Short communication: Proinflammatory gene expression relative to the collection technique of endometrial samples from cows with and without subclinical endometritis.

Uterine inflammation negatively affects reproductive performance and is an important cause of infertility and subfertility in dairy cows. Several studies have investigated the use of gene expression in endometrial samples collected by biopsy or cytology to evaluate the inflammatory response of the cow uterus. This study aimed to compare the expression of the CCL5, CXCL8, IL6, and IL1B genes in the bovine endometrium according to the site of sample collection [caruncular (C) or intercaruncular (IC)], the collection method (biopsy or cytology), and the category of inflammation based on endometrial cytology (zero, medium, or high) in subclinical endometritis. The reproductive tracts of dairy cows were collected from a slaughterhouse, and punch-biopsy samples of endometrial tissues were obtained from both regions (C and IC). Endometrial cells from these regions were collected with the cytobrush technique and then used for the analysis of mRNA expression by quantitative PCR. After counting polymorphonuclear cells (PMN) by endometrial cytology, 20 uteri with an ovary at stage I (d 1-4 of estrous cycle) were categorized into 3 groups. Uteri with 0% PMN (n = 10) were assigned to group zero, uteri with 5 to 15% PMN (n = 5) to group medium (12.2 ± 1.6% PMN), and uteri with >15% PMN (n = 5) to group high (53.8 ± 32.9% PMN). All data were analyzed with 2-way ANOVA with Bonferroni multiple comparison post test. The results from gene transcripts demonstrated that the region (C or IC) of the endometrial biopsy had no influence on any of the degrees of inflammatory reaction observed. However, gene expression was more elevated in the endometrium of cows with greater inflammation compared with those without inflammation (CCL5, CXCL8, IL6, IL1B) and those with medium inflammation (CCL5, IL6). Expression of the genes evaluated did not differ between the endometrium without inflammation and with medium inflammation. However, in the high inflammation group, all genes were comparatively more expressed in samples collected by cytology relative to those derived from biopsies for both anatomical regions. In conclusion, gene expression did not differ between the C and IC tissue. Samples collected from animals with greater inflammation had greater gene expression than those with zero or medium inflammation. In addition, cytology samples had greater gene expression than biopsy samples in the high inflammation group.

Immunophenotyping of leukocytes in bovine placenta / Imunofenotipagem de leucócitos na placenta bovina

Lymphocytes and macrophages are the main white cells involved in fetal-maternal tolerance. Little is known about these leukocytes in bovine placenta, such as the quantity and location of these cells. Thus, the objective of this study was to identify lymphocyte and macrophage populations in bovine placenta using specific markers and flow cytometry. This study analyzed samples of placentomes and intercaruncular regions of cows in the three quarters of pregnancy. In the placentomes, during the first quarter of pregnancy, mean percentage of labeled CD3+ cells was 2.34%; CD8+, 1.28%; CD14+, 1.66%; and CD335+, 0.96%. For the intercaruncular region, percentage of CD3+ cells was 0.71%; CD8+, 1.63%; CD14+, 2.81%; and CD335+, 2.81%. In the second quarter, placentomes showed 0.94% CD3+ cells; 0.77% CD8+; 0.72% CD14+; and 0.51% CD335+. In the intercaruncular region, percentage of labeled CD3+ cells was 0.50%; CD8+, 1.81%; CD14+, 2.64%; and CD335+, 0.51%. In the third quarter, placentomes showed labeling of 0.88% CD3+; 0.66% CD8+; 1.06% CD14+; and 0.74% CD335+ cells. In the intercaruncular region, percentage of labeled CD3+ cells was 0.19%; CD8+, 2.23%; CD14+, 2.43%; and CD335+, 0.16%. The results showed that there was a greater immunomarking of leukocytes CD3+ and CD335+ in the placentome when compared to the intercaruncular region during the third trimester. It can be concluded that leukocytes populations in bovine placenta is reduced, probably because of the syndesmochorial characteristic of bovine placenta. This represents a significant barrier for the immunological system of the mother, sharply decreasing the exposure of the conceptus to the mother's immune system.(AU)

Linfócitos e macrófagos são os principais leucócitos envolvidos na tolerância materno-fetal. Pouco se sabe sobre esses leucócitos na placenta bovina, como por exemplo, a quantidade e localização dessas células. Assim, o objetivo desse estudo foi identificar populações de linfócitos e macrófagos na placenta bovina utilizando marcadores específicos e citometria de fluxo. Este estudo analisou amostras de placentônios da região intercaruncular de bovinos nos três trimestres da gestação. No primeiro trimestre, nos placentônios, a porcentagem média de células CD3+ foi 2,34%; CD8+, 1,28%; CD14+, 1,66%; e CD335+, 0,96%. Na região intercaruncular, a porcentagem de células CD3+ foi 0,71%; CD8+, 1,63%; CD14+, 2,81%; e CD335+, 2,81%. No segundo trimestre, os placentônios apresentaram 0,94% de células CD3+; 0,77% de CD8+; 0,72% de CD14+e 0,51% de CD335+. Na região intercaruncular, a porcentagem de células CD3+ foi 0,50%; CD8+, 1,81%; CD14+, 2,64%; e CD335+, 0,51%. No terceiro trimestre, os placentônios apresentaram 0,88% de células marcadas CD3+; 0,66% de CD8+; 1,06% de CD14+ e 0,74% de CD335+. Na região intercaruncular, a porcentagem de células CD3+ foi 0,19%; CD8+, 2,23%; CD14+, 2,43% e CD335+, 0,16%. Os resultados mostraram que a imunomarcação de leucócitos na região do placentônio foi maior do que na região intercaruncular no terceiro trimestre. Concluiu-se que a população de leucócitos CD3+ e CD335+ na placenta bovina está reduzida, provavelmente devido à sua característica sindesmocorial. Essa característica representa uma barreira significante para o sistema imunológico da mãe, o que diminui drasticamente a exposição do concepto ao sistema de defesa da mãe.(AU)