MHC class II tetramers identify peptide-specific human CD4+ T cells proliferating in response to influenza A antigen
Erik J. Novak, … , Gerald T. Nepom, William W. Kwok
Erik J. Novak, … , Gerald T. Nepom, William W. Kwok
Published December 15, 1999
Citation Information: J Clin Invest. 1999;104(12):R63-R67. https://doi.org/10.1172/JCI8476.
View: Text | PDF
Rapid Publication

MHC class II tetramers identify peptide-specific human CD4+ T cells proliferating in response to influenza A antigen

Abstract

Antigen-specific T helper cells present in peripheral blood at very low frequencies are capable of rapid clonal expansion during antigenic challenge. The exquisite specificity of this response provides for activation and expansion of a very select cohort of T cells, a feature we have used to directly identify and quantify human epitope-specific T helper cells from peripheral blood. Soluble tetramerized class II MHC molecules, loaded with an immunodominant peptide from hemagglutinin (HA) and labeled with fluorescent dyes, were constructed and used to directly identify antigen-specific T cells from influenza-immune individuals. After 7 days of proliferation in response to stimulation by HA peptide or whole influenza vaccine, cells staining positive with the HA tetramer had undergone between 6 and 9 divisions and were CD3+, CD4+, CD25+, and CD8–, characteristic of activated T helper cells responding to antigen. The HA epitope-specific component of the complex response to whole influenza vaccine represented a major subset of proliferating T helper cells. Soluble class II tetramers allow a direct approach for the analysis of immunodominant antigenic specificities. The identification of antigen-specific T helper cells in the peripheral blood provides a means for tracking the immune response against infectious agents and in autoimmune disease.

Authors

Erik J. Novak, Andrew W. Liu, Gerald T. Nepom, William W. Kwok

×

Figure 1

Options: View larger image (or click on image) Download as PowerPoint
Specificity and HLA restriction of HA307–319 tetramer. (a) Comparison wi...
Specificity and HLA restriction of HA307–319 tetramer. (a) Comparison with thymidine incorporation at 72 hours between the T-cell clone cultured with the DRA1*0101/DRB1*0401 transfected BLS-1 pulsed with no antigen (left bar) and 10 μg/mL of HA307–319 peptide (right bar). Error bars indicate SEM of triplicates. In b and c, clonal cells were stained with PE-labeled tetramer for 3 hours at 37°C, washed, stained with anti-CD4, washed again, and analyzed by flow cytometry. Staining with tetramer loaded with HA307–319 is shown in b, and tetramer loaded with TT830-843 peptide is shown in c. In both, cells are gated on forward and side scatter, the vertical axis shows tetramer fluorescence, and the horizontal axis shows CD4 fluorescence. Percentages shown in the margins of each panel represent the percent of total cells present in each quadrant.