J Rheum Dis.  2021 Jul;28(3):133-142. 10.4078/jrd.2021.28.3.133.

Intravenous Administration of Toll-Like Receptor Inhibitory Peptide 1 is Effective for the Treatment of Systemic Lupus Erythematosus in a Mus musculus Model

• Baek WY
• Lee SM
• Lee SW
• Son IO
• Choi S
• Suh CH

• KMID: 2516949
• DOI: http://doi.org/10.4078/jrd.2021.28.3.133

Abstract

Objective
Systemic lupus erythematosus (SLE) is a common chronic autoimmune inflammatory disease. According to recent studies, signaling through Toll-like receptor (TLR) protein, which promotes the production of inflammatory cytokines, leads to the development of SLE. TLR-inhibitory peptide 1 (TIP1) has been newly identified for the treatment of autoimmune diseases.
Methods
The effect of TIP1 was analyzed in an SLE mouse model (MRL/lpr). The mice in the control treatment group (n=5) were administered an intravenous injection of phosphate-buffered saline twice weekly, whereas the mice in the TIP1 treatment group (n=6) were administered an intravenous injection of TIP1 (1 nmol/g) twice weekly. MRL/mpj mice (n=5) were selected as normal controls. The mice were injected for 4 weeks between 14 and 18 weeks of age, followed by assays of their spleen, kidneys, lymph nodes, serum, and urine.
Results
The antinuclear antibody and inflammatory cytokine (interferon-α) in the serum as well as levels of albumin in the urine of the mice in the TIP1 treatment group had decreased when compared to those of mice in the control treatment group. Kidney inflammation in mice in the TIP1 treatment group was alleviated. The mRNA expression levels of TLR7- or TLR9-related downstream signaling molecules also decreased in all organs of the mice in the TIP1 treatment group.
Conclusion
Intravenous treatment with TIP1 reduces symptoms and markers of inflammation in MRL/lpr mice. Hence, TIP1 is a promising medication for the treatment of SLE.

Keyword

Systemic lupus erythematosus; Inflammation; MRL lpr mice; Toll-like receptor

Figure

• 

  Figure 1 Effect of treatment with Toll-like receptor inhibitory peptide 1 (TIP1) in an SLE mouse model. (A) Whole body images of MRL/lpr mice and age- and sex-matched controls (MRL/mpj) are shown. These images show the (B) change in the weight of the mice during the 4-week injection period and (C) effect of TIP1 treatment on spleen hypertrophy in MRL/lpr mice. SLE: systemic lupus erythematosus. *p<0.05.

• 

  Figure 2 Toll-like receptor inhibitory peptide 1 (TIP1) treatment improved inflammation of the kidneys in lupus nephritis in MRL/lpr mice. (A) Urine albumin analysis. The results of the TIP1 treatment group and control treatment group were compared using the Mann–Whitney U-test. (B) H&E staining (×200) and (C) PAS staining (×200) in the kidney tissues of the MRL/mpj normal control group, MRL/lpr control treatment group, and MRL/lpr TIP1 treatment group. Representative micrographs obtained from H&E and PAS staining of kidneys show differences in the glomeruli and surrounding inflammatory cells in mice of each group. The shape of the glomerulus in the control treatment group was destroyed, but that was maintained in the TIP1 treatment group (circle).

• 

  Figure 3 Serological and inflammatory markers after treatment of MRL/lpr mice with Toll-like receptor inhibitory peptide 1 (TIP1). (A) Antinuclear antibody (ANA), (B) anti-ds DNA antibody, and (C) interferon-alpha (IFN-α). The results of the TIP1 treatment group and control treatment group were compared using the Mann–Whitney U-test. *p<0.05.

• 

  Figure 4 Toll-like receptor (TLR) inhibitory peptide (TIP) 1 blocks the TLR 7/9 downstream signaling pathway in major organs in mice. Western blot analysis of the expression levels the TLR 7/9 signaling pathway molecules in (A) spleen and (B) kidney of MRL/lpr mice. The bar graph shows the band intensity presented as the average value of the target protein to β-actin ratio (*p< 0.05, **p<0.01).

• 

  Figure 5 The mRNA expression level of proteins of the Toll-like receptor (TLR) 7/9 signaling pathway were analyzed using real-time polymerase chain reaction analysis in mouse (A) spleen, (B) kidney, and (C) lymph node. Using the Mann–Whitney U-test in the analysis, p-values less than 0.05 or 0.01 were considered statistically significant (*p<0.05, **p<0.01).

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