J. Exp. Med.
© The Rockefeller University Press
0022-1007/97/09/899/10 $2.00
Volume 186, Number 6, September 15, 1997 899-908

Major Histocompatibility Complex-independent Recognition of a Distinctive Pollen Antigen, Most Likely a Carbohydrate, by Human CD8⁺ / T Cells

By Silvia Corinti,^* Raffaele De Palma, Angelo Fontana,^§ Maria Cristina Gagliardi,^* Carlo Pini,^* and Federica Sallusto^*

From the ^* Laboratorio di Immunologia, Istituto Superiore di Sanità, I-00161 Roma;  Dipartimento di Internistica Clinica e Sperimentale, II Università di Napoli, I-80131 Napoli; and ^§ Istituto per la Chimica di Molecole di Interesse Biologica, Consiglio Nazionale delle Richerche, I-80072 Arco Felice, Napoli, Italy

We have isolated CD8⁺ / T cells from the blood of atopic and healthy individuals which recognize a nonpeptide antigen present in an allergenic extract from Parietaria judaica pollen. This antigen appears to be a carbohydrate because it is resistant to proteinase K and alkaline digestion, is hydrophilic, and is sensitive to trifluoromethane-sulphonic and periodic acids. In addition, on a reverse-phase high performance liquid chromatography column the antigen recognized by CD8⁺ T cells separates in a fraction which contains >80% hexoses (glucose and galactose) and undetectable amounts of proteins. Presentation of this putative carbohydrate antigen (Pj^CHOAg) to CD8⁺ T cell clones is dependent on live antigen presenting cells (APCs) pulsed for >1 h at 37°C, suggesting that the antigen has to be internalized and possibly processed. Indeed, fixed APCs or APCs pulsed at 15°C were both unable to induce T cell response. Remarkably, Pj^CHOAg presentation is independent of the expression of classical major histocompatibility complex (MHC) molecules or CD1. CD8⁺ T cells stimulated by Pj^CHOAg-pulsed APCs undergo a sustained [Ca²⁺]_i increase and downregulate their T cell antigen receptors (TCRs) in an antigen dose- and time-dependent fashion, similar to T cells stimulated by conventional ligands. Analysis of TCR V transcripts shows that six independent Pj^CHOAg-specific T cell clones carry the V8 segment with a conserved motif in the CDR3 region, indicating a structural requirement for recognition of this antigen. Finally, after activation, the CD8⁺ clones from the atopic patient express CD40L and produce high levels of interleukins 4 and 5, suggesting that the clones may have undergone a Th2-like polarization in vivo. These results reveal a new class of antigens which triggers T cells in an MHC-independent way, and these antigens appear to be carbohydrates. We suggest that this type of antigen may play a role in the immune response in vivo.

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