CD3 bright lymphocyte population reveal gammadelta T cells.

BACKGROUND

In routine CD3/CD4/CD8 T-cell analysis, a CD3 bright population of lymphocytes is frequently observed. The aim of the present study was to identify the immunological significance of such CD3 bright lymphocytes.

METHODS

We analyzed samples from 31 healthy adult volunteers, 78 human immunodeficiency virus (HIV)-positive, and 78 renal transplanted patients.

RESULTS

A clearly distinct CD3 bright (frequently CD4-/CD8-) T-cell fraction was observed in 84% of donors and was directly correlated with the fraction of gammadelta T cells (r2 = 0.64). CD3 overexpression on gammadelta T cells was confirmed by a combination of monoclonal antibody staining (CD3-ECD, gammadeltaTCR-FITC, and alphabetaTCR-PE-Cy5) or immunomagnetic purification of gammadelta T cells (i.e., MdFI 20 vs 8.86). The gammadelta T cells expressed CD8 polypeptide chains (alpha and beta) in all possible combinations. The largest proportion, surprisingly, were cells expressing CD8betabeta homodimers (43.8 +/- 16.5%). CD8alphaalpha homodimers were expressed on 14.2% (+/- 12.3) of total gammadelta T cells, whereas CD8alphabeta heterodimers were expressed on 12.2% (+/- 7.5). We also observed a bimodal distribution of the intensity of CD3 fluorescence of gammadelta T cells in immunocompromised patients with a threshold at 105 cell/microl. CD3 bright gammadelta T cells were more frequently observed in HIV patients (29%) compared with renal transplant patients (11%) and healthy donors (3%; chi2 test: P = 0.0007).

CONCLUSIONS

The simple observation of a CD3 bright T-cell subset on CD3/CD4/CD8 routine analysis suggests a high gammadelta T-cell fraction and, in our opinion, should be followed by a complementary analysis to determine precisely the number of gammadelta T cells and to identify their CD8alpha/beta phenotype. When CD3 bright T cells/microl were more than 40%, high gammadelta T cells were detected in more than 87% of cases, with a specificity of 76%. Occasionally, the CD3 bright subset appeared to be strongly homogeneous, suggesting an oligoclonal proliferation that could possibly reveal a chronic localized stimulation or an early lymphoproliferative disorder. Because the gammadelta T cells have interesting immunological peculiarities, the clinical significance of their quantitative abnormality should be clarified in diseases such as HIV, organ transplantation, autoimmunity and lymphoma.