The gamma/delta T lymphocytes represent a minority of T lymphocytes in human peripheral blood. Although there have been reports of reactivity against (myco-) bacterial antigens and heat shock proteins, their function and antigen specificity remain ill defined. The biological role of gamma/delta T cells has been related to functions within the 'first line of defense'. Similar to gamma/delta T lymphocytes in the T-cell compartment, CD5 positive B cells represent a small subset of B lymphocytes, which is thought to be involved in the maintenance of natural immunity and autoimmunity. We provide evidence for the cooperation of gamma/delta T cells and CD5 positive B cells in the proliferative response of gamma/delta T cells to bacterial antigens. Our data indicate a strong proliferation of V gamma 9 delta 2 T cells in response to gram-negative bacteria, which is dependent upon the presence of CD5 positive B-CLL or activated normal B lymphocytes. The selective stimulation of the V gamma 9 delta 2 subpopulation by gram-negative bacteria is also confirmed by analysis of different gamma/delta T-cell clones. The interaction of gamma/delta T cells with activated B cells and gram-negative bacteria may prove to be a useful model similar to the expansion of the V gamma 9 delta 2 subpopulation during development. In addition, our in vitro system should provide new insights in the interaction of CLL B cells with the immune system and the antigens recognized by gamma/delta T cells.