Primary Study On The Effect Of RALR On The Activation Of Rat Mononuclear Cells By Different Antigens In Vitro

Objective: Due to the phenomenon that ALR maybe have immune adjustment in vivo, we aim to study the possible role of ALR on the immune adjustment in vitro and evaluate the relavent mechanism simply.Methods: The mononuclear cells isolated from rat spleen were stimulated by ConA in vitro. The administrations of the different dose of rALR were done when the mononuclear cells were activated or after. The proliferation of mononuclear cells was detected with 3H-TdR method. The IL-2 levels in the supernatants from the mononuclear cells by various treatments were detected with RIA test kit. The cell apoptosis was analyzed with Annexin V kit. Adult and newborn Wistar rat hepatocytes were isolated respectively. QGY and HK2 cells were cultured and collected. Four cells were frozen and thawed repeatly to obtain the cell membraneantigen. SD rat mononuclear cells were isolatd and stimulated by cell membrane antigens above at various quantity in vitro. The proliferation of mononuclear cells at various stimulation times was detected by 3H-TdR method. The ratio of stimulation group vs control was expressed as stimulation index. rALR or CsA was added at the beginning of stimulation or 48h before ending the culture and their inhibitory effects to mononuclear cell proliferation were still evaluated by 3H-TdR method.Results: rALR inhibited the proliferation of the mononuclear cells from rat spleen stimulated by ConA in a dose-dependent manner. The pretreatment of rALR at ID50 dose for 30h had no influence on the cell reactivity of to ConA compared with control. Their proliferations were still inhibited by 30ug/ml rALR after mononuclear cells stimulated by ConA for 10h or 32h. The IL-2 level from mononuclear cell activated by ConA with intervention of rALR was lower than that withot rALR at 24h. ALR at ID50 dose did not influenced the early and total apoptosis of rest mononuclear cells vs the control. The ConA+rALR (ID50 ) group had lower early apoptosis rate at 14h and 3Oh and higher total apoptosis rate than ConA group at 30h. The lOOug/ml or 30ug/ml rALR inhibited the proliferations of mononuclear cell induced by adult rat hepatocyte (PARH) antigen and HK2 antigen. Moreover, rALR with above two concentrations also inhibited the proliferation of the cells prestimulated by PARH or HK2antigen.Conclusion: rALR could inhibit the proliferation of mononuclear cells from rat spleen induced by ConA or specific antigens in vitro and also inhibited the production of IL-2, but it couldn't influence the rest mononuclear cells. The inhibitory role of rALR could have no direct relation with apoptosis.