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Naïve, hapten-specific human T lymphocytes are primed in vitro with derivatized blood mononuclear cells
1The Schepens Eye Research Institute and Department of Dermatology, Harvard Medical School, Boston, Massachusetts 02114, USA.
Our goal is to develop an in vitro system for priming naïve, hapten-specific human T cells similar to the effector cells that mediate contact hypersensitivity in vivo. Using strategies based on results of experiments conducted with murine cells, we stimulated peripheral blood T cells from normal human beings in vitro with peripheral blood mononuclear cells that had been pretreated with granulocyte-macrophage colony stimulating factor and then derivatized with dinitrochlorobenzene. T cells activated in this manner in vitro were compared functionally with immune T cells from individuals sensitized to dinitrochlorobenzene. We found that dinitrochlorobenzene-derivatized fresh peripheral blood mononuclear cells induced significant proliferation among immune T cells. Large amounts of interferon-gamma, but no interleukin-4, were detected in supernatant of these cultures. More interestingly, a significant number of T cells were activated when nave, hapten-specific T cells were stimulated with dinitrochlorobenzene-modified, cultured autologous peripheral blood mononuclear cells. Again, copious amounts of interferon-gamma were detected in the culture supernatants. Furthermore, this subset of T cells proliferated and secreted interferon-gamma, but not interleukin-4, when restimulated in vitro with dinitrochlorobenzene-derivatized fresh peripheral blood mononuclear cells. In these regards, in vitro activated T cells functionally resembled T cells that were harvested from the blood of individuals previously sensitized with dinitrochlorobenzene. This in vitro system for activation of naive, hapten-specific T cells offers opportunities on the one hand to study the earliest T cell and antigen-presenting cell events in the induction of human contact hypersensitivity, and on the other hand to simulate in vitro therapies with molecules designed to abort or prevent contact hypersensitivity when it causes disease.
Our goal is to develop an in vitro system for priming naïve, hapten-specific human T cells similar to the effector cells that mediate contact hypersensitivity in vivo. Using strategies based on results of experiments conducted with murine cells, we stimulated peripheral blood T cells from normal human beings in vitro with peripheral blood mononuclear cells that had been pretreated with granulocyte-macrophage colony stimulating factor and then derivatized with dinitrochlorobenzene. T cells activated in this manner in vitro were compared functionally with immune T cells from individuals sensitized to dinitrochlorobenzene. We found that dinitrochlorobenzene-derivatized fresh peripheral blood mononuclear cells induced significant proliferation among immune T cells. Large amounts of interferon-gamma, but no interleukin-4, were detected in supernatant of these cultures. More interestingly, a significant number of T cells were activated when nave, hapten-specific T cells were stimulated with dinitrochlorobenzene-modified, cultured autologous peripheral blood mononuclear cells. Again, copious amounts of interferon-gamma were detected in the culture supernatants. Furthermore, this subset of T cells proliferated and secreted interferon-gamma, but not interleukin-4, when restimulated in vitro with dinitrochlorobenzene-derivatized fresh peripheral blood mononuclear cells. In these regards, in vitro activated T cells functionally resembled T cells that were harvested from the blood of individuals previously sensitized with dinitrochlorobenzene. This in vitro system for activation of naive, hapten-specific T cells offers opportunities on the one hand to study the earliest T cell and antigen-presenting cell events in the induction of human contact hypersensitivity, and on the other hand to simulate in vitro therapies with molecules designed to abort or prevent contact hypersensitivity when it causes disease.