Cited by 936T-cell and B-cell ImmunologyMonoclonal and Polyclonal Antibodies ResearchArtificial Immune Systems Applications

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A Direct Estimate of the Human αβ T Cell Receptor Diversity

Arstila, T. Petteri, Casrouge, Armanda, Baron, Véronique, Even, Jos, Kanellopoulos, Jean, Kourilsky, Philippe·1999·Science

DOI 10.1126/science.286.5441.958

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Abstract

Generation and maintenance of an effective repertoire of T cell antigen receptors are essential to the immune system, yet the number of distinct T cell receptors (TCRs) expressed by the estimated 10 12 T cells in the human body is not known. In this study, TCR gene amplification and sequencing showed that there are about 10 6 different β chains in the blood, each pairing, on the average, with at least 25 different α chains. In the memory subset, the diversity decreased to 1 × 10 5 to 2 × 10 5 different β chains, each pairing with only a single α chain. Thus, the naı̈ve repertoire is highly diverse, whereas the memory compartment, here one-third of the T cell population, contributes less than 1 percent of the total diversity.

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Bacterial colonies were lifted on nylon filters (Hybond) lysed denatured fixed and then hybridized overnight at 63°C in 6× concentrated standard saline citrate 1 mM EDTA 0.5% SDS 0.1% bovine serum albumin preboiled salmon sperm DNA (100 μg/ml) with 10 pmol [γ- 32 P]-adenosine-5′-triphosphate–labeled oligonucleotides as recommended by Amersham Pharmacia Biotech and by J. Sambrook E. F. Fritsch T. Maniatis Molecular Cloning (Cold Spring Harbor Laboratory Press Cold Spring Harbor NY ed. 2 1989). The oligonucleotides were 20 to 22 mers each with melting temperature of 64°C. The first oligonucleotide mix consisted of 12 oligonucleotides and in control experiments hybridized to 87% of a mix of bacterial clones known to bear a relevant insert. We also tested two relevant clones separately and in both cases the mix hybridized to >90% of colonies although the strength of the signal was not uniform. Of 28 different clones with unrelated CDR3 sequences none gave a positive signal. The second mix consisted of five oligonucleotides. In random sequencing 19% of the clones in the first sample bore relevant inserts; the second oligonucleotide mix gave a detectable signal in 10% of the clones. We identified one relevant clone not detected by the mix and none of 16 irrelevant clones were positive.

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Authors: Arstila, T. P., Casrouge, A., Baron, V., Even, J., Kanellopoulos, J., & Kourilsky, P.
Affiliations: Inserm (FR)
Unité de Biologie Moléculaire du Gène, INSERM U277, Institut Pasteur, 75724 Paris Cedex 15, France.
Corresponding author: Arstila, T. Petteri
ORCID: Arstila, T. Petteri
Casrouge, Armanda
Kourilsky, Philippe
Venue: Science
Publisher: American Association for the Advancement of Science (AAAS)
Published: 1999-10-29
Volume: 286
Issue: 5441
Pages: 958-961
ISSN: 0036-8075 (ISSN-L)
1095-9203
DOI: 10.1126/science.286.5441.958
OpenAlex: W2105381977
Cited by: 931
References: 30
Citations by year: 2026: 25 · 2025: 28 · 2024: 29 · 2023: 24 · 2022: 26 · 2021: 32 · 2020: 57 · 2019: 42 · 2018: 42 · 2017: 38 · 2016: 51 · 2015: 45 · 2014: 42 · 2013: 49 · 2012: 49
FWCI: 8.21
Type: journal article
Language: en
Open access: closed
Locations: Science (publishedVersion)
PubMed (publishedVersion)
Topics: Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
Life Sciences › Immunology and Microbiology › Immunology › Monoclonal and Polyclonal Antibodies Research
Physical Sciences › Computer Science › Artificial Intelligence › Artificial Immune Systems Applications
Keywords: T cell receptor diversity, alpha beta T cells, T cell receptor repertoire, T cell receptor beta chain, TCR alpha chain, T cell memory, naive T cells, T cell receptor
MeSH terms: Adult, Female, Gene Amplification, Humans, Immunologic Memory, Male, T-Lymphocytes, Genetic Variation, Gene Rearrangement, T-Lymphocyte, Receptors, Antigen, T-Cell, alpha-beta

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A Direct Estimate of the Human αβ T Cell Receptor Diversity

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