Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2001 Jan;102(1):39-43.
doi: 10.1046/j.1365-2567.2001.01148.x.

Functional activity of CD40 antibodies correlates to the position of binding relative to CD154

T A Barr  1 , A W Heath
Affiliations

Functional activity of CD40 antibodies correlates to the position of binding relative to CD154

T A Barr et al. Immunology. 2001 Jan.

Abstract

In this study we describe the characterization of a panel of 12 anti-mouse CD40 monoclonal antibodies (mAb). Characterization was performed in terms of antibody-binding site relative to the CD154 ligand, and the relationship between position and functional outcome of binding. The antibodies divided into three groups. The first were strong inhibitors of CD154 binding, and induced strong proliferative and activation signals to B cells. Two antibodies gave intermediary inhibition and comparable levels of activation. The remaining antibodies were found to bind outside the CD154 binding site and were poor inducers of B-cell activation. Data presented show a strong correlation between location of mAb binding and the resultant activation signal delivered. This correlation is shown to be independent of the isotype of the antibody involved and of its affinity. Implications of these findings are discussed.

PubMed Disclaimer

Figures

Figure 1
Figure 1
FACS histogram plot illustrating inhibition of CD154-CD8 binding to CD40 transfected fibroblast cells. The filled histogram represents the positive control (i.e. cells incubated with CD154-CD8 in the absence of blocking reagents) the thin, solid line represents the negative control (i.e. cells incubated with secondary reagents only), the thick, solid line represents complete inhibition by the 10C8 mAb and the broken line indicates partial inhibition by the mAb 1G7.
Figure 2
Figure 2
Scatter plots showing relationship between the ability of each mAb to induce expression of various B cell activation markers and its position of binding relative to the natural ligand, as indicated by percent inhibition of CD154 binding. The superimposed lines indicate curve fit as determined by Spearman's correlation coefficient. The critical rs value (P < 0·05) for a sample size of 12 is 0·576. (a) The activation/position relationship for CD23 expression (rs = 0·810). (b) Data for induction of ICAM-1 expression (rs = 0·613). (c) Data for MHC II expression (rs = 0·467, indicating a lack of correlation between these two variables). (d) Data from the B-cell proliferation assays, as determined by tritiated thymidine uptake assay (rs = 0·598).
See this image and copyright information in PMC

References

    1. Paulie S, Ehlin-Henriksson B, Mellstedt H, Koho H, Ben-Aissa H, Perlmann P. A p50 surface antigen restricted to human urinary bladder carcinomas and B lymphocytes. Cancer Immunol Immunother. 1985;20:23–8. - PMC - PubMed
    1. Armitage RJ, Fanslow WC, Strockbine L, et al. Molecular and biological characterisation of a murine ligand for CD40. Nature. 1992;357:80–2. - PubMed
    1. Allen RC, Armitage RJ, Conley ME, et al. CD40 ligand gene defects responsible for X-linked hyper–IgM syndrome. Science. 1993;259:990–3. - PubMed
    1. Korthauer U, Graf D, Mages HW, et al. Defective expression of T-cell CD40 ligand causes X-linked immunodeficiency with hyper-IgM. Nature. 1993;361:539–41. - PubMed
    1. Banchereau J, Bazan F, Blanchard D, et al. The CD40 antigen and its ligand. Ann Rev Immunol. 1994;12:881–922. - PubMed

Publication types