Source: WASHINGTON STATE UNIVERSITY submitted to
REGULATION OF EXPRESSION OF CLASSICAL AND NON-CLASSICAL MAJOR HISTOCOMPATIBILITY COMPLEX CLASS I GENES IN BOVINE TROPHOBLAST CELLS
Sponsoring Institution
National Institute of Food and Agriculture
Project Status
NEW
Funding Source
Reporting Frequency
Annual
Accession No.
0204973
Grant No.
(N/A)
Project No.
WNV-DAVIESCJ
Proposal No.
(N/A)
Multistate No.
(N/A)
Program Code
(N/A)
Project Start Date
Jul 1, 2005
Project End Date
Jun 30, 2006
Grant Year
(N/A)
Project Director
Davies, C. J.
Recipient Organization
WASHINGTON STATE UNIVERSITY
240 FRENCH ADMINISTRATION BLDG
PULLMAN,WA 99164-0001
Performing Department
ANIMAL HEALTH RESEARCH CENTER
Non Technical Summary
We have shown that inappropriate expression of major histocompatibility complex antigens by the bovine placenta is associated with the induction of immune mediated abortion. Furthermore, we have established that the bovine placenta expresses both classical and non-classical (immunosuppressive) major histocompatibility complex antigens. The purpose of this project is to establish the default state for major histocompatibility complex gene expression in the bovine placenta. In addition, we will determine if any of the bovine non-classical major histocompatibility antigens are produced in a secreted, rather than a cell associated, form.
Animal Health Component
100%
Research Effort Categories
Basic
80%
Applied
20%
Developmental
(N/A)
Classification

Knowledge Area (KA)Subject of Investigation (SOI)Field of Science (FOS)Percent
3013310103010%
3013310104010%
3013310105010%
3013310108010%
3013310109010%
3013410103010%
3013410104010%
3013410105010%
3013410108010%
3013410109010%
Goals / Objectives
This project has two objectives: (1) to establish a trophoblast cell culture system and determine whether major histocompatibility complex class I expression is induced or suppressed when trophoblast cells are put in culture; and (2) to determine if bovine trophoblast cells produce a secreted form of any of the non-classical major histocompatibility complex class I isoforms.
Project Methods
This proposal is based on the general hypothesis that appropriate regulation of classical and non-classical, major histocompatibility complex class I genes in bovine trophoblast cells is required for the establishment and maintenance of normal placentation. The project addresses two working hypotheses: (1) that when bovine trophoblast cells are placed in tissue culture, they will commence expression of both classical and non-classical major histocompatibility complex class I antigens regardless of the stage of pregnancy and region of the placenta from which they were isolated; and (2) that bovine trophoblast cells produce a secreted form of at least one non-classical major histocompatibility complex class I antigen that is analogous to the secreted forms of human HLA-G and murine Qa-2. The project's specific aims correspond directly to the two working hypotheses and are: (1) establish a trophoblast cell culture system and determine whether major histocompatibility complex class I expression is induced or suppressed when trophoblast cells are put in culture; and (2) determine if bovine trophoblast cells produce a secreted form of any of the non-classical major histocompatibility complex class I isoforms. Specific aim #1 will be addressed by: isolating cotyledonary and intercotyledonary trophoblast cells from six placentas of three different gestational ages, culturing the trophoblast cells for up to two weeks, measuring major histocompatibility complex class I protein expression by flow cytometry, and characterizing major histocompatibility complex class I message by realtime RT-PCR and microarray screening. Specific aim #2 will be addressed by expressing a bovine non-classical MHC-I allele as a recombinant protein and using this protein to develop an ELISA that will then be used to test tissue culture supernatants for secreted major histocompatibility complex class I isoforms.