Hsp65-Producing Lactococcocus lactis Prevents Antigen-Induced Arthritis in Mice

Abstract

Oral tolerance is the physiological process that enables the immune system to differentiate between harmless dietary and microbiota antigens from pathogen derived antigens. It develops at the mucosal surfaces and can result in local and systemic regulatory and anti-inflammatory effects. Translation of these benefits to the clinical practice faces limitations involving specificity and doses of antigen as well as regimens of feeding. To circumvent these problems, we developed a recombinant Hsp65 delivered by the acid lactic bacteria Lactococcus lactis NCDO 2118 directy in the intestinal mucosa. Hsp65 is a ubiquitous protein overexpressed in inflamed tissues and capable of inducing immunoregulatory mechanisms. L. lactis has probiotic properties and is commonly and safely used in dairy products. In this study, we showed that continuous delivery of HSP65 in the gut mucosa by L. lactis is a potent tolerogenic stimulus inducing regulatory CD4 + LAP + T cells that prevented collagen-induced and methylated bovine serum albumin-induced arthritis in mice. Clinical and histological signs of arthritis were inhibited as well as levels of inflammatory cytokines such as IL-17 and IFN-γ, serum titers of anti-collagen antibodies and rheumatoid factor. Oral administration of L. lactis induced alterations in microbiota composition toward an increased abundance of anaerobic bacteria such as Bifidobacterium and Lactobacillus. Tolerance to HSP65 and arthritis prevention induced by the recombinant L. lactis was associated with increase in IL-10 production by B cells and it was dependent on LAP + T cells, IL-10 and TLR2 signaling. Therefore, HSP65-producing treatment induced effective tolerance and prevented arthritis development suggesting it can be used as a therapeutic tool for autoimmune diseases.

Use of genetically modified lactic acid bacteria and bifidobacteria as live delivery vectors for human and animal health

“… 150 Two different studies used a strain of L. lactis engineered to deliver IL-5 and Hsp-65, respectively, and the GM strain was able to attenuate or prevent collagen-induced arthritis in mice. 151 , 152 …”

Oral tolerance as antigen-specific immunotherapy

“…These cells can be either FoxP3 + or FoxP3 - [ 78 ]. LAP + iTregs exert their suppressive role in a TGF-β-dependent manner [ 79 ] and neutralization of these cells by intravenous administration of anti-LAP antibody prevents oral tolerance induction in murine models of experimental autoimmune encephalomyelitis (EAE) [ 14 , 80 ] and arthritis [ 81 ]. TGF-β activation in these cells involves release of mature TGF-β from LAP which may occur via Glycoprotein A repetitions predominant (GARP).…”

“…The indirect effects of oral tolerance induction were demonstrated using OVA-fed mice in which granuloma reaction caused by the helminth Schistosoma mansoni and cutaneous scar formation were inhibited by simultaneous injection of OVA at the time of infection/inflammation induction [ 140 , 141 ]. In addition, our group has also used a recombinant Hsp65-producing Lactococcus lactis to prevent and treat inflammatory disease models such as EAE [ 14 ], DSS-induced colitis[ 142 ] and antigen-induced arthritis [ 81 ] showing that Hsp65-induced Tregs are able to inhibit inflammatory reactions towards myelin antigens, colonic antigens and collagen. It has been proposed that bystander suppression is due to the simultaneous presentation of unrelated antigens by the same APC in the draining lymph node [ 10 ].…”

“…In the same line, a recent developed recombinant Hsp65-producing Lactococcus lactis NCDO2118 is a slow-delivery strategy used by our group to prevent and control inflammatory diseases. This recombinant lactic bacteria lodges in the duodenum and induces high frequencies of CD4 + Foxp3 + and CD4 + LAP + iTreg in mesenteric lymph nodes that can be found also in distant peripheral lymphoid organs [ 14 , 81 , 142 ]. The anti-inflammatory probiotic properties of lactic bacteria such as L. lactis combined with its ability to promote the slow delivery of antigen in a highly tolerogenic segment of the intestine may render these bacteria a potential efficient alternative adjuvant for antigen-specific OIT.…”

The engineered probiotics for the treatment of chronic diseases: A systematic review

“…Regarding the role of immune response against HSP65 in the pathogenesis of CVD, arthritis, and other autoimmune diseases, EP‐HSP65 can be a novel therapeutic approach to the treatment of diseases. In fact, EPs have a high ability to induce OT, and numerous studies have used this ability to treat chronic diseases (Gomes‐Santos et al, 2017; Gusmao‐Silva et al, 2020; Ma et al, 2014). On the other hand, many studies have used EPs as vaccine, anti‐inflammatory cytokine, and bioactive compound carrier (Fensterle et al, 2008; McLean et al, 2017; Tseng et al, 2020).…”