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Updated: Oct 18, 2026
Production of Monoclonal Antibodies Targeting Aminopeptidase N in the Porcine Intestinal Mucosal Epithelium
Published on: May 18, 2021
Production and characterization of monoclonal antibodies against pigtailed macaque (Macaca nemestrina) cell adhesion
J E Hildreth1, J L Mankowski, C Zink
1Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA. jhildret@jhmi.edu
Abstract:
Our previous in vitro studies indicate a significant role for cell adhesion molecules in the biology of HIV-1 and HTLV-1. Confirmation of the involvement of these molecules in the pathogenesis of retrovirus infection in vivo will require a suitable animal model. The SIV/pigtailed macaque (Macaca nemestrina) model of acquired immunodeficiency syndrome (AIDS) is an ideal system in which to study adhesion molecules and viral pathogenesis. The monoclonal antibodies (MAbs) against human adhesion molecules previously produced in our laboratory either do not react with or fail to block function of pigtailed macaque adhesion molecules. We have used papiovirus-transformed pigtailed macaque B cells as immunogen to generate murine MAbs against macaque adhesion molecules including ICAM-1, VCAM-1, and LFA-1. The specificity of the MAbs was confirmed by immunoprecipitation from lysates of vectorially iodinated cells, flow cytometry analysis of transfected cell lines and primary cells, binding assays on recombinant soluble human VCAM-1 and ICAM-1, and by inhibition of adhesion functions. MAbs against ICAM-1 and VCAM-1 showed positive staining of fixed tissue in immunohistochemistry studies. The same antibodies also blocked the function of these two adhesion molecules. The new MAbs can be used to study the tissue expression of adhesion molecules in SIV-infected animals as well as to test the involvement of these molecules in virus infection. Thus they should prove invaluable as probes of the role of cell adhesion molecules in AIDS pathogenesis in an animal model.
Insights
New antibodies were developed to study cell adhesion molecules in the pigtailed macaque model of AIDS. These tools will help investigate the role of adhesion molecules in simian immunodeficiency virus (SIV) infection and AIDS pathogenesis.
Area of Science:
- Immunology
- Virology
- Pathogenesis
Background:
- Cell adhesion molecules play a crucial role in HIV-1 and HTLV-1 infection in vitro.
- An in vivo animal model is necessary to confirm the role of these molecules in retroviral pathogenesis.
- The pigtailed macaque model of acquired immunodeficiency syndrome (AIDS) is suitable for studying adhesion molecules and viral pathogenesis.
Purpose of the Study:
- To generate murine monoclonal antibodies (MAbs) against pigtailed macaque adhesion molecules, specifically ICAM-1, VCAM-1, and LFA-1.
- To validate the specificity and functionality of these newly generated MAbs.
- To enable the study of adhesion molecule tissue expression and function in the context of SIV infection.
Main Methods:
- Generation of murine MAbs using papiovirus-transformed pigtailed macaque B cells as immunogen.
- Specificity confirmation via immunoprecipitation, flow cytometry, binding assays, and functional adhesion inhibition assays.
- Immunohistochemistry to assess tissue staining and functional blocking studies.
Main Results:
- Successfully generated MAbs against macaque ICAM-1, VCAM-1, and LFA-1.
- Confirmed MAb specificity and function, demonstrating their ability to block adhesion.
- MAbs against ICAM-1 and VCAM-1 showed positive staining in immunohistochemistry and inhibited adhesion functions.
Conclusions:
- The developed MAbs are valuable tools for investigating the role of cell adhesion molecules in AIDS pathogenesis.
- These MAbs will facilitate studies on tissue expression and functional involvement of adhesion molecules in SIV infection.
- The new reagents are essential for advancing research in the SIV/pigtailed macaque model of AIDS.
