Gene Therapy of Arthritis with TCR Isolated from the Inflamed Paw

Abstract

In recent years, the treatment of autoimmune diseases has been significantly advanced by the use of biological agents. However, some biologics are accompanied with severe side effects, including tuberculosis and other types of infection. There is thus a critical need for nonsystemic and lesion-specific methods of delivering these therapeutic agents. We attempted to treat a mouse model of arthritis by using T cells that expressed a regulatory molecule and were specifically directed to the inflamed paw. To this end, we first identified the TCR αβ genes accumulating in the inflamed paw of mice with collagen-induced arthritis (CIA) by a combination of single-strand chain polymorphism analysis of TCR and single-cell sorting. We identified an expanded clone B47 which is autoreactive but is not specific to type II collagen. In vivo, TCR genes from B47-transduced T cells accumulated in the inflamed paw. Injection of cells cotransduced with the B47 and soluble TNFRIg genes resulted in a significant suppression of CIA. The suppression was correlated with the amount of TNFRIg transcripts in the hind paw, not with the serum concentrations of TNFRIg. Moreover, T cells cotransduced with the B47 and intracellular Foxp3 genes significantly suppressed CIA with reductions in TNF-α, IL-17A, and IL-1β expression and bone destruction. T cells cotransduced with B47 and Foxp3 genes also suppressed the progression of established CIA. Therefore, immunosuppressive therapy with autoreactive TCR is a promising therapeutic strategy for arthritis whether the TCRs are used to deliver either soluble or intracellular suppressive molecules.

CD4+CD25−LAG3+regulatory T cells controlled by the transcription factor Egr-2

“…The experimental groups consisted of CD4 ϩ T cells from BALB/c or DO11.10 mice transduced with pMIG or pMIG-Egr2. In accordance with our previous experiments (30,31), the average transduction efficiency was Ϸ50%. The GFP-positive fraction of the transduced CD4 ϩ T cells were sorted with FACSAria and transferred to immunized mice.…”

“…To examine the in vivo suppressive activity of Egr-2, we next performed the delayed-type hypersensitivity (DTH) reaction of BALB/c mice against chicken ovalbumin (OVA) by using T cells transduced with the Egr2 gene. The in vivo functions of T cells transduced with regulatory genes have been verified (30,31). In this experiment, CD4 ϩ T cells from BALB/c mice were transduced with pMIG or pMIG-Egr2.…”

Adoptive therapy with redirected primary regulatory T cells results in antigen-specific suppression of arthritis

“…In fact, recent experiments with TGF␤-induced polyclonal Tregs suggested that they were less effective than natural Tregs in ameliorating autoimmune gastritis (29). Although Ag-specific FoxP3-converted T cells failed to ameliorate arthritis to the same degree as natural Ag-specific Tregs in our hands, it is worth noting that studies carried out using a similar FoxP3-conversion technique in CIA demonstrated a significant suppression of arthritis progression (30). Furthermore, these studies identified a reduction in IL-17 transcripts within the arthritic paws of mice treated with Ag-specific FoxP3-converted T cells, however, in agreement with the data presented here, found no reduction in the draining lymph nodes.…”

Resistance to regulatory T cell-mediated suppression in rheumatoid arthritis can be bypassed by ectopic foxp3 expression in pathogenic synovial T cells

“…"switched" off by ectopic expression of foxp3, the transcription factor essential for natural Treg function? Although Tregs have received much attention because of their potential therapeutic use in autoimmune diseases, evidence suggests this will only be effective if the Treg population is specific to the relevant antigen (37)(38)(39)(40)(41). This is a problem in chronic inflammatory diseases such as RA, for which the autoantigen(s) driving disease perpetuation are not understood.…”