A new method to detect response of lymphocytes to allo-antigens.
The mixed lymphocyte reaction (MLR) is the most commonly used method of investigating the immunological response of lymphocytes. However, this conventional technique requires the use of a radioisotope and is time consuming. Furthermore, there is a possibility that MLR does not reflect the response of each lymphocyte subset individually. The purpose of this study is to determine whether detection of IL-2 mRNA by reverse transcription polymerase chain reaction (RT-PCR) can be used as a new method of studying a precise response of PBL to allo-antigens. The study was conducted using lymphocytes from healthy volunteers, a renal transplant recipient who exhibited donor specific MLR unresponsiveness, the donor and the recipient's father. A kinetics study revealed that the expression of IL-2 mRNA in the MLC reached its maximum at 24-48 h. Lymphocytes from a renal transplant recipient also exhibited the same IL-2 mRNA expression pattern in response to stimulation with PBL from a non-related 3rd party but not to that from the donor. These data suggest that the expression level of IL-2 mRNA in MLC measured by RT-PCR at 24-48 h provides accurate response of PBL to allo-antigens.
Okimoto
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2nd Department of Surgery, Hiroshima University School of Medicine, Japan.
D000328:Adult; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D007376:Interleukin-2 / Q000235:genetics; D007519:Isoantigens / Q000008:administration & dosage*; D016030:Kidney Transplantation / Q000276:immunology; D007959:Lymphocyte Culture Test, Mixed / Q000379:methods*; D008214:Lymphocytes / Q000276:immunology* / Q000378:metabolism; D008297:Male; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
D007376:Interleukin-2; D007519:Isoantigens; D012333:RNA, Messenger