Skip to main content

Open Access Detection of Transgene Product in Mouse Peripheral Blood Lymphocytes: A Minimally Invasive Microassay

Download Article:
 Download
(PDF 79.083984375 kb)
 

Abstract:

Confirmation of transgene-coded protein expression on the surface of mouse white blood cells has traditionally involved use of spleen and lymph nodes as a cell source. However, this eliminates the mice from further analysis and increases the number of animals needed for testing. A minimally invasive method of cell collection would allow these mice to remain in the colony, thereby conserving valuable resources. The goal of the study reported here was to detect transgene expression on mouse white blood cells collected from peripheral blood. Mice over-expressing the T-cell receptor (TCR) transgene product V8.2 on the surface of T lymphocytes were compared with wild-type control mice. Mononuclear cells were counted, using a hemacytometer. Peripheral blood cells were stained with anti-V8.2 fluorescent conjugate and were evaluated, using fluorescence-activated (FA) microscopy as well as flow cytometry (fluorescence-activated cell sorting [FACS]). Mean ± SD number of mononuclear cells was 5.34 ± 1.72 × 105 cells/100 ml from TCR Tg mice and 4.99 ± 1.99 × 105 cells/100 l from control mice. Expression of transgene V8.2 was confirmed by use of FA microscopy and FACS. Mean population of lymphocytes expressing V8.2 was 43.0 ± 12.9% from TCR Tg blood samples, and 9.4 ± 1.0% from control blood samples. Spleen and lymph node specimens also were compared. Results of the unpaired t test and the Mann-Whitney test indicated significant difference in the amount of expressed V8.2 between transgenic and wild-type mice. It is concluded that this microassay can serve as a quick, minimally invasive, non-lethal alternative for detecting TCR transgene-encoded antigens on the surface of mouse lymphocytes, and is potentially applicable to a number of other transgene-coded cell surface proteins.

Document Type: Research Article

Affiliations: 1: Department of Comparative Medicine, University of Washington School of Medicine, Seattle, Washington 2: Department of Comparative Medicine and Comparative Mouse Genomics Center, University of Washington, School of Medicine, Seattle, Washington

Publication date: 2002-12-01

More about this publication?
  • Comparative Medicine (CM), an international journal of comparative and experimental medicine, is the leading English-language publication in the field and is ranked by the Science Citation Index in the upper third of all scientific journals. The mission of CM is to disseminate high-quality, peer-reviewed information that expands biomedical knowledge and promotes human and animal health through the study of laboratory animal disease, animal models of disease, and basic biologic mechanisms related to disease in people and animals.

    Attention Members: To access the full text of the articles, be sure you are logged in to the AALAS website.

    Attention: please note, due to a temporary technical problem, reference linking within the content is not available at this time

  • Editorial Board
  • Information for Authors
  • Submit a Paper
  • Subscribe to this Title
  • Membership Information
  • Information for Advertisers
  • For issues prior to 1998
  • Institutional Subscription Activation
  • Ingenta Connect is not responsible for the content or availability of external websites
  • Access Key
  • Free ContentFree content
  • Partial Free ContentPartial Free content
  • New ContentNew content
  • Open Access ContentOpen access content
  • Partial Open Access ContentPartial Open access content
  • Subscribed ContentSubscribed content
  • Partial Subscribed ContentPartial Subscribed content
  • Free Trial ContentFree trial content
Cookie Policy
X
Cookie Policy
Ingenta Connect website makes use of cookies so as to keep track of data that you have filled in. I am Happy with this Find out more