Prevention of spontaneous abortion in the CBA x DBA/2 mouse model by intravaginal TGF-β and local recruitment of CD4+ 8+ FOXP3+ cells

Problem Activation of latent transforming growth factor (TGF)-beta in seminal plasma has been suggested by Robertson et al. to promote maternal tolerance to paternal antigens. A possible consequence reported by Tremellen et al. is increased pregnancy rates in women undergoing IVF. A decreased spontaneous abortion rate has also been postulated. Seminal plasma contains many factors besides TGF-beta, and a critical test of the hypothesis was required. The purpose of the present study was to directly test the effect of pure TGF-beta. Method of study Pharmaceutical grade bioactive TGF-beta 3 with a bovine serum albumin (BSA) carrier 0.1-1% in phosphate-buffered saline (PBS) was given into the vaginal tract of CBA/J female mice at the time of mating with DBA/2 males. One microgram Salmonella enteritidis lipopolysaccharide was given intraperitoneally to augment occult losses and spontaneous resorptions assessed on day 13.5 of pregnancy. The effect of TGF-beta 3 on recruitment of lymphomyeloid cells to the vaginal wall and vaginal lumen of unmated mice in estrus was assessed using immunohistochemistry and flow cytometry. Results Two nanogram of intravaginal TGF-beta 3 in 0.1% BSA-PBS or 20 ng in 1% BSA-PBS reduced abortion rates. Protection was comparable to that achieved by immunization with BALB/c spleen cells. Fraction V BSA, a binder of TGF-beta s, had some activity, and could reduce availability of added TGF-beta 3. CD11c dendritic cells, CD3(+) T cells, and CD25(+) cells were recruited to the vaginal wall by 48 hr after TGF-beta 3 treatment, and cellularity of vaginal exudates increased. Foxp3(+) cells were present in increased numbers, and appeared to be CD8(+) and CD4(+) 8(+). Semen, but not TGF-beta 3, stimulated a physiological polymorphonuclear leukocyte exudate. Conclusion Intravaginal bioactive TGF-beta 3 can enhance success of pregnancy in vivo in an established model of abortion. The result could be explained by the independent ability of TGF-beta to promote a regulatory T-cell response.