Peptides with cytokine-releasing activity and methods for characterizing cytokine-releasing profiles of peptides

An anti-inflammatory peptide (AIP) comprising an amino acid sequence of KLAKRLLLSMNQ (SEQ ID NO:

• 1), LALLAPKLLRRF (SEQ ID NO:
• 2), FAVPAKQAKKTLT (SEQ ID NO:
• 3), or an amino acid sequence that has 1, 2, or 3 amino acid substitutions, additions, or deletions relative to the amino acid sequences of KLAKRLLLSMNQ (SEQ ID NO:
• 1), LALLAPKLLRRF (SEQ ID NO:
• 2), or FAVPAKQAKKTLT (SEQ ID NO:
• 3), wherein the AIP consists of 10-16 amino acid residues.

An anti-inflammatory peptide (AIP) comprising an amino acid sequence of Formula I,

• (Formula I)
• X1-A-X3-X4-X5-X6-X7-X8-K-X10-T-X12-T,
• wherein
• X1 is F or H;
• X3 is V, Y, W, or F;
• X4 is P, Y, W, V, T, R, M, K, I, H, F, or E;
• X5 is A or W;
• X6 is K, Y, W, M, L, I, or F;
• X7 is Q or M;
• X8 is A or N;
• X10 is K, W, F, or C;
• and
• X12 is L, T, Q, E, or A.

An anti-inflammatory peptide (AIP) comprising an amino acid sequence of Formula II,

• (Formula II)
• L-X2-X3-X4-X5-X6-K-L-X9-X10-X11-X12
• wherein
• X2 is A, Y, W, V, R, Q, M, L, K, I, H, or F;
• X3 is L, W, or F;
• X4 is L, W, N, or C;
• X5 is A, W, or F;
• X6 is P, Y, W, V, T, S, R, Q, N, M, L, I, H, G, F, D, C, or A;
• X9 is L or W;
• X10 is R or S;
• X11 is R or Q;
• and
• X12 is F, V, T, S, R, Q, P, N, K, I, G, E, D, C, or A.

A method for characterizing a cytokine profile induced by a peptide, comprising:

• (a) isolating naïve CD4 positive T cells from peripheral blood mononuclear cells (PBMCs);
• (b) expanding the isolated naïve CD4 positive T cells;
• (c) co-culturing the expanded naïve CD4 positive T cells with PBMCs and stimulating the expanded naïve CD4 positive T cells one or more times with the peptide;
• and
• (d) detecting a signal corresponding to a level of one or more cytokines produced by the expanded naïve CD4 positive T cells.