Partial characterization of ovine intrauterine suppressor cells

Ovine uterine cells that represented Day 14 cyclic and pregnant endometrium were fractionated with Percoll and evaluated for suppression of cocultured phytohemagglutinin (PHA)-induced peripheral blood lymphocyte (PBL) proliferation and for the presence of Tlymphocyte markers. Uterine cells were then evaluated for suppressor activity following the depletion of conventional lymphocyte classes (i.e., T-, B-, and NK-like) with complement + antibody treatment. In addition, supernatant (derived from cultured uterine cells) was tested for transforming growth factor (beta) (TGFbeta) activity using neutralization antibodies to TGFbeta. Fractionated uterine cells (density range of 1.002-1.056 g/ml) from cyclic and pregnant ewes suppressed PHA-induced proliferation of PBL, and the majority (69.5%) of these cells were < or = 5.2 microm in diameter. Percentages of CD5+, CD4+, and CD8+ lymphocytes recovered from endometrial curettage were less for cells in this density range than for cells with greater densities. Uterine cells released suppressor factor(s) into the culture medium (supernatant); however, suppressor activity was unaffected by either anti-TGFbeta or complement + antibody treatment. In conclusion, low-density uterine cells from Day 14 cyclic and pregnant ewes suppressed the proliferation of cocultured PBL and released a suppressor factor(s) into the medium that did not exhibit TGFbeta activity. It is unlikely that the suppressor cells comprise conventional T-, B-, or NK-like lymphocyte lineages.