MAB1044 Sigma-AldrichAnti-Interleukin-8 Antibody, clone 8M6

After ovulation in the cow, the corpus luteum (CL) rapidly develops within a few days with angiogenesis and progesterone production. CL formation resembles an inflammatory response due to the influx of immune cells. Neutrophils play a role in host defense and inflammation, and secrete chemoattractants to stimulate angiogenesis. We therefore hypothesized that neutrophils infiltrate in the developing CL from just after ovulation and may play a role in angiogenesis of the CL.Polymorphonuclear neutrophils (PMN) were detected in CL tissue by Pas-staining, and interleukin-8 (IL-8, a neutrophil-specific chemoattractant) was measured in supernatant of the CL tissue culture: considerable amounts of PMNs and the high level of IL-8 were observed during the early luteal phase (days 1-4 of the estrous cycle). PMNs and IL-8 were low levels in the mid and late luteal phases, but IL-8 was increased during luteal regression. The PMN migration in vitro was stimulated by the supernatant from the early CL but not from the mid CL, and this activity was inhibited by neutralizing with an anti-IL-8 antibody, indicating the major role of IL-8 in inducing active PMN migration in the early CL. Moreover, IL-8 stimulated proliferation of CL-derived endothelial cells (LECs), and both the supernatant of activated PMNs and IL-8 stimulated formation of capillary-like structures of LECs.PMNs migrate into the early CL partially due to its major chemoattractant IL-8 produced at high levels in the CL, and PMNs is a potential regulator of angiogenesis together with IL-8 in developing CL in the cow.

Leukocyte infiltration and increased synthesis of cytokines in response to insemination is considered to enhance reproductive success. The present study investigated the inflammatory response to whole semen, spermatozoa and seminal plasma, with and without the addition of antibiotics, in the ovine uterus at oestrus and dioestrus. Seminal plasma and spermatozoa both contributed to increased IL-8 secretion (P < 0.01) by endometrial epithelial cells and a concurrent infiltration by neutrophils (P < 0.01). Increased GM-CSF secretion (P < 0.01) occurred in response to whole semen and spermatozoa when antibiotics were not used. Macrophages and eosinophils increased (P < 0.05) in the endometrial stroma when antibiotics were not used, and fewer mast cells were detected in the deep endometrial stroma after treatments containing antibiotics (P < 0.05). Neutrophil and IL-8 responses to insemination were greater at oestrus (P < 0.01) than at dioestrus and the GM-CSF response followed a similar trend. Eosinophil numbers were increased at oestrus (P < 0.01) but minimally affected by insemination. More macrophages were located in the superficial endometrial stroma at oestrus. These results indicate that spermatozoa, seminal plasma and possibly bacteria contribute to the post-insemination inflammatory response, and that leukocytes, GM-CSF and IL-8 secretion in the ovine uterus are influenced by ovarian hormones.

Cytokines produced in the female reproductive tract after mating may enhance reproductive success. The present study investigated the distribution of granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin (IL)-8 in tissues and luminal secretions from different sites in the reproductive tract of the ewe following oestrus and after natural mating. Fifteen ewes were mated with a ram for 1 h and their reproductive tracts collected 3, 6, 18, 24 or 48 h later. Another 15 ewes were used as oestrous controls. Luminal secretions and tissue samples were collected from seven sites in each reproductive tract. Secretions were analysed by enzyme-linked immunosorbent assay and tissues were stained immunohistochemically using anti-sheep GM-CSF and anti-sheep IL-8 antibodies. Both cytokines were found in luminal and glandular endometrial epithelium and, to a lesser extent, in cervical epithelium; neither was found in the vaginal epithelium. Twice as many (P < 0.05) luminal samples from mated ewes than non-mated ewes were positive for GM-CSF. The vaginal lumen contained significantly higher (P < 0.01) concentrations of IL-8 compared with other sites, irrespective of mating status. Significant differences (P < 0.05) were found in staining intensity of GM-CSF and IL-8 from different sites. Production of GM-CSF and IL-8 by reproductive tissues is likely to contribute to leucocyte infiltration into the ovine reproductive tract.