[PMC free article] [PubMed] [Google Scholar] 41

[PMC free article] [PubMed] [Google Scholar] 41. of a monoclonal CD3 antibody (e.g. clone 145-2C11)29, 30. This antibody recognizes the Rabbit Polyclonal to ZNF691 CD3 molecule and activates T cells in the absence of antigen, since it evades the TCR antigen-specific acknowledgement mechanism31, 32. Herein, we hypothesized that this administration of a monoclonal CD3 antibody (clone 145-2C11) in late gestation will cause pathological inflammation by initiating innate and adaptive immune responses which, in turn, could lead to preterm labor and birth. The aim of this study was to determine whether T cell activation via a monoclonal CD3 antibody induces preterm labor and birth. Also, we examined whether administration of this antibody would cause fetal death or fetal compromise using Doppler ultrasound. MATERIALS AND METHODS Animals C57BL/6J (B6) mice were purchased from your Jackson Laboratory (Bar Harbor, ME, USA) and bred in the animal care facility at the C.S. Mott Center for Human Growth and Morin hydrate Development at Wayne State University or college, Detroit, Michigan, USA. Mice were housed under a circadian cycle (light: dark=12:12 h). Eight- to 12-week-old females were mated with males of confirmed fertility. Females were examined daily between 8:00 a.m. and 9:00 a.m. for the presence of a vaginal plug, which indicated 0.5 days (dpc). Upon observation of a vaginal plug, females were housed separately from males, their excess weight was monitored, and a gain of two or more grams by 12.5 dpc confirmed pregnancy. Procedures were approved by the Institutional Animal Care and Use Committee at Wayne State University (Protocol No. A 09-08-12). Intraperitoneal administration of Morin hydrate a monoclonal CD3 antibody Pregnant B6 mice were intraperitoneally injected with 10g of a purified anti-mouse CD3 (CD3) (BD Biosciences, San Jose, CA, USA, Clone 145-2C11; n=12) dissolved in 200L of sterile 1X phosphate-buffered saline (PBS) on 16.5 dpc. Controls were injected with 10g of isotype (IgG1 Isotype; BD Biosciences, Clone A19-3; n=8) dissolved in 200L of sterile PBS on 16.5 dpc. Following injection, mice were monitored using a video video camera with an infrared light (Sony Corporation, Tokyo, Japan) until delivery. End result variables Preterm labor/birth was defined as delivery occurring before 18.0 dpc, and its rate was represented by the percentage of females delivering preterm among those delivering at term (19.5 0.5 dpc). Gestational age was defined as the time elapsed from your detection of the vaginal plug (0.5 dpc) through the delivery of the first pup. The rate of pup mortality at birth was defined as the percentage of pups found dead among the total litter size. imaging by ultrasound Pregnant B6 mice were intraperitoneally injected with a monoclonal CD3 antibody or its isotype control on 16.5 dpc (n=12-13 each). Sixteen hours post-injection (prior to preterm labor/birthin mice injected with CD3), ultrasound was performed, as previously described.33,34 Mice were anesthetized by inhalation of 2%-3% of isoflurane (Aerrane; Baxter Healthcare Corporation, Deerfield, IL, USA) and of 1-2 L/min of oxygen in an induction chamber. Anesthesia was managed with a mixture of 1.5%-2% of isoflurane and 1.5-2 L/min of oxygen. Mice were positioned on a heated platform and stabilized using adhesive tape. Fur was removed from the stomach and thorax following the application of Nair cream (Church & Morin hydrate Dwight Co., Inc., Ewing, NJ, USA) to those areas. Body temperature was managed at 371C and monitored using a rectal probe. Respiratory and heart rates were monitored by electrodes embedded in the heated platform. An ultrasound probe was fixed and mobilized with a mechanical holder, and the transducer was slowly relocated toward the stomach. Fetal heart rate and umbilical artery pulsatility index (PI) were examined with the 55MHz linear ultrasound probe (VisualSonics Inc., Toronto, ON, Canada). Umbilical artery PI was calculated using the following formula: PI = (systolic velocity – diastolic velocity / mean velocity). Ultrasound signals were processed, displayed, and stored using the Vevo Imaging Station.