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Oligoclonality of V1 and V2 Cells in Human Peripheral Blood Mononuclear Cells: TCR Selection Is Not Altered by Stimulation with Gram-Negative Bacteria

Jian Shen^*, David M. Andrews^*, Franco Pandolfi, Lenora A. Boyle^*, Catalina M. Kersten^*, Robert N. Blatman and James T. Kurnick^1,*

^* Pathology Research Laboratory, Massachusetts General Hospital, Charlestown, MA 02129; Chair of Semeiotica Medica, Catholic University, Rome, Italy; Vincent Memorial Obstetrical Service, Massachusetts General Hospital, Boston, MA 02114

Despite the enormous potential repertoire of T cells, there are several observations which suggest that the expressed repertoire in the periphery of normal individuals is often quite restricted. To assess selective expansions among T cells from both adult and newborn blood samples, PBMC from 12 normal adults and cord blood from 15 normal newborns were analyzed for TCRDV1 and TCRDV2 junctional diversity by CDR3 size spectratyping and single-strand conformational polymorphism. Although TCRBV usage showed extensive heterogeneity in adults and newborns, both populations often showed CDR3 region restriction for TCRDV1 and TCRDV2. Analysis of the CDR3 spectratype patterns of newborn twins suggested that clonal selection for TCRDV is independent of genetic background. The possible role of Gram-negative bacteria in driving selective responsiveness of T cells in PBMCs from adults was examined by in vitro stimulation with Escherichia coli and Pseudomonas aeruginosa. Donors whose TCRDV repertoire was highly clonal in the unstimulated blood cells showed the same predominant clones among the bacteria-stimulated cultures. In individuals whose T cells were less restricted, in vitro stimulation did not select for clonality; rather, the TCRDV repertoires were similar before and after bacterial stimulation. Together, these data indicate that T cells are often clonally restricted in adults as well as in newborns and suggest that the prominent stimulatory activity of Gram-negative bacteria does not by itself account for the restriction or diversity of the T cell repertoire.