Th1 Cytokine ( IFN-gamma ) Secretion Pattern of Peripheral Blood Mononuclear Cells Response to Trophoblast Antigen in Women with Unexplained Recurrent Spontaneous Abortion and Normal Fertile Controls

Yoo, Keun Jai; Song, In Ok; Choi, Bum Chae; Kang, Inn Soo; Park, In Sou; Byun, Hye Kyung; Lee, Ji Ae; Kim, Jeong Wook; Kim, Hyun Joo

Abstract

OBJECTIVE
A dichotomous Thl and Th2 cytokine profile has been associated with reproductive failure and success, respectively. The purpose of our study was to determine the levels of Thl cytokine (IFN- y ) secreted by peripheral blood mononuclear cells (PBMCs) form women with unexplained recurrentabortion (URA) and fertile controls in response to trophoblast antigen.
METHODS
PBMCs were isolated from 30 nonpregnant women with URA and from 10 nonpregnant fertile controls. Following 4 days of culture (1 * 10(6) cells/mL) with and without a protein extract derived from a trophoblast cell line (30 ug/mL, protein). None of the women had allergies, atopy or recent infection. Cytokines were measured in supernatants with enzyme-linked immunosorbent assay (ELISA) kits. IFN- r kit was obtained from BOISOURCE (lower limit of sensitivity, 15.6 pg/mL for IFN- r ). All values below the lowest limit of sensitivity as determined by test kit standards were considered negative. The cytokine stimulation test is considered positive if the IFN- r concentration increases by 200% or more with the trophoblast antigen stimulation. Datas are presented as mean+ SEM. Nonparametric testing (Mann-Whitney U) was used for analysis with P<0.05 considered statistically significant.
RESULTS
The Thl-type cytokine (IFN- r ) was detected in 20(67%) of 30 supernatants from women with URA. In contrast, 2 (20%) of trophoblast-activated PBMC culture supernatants from the 10 parus women with normal reproductive histories was detected IFN- r and but were significantly lower than levels in women with URA who had secreted IFN- r upon trophoblast stimulation (99.80+ 18.17 pg/mL versus 166.47 + 36.96 pg/mL, p<0.05). Spontaneous secretion of IFN- r was significantly higher in culture supernatants from women with URA than in supernatants from women with successful reproductive histories (41.36.09+6.99 pg/mL versus 25.89+9.34 pg/mL, p<0.05).
CONCLUSION
These data indicate that there are significant differences between women with URA and women with normal reproductive histories in their regulation of the Thl-cytokine (IFN- r) in response to trophoblast. Thl-type immunity to trophoblast is associated with URA and may play a role in reproductive failure.