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AILIM/ICOS: its expression and functional analysis with monoclonal antibodies
S Sakamoto1, K Tezuka, T Tsuji
1Pharmaceutical Frontier Research Laboratories, JT Inc., 13-2, Fukuura 1-chome, Kanazawa-ku, Yokohama 236-0004, Japan. shinji.sakamoto@ims.jti.co.jp
Hybridoma and Hybridomics
|February 13, 2002
Summary
Researchers developed monoclonal antibodies to study Activation-inducible lymphocyte immuno-mediatory molecule (AILIM/ICOS). They found AILIM/ICOS expression varies across species and cell types, with significant co-stimulatory activity in human T-cell proliferation.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- Activation-inducible lymphocyte immuno-mediatory molecule (AILIM/ICOS) is a co-stimulatory molecule in the CD28/CTLA-4 family.
- AILIM/ICOS is an inducible cell surface glycoprotein expressed on activated lymphocytes.
- Understanding AILIM/ICOS expression is crucial for deciphering immune responses.
Purpose of the Study:
- To generate monoclonal antibodies (MAbs) against human, rat, and mouse AILIM/ICOS.
- To determine the expression profile of AILIM/ICOS across different species and cell types.
- To investigate the co-stimulatory function of AILIM/ICOS in T-cell proliferation.
Main Methods:
- Generation of species-specific monoclonal antibodies against AILIM/ICOS.
- Flow cytometry analysis of AILIM/ICOS expression on human PBMCs, rat and mouse splenocytes, lymph node cells, and thymocytes.
- Inhibition assays using MAbs to block AILIM/ICOS binding to its ligand (B7h-Ig).
- Assessment of T-cell proliferation assays stimulated by CD3 and AILIM/ICOS MAbs compared to CD3 and CD28.
Main Results:
- Generated specific MAbs for human, rat, and mouse AILIM/ICOS, demonstrating species-specificity.
- Low basal expression of AILIM/ICOS observed in human PBMCs, rat, and mouse splenocytes, primarily on T cells (human, mouse) and B cells (rat).
- AILIM/ICOS expression was detected on CD4+/CD8+ double positive T cells in rat thymocytes.
- All generated MAbs, except SG430, inhibited AILIM/ICOS binding to B7h-Ig.
- AILIM/ICOS demonstrated substantial co-stimulatory activity for human T-cell proliferation, comparable to CD28.
- Co-stimulatory potency of AILIM/ICOS in rats and mice was lower than that mediated by CD28.
Conclusions:
- The study successfully generated species-specific MAbs for AILIM/ICOS, enabling detailed expression profiling.
- AILIM/ICOS exhibits distinct expression patterns across different immune cell populations and species.
- AILIM/ICOS plays a significant role in human T-cell co-stimulation, with varying potency in rodents compared to CD28.