Core - Technical Support Facility

Noelle, Randolph J.

Abstract

The purpose of the Technical Support Core Facility is to provide the technological expertise to: quantitate cellular subsets in cell suspensions derived from reproductive tract tissues, determine the expression of cytokines in and their receptors on cells derived from reproductive tract tissues and analyze the expression of cytokine mRNA and cell surface antigens in tissue sections from the reproductive tract. This Core Facility is composed of three units: (1) Flow cytometry service; (2) Cytokine analysis service and (3) In situ analysis/image analysis service. This facility has been developed as a central service for Program Investigators to provide standardized technology to meet specific scientific needs. The functions of the 3 components of the facility can be summarized as follows: 1. Flow Cytometry Service: The function of this component is to perform comparative multi-parameter flow cytofluorometric analysis on single cell suspensions from reproductive tract tissues and peripheral blood cells. This facility will maintain a panel of monoclonal antibodies (mabs) and, using titered amounts of these antibodies, stain and analyze samples on the cytometer, and report data. In addition, this facility will prepare samples for sterile and non=sterile sorting and sort cells into required sub-populations. 2. Cytokine Analysis Service: The purpose of this component is to provide a multi-tiered approach to the quantitation of cytokine protein and mRNA. Reverse PCR will be applied as a semi-quantitative means to estimate mRNA levels for cytokines and cytokine receptors. Bioassays that specifically measure biologically active cytokine will be used for the detection of cytokines in tissue culture fluids. Finally, flow cytofluorometric analysis of cytokine-expressing cells will be provided. 3. Immunohistochemistry and In SITU Analysis Service: This component of the Core will provide the technology for the immunohistochemical and in situ analysis of cell surface antigen expression and the identification of cytokine-producing cells. With the recent success in confocal image analysis, expanded support for this technology will be provided.

Funding Agency

• Agency: National Institute of Health (NIH)
• Institute: National Institute of Allergy and Infectious Diseases (NIAID)
• Type: Research Program Projects (P01)
• Project #: 5P01AI034478-08
• Application #: 6421833
• Project Start: 2001-03-01
• Project End: 2003-02-28
• Support Year: 8
• Fiscal Year: 2001

Institution

• Name: Dartmouth College
• DUNS #: 041027822
• City: Hanover
• State: NH
• Country: United States
• Zip Code: 03755

Publications

Haddad, Severina N; Wira, Charles R (2014) Estradiol regulation of constitutive and keratinocyte growth factor-induced CCL20 and CXCL1 secretion by mouse uterine epithelial cells. Am J Reprod Immunol 72:34-44

Wira, Charles R; Rossoll, Richard M; Young, Roger C (2005) Polarized uterine epithelial cells preferentially present antigen at the basolateral surface: role of stromal cells in regulating class II-mediated epithelial cell antigen presentation. J Immunol 175:1795-804

Fahey, John V; Rossoll, Richard M; Wira, Charles R (2005) Sex hormone regulation of anti-bacterial activity in rat uterine secretions and apical release of anti-bacterial factor(s) by uterine epithelial cells in culture. J Steroid Biochem Mol Biol 93:59-66

Asin, Susana N; Fanger, Michael W; Wildt-Perinic, Dunja et al. (2004) Transmission of HIV-1 by primary human uterine epithelial cells and stromal fibroblasts. J Infect Dis 190:236-45

Yeaman, Grant R; Asin, Susana; Weldon, Sally et al. (2004) Chemokine receptor expression in the human ectocervix: implications for infection by the human immunodeficiency virus-type I. Immunology 113:524-33

Yeaman, Grant R; Howell, Alexandra L; Weldon, Sally et al. (2003) Human immunodeficiency virus receptor and coreceptor expression on human uterine epithelial cells: regulation of expression during the menstrual cycle and implications for human immunodeficiency virus infection. Immunology 109:137-46

Wira, Charles R; Fahey, John V; Abrahams, Vikki M et al. (2003) Influence of stage of the reproductive cycle and estradiol on thymus cell antigen presentation. J Steroid Biochem Mol Biol 84:79-87

Wira, Charles R; Rossoll, Richard M (2003) Oestradiol regulation of antigen presentation by uterine stromal cells: role of transforming growth factor-beta production by epithelial cells in mediating antigen-presenting cell function. Immunology 109:398-406

Abrahams, Vikki M; Collins, Jane E; Wira, Charles R et al. (2003) Inhibition of human polymorphonuclear cell oxidative burst by 17-beta-estradiol and 2,3,7,8-tetrachlorodibenzo-p-dioxin. Am J Reprod Immunol 50:463-72

Fahey, John V; Wira, Charles R (2002) Effect of menstrual status on antibacterial activity and secretory leukocyte protease inhibitor production by human uterine epithelial cells in culture. J Infect Dis 185:1606-13