml was superior to PPD used at a concentration of 100 or 150 .mu.g/ml; 68-h incubation was significantly better than 92-h incubation. The magnitude of the in vitro transformation response to PPD was greater at 6 wk after sensitization than at 2 or 4 wk and it was not directly related to the magnitude of the in vivo wattle test response in sensitized chickens.

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