TCR expression and clonality analysis in peripheral blood and lymph nodes of HIV-infected patients.
We compared the T cell receptor (TCR) V beta gene family repertoire in peripheral blood mononuclear cells (PBMC) and lymph node (LN) cells from 7 human immunodeficiency virus (HIV)-infected patients and 3 seronegative healthy controls. Virtually all the V beta family specificities were represented in patient PBMC and LN cells, and mean values for each specificity were comparable to figures in seronegative controls. In 4 patients, however, some V beta gene segment transcripts were overrepresented in the LN compartment, compared to the peripheral blood counterpart. To ascertain whether this phenomenon was due to polyclonal or oligoclonal expansion of T cells bearing the relevant V beta gene product, we sequenced the entire CDR3 region of a panel of 238 PCR clones corresponding to the V beta transcripts expanded in LN; as control, the same regions were cloned and sequenced in patient's PBMC, and in PBMC and LN cells from seronegative individuals. This analysis disclosed preferential usage of J beta 2 genes in PBMC and LN cells from both seropositive patients and controls, regardless of the V beta gene segment considered, thus indicating that this skewness in the V beta-J beta repertoire could be a consistent feature of at least a part of the V beta repertoire in different lymphoid compartments, regardless of the pathologic conditions. In addition, in LN from HIV seropositive patients we found the presence of recurrent TCR rearrangements, accounting for 8-23% of the generated clones, in each of the 4 V beta specificities analyzed; recurrent sequences were not found in PBMC from patients nor in PBMC and LN cells from seronegative controls. These findings suggest that antigen-driven oligoclonal T cell expansions may occur in vivo in lymphoid organs of HIV seropositive patients.
Mion
M
M
Department of Oncology and Surgical Sciences, University of Padova, Italy.
Indraccolo
S
S
Feroli
F
F
Minuzzo
S
S
Masiero
S
S
Zamarchi
R
R
Barelli
A
A
Borri
A
A
Chieco-Bianchi
L
L
Amadori
A
A
D018791:CD4 Lymphocyte Count; D002999:Clone Cells; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D015333:Gene Rearrangement, beta-Chain T-Cell Antigen Receptor; D019674:Genes, T-Cell Receptor beta; D015658:HIV Infections / Q000097:blood* / Q000276:immunology*; D018023:HIV Seronegativity; D006801:Humans; D008198:Lymph Nodes / Q000276:immunology*; D016133:Polymerase Chain Reaction; D012313:RNA / Q000235:genetics; D012367:RNA, Viral / Q000032:analysis; D017423:Sequence Analysis, RNA; D013601:T-Lymphocytes / Q000276:immunology*; D014158:Transcription, Genetic
D018076:DNA, Complementary; D012367:RNA, Viral; D012313:RNA