Luis Jiménez-Zamudio

Quantitative and qualitative peritoneal immune profiles, T-cell apoptosis and oxidative stress-associated characteristics in women with minimal and mild endometriosis

Please cite this paper as: Mier‐Cabrera J, Jiménez‐Zamudio L, García‐Latorre E, Cruz‐Orozco O, Hernández‐Guerrero C. Quantitative and qualitative peritoneal immune profiles, T‐cell apoptosis and oxidative stress‐associated characteristics in women with minimal and mild endometriosis. BJOG 2011;118:6–16.

Women with endometriosis improved their peripheral antioxidant markers after the application of a high antioxidant diet.

BackgroundOxidative stress has been identified in the peritoneal fluid and peripheral blood of women with endometriosis. However, there is little information on the antioxidant intake for this group of women. The objectives of this work were 1) to compare the antioxidant intake among women with and without endometriosis and 2) to design and apply a high antioxidant diet to evaluate its capacity to reduce oxidative stress markers and improve antioxidant markers in the peripheral blood of women with endometriosis.MethodsWomen with (WEN, n = 83) and without endometriosis (WWE, n = 80) were interviewed using a Food Frequency Questionnaire to compare their antioxidant intake (of vitamins and minerals). Then, the WEN participated in the application of a control (n = 35) and high antioxidant diet (n = 37) for four months. The high antioxidant diet (HAD) guaranteed the intake of 150% of the suggested daily intake of vitamin A (1050 μg retinol equivalents), 660% of the recommended daily intake (RDI) of vitamin C (500 mg) and 133% of the RDI of vitamin E (20 mg). Oxidative stress and antioxidant markers (vitamins and antioxidant enzymatic activity) were determined in plasma every month.ResultsComparison of antioxidant intake between WWE and WEN showed a lower intake of vitamins A, C, E, zinc, and copper by WEN (p < 0.05, Mann Whitney Rank test). The selenium intake was not statistically different between groups. During the study, the comparison of the 24-hour recalls between groups showed a higher intake of the three vitamins in the HAD group. An increase in the vitamin concentrations (serum retinol, alpha-tocopherol, leukocyte and plasma ascorbate) and antioxidant enzyme activity (superoxide dismutase and glutathione peroxidase) as well as a decrease in oxidative stress markers (malondialdehyde and lipid hydroperoxides) were observed in the HAD group after two months of intervention. These phenomena were not observed in the control group.ConclusionWEN had a lower intake of antioxidants in comparison to WWE. Peripheral oxidative stress markers diminished, and antioxidant markers were enhanced, in WEN after the application of the HAD.

TLRs and NODs mRNA expression pattern in healthy mouse eye.

Aims: To look for TLR and NOD mRNA expression in the healthy eye and in other immune privileged and non-immune privileged mouse organs. Methods: Semiquantitative RT-PCR was performed to look for TLR1–9 and NOD1 and NOD2 mRNA expressions in the whole eye, in the anterior (AP) and posterior (PP) portions of the eye, in corneal fibroblasts (CF) and in ovary, brain, testis, heart, lung, and spleen. Results: All the TLR mRNAs were expressed in the whole eye of Balb/c mice. NIH and C57BL/6 did not express TLR9 and TLR8, respectively. NIH expressed higher levels of TLR1, 2, 3, and 6 than the other strains. C57BL/6 expressed the lowest levels of all TLRs. TLR9, 5, and 4 were the less expressed in all strains. All TLRs were expressed in Balb/c PP and TLR1 was not expressed in AP. In NIH and Balb/c CF the majority of TLRs were overexpressed with LPS. In testis, expression of most TLRs was absent. Non-immune privileged organs expressed most of the TLRs. All the organs expressed NOD1 and NOD2. In PP NOD2 was not expressed. Conclusion: TLRs and NODs are expressed in the eye, and could have an important role in the innate immunity.

The Predominant Th1 Cytokine Profile in Maternal Plasma of Preeclamptic Women Is Not Reflected in the Choriodecidual and Fetal Compartments

Objective: Human pregnancy disorders such as preeclampsia are thought to involve variations in cytokine levels. It has been proposed that, in preeclamptic women, a balance favoring the Th1-type over the Th2-type cytokine profile determines local or systemic immunologic responses to pregnancy and that this may cause defective placental implantation and placental ischemia, which activate systemic endothelial cells. The purpose of this study was to determine whether cytokine expression differs in the maternal, choriodecidual, and fetal compartments, and between women with or without preeclampsia. Methods: Plasma concentrations of interferon gamma (IFNy), interleukin (IL)-2, IL-4, and IL-10 were measured by enzyme-linked immunosorbent assay (ELISA) in samples obtained from maternal peripheral blood (MPB), choriodecidual (CD), and fetal cord (FC) blood compartments of 17 women with preeclampsia and in 15 normotensive women. Intracellular concentrations of IFNy and IL-2 in T lymphocytes were assessed by flow cytometry. Results: Plasma IFNy concentrations in both MPB and CD compartments were significantly higher in preeclamptic than in normotensive women. Maternal plasma IL-4 concentration was significantly lower in preeclamptic than in normotensive women. Intracellular IFNy and IL-2 concentrations did not differ signficantly between preeclamptic and normotensive women. Conclusions: The dominant Th1-type over Th2-type cytokine profile is evident in MPB, but not in the CD and FC blood compartments. This might reflect the complex cytokine networks in the fetal-placental interface and might involve trophoblasts or decidual and endothelial cells, which could account for the increased plasma IFNy concentration and T-helper cell number.

Toxoplasma gondii: effect of maternal infection in the development of lymphoid organs of BALB/c neonates.

Toxoplasmosis is one of the worldwide parasitic zoonoses. Alterations in the lymphopoietic system are still poorly studied. We analyzed lymphoid organs of BALB/c mice neonates from Toxoplasma gondii-intraperitoneally-infected mothers on 19th day of gestation, with 30 tachyzoites of strain RH. Normal non-infected pregnant females were used as controls. At 7 days after birth, animals were classified as neonates from infected (NIM) and neonates from non-infected mothers (NNIM). Weight of the thymus and number of thymic cells in NIM were decreased, percentage of apoptosis was significantly increased. Decrease in lymphocytes and monocytes and an increase of plasma cells were observed in bone marrow of NIM. Peripheral blood of NIM showed an increase of monocytes and neutrophils and a decrease in lymphocytes. Infection of the mother during the last day of gestation provokes in the neonates changes in the lymphoid organs that could explain survival of 75% of them.

Prooxidant and antioxidant sex-linked response and its relationship to mixed oxidase function enzymes in liver of Ameca splendens, an endangered goodeid fish exposed to PCBs

The butterfly split-fin goodeid Ameca splendens is an endemic and endangered Mexican fish species whose habitat is polluted with persistent xenobiotics, such as polychlorinated biphenyls (PCB). These toxicants are potent inducers of mixed oxidase function (MOF) enzymes including cytochrome P450 (CYP450) and other cytoplasmatic enzymes such as alcohol dehydrogenase (ADH). During PCB biotransformation by MOF isoenzymes, reactive oxygen species (ROS) are generated that are able to produce oxidative stress. However, in A. splendens the linkage between the induction of MOF and the oxidative stress response produced by PCB remains unknown. To clarify the possible relationships between the induction of CYP450 and the ADH activity with the oxidative stress response, the induction of lipid peroxidation (LPOX) and superoxide dismutase (SOD) and the catalase (CAT) activity produced by exposure to PCB mixture were examined for 1, 2, 4, 8, or 16 days. In both genders, the response pattern induced by PCB was different. PCB induced LPOX and CAT, while a decrease in SOD activity was observed. It was not possible to explain the toxicity of PCB based on toxic equivalency factors. The response seems to be dependent on several factors including species, gender, and interaction of PCB in the mixture. A correlation was found between LPOX and ADH but not with CYP450. A correlation between ADH and SOD was observed in both genders but not with CAT. In males, a correlation between CYP450 with SOD was detected. Data suggest that the redox process involving ADH possesses a dual function, whereby in an indirect manner there is induction of LPOX, which generates energy for SOD and CYP450.

Measurement of Suppressor Activity of T CD4+CD25+ T Reg Cells Using Bromodeoxyuridine Incorporation Assay

The suppressor effect of T regulatory lymphocytes in co-cultures with T effector cells obtained by magnetic columns from healthy donors and activated by CD3/CD28 was measured by a proliferation assay using BrdU incorporation and an ELISA test. Tritiated thymidine incorporation was used as a reference since it is the gold standard for proliferation assays. Both methods were used simultaneously in the same samples in order to compare them. Correlation between them was statistically significant (p < 0.001). The purification using magnetic columns was very efficient since CD4+CD25+ cells were also FOXP3+ therefore; they were identified as suppressor T cells. The use of BrdU incorporation in suppression assays is an excellent method that avoids the use of radioactive contaminating materials.

Increased NHC Cells in the Peritoneal Cavity of Plasmacytoma Susceptible BALB/c Mouse.

BALB/c strain mice are unique in that they develop murine plasmacytoma (MPC) as a consequence of the inflammation induced by pristane oil injection in the peritoneal cavity. In this work the Treg, Th17, B1, B2, and NHC lymphocyte populations from the peritoneal environment of BALB/c, the susceptible strain, and C57BL/6 mice, which do not develop MPC after oil treatment, were studied. Both oil-treated strains showed decreased levels of Th17 lymphocytes, no significant variation in Treg lymphocytes, and a drastic decrease of all B lymphocyte populations. However, only oil-induced BALB/c showed increased levels of natural helper cells (NHC) which could be important in the myeloma induction.