Method of determining the suppressor cell component of the...

C - Chemistry – Metallurgy – 12 – Q

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C12Q 1/02 (2006.01) G01N 33/50 (2006.01) G01N 33/53 (2006.01) G01N 33/68 (2006.01) G01N 33/74 (2006.01)

Patent

CA 2185795

The method for determining a suppressor component of human immune status comprises collecting peripheral blood, obtaining a suspension of mononuclear cells (MNCs), dividing said suspension into two equal portions, cultivating MNCs in the first portion without a suppressor activator, cultivating MNCs in the second portion with said suppressor activator (TBG), washing MNCs out of culture medium, blocking the proliferation, adding newly isolated MNCs from a normal donor, which MNCs have been stimulated with phytohemagglutinin, into each of MNC portions in equal proportions to obtain test cultures, cultivating said test cultures, further evaluating the proliferation in said test cultures and determining the suppression value on the basis of the ratios of proliferation levels in test cultures.

La technique selon l'invention consiste à prélever un échantillon de sang périphérique, à préparer une suspension de cellules mononucléaires divisées en deux fractions égales, à cultiver la première fraction sans activateur de cellules suppressives, et la seconde avec un tel activateur (glycoprotéine béta-1 trophoblastique), à éliminer par lavage le milieu de culture des cellules mononucléaires et à bloquer la prolifération, à ajouter à chaque fraction de cellules mononucléaires des cellules mononucléaires fraîchement séparées, prélevées sur un donneur sain et stimulées par l'action de phytohémagglutinine, afin d'obtenir une culture-test, à cultiver ces cellules mononucléaires, à évaluer ensuite la prolifération et à déterminer le niveau de suppression par rapport aux niveaux de prolifération dans les cutures-test.

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