Endometriotic haptoglobin binds to peritoneal macrophages and alters their function in women with endometriosis Kathy L Sharpe-Timms, Ph.D., Randy L.

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Endometriotic haptoglobin binds to peritoneal macrophages and alters their function in women with endometriosis Kathy L Sharpe-Timms, Ph.D., Randy L Zimmer, M.S., Emily A Ricke, M.S., Marta Piva, Ph.D., Gary M Horowitz, M.D. Fertility and Sterility Volume 78, Issue 4, Pages 810-819 (October 2002) DOI: 10.1016/S0015-0282(02)03317-4

FIGURE 1 Representative immunoreactivity with CD68 and haptoglobin of peritoneal macrophages from women with endometriosis (n=11) and without (n=12) endometriosis. (A), Peritoneal macrophages from women with endometriosis, tested for immunoreactivity with CD68. (B), Peritoneal macrophages from women without endometriosis, tested for immunoreactivity with CD68. Immunoreactivity with the macrophage marker CD68 was equivalent in the two groups. (C), Peritoneal macrophages from women with endometriosis, tested for immunoreactivity with the haptoglobin antibody. (D), Peritoneal macrophages from women without endometriosis, tested for immunoreactivity with the haptoglobin antibody. Significantly more peritoneal macrophages from women with endometriosis showed immunoreactivity with the haptoglobin antibody. See Table 1 for statistical analyses. Sharpe-Timms. Haptoglobin alters macrophage function. Fertil Steril 2002. Fertility and Sterility 2002 78, 810-819DOI: (10.1016/S0015-0282(02)03317-4)

FIGURE 2 Adhesion in women with endometriosis compared to women without endometriosis. The adherence index (mean [±SD]) of the freshly isolated, untreated peritoneal macrophages was significantly reduced in women with endometriosis (open bars; n=6 per bar) compared with controls (closed bars; n=5 per bar) (∗P<.043, β=.437), an effect that was observed at all time points (P=.713, β=.500). Sharpe-Timms. Haptoglobin alters macrophage function. Fertil Steril 2002. Fertility and Sterility 2002 78, 810-819DOI: (10.1016/S0015-0282(02)03317-4)

FIGURE 3 The effect of haptoglobin treatment on the index (mean [±SD]) of peritoneal macrophages from women with endometriosis and those without endometriosis. Addition of exogenous haptoglobin adhesion (right bar of each time point pair) had no effect on the peritoneal macrophage adhesion index in women with endometriosis (open bars; n=5 per bar; P=.316; β=.613) at any time point (P=.118; β=.284). However, haptoglobin (10 μg/mL) significantly reduced the peritoneal macrophage adhesion index in women without endometriosis (striped bars; n=5 per bar; P=.032; β=.664) at all times tested (∗P<.001; β=.518). Sharpe-Timms. Haptoglobin alters macrophage function. Fertil Steril 2002. Fertility and Sterility 2002 78, 810-819DOI: (10.1016/S0015-0282(02)03317-4)

FIGURE 4 Interleukin-6 (IL-6) production by peritoneal macrophages after haptoglobin administration. Haptoglobin at 1.0 and 10 μg/mL significantly increased IL-6 production by peritoneal macrophages from women without endometriosis compared to 0 and 0.1 μg/mL (n=3; six replicates per treatment). a versus b values shown as mean (±SD) P=.028; β=.569. Sharpe-Timms. Haptoglobin alters macrophage function. Fertil Steril 2002. Fertility and Sterility 2002 78, 810-819DOI: (10.1016/S0015-0282(02)03317-4)

FIGURE 5 Interleukin-6 (IL-6) production by peritoneal macrophages from women with (n=6, Endo) and without (n=6, control) endometriosis after haptoglobin administration. Because these data were not normally distributed, they are presented as the median with 25th and 75th percentiles. Haptoglobin treatment significantly increased IL-6 production by peritoneal macrophages from women with endometriosis compared with untreated peritoneal macrophages from women with endometriosis or peritoneal macrophages from controls that were or were not treated with haptoglobin (a versus b P=.023; β=.800). Sharpe-Timms. Haptoglobin alters macrophage function. Fertil Steril 2002. Fertility and Sterility 2002 78, 810-819DOI: (10.1016/S0015-0282(02)03317-4)

FIGURE 6 Model of the effects of endometriotic haptoglobin on peritoneal macrophage function. Endometriotic haptoglobin is a differentially glycosylated form of haptoglobin, synthesized and secreted by endometriotic lesions, that binds to peritoneal macrophages and reduces their phagocytic capacity by blocking adherence while up-regulating macrophage interleukin-6 (IL-6) production. Hence, endometriotic lesions that express ENDO-I may both avoid phagocytosis by peritoneal macrophages while stimulating them to produce cytokines that propagate ENDO-I expression by the ectopic endometrial cells. Sharpe-Timms. Haptoglobin alters macrophage function. Fertil Steril 2002. Fertility and Sterility 2002 78, 810-819DOI: (10.1016/S0015-0282(02)03317-4)