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The frequency of CD25+CD4+ and FOXP3+ regulatory T cells in ectopic endometrium and ectopic decidua.

Basta P, Majka M, Jozwicki W, Lukaszewska E, Knafel A, Grabiec M, Stasienko E, Wicherek L - Reprod. Biol. Endocrinol. (2010)

Bottom Line: We then compared these findings to what we observed in the normal eutopic endometrium of women during the secretory phase of the menstrual cycle (with immune cells under individual control).In our study, the percentages of FOXP3+ cells within the subpopulation of CD4+ T lymphocytes found in the decidua of the patients treated for Fallopian tube pregnancies were statistically significantly lower than both those observed in the ovarian endometriosis tissue samples and those found in the secretory eutopic endometrium samples of the control group.The disturbance in the immunological equilibrium observed in ectopic endometrium and decidua would seem to be related to the alteration in the Treg cell population that occurs in these ectopic tissues.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Gynecology and Oncology of the Jagiellonian University, Krakow, Poland.

ABSTRACT

Background: The presence of regulatory T (Treg) cells in human endometrium is crucial for maintaining immunological homeostasis within the uterus. For this study we decided to evaluate the subpopulations of Treg cells in conditions where a disturbance in the immunological equilibrium in ectopic endometrium and decidua has been observed, such as in cases of ovarian endometriosis (involving local immune cell suppression) and ectopic pregnancy (involving an increase in local immune system activity). We then compared these findings to what we observed in the normal eutopic endometrium of women during the secretory phase of the menstrual cycle (with immune cells under individual control).

Methods: The endometrium tissue samples evaluated in our study were obtained from 47 women during one of two kinds of laparoscopic procedures. 16 of the women underwent laparoscopies due to Fallopian tube pregnancies (EP), and 16 due to ovarian endometrioma, while 15 women made up a control group. The presence of regulatory T cells in these tissue samples was evaluated by FACS.

Results: In our study, the percentages of FOXP3+ cells within the subpopulation of CD4+ T lymphocytes found in the decidua of the patients treated for Fallopian tube pregnancies were statistically significantly lower than both those observed in the ovarian endometriosis tissue samples and those found in the secretory eutopic endometrium samples of the control group.

Conclusion: The disturbance in the immunological equilibrium observed in ectopic endometrium and decidua would seem to be related to the alteration in the Treg cell population that occurs in these ectopic tissues.

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Related in: MedlinePlus

Treg cell gating strategy. The characterization of the subpopulation of Treg cells (CD4+CD25+FOXP3+) within the subpopulation of T lymphocytes in eutopic endometrium during the secretory cycle phases (E), ovarian endometriosis (OE), and Fallopian tube pregnancy (EP). Left panel shows the gating strategy for CD4-positive cells (P1). Middle panel is used to set the gate on double positive CD4+CD25+ cells (P2). On the right panel the histogram representing FoxP3 staining is shown (FoxP3+).
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Figure 1: Treg cell gating strategy. The characterization of the subpopulation of Treg cells (CD4+CD25+FOXP3+) within the subpopulation of T lymphocytes in eutopic endometrium during the secretory cycle phases (E), ovarian endometriosis (OE), and Fallopian tube pregnancy (EP). Left panel shows the gating strategy for CD4-positive cells (P1). Middle panel is used to set the gate on double positive CD4+CD25+ cells (P2). On the right panel the histogram representing FoxP3 staining is shown (FoxP3+).

Mentions: The cell phenotype was analyzed with the panel of mAb-CD4 FITC, CD25 APC, and FOXP3 PE (Pharmingen). Briefly stated, to the 1 × 106 cells suspended in 60 μl of staining buffer (PBS, 2% FBS) 20 μl of each mAb (CD4, CD25) was added. Next the cells were incubated in the dark for 30 min at 4°C. After incubation the cells were washed twice in PBS and were permeabilized with FoxP3 permeabilization buffer (Becton Dickinson; USA) for 10 min at room temperature in the dark. Following this, they were stained with anti-FOXP3 antibodies for 30 min at 4°C in the dark. The stained cells were then washed and collected using the FACSC anto-cytometer (Becton Dickinson; USA), and finally were analyzed with FACS Diva software (Becton Dickinson; USA). Each time 3 × 104 events were saved for analysis. Logical gates were used to analyze particular populations of cells. The cells were gated first according to SSC and CD4FITC parameters (gate P1). After this, the cells from gate P1 were analyzed according to CD25PE and CD4FITC parameters, and gate P2 was established on double positive CD4FITC CD25PE cells. The double positive cells from gate P2 were analyzed for the presence of FoxP3 antigen. The gate FoxP3+ was set on cells which had stained positively with FoxP3 APC antibodies (Figure 1).


The frequency of CD25+CD4+ and FOXP3+ regulatory T cells in ectopic endometrium and ectopic decidua.

Basta P, Majka M, Jozwicki W, Lukaszewska E, Knafel A, Grabiec M, Stasienko E, Wicherek L - Reprod. Biol. Endocrinol. (2010)

Treg cell gating strategy. The characterization of the subpopulation of Treg cells (CD4+CD25+FOXP3+) within the subpopulation of T lymphocytes in eutopic endometrium during the secretory cycle phases (E), ovarian endometriosis (OE), and Fallopian tube pregnancy (EP). Left panel shows the gating strategy for CD4-positive cells (P1). Middle panel is used to set the gate on double positive CD4+CD25+ cells (P2). On the right panel the histogram representing FoxP3 staining is shown (FoxP3+).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2958978&req=5

Figure 1: Treg cell gating strategy. The characterization of the subpopulation of Treg cells (CD4+CD25+FOXP3+) within the subpopulation of T lymphocytes in eutopic endometrium during the secretory cycle phases (E), ovarian endometriosis (OE), and Fallopian tube pregnancy (EP). Left panel shows the gating strategy for CD4-positive cells (P1). Middle panel is used to set the gate on double positive CD4+CD25+ cells (P2). On the right panel the histogram representing FoxP3 staining is shown (FoxP3+).
Mentions: The cell phenotype was analyzed with the panel of mAb-CD4 FITC, CD25 APC, and FOXP3 PE (Pharmingen). Briefly stated, to the 1 × 106 cells suspended in 60 μl of staining buffer (PBS, 2% FBS) 20 μl of each mAb (CD4, CD25) was added. Next the cells were incubated in the dark for 30 min at 4°C. After incubation the cells were washed twice in PBS and were permeabilized with FoxP3 permeabilization buffer (Becton Dickinson; USA) for 10 min at room temperature in the dark. Following this, they were stained with anti-FOXP3 antibodies for 30 min at 4°C in the dark. The stained cells were then washed and collected using the FACSC anto-cytometer (Becton Dickinson; USA), and finally were analyzed with FACS Diva software (Becton Dickinson; USA). Each time 3 × 104 events were saved for analysis. Logical gates were used to analyze particular populations of cells. The cells were gated first according to SSC and CD4FITC parameters (gate P1). After this, the cells from gate P1 were analyzed according to CD25PE and CD4FITC parameters, and gate P2 was established on double positive CD4FITC CD25PE cells. The double positive cells from gate P2 were analyzed for the presence of FoxP3 antigen. The gate FoxP3+ was set on cells which had stained positively with FoxP3 APC antibodies (Figure 1).

Bottom Line: We then compared these findings to what we observed in the normal eutopic endometrium of women during the secretory phase of the menstrual cycle (with immune cells under individual control).In our study, the percentages of FOXP3+ cells within the subpopulation of CD4+ T lymphocytes found in the decidua of the patients treated for Fallopian tube pregnancies were statistically significantly lower than both those observed in the ovarian endometriosis tissue samples and those found in the secretory eutopic endometrium samples of the control group.The disturbance in the immunological equilibrium observed in ectopic endometrium and decidua would seem to be related to the alteration in the Treg cell population that occurs in these ectopic tissues.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Gynecology and Oncology of the Jagiellonian University, Krakow, Poland.

ABSTRACT

Background: The presence of regulatory T (Treg) cells in human endometrium is crucial for maintaining immunological homeostasis within the uterus. For this study we decided to evaluate the subpopulations of Treg cells in conditions where a disturbance in the immunological equilibrium in ectopic endometrium and decidua has been observed, such as in cases of ovarian endometriosis (involving local immune cell suppression) and ectopic pregnancy (involving an increase in local immune system activity). We then compared these findings to what we observed in the normal eutopic endometrium of women during the secretory phase of the menstrual cycle (with immune cells under individual control).

Methods: The endometrium tissue samples evaluated in our study were obtained from 47 women during one of two kinds of laparoscopic procedures. 16 of the women underwent laparoscopies due to Fallopian tube pregnancies (EP), and 16 due to ovarian endometrioma, while 15 women made up a control group. The presence of regulatory T cells in these tissue samples was evaluated by FACS.

Results: In our study, the percentages of FOXP3+ cells within the subpopulation of CD4+ T lymphocytes found in the decidua of the patients treated for Fallopian tube pregnancies were statistically significantly lower than both those observed in the ovarian endometriosis tissue samples and those found in the secretory eutopic endometrium samples of the control group.

Conclusion: The disturbance in the immunological equilibrium observed in ectopic endometrium and decidua would seem to be related to the alteration in the Treg cell population that occurs in these ectopic tissues.

Show MeSH
Related in: MedlinePlus