CD40 Ligand in Pathogenesis of Autoimmune Ovarian Disease of Day 3-Thymectomized Mice: Implication for CD40 Ligand Antibody Therapy

Abstract

The blockade of CD40 ligand (CD40L) is effective in autoimmune disease prevention. Recently, a brief period of CD40L mAb treatment was reported to induce tolerance and enhancement of CD4+CD25+ regulatory T cell activity. We therefore determined the efficacy of CD40L mAb treatment in autoimmunity that resulted from CD4+CD25+ regulatory T cell deficiency. Autoimmune ovarian disease (AOD) and oocyte autoantibody response of day 3-thymectomized (d3tx) mice were inhibited by continuous CD40L mAb treatment from day 3, or from days 10–14, whereas CD40L mAb treatment confined to the neonatal week was ineffective. The enhanced expression of memory markers (CD44 and CD62Llow) on CD4+ T cells of the d3tx mice was unaffected by CD40L mAb treatment. In contrast, their increased T cell activation markers (CD69 and CD25) were eliminated by CD40L mAb treatment. Moreover, ex vivo activated T cells of d3tx mice expressed elevated intracellular IFN-γ, and this was also blocked by CD40L mAb. The memory T cells, although nonpathogenic in CD40L mAb-positive environment, transferred severe AOD to CD40L mAb− neonatal recipients. Most importantly, CD40L mAb treatment inhibited AOD in recipients of T cells from d3tx donors with severe AOD and led to regression of AOD in d3tx mice documented at 4 wk. Therefore, 1) the continuous presence of CD40L mAb both prevents and causes regression of AOD in the d3tx mice; and 2) the multiple steps of the d3tx autoimmune disease, including T cell activation, cytokine production, T cell-mediated inflammation, and tissue injury, are CD40L dependent.

Continuous control of autoimmune disease by antigen-dependent polyclonal CD4+CD25+ regulatory T cells in the regional lymph node

“…Serum diluted 1:50 in PBS, was incubated with frozen sections of adult mouse ovaries prefixed in 95% ethanol and blocked by goat serum, followed by incubation with FITC-conjugated goat anti–mouse IgG (Jackson ImmunoResearch Laboratories). Inflammatory cells in normal and diseased ovaries were identified by immunofluorescence using the TSA Biotin System (PerkinElmer), with antibody to CD5 (53–7.313), CD4 (GK1.5), CD8 (53–6.7) T cells, macrophage (F4/80), B cell (B220), and MHC class II (M5/114.15-2) for activated macrophages and dendritic cells, as described previously ( 56 ).…”

SHP‐1 and SHP‐2 in T cells: two phosphatases functioning at many levels

“…CD4 + CD25 + Treg cells produce IL‐10 and TGF‐β; however at least in in vitro assays, cytokine secretion is not essential for suppressive activity (194). In vivo , Treg cells have been implicated in the control of a variety of autoimmune diseases, such as intestinal inflammation, autoimmune gastritis, diabetes, ovarian disease, arthritis, and experimental autoimmune encephalomyelitis (193, 221–225). While many studies have addressed the in vivo and in vitro function of Treg cells, the intracellular signaling of Treg cells are still relatively poorly understood.…”

Neonatal Autoimmune Disease: Influence of CD4+CD25+Regulatory T Cells

“…Selection of the target organ was determined by the non-major histocompatibility complex (MHC) genetic background of the inbred strains [9,10]. The autoimmune disease in d3tx mice is mediated by CD4 þ T cells [11] that respond to endogenous antigenic stimulation [12] and are crucially dependent on CD40 ligand costimulation operative at multiple steps of disease pathogenesis [13]. Because d3tx disease is inhibited by normal adult spleen cells or purified adult CD4 þ CD25 þ T cells, it is surmised that the naturally occurring CD4 þ CD25 þ T cells in normal mice prevent spontaneous autoimmunity that is mediated by naturally occurring pathogenic T cells.…”