Lipoteichoic acid inhibits interleukin-2 (IL-2) function by direct binding to IL-2.

Lipoteichoic acid (LTA) is associated with the cell envelope of most gram-positive bacteria. Although previously thought to act mainly as a virulence factor by virtue of its adhesive nature, evidence is now provided that LTA can also suppress the function of interleukin-2 (IL-2), an autocrine growth factor for T cells. LTA from four separate bacterial strains lowered the levels of detectable IL-2 during a peripheral blood mononuclear cell response to the antigen tetanus toxoid (TT). T-cell proliferation in response to TT was similarly inhibited by LTA. In contrast, levels of detectable gamma interferon increased. In addition, LTA inhibited IL-2 detection by enzyme-linked immunosorbent assay (ELISA) and blocked the proliferative response of an IL-2-dependent T-cell line to soluble IL-2. Further studies using ELISA demonstrated that LTA blocks IL-2 detection and function by binding directly to IL-2. Flow cytometric analysis revealed that IL-2 binding to T cells is inhibited in the presence of purified LTA but not LTA plus anti-LTA monoclonal antibody. In summary, these studies demonstrate a novel effect of LTA on the immune response through direct binding to IL-2 and inhibition of IL-2 function. Importantly, gram-positive organisms from which LTA is obtained not only play an important role in the pathology of diseases such as bacterial endocarditis, septic shock, acute respiratory distress syndrome, and multiple organ failure but also comprise a significant portion of commensal populations within the human host. Inhibition of IL-2 function by LTA may represent yet another mechanism by which gram-positive bacteria dampen the host immune response and facilitate survival. Thus, LTA provides a potential target for therapeutic intervention when gram-positive organisms are involved.

Two distinct stages in the transition from naive CD4 T cells to effectors, early antigen-dependent and late cytokine-driven expansion and differentiation.

Efficient peptide presentation by professional APC to naive and effector CD4 T cells in vitro is limited to the first 1-2 days of culture, but is nonetheless optimum for effector expansion and cytokine production. In fact, prolonging Ag presentation leads to high levels of T cell death, decreased effector expansion, and decreased cytokine production by recovered effectors. Despite the absence of Ag presentation beyond day 2, T cell division continues at a constant rate throughout the 4-day culture. The Ag-independent later stage depends on the presence of IL-2, and we conclude optimum effector generation depends on an initial 2 days of TCR stimulation followed by an additional 2 days of Ag-independent, cytokine driven T cell expansion and differentiation.

Differences in IgG subclass do not effect immune complex-enhanced T cell activation despite differential binding to antigen presenting cells.

Ag presentation to CD4 T cells is a critical event in the generation of protective immunity. IgG, in the form of IgG-pathogen (Ag) complexes, is capable of mediating FcgammaR-dependent Ag presentation, and thereby enhanced T cell activation. Therefore, it is important to understand the ability of the individual human IgG subclasses to function in enhanced T cell activation. We hypothesized that increased delivery of Ag to monocyte FcgammaR by high affinity human IgG subclasses, IgG1 and IgG3, would lead to increased Ag presentation, as compared to low affinity IgG subclasses, IgG2 and IgG4. To create immune complexes, we linked biotinylated IgG subclasses to biotinylated Ag via an avidin bridge, and examined T cell responses to them. Although IgG2- and IgG4-Ag complexes bound to monocytes at significantly lower levels than those made with IgG1 and IgG3, we observed no significant difference in the ability of the four human IgG subclasses to mediate enhanced T cell activation. Studies suggest the explanation for this dichotomy lies within the first 24 h of Ag processing, and that processing efficiency may vary with IgG subclass. They also suggest the existence of a highly efficient, and selective processing pathway, which is dependent on IgG subclass, and can compensate for low level production and FcgammaR binding of IgG2- and IgG4-Ag complexes.

Inhibition of interleukin-2 by a Gram-positive bacterium, Streptococcus mutans.

Generation of an effective cellular immune response is key to the successful development of both humoral and cellular immune defences against most pathogens. However, while the type of cellular immune response elicited by any given pathogen is dictated by the entire array of antigens and molecules which comprise that pathogen, most studies of human immune responses to bacterial pathogens tend to focus on selected antigens. This is a result, in part, of a desire to find those antigens that will generate a desired immune response, as well as limited technology for monitoring the complex array of responses generated by an intact organism. Utilizing Streptococcus mutans as a model Gram-positive organism, a novel flow cytometric assay that permits the identification of individual cells within a responding population, and highly sensitive cytokine assays, we show for the first time that CD8 T cells and natural killer (NK) cells comprise a significant component of the response to this organism in humans. This is despite the fact that CD8 T cells are traditionally thought to respond to endogenously derived antigens only. In addition, we provide the first evidence that a Gram-positive organism can actively inhibit interleukin-2 (IL-2), an important autocrine growth factor for T cells. The latter observation could represent an additional mechanism by which Gram-positive organisms evade host defences.