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Infection and Immunity, August 1999, p. 3800-3809, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.

In Vitro Expansion of T-Cell-Receptor Valpha 2.3+ CD4+ T Lymphocytes in HLA-DR17(3), DQ2+ Individuals upon Stimulation with Mycobacterium tuberculosis

Semih Esin,1,2,* Giovanna Batoni,2,3 Güher Saruhan-Direskeneli,4 Robert A. Harris,1 Johan Grunewald,1 Manuela Pardini,2 Stefan B. Svenson,3,dagger Mario Campa,2 and Hans Wigzell1

Microbiology and Tumorbiology Center, Karolinska Institute, S-17177 Stockholm,1 and Swedish Institute for Infectious Disease Control, S-10521 Stockholm,3 Sweden; Dipartimento di Patologia Sperimentale, Biotecnologie Mediche, Infettivologia ed Epidemiologia, Università degli Studi di Pisa, I-56127 Pisa, Italy2; and Electro-Neurophysiology Center, Department of Physiology, Faculty of Medicine, University of Istanbul, 34390 Çapa, Istanbul, Turkey4

Received 6 November 1998/Returned for modification 8 January 1999/Accepted 11 May 1999

The T-cell receptor (TCR) Valpha /beta gene product expression upon in vitro stimulation with mycobacteria was investigated to assess whether T-cell proliferation was associated with any specific TCR V gene usage. T-cell-enriched populations from peripheral blood of Mycobacterium bovis BCG-vaccinated healthy blood donors were stimulated in vitro with live or killed M. tuberculosis or with a soluble extract thereof. TCR Valpha /beta repertoire analysis of reactive CD4+ and CD8+ T cells revealed a selective HLA-DR17(3), DQ2-restricted expansion of Valpha 2.3+ CD4+ T cells upon stimulation with live M. tuberculosis or its soluble extract. Third-complementarity-determining-region (CDR3) length analysis of the expanded Valpha 2.3+ T cells indicated an oligoclonal pattern with short CDR3 lengths in six of seven HLA-DR17(3), DQ2+ individuals tested. In addition, Valpha /Vbeta repertoire analysis of T lymphocytes from a DR17(3), DQ2+ donor before and after BCG vaccination revealed that positivity of skin test reactivity was associated with expansion of Valpha 2.3+ CD4+ T lymphocytes with preferential use of a short CDR3 peak length after in vitro stimulation. Separation of M. tuberculosis soluble extract by fast protein liquid chromatography (FPLC) purification indicated that fractions corresponding to molecular masses of 60 to 70 and 15 to 25 kDa were particularly effective in eliciting Valpha 2.3+ CD4+ T-cell expansion.

* Corresponding author. Present address: Dipartimento di Patologia Sperimentale, Biotecnologie Mediche, Infettivologia ed Epidemiologia, Università degli Studi di Pisa, Via San Zeno 35-39, I-56127 Pisa, Italy. Phone: 39050836565. Fax: 39050836570. E-mail: Semih.Esin{at}mtc.ki.se.

dagger Present address: Department of Veterinary Microbiology, Section of Bacteriology, Swedish University of Agricultural Sciences, S-75123 Uppsala, Sweden.

Infection and Immunity, August 1999, p. 3800-3809, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.


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