Lawrence W Chamley, Ph. D. , Barbara Konarkowska, M. Sc

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Is interleukin-3 important in antiphospholipid antibody-mediated pregnancy failure? Lawrence W Chamley, Ph.D., Barbara Konarkowska, M.Sc., Andrea M Duncalf, B.Sc., Murray D Mitchell, D.Sc., Peter M Johnson, D.Sc. Fertility and Sterility Volume 76, Issue 4, Pages 700-706 (October 2001) DOI: 10.1016/S0015-0282(01)01984-7

FIGURE 1 Reverse transcriptase-polymerase chain reaction reactions using IL-3 α-chain primers. Lane 1: normal villous placenta; lane 2: Percoll-purified trophoblasts; lane 3: immuno-purified trophoblasts; lane 4: BeWo; lane 5: Jar; lane 6: Jeg-3. Chamley. Interleukin-3 important antiphospholipid. Fertil Steril 2001. Fertility and Sterility 2001 76, 700-706DOI: (10.1016/S0015-0282(01)01984-7)

FIGURE 2 The effect of hrIL-3 on the proliferation of trophoblast cells purified from normal term placentae by enzymatic digestion, density gradient centrifugation and immunomagnetic bead depletion of contaminating cells was examined by [3H] thymidine incorporation assay. The proliferation of cells treated with hrIL-3 is expressed as a percentage of the proliferation of control untreated cells. The addition of IL-3 had no significant effect on the proliferation of trophoblast at any of the concentrations tested. Experiments were conducted in quadruplicate on three separate occasions. Bars represent standard errors. Chamley. Interleukin-3 important antiphospholipid. Fertil Steril 2001. Fertility and Sterility 2001 76, 700-706DOI: (10.1016/S0015-0282(01)01984-7)

FIGURE 3 The ability of hrIL-3 to reverse the inhibitory effect of the mAB JAB-1 on the proliferation of the choriocarcinoma cell line Jar was examined by [3H] thymidine incorporation assay. The proliferation of cells treated with hrIL-3 and JAB-1 is expressed as a percentage of the proliferation of control cells treated with JAB-1 alone. A significant (∗P<.001; Dunnett’s t-test) inhibition of the proliferation of these cells was observed. Experiments were conducted in quadruplicate on two separate occasions. Bars represent standard errors. Chamley. Interleukin-3 important antiphospholipid. Fertil Steril 2001. Fertility and Sterility 2001 76, 700-706DOI: (10.1016/S0015-0282(01)01984-7)

FIGURE 4 The effect of hrIL-3 on the secretion of β2 glycoprotein I by trophoblast cells was examined by incubating increasing concentrations of hrIL-3 with partially purified trophoblast cells from normal term placentae for up to 4 days. The relative amounts of β2 glycoprotein I secreted by the trophoblast cells examined by dot blot of 5-μL samples of conditioned medium probed with a mixture of the mABs ID2 and IIC5. This analysis demonstrated a time-dependent increase in the secretion of β2 glycoprotein I, whereas addition of hrIL-3 had no effect on the secretion of β2 glycoprotein I. (A) Dilution series of purified β2 GPI (in micrograms per milliliter), (B) 24 hours, (C) 48 hours, (D) 72 hours, (E) 96 hours. Chamley. Interleukin-3 important antiphospholipid. Fertil Steril 2001. Fertility and Sterility 2001 76, 700-706DOI: (10.1016/S0015-0282(01)01984-7)