Howard Gebel

Basal and stimulated secretion of cytokines by peritoneal macrophages in women with endometriosis

OBJECTIVEnTo evaluate basal (constitutive) and stimulated synthesis of tumor necrosis factor alpha (TNF alpha), interleukin (IL)-8, IL-10 by peritoneal macrophages (PM) in women with endometriosis.nnnDESIGNnPeritoneal macrophages were cultured in the presence or absence of lipopolysaccharide (LPS) for 24 hours. Peritoneal fluids (PF) and PM supernatants were assayed for cytokines using ELISA.nnnSETTINGnInstitute for the Study and Treatment of Endometriosis and university-based research laboratories.nnnSUBJECTSnFertile controls undergoing tubal ligation (n = 8) and women with endometriosis (n = 17).nnnINTERVENTIONnPeritoneal fluid samples were obtained at the time of diagnostic laparoscopy (endometriosis group) or laparoscopy for tubal ligation; both were performed in the midluteal phase of the cycle.nnnRESULTSnBoth basal and LPS stimulated production of TNF alpha, IL-8, and IL-10 by the PM were elevated significantly in women with endometriosis as compared with the fertile controls.nnnCONCLUSIONSnThis study demonstrated that cytokines TNF alpha, IL-8, and IL-10 are synthesized at greater than normal levels by basal and stimulated PM from women with endometriosis. The levels of TNF alpha and IL-8 correlated with the levels in the PF, suggesting that PM are the principal source of these cytokines in the PF.

Spontaneous Apoptosis of Endometrial Tissue is Impaired in Women with Endometriosis

OBJECTIVEnTo evaluate spontaneous apoptosis in single-cell suspensions of eutopic and ectopic endometrium from women with endometriosis and in eutopic endometrium from fertile controls without endometriosis.nnnDESIGNnPaired specimens of eutopic and ectopic endometrial tissue from patients with endometriosis and eutopic endometrium from controls were assessed for spontaneous apoptosis.nnnSETTINGnInstitute for the Study and Treatment of Endometriosis and university-based research laboratories.nnnPATIENT(S)nFertile controls (n = 10) and women with untreated endometriosis (n = 16).nnnINTERVENTION(S)nNone.nnnMAIN OUTCOME MEASURE(S)nSpontaneous apoptosis assessed with an ELISA-based cell death detection kit.nnnRESULT(S)nSpontaneous apoptosis (monitored by absorbance) of eutopic endometrium from patients with endometriosis and fertile controls was 0.63 +/- 0.1 and 1.43 +/- 0.11, respectively. Among patients with endometriosis, spontaneous apoptosis of ectopic endometrium was 0.26 +/- 0.06. Decreased apoptosis of ectopic versus eutopic endometrium was observed independent of cycle phase.nnnCONCLUSION(S)nThe susceptibility of endometrial tissue to spontaneous apoptosis is significantly lower in women with endometriosis than in fertile controls. We suggest that decreased susceptibility of endometrial tissue to apoptosis contributes to the etiology or pathogenesis of endometriosis.

The development of cytotoxicity in peritoneal macrophages from women with endometriosis

OBJECTIVEnTo assess the activation status of peritoneal macrophages from women with endometriosis.nnnDESIGNnPeritoneal macrophages from patients undergoing laparoscopy were tested for cytotoxic activity against a cultured hepatoma cell line.nnnSETTINGnPatients were tested at initial laparoscopy or at the completion of therapy.nnnPATIENTS AND PARTICIPANTSnFertile controls (n = 27), infertile controls (n = 20), untreated endometriosis (n = 43), danazol-treated endometriosis (n = 22), and gonadotropin-releasing hormone agonist (GnRH-a)-treated endometriosis (n = 13) were tested.nnnINTERVENTIONSnDanazol (800 mg/d) or GnRH-a therapy for 6 months.nnnRESULTSnCytotoxicity was elevated in stage I and II endometriosis (P less than 0.02) and in infertile controls (P less than 0.05) compared with fertile controls. Cytotoxicity in stage III and IV endometriosis was lower (P less than 0.02) than in stage I and II endometriosis. Indomethacin in vitro increased cytotoxicity (P less than 0.05) in stage III and IV endometriosis but not in the other groups tested. Cytotoxicity in danazol or GnRH-a-treated patients was increased (P less than 0.05 or greater) compared with untreated patients with comparable stage of disease.nnnCONCLUSIONSnPeritoneal macrophage cytotoxicity in women with endometriosis is affected by (1) the extent of endometriosis, (2) prostaglandin metabolism, and (3) treatment with danazol or GnRH-a.

Monocyte-mediated enhancement of endometrial cell proliferation in women with endometriosis * †

OBJECTIVEnTo investigate the capacity of monocytes from women with endometriosis to influence endometrial cell proliferation.nnnDESIGNnUterine endometrial cells were cultured in the presence and absence of autologous blood monocytes for 72 hours before assessment of endometrial cell proliferation by thymidine incorporation.nnnSETTINGnPatients were tested at initial presentation for evaluation of infertility and/or endometriosis.nnnPATIENTS, PARTICIPANTSnFertile controls, n = 17; infertile controls, n = 9; untreated endometriosis, n = 29.nnnINTERVENTIONSnNone.nnnRESULTSnEndometrial cell proliferation was enhanced significantly by blood monocytes in patients with endometriosis but was suppressed significantly by blood monocytes in fertile controls. Endometrial cell proliferation was not affected significantly by blood monocytes in infertile controls analyzed as a group, but a subset of infertile patients also showed enhancement of endometrial cell proliferation by blood monocytes.nnnCONCLUSIONSnBlood monocytes from patients with endometriosis and a subset of patients with unexplained infertility enhance autologous endometrial cell proliferation, whereas blood monocytes from fertile patients suppress endometrial cell proliferation. The capacity of monocytes to enhance endometrial cell proliferation appears to require both monocyte-derived factors that stimulate endometrial cell proliferation and endometrial cells capable of responding to those stimulatory factors. If either of these factors is absent, monocytes either suppress or have no effect on endometrial cell proliferation.

Effect of danazol in vitro and in vivo on monocyte-mediated enhancement of endometrial cell proliferation in women with endometriosis *†

OBJECTIVEnTo investigate danazols effect in vitro and in vivo on the ability of peripheral blood monocytes (PBM) from women with endometriosis to stimulate endometrial cell proliferation.nnnDESIGNnUterine endometrial cells from untreated or danazol-treated patients with endometriosis were cultured with or without autologous or heterologous PBM in the presence of different concentrations of danazol for 72 hours before assessment of endometrial cell proliferation by thymidine incorporation.nnnSETTINGnNot for profit clinical research institute and academic cell culture laboratory.nnnPATIENTSnWomen of reproductive age undergoing laparoscopy for endometriosis, 19 untreated patients and 17 danazol-treated patients.nnnINTERVENTIONSnPeripheral blood monocytes and endometrial biopsies obtained at laparoscopy. Danazol (800 mg/d) administered for 2 to 6 months (treated group) or added to cell cultures in concentrations of 10(-6), 10(-7), or 10(-9) M.nnnRESULTSnEndometrial cell proliferation was enhanced by autologous or heterologous PBM from untreated patients with endometriosis but was unaffected or suppressed by PBM from danazol-treated patients. Danazol in vitro reduced PBM-enhanced endometrial cell proliferation. Endometrial cell proliferation from danazol-treated patients was not enhanced by PBM from untreated patients with endometriosis.nnnCONCLUSIONSnDanazol treatment in vitro or in vivo suppresses PBM-mediated enhancement of endometrial cell proliferation. The effects are against both PBM and endometrial cells, suggesting that danazol affects monocyte-derived growth-stimulating factors and endometrial cell response to growth-stimulating factors.

Cytolysis of Eutopic and Ectopic Endometrial Cells by Peripheral Blood Monocytes and Peritoneal Macrophages in Women with Endometriosis

OBJECTIVEnTo compare the ability of peripheral blood monocytes (PBM) and peritoneal macrophages (PM) to mediate the in vitro cytolysis of endometrial cells from eutopic and ectopic endometrium in women with endometriosis.nnnDESIGNnProspective study of immune function.nnnSETTINGnInstitute for the Study and Treatment of Endometriosis and university-based research laboratories.nnnPATIENT(S)nTwenty-four women with endometriosis (15 in stage I/II, 9 in stage III/IV) and 4 patients treated with GnRH agonists.nnnINTERVENTION(S)nPeritoneal fluid and peripheral blood were sampled and eutopic and ectopic endometrium were biopsied during diagnostic laparoscopy.nnnMAIN OUTCOME MEASURE(S)nLysis of autologous endometrial cells.nnnRESULT(S)nPeripheral blood monocytes were significantly more cytolytic than peritoneal macrophages against autologous uterine endometrial cells. However, PBM and PM displayed a similar degree of cytolysis against a hepatoma cell line. Ectopic endometrial cells were significantly more resistant to cytolysis by autologous PBMC than were matched eutopic endometrial cells, and were completely resistant to cytolysis by autologous PM.nnnCONCLUSION(S)nThe reduced capacity of PM from women with endometriosis to mediate the destruction of endometrial cells coupled with the increased resistance of ectopic endometrial cells to macrophage-mediated cytolysis may facilitate the survival of these cells within the peritoneal cavity of women with endometriosis.

Mitogen induced production of polyclonal IgG is decreased in women with severe endometriosis

PROBLEM: The etiology and/or pathogenesis of endometriosis may involve aspects of both humoral and cellular immunity.

Differential Expression of VLAβ1 (CD29) on Monocytes From Patients With Endometriosis

PROBLEM: Previous studies have established that in vitro proliferation of endometrial cells is enhanced by peripheral blood monocytes (PBM) and suppressed by peritoneal macrophages (PM) from patients with endometriosis but only suppressed by PBM and PM obtained from normal subjects. The functional activity of PBM and PM is influenced by the engagement of numerous cell surface receptors with their respective physiological ligands.