Animal study on CD4+ CD25+ regulatory T cells for treating female mouse with recurrent spontaneous abortion

Abstract

Objective: To explore immunotherapy effectiveness of the CD4+ CD25+ regulatory T cells for treating female mouse with recurrent spontaneous abortion (RSA) by animal experiments.

Methods: Mononuclear lymphocytes were isolated from the blood (instead of cord blood) of new-born baby of KunMing Bai mouse or BALB/c male mouse with normal birth ability (as unrelated third party blood source) by density gradient centrifugation method. The CD4+ CD25+ regulatory T cells were selected by magnetic-activated cell sorting from mononuclear cells of cord blood cells. CBA/J female mouse copulated with DBA/2J male mouse was utilized as RSA animal model. Pregnant RSA mice were injected different types of lymphocytes through tail vein. Independent sample t-test was used to analyze the data from each group.

Results: The proportion of CD4+ CD25+ T cells in CD4+ T cells was (17.49 ± 0.60)% in CD4+ CD25+ regulatory T cells injection group, which was statistical significant higher than that of mononuclear lymphocyte injection group (14.68 ± 0.83)%, sterile PBS group (9.54 ± 0.85)% or no injection group (9.28 ± 0.68)% (p < .05, t-value was 4.754, 13.242 and 15.621, respectively). The Foxp3 relative protein expression level of CD4+ CD25+ regulatory T cells injected group was 5.85 ± 0.45, which was also significant higher than that of mononuclear lymphocyte injection (2.86 ± 0.54), sterile PBS group (1.08 ± 0.16) or no injection group (1.00 ± 0.00) (p <. 05, t-value was 7.276, 17.227 and 18.635, respectively). Finally, two times of CD4+ CD25+ T cell injected group at the 4th and 8th day had well effect for RSA mouse, and embryo sorption rate was (4.92 ± 0.08)%, which significant lower than that of two times of mononuclear lymphocyte injected group (13.07 ± 0.06)%, sterile PBS group (23.11 ± 0.12)%, or no injection group (25.47 ± 0.11)% (p <. 05, t-value was -2.603, -4.012 and -4.700, respectively).

Conclusions: Pregnant mouse with RSA injected CD4+ CD25+ T cells several times for immunotherapy can get better effectiveness than that of pregnant mouse injected traditional mononuclear cells.