Lactobacillus spp. act in synergy to attenuate splenomegaly and lymphadenopathy in lupus-prone MRL/lpr mice

Abstract

Commensal bacteria and the immune system have a close and strong relationship that maintains a balance to control inflammation. Alterations of the microbiota, known as dysbiosis, can direct reactivity to self-antigens not only in the intestinal mucosa but also at the systemic level. Our laboratory previously reported gut dysbiosis, particularly lower abundance of bacteria in the family <i>Lactobacillaceae</i>, in lupus-prone MRL/<i>lpr</i> mice, a model of systemic autoimmunity. Restoring the microbiota with a mix of 5 different <i>Lactobacillus</i> species (spp.), <i>L. reuteri, L. oris, L. johnsonii, L. gasseri</i> and <i>L. rhamnosus</i>, attenuated lupus-liked clinical signs, including splenomegaly and lymphadenopathy. However, our understanding of the mechanism was limited. In this study, we first investigated the effects of individual species. Surprisingly, none of the species individually recapitulated the benefits of the mix. Instead, <i>Lactobacillus</i> spp. acted synergistically to attenuate splenomegaly and renal lymphadenopathy through secreted factors and a CX₃CR1-dependent mechanism. Interestingly, oral administration of MRS broth exerted the same benefits likely through increasing the relative abundance of endogenous <i>Lactobacillus</i> spp. Mechanistically, we found increased percentages of FOXP3-negative type 1 regulatory T cells with administration of the mix in both spleen and mesenteric lymph nodes. In addition, oral gavage of <i>Lactobacillus</i> spp. decreased the percentage of central memory T cells while increasing that of effector memory T cells in the lymphoid organs. Furthermore, a decreased percentage of double negative T cells was observed in the spleen with the mix. These results suggest that <i>Lactobacillus</i> spp. might act on T cells to attenuate splenomegaly and lymphadenopathy. Together, this study advances our understanding of how <i>Lactobacillus</i> spp. attenuate lupus in MRL/<i>lpr</i> mice. The synergistic action of these bacteria suggests that multiple probiotic bacteria in combination may dampen systemic autoimmunity and benefit lupus patients.