Lymph node, spleen and peripheral blood lymphocytes as stimulators of alloreactivity [Limfociti limfnog čvora, slezene i periferne krvi kao stimulatori aloreaktivnosti]

Puc, Maja and Humar, Ines and Bulić-Jakuš, Floriana (2011) Lymph node, spleen and peripheral blood lymphocytes as stimulators of alloreactivity [Limfociti limfnog čvora, slezene i periferne krvi kao stimulatori aloreaktivnosti]. Collegium Antropologicum, 35 (2). pp. 369-376. ISSN 0350-6134

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Abstract

Before and after kidney transplantations, in vitro tests that measure the level of reactivity between donor and recipient lymphocytes are performed for better organ selection and as indicator of possible organ rejection. In these tests, donor's and recipient's lymphocytes are stimulated for proliferation, which intensity is measured and accordingly organ recipient reactivity towards graft is determined. Lymph node, spleen and peripheral blood lymphocytes are used for those purposes. For better interpretation of these in vitro tests it should be important to determine mitogenic ability of lymphocytes of different origin and to choose the most adequate cells. To compare mitogenic ability of deceased donor lymph node, spleen and peripheral blood lymphocytes one-way mixed lymphocyte culture (MLC) was used. As stimulators irradiated lymphocytes from spleen, lymph node and peripheral blood samples of 12 deceased donors were used while as responders lymphocytes from peripheral blood of healthy individuals, chosen according HLA-DRB1 alleles (stimulators and responders were HLA-DRB1 identical, semi-identical or different), were used. Spleen lymphocyte activity was the best with different cells and the weakest with identical cells. Impact of polyclonal mitogens (PHA - phytohemagglutinin, Con A - concanavalin A and PWM - pokeweed mitogen) on lymphocyte proliferation was tested on lymphocytes from spleen and lymph node of deceased donors. Results obtained in culture in vitro showed that spleen cells had exerted the best mitogenic potential and PHA had the greatest impact upon lymphocyte proliferation. This investigation is of importance for establishing the best model to reflect in vivo situation in transplanted patient.

Abstract in Croatian

Prije i nakon transplantacije bubrega rade se in vitro testovi kojima se mjeri razina reaktivnosti između limfocita primatelja i davatelja radi što boljeg odabira davatelja organa te radi utvrđivanja mogućeg odbacivanja organa. U tim testovima limfociti primatelja i davatelja se potiču na proliferaciju, intenzitet proliferacije se mjeri te se na temelju toga određuje reaktivnost primatelja organa prema presatku. Pritom se koriste limfociti slezene, limfnog čvora ili periferne krvi. Za bolju interpretaciju tih in vitro testova važno je odrediti mitogenu sposobnost limfocita iz različitih izvora (periferna krv, slezena, limfni čvor) te izabrati najprikladnije stanice za upotrebu što boljeg in vitro modela. Kako bismo usporedili mitogenu sposobnost limfocita limfnog čvora, slezene i periferne krvi koristili smo jednosmjerni test miješane kulture limfocita (MLC). Kao stimulatore smo koristili ozračene limfocite iz uzoraka limfnog čvora, slezene i periferne krvi 12 mrtvih davatelja, dok su nam responderi bili limfociti periferne krvi zdravih osoba izabranih prema HLA-DRB1 alelima (tako da su stimulatori i responderi bili HLA-DRB1 identični, poluidentični i različiti). Utjecaj poliklonskih mitogena (PHA – fitohemaglutinin, Con A – konkanavalin, PWM – korovski mitogen) na proliferaciju limfocita smo testirali na limfocitima slezene i limfnog čvora mrtvih davatelja. Rezultati dobiveni u kulturi in vitro pokazali su da stanice slezene imaju najsnažniji mitogeni potencijal. Od svih testiranih poliklonskih mitogena, PHA potiče najobilniju proliferaciju. Ovo istraživanje je važno za uspostavu najboljeg in vitro modela koji prikazuje in vivo situaciju u transplantiranih bolesnika.

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