Journal: Scientific Reports

Article Title: Suppressor of cytokine signaling-1 mimetic peptides attenuate lymphocyte activation in the MRL/lpr mouse autoimmune model

doi: 10.1038/s41598-021-86017-4

Figure Lengend Snippet: SOCS1-KIR mimetic peptides mitigate MRL/lpr T lymphocyte activation in vitro. Single cell suspensions of pooled MRL/lpr lymph nodes were stimulated with anti-CD3/anti-CD28 in the presence or absence of SOCS1-KIR monomer mimetic, SOCS1-KIR dimer mimetic, or pJAK2 peptides for 5 days. ( A – F ) Quantification (left) of representative flow cytometric analysis (right) depicting changes in CD25, CD69, and CD44 surface expression on MRL/lpr T lymphocytes. Data are from two independent experiments, with 4 animals per group (graphs display average frequency, error bars, SD). *p < 0.05, **p < 0.01, and ***p < 0.001 (One-way ANOVA with Dunnett’s multiple comparison test). The figure was prepared in Graphpad prism v9 [ https://www.graphpad.com/scientific-software/prism/ ].

Article Snippet: The SOCS1-KIR mimetic peptide ( 53 DTHFRTFRSHSDYRRI), SOCS1-KIR dimeric variant (DTHFRTFRSHSDYRRIGGGGGDTHFRTFRSHSDYRRI), or pJAK2 ( 1001 LPQDKEYYKVKEP) was generated in-house using Applied Biosystems 431a automated peptide synthesizer (Applied Biosystems, Carlsbad, CA) by conventional fluorenylmethylcarbonyl chemical methods as described , , or purchased from GenScript (Piscataway, NJ) at 95% purity.

Techniques: Activation Assay, In Vitro, Expressing, Comparison, Software

Journal: Scientific Reports

Article Title: Suppressor of cytokine signaling-1 mimetic peptides attenuate lymphocyte activation in the MRL/lpr mouse autoimmune model

doi: 10.1038/s41598-021-86017-4

Figure Lengend Snippet: SOCS1-KIR mimetic peptides mitigate MRL/lpr T lymphocyte activation in vitro. Single cell suspensions of pooled MRL/lpr lymph nodes were stimulated with anti-CD3/anti-CD28 in the presence or absence of SOCS1-KIR monomer mimetic, SOCS1-KIR dimer mimetic, or pJAK2 peptides for 5 days. ( A – F ) Quantification (left) of representative flow cytometric analysis (right) depicting changes in CD25, CD69, and CD44 surface expression on MRL/lpr T lymphocytes. Data are from two independent experiments, with 4 animals per group (graphs display average frequency, error bars, SD). *p < 0.05, **p < 0.01, and ***p < 0.001 (One-way ANOVA with Dunnett’s multiple comparison test). The figure was prepared in Graphpad prism v9 [ https://www.graphpad.com/scientific-software/prism/ ].

Article Snippet: The SOCS1-KIR mimetic peptide ( 53 DTHFRTFRSHSDYRRI), SOCS1-KIR dimeric variant (DTHFRTFRSHSDYRRIGGGGGDTHFRTFRSHSDYRRI), or pJAK2 ( 1001 LPQDKEYYKVKEP) was generated in-house using Applied Biosystems 431a automated peptide synthesizer (Applied Biosystems, Carlsbad, CA) by conventional fluorenylmethylcarbonyl chemical methods as described , , or purchased from GenScript (Piscataway, NJ) at 95% purity.

Techniques: Activation Assay, In Vitro, Expressing, Comparison, Software

Journal: Scientific Reports

Article Title: Suppressor of cytokine signaling-1 mimetic peptides attenuate lymphocyte activation in the MRL/lpr mouse autoimmune model

doi: 10.1038/s41598-021-86017-4

Figure Lengend Snippet: SOCS1-KIR treatment reduces interferon gamma production by activated T cell subsets. LN and spleen were isolated from MRL/lpr mice and cultured for 5 days under indicated conditions. ( A , B ) Quantification of flow cytometric analysis. Cell populations were gated for the co-expression of CD4 or CD8 with CD25 and IFN-γ. ( C ) Fold change of ifng mRNA expression relative to GAPDH after 5 days of culture. ( D ) IFN-γ ELISA performed on supernatants after cell culture for 5 days. ( E ) Fold change of H2-Aa mRNA expression relative to GAPDH after culture for 5 days. The results were compiled from two independent experiments with 3–5 animals per treatment group. RT-qPCR and ELISA assays were performed in triplicate (error bars, SD). *p < 0.05, **p < 0.01, and ***p < 0.001 (One-way ANOVA with Dunnett’s multiple comparison test). The figure was prepared in Graphpad prism v9 [ https://www.graphpad.com/scientific-software/prism/ ].

Article Snippet: The SOCS1-KIR mimetic peptide ( 53 DTHFRTFRSHSDYRRI), SOCS1-KIR dimeric variant (DTHFRTFRSHSDYRRIGGGGGDTHFRTFRSHSDYRRI), or pJAK2 ( 1001 LPQDKEYYKVKEP) was generated in-house using Applied Biosystems 431a automated peptide synthesizer (Applied Biosystems, Carlsbad, CA) by conventional fluorenylmethylcarbonyl chemical methods as described , , or purchased from GenScript (Piscataway, NJ) at 95% purity.

Techniques: Isolation, Cell Culture, Expressing, Enzyme-linked Immunosorbent Assay, Quantitative RT-PCR, Comparison, Software

Journal: Scientific Reports

Article Title: Suppressor of cytokine signaling-1 mimetic peptides attenuate lymphocyte activation in the MRL/lpr mouse autoimmune model

doi: 10.1038/s41598-021-86017-4

Figure Lengend Snippet: Peptide administration decreases the frequency of memory T lymphocytes within peripheral blood and secondary lymphoid organs, while decreasing splenomegaly and lymphadenopathy in vivo. ( A ) Line graph displaying weight of animals throughout the study (left). Total weight of lymph nodes and spleens extracted from treated and untreated animals at sacrifice (middle, right). Each symbol represents an individual mouse. ( B ) Bar graph showing the percentage of CD4 + and CD8 + cells that are double positive for CD44 and IFN-γ within peripheral blood of SOCS1-KIR treated and untreated mice over time. ( C ) Bar graphs showing the frequency of CD4 + , CD8 + , or CD4 − CD8 − that are also CD44 + cells within the LN and spleen of treated or untreated mice at 15 weeks. Data are from two independent experiments with four animals per group (error bars, SD). *p < 0.05, **p < 0.01, and ***p < 0.001 (two-way ANOVA with sidak’s multiple comparison test ( A ) and unpaired student’s t-test ( B , C )). The figure was prepared in Graphpad prism v9 [ https://www.graphpad.com/ ].

Article Snippet: The SOCS1-KIR mimetic peptide ( 53 DTHFRTFRSHSDYRRI), SOCS1-KIR dimeric variant (DTHFRTFRSHSDYRRIGGGGGDTHFRTFRSHSDYRRI), or pJAK2 ( 1001 LPQDKEYYKVKEP) was generated in-house using Applied Biosystems 431a automated peptide synthesizer (Applied Biosystems, Carlsbad, CA) by conventional fluorenylmethylcarbonyl chemical methods as described , , or purchased from GenScript (Piscataway, NJ) at 95% purity.

Techniques: In Vivo, Comparison

Journal: Scientific Reports

Article Title: Suppressor of cytokine signaling-1 mimetic peptides attenuate lymphocyte activation in the MRL/lpr mouse autoimmune model

doi: 10.1038/s41598-021-86017-4

Figure Lengend Snippet: SOCS1 KIR mimetic peptide treatment enhances Foxp3 expression in regulatory T cells, attenuates T cell activation, and reduces germinal center B cells in vivo. ( A ) CD4 + Foxp3 + T regulatory cells and Foxp3 MFI. ( B ) CD4 + T follicular regulatory cells [CD4 + PD1 + CXCR5 + Bcl6 + Foxp3 + ] were analyzed by flow cytometry. ( C ) Ratio of CD4 + and CD8 + effector memory (CD44 + CD62L − ) to naïve T cells (CD44 − CD62L + ). ( D ) Frequency of Germinal Center B cells (CD19 + GL7 + ) and CD80 + Plasma cells (CD19 − CD138 + ). Each cohort had 9–10 animals. (Error bars, SD) *p < 0.05, **p < 0.01, ***p < 0.001 (One-way ANOVA followed by dunnett’s multiple comparison test)]. The histograms were prepared in FlowJo v10 [ https://www.flowjo.com/solutions/flowjo ]. All other graphs were prepared in Graphpad prism v9 [ https://www.graphpad.com/scientific-software/prism/ ].

Article Snippet: The SOCS1-KIR mimetic peptide ( 53 DTHFRTFRSHSDYRRI), SOCS1-KIR dimeric variant (DTHFRTFRSHSDYRRIGGGGGDTHFRTFRSHSDYRRI), or pJAK2 ( 1001 LPQDKEYYKVKEP) was generated in-house using Applied Biosystems 431a automated peptide synthesizer (Applied Biosystems, Carlsbad, CA) by conventional fluorenylmethylcarbonyl chemical methods as described , , or purchased from GenScript (Piscataway, NJ) at 95% purity.

Techniques: Expressing, Activation Assay, In Vivo, Flow Cytometry, Clinical Proteomics, Comparison, Software

Journal: Scientific Reports

Article Title: Suppressor of cytokine signaling-1 mimetic peptides attenuate lymphocyte activation in the MRL/lpr mouse autoimmune model

doi: 10.1038/s41598-021-86017-4

Figure Lengend Snippet: SOCS1-KIR mediated lymphocyte changes correlated to reduced anti-DNA IgG production. Linear correlation was evaluated for the relative frequency of CD8+ naïve T cells and anti-dsDNA IgG. Pearson’s correlation coefficient (R) and p values are indicated on the graphs. The figure was prepared in Graphpad prism v9 [ https://www.graphpad.com/scientific-software/prism/ ].

Article Snippet: The SOCS1-KIR mimetic peptide ( 53 DTHFRTFRSHSDYRRI), SOCS1-KIR dimeric variant (DTHFRTFRSHSDYRRIGGGGGDTHFRTFRSHSDYRRI), or pJAK2 ( 1001 LPQDKEYYKVKEP) was generated in-house using Applied Biosystems 431a automated peptide synthesizer (Applied Biosystems, Carlsbad, CA) by conventional fluorenylmethylcarbonyl chemical methods as described , , or purchased from GenScript (Piscataway, NJ) at 95% purity.

Techniques: Software