Engineered IL-2 FC fusion protein

Abstract

The present invention provides an IL-2-Fc fusion protein comprising an IL-2 variant.

Description

[0001] Priority claim

[0002] This application claims the benefit of U.S. Provisional Application No. 62 / 607,850, filed December 19, 2017, and U.S. Provisional Application No. 62 / 675,070, filed May 22, 2018, which are hereby expressly incorporated by reference in their entireties, particularly with reference to the drawings, legends, and claims therein.

[0003] Sequence Listing

[0004] This application contains a sequence listing that has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. The ASCII copy, created on November 30, 2018, is named 067461-5217-WO_ST25.txt and is 605,038 bytes in size. Background Art

[0005] Immune system homeostasis depends on a good balance between multiple immune cell populations, including CD8+ T cells and CD4+ T cells (CD3+CD25-FOXP3-) and regulatory T cells (Treg; CD3+CD4+CD25+FOXP3+). Disruption of this balance may lead to diseases such as autoimmune diseases in which T cells remain unregulated and attack the body's own tissues. Under normal conditions, Treg regulates T cell differentiation and effector and cytotoxic functions. Therefore, the main premise in this regard is that defects in Treg cell number and / or function are contributing factors to the disease state. Thus, the ability to change the balance between cytotoxicity and regulation by fine-tuning T cell responses has great potential in the treatment of autoimmune diseases and other diseases.

[0006] IL-2 contributes to the proliferation and differentiation of B cells, T cells, and NK cells. IL-2 is also essential for Treg function and survival. IL-2 exerts its cell signaling function by binding to a high-affinity trimeric receptor complex composed of three different proteins: the common gamma chain (γc; CD132) and the IL-2 receptor B chain (IL-2Rβ; CD122) shared with IL-15, and a unique alpha chain receptor (IL-2Rγ; CD25). IL-2 can also exert its cell signaling function by binding to an intermediate-affinity dimeric receptor complex composed only of IL-2Rβ and γc (IL-2Rβγ).

[0007] Since the concentration of IL-2 normally present in tissues is low, IL-2 preferentially activates cells expressing a high-affinity receptor complex (CD25:CD122:CD132; IL-2Rγβγ) and therefore prefers FOXP3+ Tregs that constitutively express CD25. However, IL-2 can also activate and induce the proliferation of FOXP3- T cells that express an intermediate-affinity receptor complex (CD122:CD132; IL-2Rβγ). FOXP3- T cells such as CD4+ T cells or CD8+ T cells can cause inflammation, autoimmunity, organ transplant rejection, or graft-versus-host disease. Since IL-2 promotes or reduces the potential of both T cells and Tregs under limited selection, there is a strong need in the art to produce more selective Treg regulators. In addition, as a potential drug, IL-2 is cleared very quickly, with a half-life of several minutes, which hinders advantageous administration. The present invention solves these two problems by providing a novel IL-2-Fc fusion protein.

[0008] Therefore, there is a need to provide useful IL-2 variants and Fc fusion proteins. Summary of the Invention

[0009] Thus, in some aspects, the present disclosure provides compositions comprising a variant human IL-2 protein (compared to SEQ ID NO: 2), wherein the variant IL-2 protein comprises one or more amino acid substitutions selected from the group consisting of: T3A, R38A; R38D; R38E; R38F; R38G; R38H; R38I; R38K; R38L; R38M; R38N; R38P; R38Q; R38S; R38T; R38V; R38W; R38Y; T41A; T41D; T41E; T41F; T41G; T41H; T41I; T41K; T41L; T41M; T41N; T41P; T41Q; T41R; T41S; T41V; T41W; T41Y; F42A; F42D; F42E ;F42G;F42H;F42I;F42K;F42L;F42M;F42N;F42P;F42Q;F42R;F42S;F42T ;F42V; F42W; F42Y; R38Q / T41K; R38Q / T41Q; R38E / T41K; R38Q / T41R; R38N / T41Q; R38Q / T41V; R38N / T41V; R38Q / T41M; R38Q / T41S; R38Q / T41L; R38N / T41M; T41I / F42Y; T41E / F42Y′T41D / F42Y; T41M / F42Y; 41Q / F42Y; T41E / F4 2H; T41E / F42L; T41E / F42P; R38Q / F42Y; R38N / T41R; R38N / T41K; R38V / T4 1R; R38P / T41R; T41E / F42K; T41D / F42K; T41M / F42K; T41Q / F42K; R38Q / F4 2K; T41I / F42K; R38N / F42K; T41H / F42K; R38Q / T41K / F42Y; R38Q / T41R / F4 2Y; R38Q / T41Q / F42Y; R38Q / T41V / F42Y; R38N / T41K / F42K; R38Q / T41H / F42 K; R38Q / T41K / F42K; R38Q / T41Q / F42K; 38Q / T41V / F42K; R38Q / T41R / F42K ;Q11E; L12D; Q13E; E15Q; H16Y; L19D; D20N; N29S / Y31H / K35R / T37A / R38L / K48E / V69A / N71R / Q74P / N88D / I89V / Q126T; Q22E; K35R; T37S; K43R; F44Y ;Y45F;K48R;K49E;E61Q;E62Q;K64R;E68Q;V69L;L72I;R81D;D84N;S87T;N88D;V91L;I92L;E95Q;Y107F;E116R;N119D;R120D;T123S;C125S / Q126E;C125S / S127T;C125S / I129L;C125S / S130T;C125S / T133S;T3A;F42A / Y45A / L72G;N29S / Y31H / K35R / T37A / K48E / V69A / N71R / Q74P / N88D / I89V;V69A / Q74P / I128T;N29S / Y31H / K35R / T37A / K48E / V69A / N71R / Q74P / N88D / I89V / Q126T;C125S / Q126T;N88R;R38I;L80F / R81D / L85V / I92F;L18R / L80F / R81D / L85V / I92F / Q126T;L18R / L80F / R81D / L85V / I92F / Q126T / S130R;F42A / Y45A / L72G / N88R;F42A / Y45A / L72G / Q126T;F42A / Y45A / L72G / N88R / Q126T;L19D;D20N;N88D;N88K;N88R;N88R;N88R;F42A / Y45A / L72G;N29S / Y31H / K35R / T37A / K48E / N71R / N88D / I89V;L19D / N29S / Y31H / K35R / T37A / K48E / N71R;D20N / N29S / Y31H / K35R / T37A;K48E / N71R;L19D / N29S / Y31H / K35R / T37A / K48E;D20N / N29S / Y31H / K35R / T37A;K48E;L19D K35R;L19D / T37R;D20N / T37R;L19D / N71K;D20N / N71K;D20N / R38I;D20N / T37R;38I;D20N / R38I / N71K;D20N / N71K;D20N;D20N / T37R;D20N / R38I;D20N / T37R R38I;D20N / R38I / N71K;D20N;D20N / T37R;D20N / N71K;D20N / R38I;D20N / T37RR38I;D20N / R38I / N71K;D20N;D20N / T37R;D20N / N71K;D20N / R38I;D20N / T37R / R38I;D20N / R38I / N71K;N29S / Y31H / K35R / T37A / K48E / V69A / N71R / Q74P / N88D / I89V / Q126T,R38A / C125S;R38D / C125S;R38E / C125S;R38F / C125S;R38G / C125S;R38H / C125S;R38I / C125S;R38K / C125S;R38L / C125S;R38M / C125S;R38N / C125S;R38P / C125S;R38Q / C125S;R38S / C125S;R38T / C125S;R38V / C125S;R38W / C125S;R38Y / C125S;T41A / C125S;T41D / C125S;T41E / C125S;T41F / C125S;T41G / C125S;T41H / C125S;T41I / C125S;T41K / C125S;T41L / C125S;T41M / C125S;T41N / C125S;T41P / C125S;T41Q / C125S;T41R / C125S;T41S / C125S;T41V / C125S;T41W / C125S;T41Y / C125S;F42A / C125S;F42D / C125S;F42E / C125S;F42G / C125S;F42H / C125S;F42I / C125S;F42K / C125S;F42L / C125S;F42M / C125S;F42N / C125S;F42P / C125S;F42Q / C125S;F42R / C125S;F42S / C125S;F42T / C125S;F42V / C125S;F42W / C125S;F42Y / C125S;R38Q / T41K / C125S;R38Q / T41Q / C125S;R38E / T41K / C125S;R38Q / T41R / C125S;R38N / T41Q / C125S;R38Q / T41V / C125S;R38N / T41V / C125S;R38Q / T41M / C125S;R38Q / T41S / C125S;R38Q / T41L / C125S;R38N / T41M / C125S;T41I / F42Y / C125S;T41E / F42Y / C125S′T41D / F42Y / C125S;T41M / F42Y / C125S;41Q / F42Y / C125S;T41E / F42H / C125S;T41E / F42L / C125S;T41E / F42P / C125S;R38Q / F42Y / C125S;R38N / T41R / C125S;R38N / T41K / C125S;R38V / T41R / C125S;R38P / T41R / C125S;T41E / F42K / C125S;T41D / F42K / C125S;T41M / F42K / C125S;T41Q / F42K / C125S;R38Q / F42K / C125S;T41I / F42K / C125S;R38N / F42K / C125S;T41H / F42K / C125S;R38Q / T41K / F42Y / C125S;R38Q / T41R / F42Y / C125S;R38Q / T41Q / F42Y / C125S;R38Q / T41V / F42Y / C125S;R38N / T41K / F42K / C125S;R38Q / T41H / F42K / C125S;R38Q / T41K / F42K / C125S;R38Q / T41Q / F42K / C125S;38Q / T41V / F42K / C125S;R38Q / T41R / F42K / C125S;N29S / Y31H / K35R / T37A / R38L / K48E / V69A / N71R / Q74P / N88D / I89V / C125S / Q126T;Q11E / C125S;L12D / C125S;Q13E / C125S;E15Q / C125S;H16Y / C125S;L19D / C125S;D20N / C125S;Q22E / C125S;K35R / C125S;T37S / C125S;K43R / C125S;F44Y / C125S;Y45F / C125S;K48R / C125S;K49E / C125S;E61Q / C125S;E62Q / C125S;K64R / C125S;E68Q / C125S;V69L / C125S;L72I / C125S;R81D / C125S;D84N / C125S;S87T / C125S;N88D / C125S;V91L / C125S;192L / C125S;E95Q / C125S;Y107F / C125S;E116R / C125S;N119D / C125S;R120D / C125S;T123S / C125S;C125S / Q126E;C125S / S127T;C125S / I129L;C125S / S130T;C125S / T133S;T3A / C125S;T3A / F42A / Y45A / L72G / C125A:N29S / Y31H / K35R / T37A / K48E / V69A / N71R / Q74P / N88D / I89V / C125S;V69A / Q74P / I128T / C125S;N29S / Y31H / K35R / T37A / K48E / V69A / N71R / Q74P / N88D / I89V / C125S / Q126T;C125S / Q126T;N88R / C125S;R38I / C125S;L80F / R81D / L85V / I92F / C125S;L18R / L80F / R81D / L85V / I92F / C125S / Q126T;L18R / L80F / R81D / L85V / 192F / C125S / Q126T / S130R;T3A / F42A / Y45A / L72G / N88R / C125A;T3A / F42A / Y45A / L72G / C125A / Q126T;T3A / F42A / Y45A / L72G / N88R / C125A / Q126T;T3A / L19D / C125S;T3A / D20N / C125S;T3A / N88D / C125S;T3A / N88K / C125S;N88R / C125S;N88R / C125S;N88R / C125S;T3A / F42A / Y45A / L72G / C125A;N29S / Y31H / K35R / T37A / K48E / N71R / N88D / I89V / C125S;L19D / N29S / Y31H / K35R / T37A / K48E / N71R / C125S:D20N / N29S / Y31H / K35R / T37A;K48E / N71R / C125S;L19D / N29S / Y31H / K35R / T37A / K48E / C125S;D20N / N29S / Y31H / K35R / T37A;K48E / C125S;T3A / L19D K35R / C125S;T3A / L19D / T37R / C125S;T3A / D20N / T37R / C125S;T3A / L19D / N71K / C125S;T3A / D20N / N71K / C125S;T3A / D20N / R38I / C125S;T3A / D20N / T37R;38I / C125S;T3A / D20N / R381 / N71K / C125S;F3A / D20N / N71K / C125S;T3A / D20N / C125S;T3A / D20N / T37R / C125S;T3A / D20N / R38I / C125S;T3A / D20N / T37R R38I / C125S;T3A / D20N / R38I / N71K / C125S;T3A / D20N / C125S;T3A / D20N / T37R / C125S;T3A / D20N / N71K / C125S;T3A / D20N / R38I / C125S;T3A / D20N / T37R R38I / C125S;T3A / D20N / R38I / N71K / C125S;T3A / D20N / C125S;T3A / D20N / T37R / C125S;T3A / D20N / N71K / C125S; T3A / D20N / R38I / C125S; T3A / D20N / T37R / R38I / C125S; T3A / D20N / R38I / N71K / C125S; and N29S / Y31H / K35R / T37A / K48E / V69A / N71R / Q74P / N88D / I89V / C125S / Q126T.

[0010] In another aspect, the present disclosure provides an IL-2-Fc fusion dimer protein comprising: a) a first monomer comprising, from N-terminus to C-terminus: i) a variant IL-2 protein according to any one of claims A1 to A4; ii) a first domain linker; and iii) a first variant Fc domain; and b) a second monomer comprising a second variant Fc domain.

[0011] In another aspect, the present invention provides an IL-2-Fc fusion dimer protein, wherein the second monomer comprises, from N-terminus to C-terminus: a) a variant IL-2 protein according to any one of claims A1 to A4; b) a second domain linker; and c) the second variant Fc domain.

[0012] In another aspect, the IL-2-Fc fusion dimer protein has a first variant Fc domain and a second variant Fc domain, wherein the first variant Fc domain and the second variant Fc domain contain heterodimerization variants selected from the group consisting of: L368D / K370S:S364K / E357Q; L368D / K370S:S364K; L368E / K370S:S364K; / T411E / K360E / Q362E:D401K; and T366S / L368A / Y407V:T366W. In some cases, the IL-2-Fc fusion protein further includes an ablated variant comprising P233P / L234V / L235A / G236_ / S267K. In some aspects, the fusion protein further comprises an Fc domain having amino acid substitutions M428L / N434S or M428L / N434A. In some aspects, the domain linker is an IGG1 hinge, and in other aspects, it can comprise a linker selected from (GS)n, (GSGGS)n, (GGGGS)n, and (GGGS)n, wherein n is an integer of at least one.

[0013] In another aspect, the present invention includes a polypeptide composition comprising a variant human IL-2 protein, wherein the variant IL-2 protein comprises one or more amino acid substitutions selected from the group consisting of T3A / D20N / T37R and T3A / D20N / N71K compared to SEQ ID NO: 2. In some cases, the variant IL-2 protein further comprises a C125S variant or a C125A variant.

[0014] In other aspects, the polypeptide composition is a homodimeric protein complex, wherein each protein monomer comprises the variant IL-2 protein covalently linked to an Fc domain. In some aspects, each Fc domain in the Fc domain is a variant Fc domain.

[0015] In another aspect, the polypeptide composition is a heterodimeric protein complex comprising a first protein monomer comprising the variant IL-2 protein covalently linked to a first variant Fc domain and a second protein monomer comprising a second variant Fc domain.

[0016] In a further aspect, the polypeptide composition has a variant Fc domain, which is a variant human IgG1 Fc domain comprising the amino acid substitutions M428L / N434S.

[0017] In a further aspect, the polypeptide composition has a variant Fc domain, which is a variant human IgG1 Fc domain comprising the amino acid substitutions E233P / L234V / L235A / G236del / S267K.

[0018] In additional aspects, the polypeptide composition has a first variant Fc domain and a second variant Fc domain, wherein the first variant Fc domain and the second variant Fc domain comprise a heterodimerization variant set selected from the group consisting of the heterodimerization variants depicted in Figure 2. In some embodiments, the heterodimerization variant set is selected from the group consisting of: L368D / K370S:S364K / E357Q; L368D / K370S:S364K; L368E / K370S:S364K; T411E / K360E / Q362E:D401K; and T366S / L368A / Y407V:T366W.

[0019] In further aspects, the polypeptide composition is selected from the group consisting of XENP27564 (SEQ ID NOs: 297 and 298), XENP27563 (SEQ ID NOs: 295 and 296), XENP26105 (SEQ ID NOs: 245 and 246), and XENP26109 (SEQ ID NOs: 249 and 250).

[0020] Also provided are nucleic acid compositions comprising: a) a first nucleic acid encoding the first protein monomer of any one of claims 6 to 13; and b) a second nucleic acid encoding the second protein monomer of any one of claims 6 to 13, respectively. Also provided are expression vector compositions comprising: a) a first expression vector comprising the first nucleic acid; and b) a second expression vector comprising the second nucleic acid, and a host cell comprising the nucleic acid composition and / or expression vector composition. Further provided are methods for preparing polypeptide compositions comprising culturing the host cells of the present invention under conditions that produce the composition and recovering the composition.

[0021] In another aspect, the IL-2-Fc fusion dimer protein is selected from the group consisting of: XENP24635; XENP24636; XENP24637; XENP24638; XENP24639; XENP24640; XENP24641; XENP24642; XENP24643; XENP24725; XENP24728; XENP2472 ... ENP24730; XENP24731; XENP24732; XENP25717; 910;XENP25911;XENP25912;XENP26086; ENP26096; XENP26104; XENP26105; 840; XENP26841; XENP26986; XENP26987; ENP25907; XENP25908; XENP25909; XENP26992; XENP26993; XENP26994; XENP26995; XENP26996; XENP27001; XENP27002; XENP27003; XENP27004; XENP27005; XENP27006 and XENP27007.

[0022] Additional aspects provide methods for activating CD25+ cells, comprising contacting the cells with an IL-2-Fc fusion dimer protein of the present invention; and methods for treating autoimmune diseases, comprising administering the protein composition herein to a patient in need thereof. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 depicts the amino acid sequences (and GenBank accession numbers) of human IL-2 and its receptors: IL-2Rα (also known as CD25), IL-2Rβ (also known as CD122), and the common γ chain (also known as IL-2Rγ or CD132).

[0024] Figure 2 depicts a useful set of Fc heterodimerization variant pairs (including skew variants and pi variants). In Figure 2, there are variants without corresponding "monomer 2" variants; these are pi variants that can be used alone on either monomer.

[0025] Figure 3Depicts a list of isosteric variant antibody constant regions and their corresponding substitutions, with pI (-) indicating lower pI variants and pI -(+) indicating higher pI variants. These can be optionally and independently combined with other heterodimerization variants of the present invention (as well as other variant types, as outlined herein).

[0026] Figure 4 Useful ablated variants (sometimes referred to as "knockout" or "KO" variants) that ablate FcγR binding are depicted. Typically, the ablated variant is found on both monomers, but in some cases it may be located on only one monomer.

[0027] Figure 5 shows a particularly useful embodiment of a "non-cytokine" component of the present invention.

[0028] Figure 6 shows the sequences of several useful IL-2-Fc fusion format backbones based on human IgG, rather than cytokine sequences. Backbone 1 is based on human IgG1 (356E / 358M allotype) and contains: C220S on both chains, S364K / E357Q:L368D / K370S skew variant, Q295E / N384D / Q418E / N421D pI variant on one chain with L368D / K370S skew variant, and E233P / L234V / L235A / G236del / S267K ablated variant on both chains. Backbone 2 is based on human IgG1 (356E / 358M allotype) and comprises: C220S on both chains, S364K / E357Q:L368D / K370S skew variant, Q295E / N384D / Q418E / N421D pI variant on one chain with L368D / K370S skew variant, and E233P / L234V / L235A / G236del / S267K ablation variant on both chains. Backbone 3 is based on human IgG1 (356E / 358M allotype) and comprises: C220S on both chains, S364K / E357Q:L368E / K370S skew variant, Q295E / N384D / Q418E / N421D pI variant on one chain with L368E / K370S skew variant, and E233P / L234V / L235A / G236del / S267K ablation variant on both chains. Backbone 4 is based on human IgG1 (356E / 358M allotype) and comprises: C220S on both chains, D401K: K360E / Q362E / T411E skew variant, Q295E / N384D / Q418E / N421D pI variant on one chain with K360E / Q362E / T411E skew variant, and E233P / L234V / L235A / G236del / S267K ablation variant on both chains. Backbone 5 is based on human IgG1 (356D / 358L allotype) and comprises: C220S on both chains, S364K / E357Q:L368D / K370S skew variant, Q295E / N384D / Q418E / N421D pI variant on one chain with L368D / K370S skew variant, and E233P / L234V / L235A / G236del / S267K ablation variant on both chains.Backbone 6 is based on human IgG1 (356E / 358M allotype) and contains: C220S on both chains, S364K / E357Q:L368D / K370S skew variant, Q295E / N384D / Q418E / N421D pI variant on both chains with L368D / K370S skew variant, and E233P / L234V / L235A / G236del / S267K ablation variant on both chains, and N297A variant on both chains. Backbone 7 is identical to Backbone 6 except that the mutation is N297S. Alternative formats of Backbone 6 and Backbone 7 can exclude the ablation variant E233P / L234V / L235A / G236del / S267K on both chains. Backbone 8 is based on human IgG4 and comprises: S364K / E357Q:L368D / K370S skew variant, Q295E / N384D / Q418E / N421D pI variant on the chain with the L368D / K370S skew variant, and S228P (EU numbering, this is S241P in Kabat) variant on both chains to ablate Fab arm exchange as known in the art. Backbone 9 is based on human IgG2 and comprises: S364K / E357Q:L368D / K370S skew variant, Q295E / N384D / Q418E / N421D pI variant on the chain with the L368D / K370S skew variant. Backbone 10 is based on human IgG2 and comprises: S364K / E357Q:L368D / K370S skew variant, Q295E / N384D / Q418E / N421D pI variant on one chain with the L368D / K370S skew variant, and S267K variant on both chains. Backbone 11 is identical to Backbone 1 except that it comprises M428L / N434S Xtend mutations. Backbone 12 is based on human IgG1 (356E / 358M allotype) and comprises: C220S on both identical chains, E233P / L234V / L235A / G236del / S267K ablation variant on both identical chains. Backbone 13 is based on human IgG1 (356E / 358M allotype) and comprises: C220S on both chains, S364K / E357Q:L368D / K370S skew variant, P217R / P229R / N276K pI variant on one chain with S364K / E357Q skew variant, and E233P / L234V / L235A / G236del / S267K ablation variant on both chains.

[0029] As will be appreciated by those skilled in the art and as outlined below, any IL-2 variant can be incorporated into the backbones of Figure 6 in any combination. Each of these backbones comprises within the backbone a sequence that is 90%, 95%, 98%, and 99% identical to the recited sequence (as defined herein), and / or contains 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 additional amino acid substitutions (as compared to the "parent" of the accompanying figures, which, as will be appreciated by those skilled in the art, are already containing multiple amino acid modifications compared to the parent human IgG1 (or IgG2 or IgG4, depending on the backbone)). That is, in addition to the skew variants, pI variants, and ablated variants contained within the backbones of this figure, the recited backbones can contain additional amino acid modifications (typically amino acid substitutions). Specifically, FcRn variants such as M428L / N434S can also be included.

[0030] Figure 7 Several exemplary variable length linkers are depicted. In some embodiments, these linkers can be used to connect the C-terminus of IL-2 to the N-terminus of the Fc region (including the hinge domain in some cases).

[0031] Figure 8A and 8B Depicted are A) a structural model of IL-2 in complex with the high-affinity IL-2 receptor (IL-2Rαβγ), and B) the positions of three IL-2 residues that contact IL-2Rα and at which substitutions are predicted to weaken the pH-dependent binding of IL-2 to IL-2Rα.

[0032] Figure 9 depicts the amino acid sequences of illustrative IL-2 variants engineered to reduce pH-dependent binding to IL-2Rα. It is important to note that these variants were generated using a polyhistidine (Hisx8 or HHHHHHHH) C-terminal tag that was removed from the sequences depicted here.

[0033] FIG10 depicts the association rate (k) of IL-2 variants to IL-2Rα at pH 6.0. a ), dissociation rate (k d ) and the dissociation constant (K D ), and fold improvement in kd and KD compared to XENP14135 (wild-type IL-2 with C125S mutation). NB indicates no measurable binding.

[0034] FIG11 depicts the association rate (k) of IL-2 variants to IL-2Rα at pH 7.4. a ), dissociation rate (k d ) and the dissociation constant (K D), and fold improvement in kd and KD compared to XENP14135 (wild-type IL-2 with C125S mutation). NB indicates no measurable binding.

[0035] Figure 12 The IL-2 dissociation rate (k) relative to IL-2Rα at pH 6.0 achieved by various point mutations is depicted. d ) of the increase multiple.

[0036] Figure 13 The IL-2 dissociation rate (k) relative to IL-2Rα at pH 7.4 achieved by various point mutations is depicted. d ) of the increase multiple.

[0037] Figure 14 Depicted are Biacore sensorgrams of A) XENP14135 (wild-type IL-2 with C125S mutation) and B) XENP14142 (variant IL-2 with R38I and C125S).

[0038] Figure 15 Depicted are the amino acid sequences of additional prior art IL-2 variants (mutants 2-4 with Q126T as described in WO 2009 / 061853 published May 14, 2009) that do not contain R38L (XENP14277) and contain R38L (XENP14381) to reduce pH-dependent binding to IL-2Rα. It is important to note that these variants were generated using a polyhistidine (Hisx8 or HHHHHHHH) C-terminal tag that was removed from the sequences depicted here.

[0039] Figure 16 Depicted are the association rates (k) of IL-2 variants binding to IL-2Rα with and without the R38L substitution at pH 7.4 and pH 6.0. a ), dissociation rate (k d ) and the dissociation constant (K D ).

[0040] Figure 17 depicts the amino acid sequences of illustrative IL-2 variants engineered to alter binding to IL-2Rα, IL-2Rβ, γc, or IL-2Rβγ. It is important to note that these variants were generated using a polyhistidine (Hisx8 or HHHHHHHH) C-terminal tag that was removed from the sequences depicted here.

[0041] Figure 18 depicts the normalized BLI responses (relative to XENP14135) of illustrative IL-2 variants of various IL-2 receptors as determined by Octet. The aim was to increase binding to IL-2Rα, or to reduce binding to the interface of IL-2Rβ and IL-2Rγ or to IL-2Rβγ, or both.

[0042] Figure 19 Several formats of the IL-2-Fc fusions of the present invention are depicted. Monovalent IL-2-Fc or "monovIL-2-Fc" ( Figure 19 A) includes IL-2 recombinantly fused to the N-terminus of the heterodimeric Fc region, with the other side of the molecule being "Fc only" or "null Fc". Bivalent IL-2-Fc or "bivIL-2-Fc" ( Figure 19 B) comprises IL-2 recombinantly fused to the N-termini on either side of the homodimeric Fc region. Monovalent IL-2-Fc with linker or "monovIL-2-Fc (with linker)" ( Figure 19 C) includes IL-2 recombinantly fused to the N-terminus of the heterodimeric Fc region via a domain linker, with the other side of the molecule being "Fc only" or "empty Fc". Bivalent IL-2-Fc with linker or "bivIL-2-Fc (with linker)" ( Figure 19 D) includes IL-2 recombinantly fused to the N-termini on both sides of the homodimeric Fc region via domain linkers. Figure 7 Non-limiting examples of domain linkers suitable for use in the monovIL-2-Fc (with linker) and bivIL-2-Fc (with linker) formats are depicted in .

[0043] Figure 20 depicts the amino acid sequence of an illustrative monovIL-2-Fc fusion of the invention. The slash ( / ) indicates the boundary between IL-2 and the Fc region (in this case, the Fc region includes the hinge and the C220S variant).

[0044] Figure 21 Depicted are the affinities (K) of illustrative IL-2-Fc fusions for IL-2Rα, IL-2Rβ, and IL-2Rβγ as determined by Octet. D ), association rate (k a ) and dissociation rate (k d ).

[0045] Figures 22A-22F Depicts the induction of CD4 by A) XENP24636, B) XENP24638, C) XENP24641, D) XENP24642, E) XENP24643, and F) XENP24731. + CD45RA -T cells, CD4 + CD45RA + T cells, CD8 + CD45RA - T cells, CD8 + CD45RA + STAT5 phosphorylation on T cells and Tregs.

[0046] Figure 23 The amino acid sequence of an illustrative bivIL-2-Fc fusion of the invention is depicted. The slash ( / ) indicates the boundary between IL-2 and the Fc region, which again comprises the hinge domain of IgG1 with the C220S variant.

[0047] Figure 24A and 24B Depicts the induction of CD4 by A) XENP25906 and B) XENP25907. + CD45RA - T cells, CD4 + CD45RA + T cells, CD8 + CD45RA - T cells, CD8 + CD45RA + STAT5 phosphorylation on T cells and Tregs.

[0048] Figure 25 depicts the amino acid sequence of an illustrative IL-2-Fc fusion with a domain linker. The slash ( / ) indicates the boundary between IL-2, the linker, and the Fc region, again containing the hinge domain of IgG1 with the C220S variant. The linker is double underlined.

[0049] Figures 26A-26E Depicts the induction of CD4 by A) XENP25908, B) XENP25909, C) XENP25910, D) XENP25911, and E) XENP25912. + CD45RA - T cells, CD4 + CD45RA + T cells, CD8 + CD45RA - T cells, CD8 + CD45RA + STAT5 phosphorylation on T cells and Tregs.

[0050] Figure 27 depicts the amino acid sequence of an additional IL-2-Fc fusion with IL-2 engineered to increase affinity for CD25 and reduce affinity for CD 122. The slash ( / ) indicates the boundary between IL-2 and the Fc region, again containing the hinge domain of IgG1 with the C220S variant.

[0051] Figures 28A-28T Depicts the effect of the following on CD8 + T cells and CD4 + Phosphorylation of STAT5 on T cells and Tregs as an indicator of activation: A) XENP24635, B) XENP24636, C) XENP24637, D) XENP24638, E) XENP24642, F) XENP25717, G) XENP25720, H) XENP25725, I) XENP25727, J) XENP26086, K) XENP26088, L) XENP26089, M) XENP26092, N) XENP26093, O) XENP26096, P) XENP26104, Q) XENP26105, R) XENP26108, S) XENP26109, and T) recombinant human IL-2.

[0052] Figure 29A and 29B Depicted are the effects of illustrative IL-2-Fc fusions on A) Treg and B) CD4 + STAT5 is phosphorylated on (CD45RA-) T cells as an indicator of activation.

[0053] Figure 30 depicts the amino acid sequences of additional IL-2-Fc fusions. Slashes ( / ) indicate the boundary between IL-2, the domain linker (double underlined), and the Fc region, which again comprises the hinge domain of IgG1 with the C220S variant.

[0054] Figure 31 depicts some preferred embodiments.

[0055] Figure 32 Depicted are some preferred examples of variants engineered with A) pH switch substitutions, B) Treg-selective substitutions, and C) a combination of pH switch substitutions and Treg-selective substitutions.

[0056] Figure 33 Depicted are the affinities (K) of illustrative IL-2-Fc fusions for IL-2Rα, IL-2Rβ, and IL-2Rβγ as determined by Octet. D ), association rate (k a ) and dissociation rate (kd ). NB indicates no binding.

[0057] Figure 34 Depicted is the sequence of the Fc-IL-2 (V91K / C125A) fusion XENP27193 engineered to increase the ratio of Treg to non-regulatory T cells as described in WO 2014 / 153111. Slashes ( / ) indicate the boundaries between IL-2, the domain linker, and the Fc region (again, wherein the Fc region comprises the hinge domain of IgG1 with the C220S variant); and the linker is double underlined.

[0058] Figure 35A A) CD4 induction by variant monovIL-2-Fc fusions XENP24638, XENP24642, XENP26105, XENP26109, XENP26835, XENP26839, XENP26991, and XENP25702 are depicted. + CD45RA - T cells, B) CD8 + CD45RA - The data show that compared with recombinant IL-2 and monovalent WT IL-2-Fc fusion (XENP24635) and the prior art bivalent IL-2-Fc fusion (XENP27193) described in WO 2014 / 153111, STAT5 phosphorylation on CD4 T cells, C) NK cells, D) γδ T cells and E) Tregs. + Memory T cells (CD45RA-), CD8 + Memory T cells (CD45RA-), NK cells, and γδ T cells, the variant monovIL-2-Fc fusion preferentially induced STAT5 phosphorylation on Tregs.

[0059] Figure 36 Depicts the induction of CD4 by IL-2 variants with N88R / C125S substitutions formatted as monovIL-2-Fc without a linker (XENP24642) and bivIL-2-Fc with a linker (XENP25908). + CD45RA - STAT5 phosphorylation on T cells and Tregs.

[0060] Figure 37Depicted are the induction of CD4 T cells by IL-2 variants with T3A / D20N / C125S substitutions formatted as monovIL-2-Fc without a linker (XENP25720), bivIL-2-Fc without a linker (XENP26992), and bivIL-2-Fc with a linker (XENP27002). + CD45RA - STAT5 phosphorylation on T cells and Tregs.

[0061] Figure 38 Figure 3 shows the induction of CD4 T cells by IL-2 variants with T3A / D20N / T37R / C125S substitutions formatted as monovIL-2-Fc without a linker (XENP26105), bivIL-2-Fc without a linker (XENP26993), and bivIL-2-Fc with a linker (XENP27003). + CD45RA - STAT5 phosphorylation on T cells and Tregs.

[0062] Figure 39 Depicts the induction of CD4 T cells by IL-2 variants with T3A / D20N / N71K / C125S substitutions formatted as monovIL-2-Fc without a linker (XENP26109), bivIL-2-Fc without a linker (XENP26994), and bivIL-2-Fc with a linker (XENP27004). + CD45RA - STAT5 phosphorylation on T cells and Tregs.

[0063] Figure 40 Figure 3 shows the induction of CD4 T cells by IL-2 variants with T3A / D20N / R38I / C125S substitutions formatted as monovIL-2-Fc without a linker (XENP26835), bivIL-2-Fc without a linker (XENP26995), and bivIL-2-Fc with a linker (XENP27005). + CD45RA - STAT5 phosphorylation on T cells and Tregs.

[0064] Figure 41Depicted are the induction of CD4 T cells by IL-2 variants with T3A / D20N / T37R / R38I / C125S substitutions formatted as monovIL-2-Fc without a linker (XENP26839), bivIL-2-Fc without a linker (XENP26996), and bivIL-2-Fc with a linker (XENP27006). + CD45RA - STAT5 phosphorylation on T cells and Tregs.

[0065] Figure 42 Depicts the induction of CD4 T cells by IL-2 variants with T3A / D20N / R38I / N71K / C125S substitutions formatted as monovIL-2-Fc with a linker (XENP26991), bivIL-2-Fc without a linker (XENP27001), and bivIL-2-Fc with a linker (XENP27007). + CD45RA - STAT5 phosphorylation on T cells and Tregs.

[0066] Figure 43 depicts the amino acid sequence of an illustrative bivalent IL-2-Fc fusion comprising the Xtend(M428L / N434S) Fc of the invention (again including the hinge and C220S variants). The slash ( / ) indicates the boundary between the IL-2 and Fc regions.

[0067] Figure 44 depicts the amino acid sequence of an illustrative monovalent IL-2-Fc fusion comprising Xtend(M428L / N434S)Fc of the invention. The slash ( / ) indicates the boundary between the IL-2 and Fc regions.

[0068] Figure 45A Figures 1 through 2 depict the induction of STAT5 phosphorylation on various lymphocyte populations by A) XENP26105, B) XENP26109, C) XENP24635, D) XENP25908, and E) XENP27193.

[0069] Figure 46 The immunophenotype of rapamycin Treg cultures treated with XENP27564 or recombinant IL-2 is depicted. The data show higher CD25 expression with XENP27564 treatment.

[0070] The histogram shows the difference between the cells treated with XENP27564 or recombinant IL-2. Figure 47Rapamycin Tregs were treated with XENP27564IL-2-Fc to assess the relative expression of CD25. Data showed that rapamycin Treg cultures treated with XENP27564IL-2-Fc displayed higher CD25 expression.

[0071] Figure 48 Depicts individual CD4 Treg-containing cells after incubation with rapamycin and XENP27564 or recombinant IL-2. + The data showed that cultures expanded with XENP27564 displayed a larger effector Treg population (CD45RA-FoxP3 mid-high ).

[0072] Figure 49A A and B depict that rapamycin Treg cultures expanded with XENP27564 or recombinant IL-2 inhibit A) CD8 responder T cell proliferation and B) CD4 responder T cell proliferation. The data suggest that Tregs expanded by XENP27564 may have enhanced suppressive function.

[0073] FIG50 depicts the expression of CD25 on Tregs in the suppression assay depicted in FIG50 , as determined by A) CD25 MFI on Tregs and B) CD25 + The percentages of Tregs are indicated.

[0074] Figure 51A A and B depict the expression of CD127 on Tregs in the suppression assay depicted in panel X, as indicated by A) CD127 MFI on Tregs and B) the percentage of CD127+ Tregs.

[0075] Figure 52A Figures 3 through 4 depict the proliferation of various lymphocyte populations (as determined by CFSE or Tag-it Violet dilution) following incubation of PBMCs and Tregs with A) XENP27563, B) XENP27564, C) XENP24635, D) recombinant IL-2, and E) recombinant IL-15. The data show that XENP27563 and XENP27564 exhibit Treg selectivity.

[0076] Figure 53AFigures 2 through 3 depict the proliferation of CD8+ T cells following incubation of PBMCs and Tregs with XENP27563, XENP27564, XENP24635, recombinant IL-2, recombinant IL-15, and negative control anti-RSV mAb XENP15074, as indicated by A) proliferating cells (as determined by CFSE or Tag-it Violet dilution) and B) total cell counts. The data show that XENP27563 and XENP27564 are less effective in inducing CD8+ T cell proliferation than recombinant IL-2 and IL-15, as well as an IL-2-Fc fusion including WT IL-2 (with a C125S mutation).

[0077] Figure 54 depicts the proliferation of CD4+ T cells after incubation of PBMCs and Tregs with XENP27563, XENP27564, XENP24635, recombinant IL-2, recombinant IL-15, and negative control anti-RSV mAb XENP15074 as indicated by A) proliferating cells (as determined by CFSE or Tag-it Violet dilution) and B) total cell counts. The data show that XENP27563 and XENP27564 are less effective in inducing CD4+ T cell proliferation than recombinant IL-2 and IL-15, as well as IL-2-Fc fusions including WT IL-2 (with a C125S mutation).

[0078] Figure 55 Depicted are proliferation of CD8 T cells following incubation of PBMCs with the indicated concentrations of the indicated test articles and A) 5 ng / ml, B) 10 ng / ml, or C) 20 ng / ml plate-bound anti-CD3 (OKT3). + T cells (as indicated by the percentage of cells expressing Ki67). The data show that XENP27563 and XENP27564 (IL-2-Fc fusions engineered to be selective for CD25) are effective in inducing CD8 + Impaired function in T cell proliferation.

[0079] Figure 56 Depicted are proliferation of CD4 T cells following incubation of PBMCs with the indicated concentrations of the indicated test articles and A) 5 ng / ml, B) 10 ng / ml, or C) 20 ng / ml plate-bound anti-CD3 (OKT3). + T cells (as indicated by the percentage of cells expressing Ki67). The data show that XENP27563 and XENP27564 (IL-2-Fc fusions engineered to be selective for CD25) are effective in inducing CD4 + Impaired function in T cell proliferation.

[0080] Figure 57 Depicted are proliferation of CD8 T cells following incubation of PBMCs with the indicated concentrations of the indicated test articles and A) 5 ng / ml, B) 10 ng / ml, or C) 20 ng / ml plate-bound anti-CD3 (OKT3). + CD45RA - T cells (as indicated by the percentage of cells expressing Ki67). The data show that XENP27563 and XENP27564 (IL-2-Fc fusions engineered to be selective for CD25) are effective in inducing CD8 + CD45RA - Impaired function in T cell proliferation.

[0081] Figure 58 Depicted are proliferation of CD8 T cells following incubation of PBMCs with the indicated concentrations of the indicated test articles and A) 5 ng / ml, B) 10 ng / ml, or C) 20 ng / ml plate-bound anti-CD3 (OKT3). + CD45RA + T cells (as indicated by the percentage of cells expressing Ki67). The data show that XENP27563 and XENP27564 (IL-2-Fc fusions engineered to be selective for CD25) are effective in inducing CD8 + CD45RA + Impaired function in T cell proliferation.

[0082] Figure 59 Depicted are proliferation of CD4 T cells following incubation of PBMCs with the indicated concentrations of the indicated test articles and A) 5 ng / ml, B) 10 ng / ml, or C) 20 ng / ml plate-bound anti-CD3 (OKT3). + CD45RA - T cells (as indicated by the percentage of cells expressing Ki67). The data show that XENP27563 and XENP27564 (IL-2-Fc fusions engineered to be selective for CD25) are effective in inducing CD4 + CD45RA - Impaired function in T cell proliferation.

[0083] Figure 60 Depicted are proliferation of CD4 T cells following incubation of PBMCs with the indicated concentrations of the indicated test articles and A) 5 ng / ml, B) 10 ng / ml, or C) 20 ng / ml plate-bound anti-CD3 (OKT3). + CD45RA +T cells (as indicated by the percentage of cells expressing Ki67). The data show that XENP27563 and XENP27564 (IL-2-Fc fusions engineered to be selective for CD25) are effective in inducing CD4 + CD45RA + Impaired function in T cell proliferation.

[0084] Figure 61 Depicted are the proliferation of NK cells (as indicated by the percentage of cells expressing Ki67) after incubation of PBMCs with the indicated concentrations of the indicated test articles and A) 5 ng / ml, B) 10 ng / ml, or C) 20 ng / ml plate-bound anti-CD3 (OKT3). The data show that XENP27563 and XENP27564 (IL-2-Fc fusions engineered for CD25 selectivity) are functionally impaired in inducing NK cell proliferation.

[0085] Figure 62 Depicted are the proliferation of Tregs (as indicated by the percentage of cells expressing Ki67) after incubation of PBMCs with the indicated test articles at the indicated concentrations and A) 5 ng / ml, B) 10 ng / ml, or C) 20 ng / ml plate-bound anti-CD3 (OKT3). The data show that XENP27563 and XENP27564 (IL-2-Fc fusions engineered for CD25 selectivity) induce the proliferation of Tregs.

[0086] Figure 63 Depicted are proliferation of CD8 T cells after treatment with the indicated concentrations of A) XENP27563, B) XENP27564, C) XENP24635, D) IL-2, and E) IL-15 and 5 ng / ml plate-bound anti-CD3 (OKT3). + T cells, CD8 + CD45RA - T cells, CD8 + CD45RA + T cells, CD4 + T cells, CD4 + CD45RA - T cells, CD4 + CD45RA + T cells, NK cells, and Tregs (as indicated by the percentage of cells expressing Ki67).

[0087] Figure 64Depicted are proliferation of CD8 T cells after treatment with the indicated concentrations of A) XENP27563, B) XENP27564, C) XENP24635, D) IL-2, and E) IL-15 and 10 ng / ml plate-bound anti-CD3 (OKT3). + T cells, CD8 + CD45RA - T cells, CD8 + CD45RA + T cells, CD4 + T cells, CD4 + CD45RA - T cells, CD4 + CD45RA + T cells, NK cells, and Tregs (as indicated by the percentage of cells expressing Ki67).

[0088] Figure 65 Depicted are proliferation of CD8 T cells after treatment with the indicated concentrations of A) XENP27563, B) XENP27564, C) XENP24635, D) IL-2, and E) IL-15 and 20 ng / ml plate-bound anti-CD3 (OKT3). + T cells, CD8 + CD45RA - T cells, CD8 + CD45RA + T cells, CD4 + T cells, CD4 + CD45RA - T cells, CD4 + CD45RA + T cells, NK cells, and Tregs (as indicated by the percentage of cells expressing Ki67).

[0089] FIG66 depicts A) CD4 + CD45RA - T cells, B) CD8 + CD45RA - T cells, C) CD8α - CD16 + NK cells and D) FoxP3 + Expansion of Tregs. The data showed that both XENP27563 and XENP27564 selectively expanded Tregs and both test articles promoted similar pharmacological effects.

[0090] Figure 67 depicts the change in serum concentration levels over time in cynomolgus monkeys dosed with A) 3X dose of XENP27563 and B) 3X dose of XENP27564. The data indicate that both test articles exhibited similar pharmacokinetic profiles.

[0091] Figure 68 Depicts changes in serum albumin concentrations in cynomolgus monkeys administered 3X doses of XENP27563 and 3X doses of XENP27564. The data show that in one animal administered XENP27563, a sustained decrease in albumin was detected after both the first and second doses; and in one animal administered XENP27564, a transient decrease in albumin was detected only after the second dose, but it quickly returned to baseline levels.

[0092] Figure 69 depicts blood pressure telemetry data for: A) a first cynomolgus monkey dosed with XENP27563 on days 0 and 15, B) a second cynomolgus monkey dosed with XENP27563 on days 0 and 15, and C) a third cynomolgus monkey dosed with XENP27564 on days 0 and 15. The data show that the blood pressure of the first monkey dropped dramatically one day after the second dose and the blood pressure of the second monkey dropped dramatically one day after the first dose, while the blood pressure of the third monkey remained stable for the duration of the study.

[0093] FIG70 depicts A) CD4 T cells over time in cynomolgus monkeys at 1X, 3X, and 10X doses of XENP27564. + CD45RA - T cells, B) CD8 + CD45RA - T cells, C) CD8α - CD16 + Expansion of NK cells and D) Tregs.

[0094] Figure 71 Depicted are the expansion of Tregs on days 7 and 14 in cynomolgus monkeys administered 1X and 3X doses of XENP27564. The data show that the 1X and 3X doses drive similar pharmacological effects in the monkeys and suggest that maximal effect is achieved at lower doses.

[0095] Figure 72 Depicted are changes in serum albumin concentration (as an indicator of vascular leakage) in cynomolgus monkeys following administration of 1X, 3X, and 10X doses of XENP27564. The data show that higher doses of XENP27564 increased toxicity.

[0096] Figure 73Depicted are changes in serum C-reactive protein concentrations (as an indicator of vascular leakage) in cynomolgus monkeys following administration of 1X, 3X, and 10X doses of XENP27564. The data show that higher doses of XENP27564 significantly increased toxicity.

[0097] Figure 74 depicts changes in A) sodium concentration, B) chloride concentration, C) eosinophil count, and D) basophil count in cynomolgus monkeys administered 1X, 3X, or 10X doses of XENP27564. The data show that higher doses of XENP27564 increased toxicity, while lower doses were well tolerated in cynomolgus monkeys.

[0098] Figure 75 Depicted are the changes in serum concentration levels over time in cynomolgus monkeys administered a 1X dose, a 3X dose, or a 10X dose of XENP27564. The data demonstrate sustained pharmacokinetic effects in cynomolgus monkeys for up to several days.

[0099] Figure 76 depicts the changes in A) eosinophil counts and B) basophil counts in cynomolgus monkeys administered a 3X dose of XENP27563 or a 3X dose of XENP27564 on Days 0 and 15. The data show that repeated dosing of XENP27564 was well tolerated in cynomolgus monkeys.

[0100] FIG77 depicts the induction of mouse CD4 T cells by A) XENP26105, B) XENP26109, and C) recombinant human IL-2. + CD44 hi cells, CD8 + CD44 hi STAT5 phosphorylation on cells and Tregs (in splenocytes of B6 mice). The data indicate that the engineered IL-2-Fc fusion is also selective and potent for Tregs in mice, making it suitable for studying autoimmune diseases using preclinical mouse models.

[0101] Figure 78 Depicts the induction of human CD4 + CD45RA - T cells, CD4 + CD45RA + T cells, CD8 + CD45RA - T cells, CD8 + CD45RA + T cells, CD56 + STAT5 phosphorylation on NK cells, γδT cells, and Tregs.

[0102] Figure 79 Depicts the induction of CD4 T cells in cynomolgus monkeys by A) XENP27563 and B) XENP27564. + CD45RA - T cells, CD4 + CD45RA + T cells, CD8 + CD45RA - T cells, CD8 + CD45RA + T cells, CD16 + NK cells, CD56 + STAT5 phosphorylation on NK cells, γδ T cells, and Tregs. The data indicate that the engineered IL-2-Fc fusion is also selective and potent for Tregs in cynomolgus monkeys, making it suitable for use in preclinical mouse models. DETAILED DESCRIPTION

[0103] A. Introduction

[0104] The present invention relates to compositions and methods for engineered IL-2 Fc fusions for the treatment of autoimmune diseases. Autoimmune diseases can be treated using a mechanism that preferentially activates regulatory T cells (commonly referred to as "Treg cells" or "Tregs"). Tregs are a subpopulation of immune T cells that are immunosuppressive and regulate the immune system by maintaining tolerance to self-antigens, thereby preventing autoimmune diseases. Tregs typically downregulate the proliferation of effector T cells. Tregs express biomarkers including CD4, FOXP3, and CD25 (CD25 is also known as IL-2Rα protein).

[0105] Treg can be regulated by IL-2, which is essential for Treg function and survival. Since IL-2 promotes or reduces the potential of both T cells and Treg under limited selection, there is a strong need in the art to produce more selective Treg regulators. In addition, as a potential drug, IL-2 has a very fast clearance rate and a half-life of several minutes, which hinders favorable administration. The present invention solves these two problems by providing a novel IL-2-Fc fusion protein.

[0106] Therefore, the present invention provides IL-2 proteins engineered in two different ways. The first way is that the IL-2 variants of the present invention preferentially activate CD25+ cells such as Tregs compared to other T cells that are CD25-, to provide Treg selectivity increased compared to other T cells, and thus cause the composition to suppress immune function, and thus allow the treatment of autoimmune diseases. This is usually done by increasing binding to IL-2Rα, or reducing binding to IL-2Rβ (and / or IL-2R7) or binding to the interface of IL-2Rβγ, or both.

[0107] In addition to the selective engineering described above, the present invention also provides IL-2 proteins with increased serum half-life, which is accomplished using Fc fusions. In this case, as is generally known in the art, the addition of an Fc domain will increase the half-life of the IL-2 molecule. However, the present invention provides two additional methods for increasing serum half-life.

[0108] The first approach involves the FcRn receptor. In IgG, a site on the Fc between the Cγ2 and Cγ3 domains mediates interaction with the neonatal receptor FcRn. Binding to FcRn allows endocytosed antibodies to be recycled from endosomes back into the bloodstream (Raghavan et al., 1996, Annu Rev Cell Dev Biol 12:181-220; Ghetie et al., 2000, Annu Rev Immunol 18:739-766, both of which are incorporated by reference in their entirety). This process, combined with the exclusion of kidney filtration due to the large size of the full-length molecule, results in a favorable antibody serum half-life ranging from one to three weeks. To increase the retention of Fc proteins in vivo, an increase in binding affinity must be achieved at around pH 6 while maintaining a lower affinity around pH 7.4. Although still under review, it is believed that the Fc region has a longer half-life in vivo because the binding to FcRn at pH 6 in the endosome sequesters the Fc (Ghetie and Ward, 1997. Immunol Today. 18(12):592-598, which is incorporated by reference in its entirety. The endosomal compartment then recycles the Fc to the cell surface. Once the compartment opens to the extracellular space, the higher pH (~7.4) induces the release of the Fc back into the blood.

[0109] Thus, increased serum half-life can utilize Fc variants that increase binding to FcRn and, in many cases, increase half-life.

[0110] Another approach to increasing the serum half-life of IL-2 Fc fusion molecules is based on pH engineering to remove the endosomal sorting pathway. As is known in the art, endocytosis of cytokines such as IL-2 into endosomes results in endocytic sorting, where the cytokine is either degraded or recycled back into the bloodstream (see Fallon et al., Journal of Biological Chemistry (JBC) 275(10):6790, 2000, which is incorporated herein by reference in its entirety). Following internalization into endosomes, IL-2, IL-2Rβ and γc are degraded, while IL-2Rα is constitutively recycled to the cell surface. Since the pH of blood is approximately 7.2 to 7.4, and the pH of endosomes is around pH 6, engineering IL-2 to increase binding to the IL-2Rα ligand at pH 6 results in IL-2 / IL-2Rα recycling rather than degradation, thereby increasing serum half-life.

[0111] In addition, the effectiveness of the IL-2 Fc fusion molecules of the present invention may also depend on other factors. For example, the present invention provides Figure 19 B depicts a bivalent IL-2 construct wherein homodimers of variant IL-2-Fc fusions are prepared, thereby providing bivalent binding to the receptor. Alternatively, the present invention provides Figure 19 A depicts a monovalent IL-2 construct in which a heterodimer is generated in which one monomer is a variant IL-2-Fc fusion and the other is an "empty arm" Fc monomer. In addition, the presence of an additional flexible linker can in some cases increase the Figure 19 The monovalent construct shown in C and Figure 19 D shows the efficacy of the bivalent construct.

[0112] Thus, the present invention provides engineered IL-2 variants and engineered IL-2 Fc fusion proteins that show preferential activation of CD25+ cells, such as Tregs, over CD25- T cells and exhibit increased serum half-life.

[0113] B. Definition

[0114] In order that this application may be more thoroughly understood, several definitions are set forth below. Such definitions are intended to encompass grammatical equivalents.

[0115] As used herein, "ablation" means reducing or removing an activity. Thus, for example, "ablation of FcγR binding" means that an Fc region amino acid variant has less than 50% of the initial binding, preferably less than 70%-80%-90%-95%-98% loss of activity compared to an Fc region not containing the specific variant, and typically the activity is below the level of detectable binding in a Biacore assay. Specific uses of ablation of FcγR binding include Figure 4 for the specific use indicated.

[0116] As used herein, "ADCC" or "antibody-dependent cell-mediated cytotoxicity" refers to a cell-mediated reaction in which nonspecific cytotoxic cells expressing FcγRs recognize bound antibody on a target cell and subsequently cause lysis of the target cell. ADCC is associated with binding to the Fc region of FcγRIIIa; increased binding to FcγRIIIa increases ADCC activity. As discussed herein, many embodiments of the present invention completely ablate ADCC activity.

[0117] As used herein, "modification" refers to amino acid substitutions, insertions, and / or deletions in a polypeptide sequence or alterations to moieties chemically linked to a protein. For example, a modification can be an altered carbohydrate or PEG structure linked to a protein. As used herein, "amino acid modification" refers to amino acid substitutions, insertions, and / or deletions in a polypeptide sequence. For clarity, unless otherwise indicated, amino acid modifications always apply to amino acids encoded by DNA, such as the 20 amino acids for which codons exist in DNA and RNA.

[0118] As used herein, "amino acid substitution" or "substitution" means replacing an amino acid at a specific position in a parent polypeptide sequence with a different amino acid. Specifically, in some embodiments, a substitution is made for an amino acid that does not naturally occur at a specific position or that does not naturally occur in an organism or in any organism. For example, the substitution S364K refers to a variant polypeptide, in this case an Fc variant in which the serine at position 364 is replaced by lysine. The numbering is based on the numbering of the parent polypeptide. For example, R38W in the context of IL-2 numbering. For clarity, a protein that has been engineered to change the nucleic acid coding sequence but does not change the starting amino acid (e.g., replacing CGG (encoding arginine) with CGA (still encoding arginine) to increase expression levels in the host organism) is not an "amino acid substitution"; that is, although a new gene encoding the same protein is generated, if the protein has the same amino acid at its starting specific position, the protein is not an amino acid substitution.

[0119] As used herein, "amino acid insertion" or "insertion" means adding an amino acid sequence at a specific position in a parent polypeptide sequence. For example, -233E or 233E refers to the insertion of glutamic acid after position 233 and before position 234. In addition, -233ADE or A233ADE refers to the insertion of AlaAspGlu after position 233 and before position 234.

[0120] As used herein, "amino acid deletion" or "deletion" means the removal of an amino acid sequence at a specific position in a parent polypeptide sequence. For example, E233- or E233#, E233(), or E233del refers to the deletion of glutamic acid at position 233. In addition, EDA233- or EDA233# refers to the deletion of the sequence GluAspAla starting at position 233.

[0121] As used herein, "protein" herein means at least two covalently linked amino acids, including proteins, polypeptides, oligopeptides and peptides. The peptidyl group can include naturally occurring amino acids and peptide bonds or synthetic peptidomimetic structures, i.e., "analogs," such as peptoids (see Simon et al., Proceedings of the National Academy of Sciences of the United States of America (PNAS USA) 89 (20): 9367 (1992), which is incorporated by reference in its entirety). Amino acids can be naturally occurring or synthetic (e.g., amino acids not encoded by DNA), as will be understood by those skilled in the art. Typically, the proteins of the present invention utilize naturally occurring amino acids. In addition, variant polypeptides may include synthetic derivatization of one or more side chains or termini, glycosylation, pegylation, cyclic transformations, cyclization, linkers to other molecules, fusions to proteins or protein domains, and the addition of peptide tags or labels.

[0122] As used herein, "residue" refers to a position in a protein and its associated amino acid identity. For example, arginine 38 (also referred to as Arg38 or R38) is the residue at position 38 (numbered from the mature sequence) in the human IL-2 protein.

[0123] As used herein, "variant protein" or "protein variant" or "variant" means a protein that is different from a parent protein by virtue of at least one amino acid modification. Protein variants may refer to the protein itself, a composition comprising the protein, or an amino acid sequence encoding the protein. Preferably, the protein variant has at least one amino acid modification compared to the parent protein, for example, from about one to about seventy amino acid modifications compared to the parent, and preferably from about one to about five amino acid modifications. As described below, in some embodiments, the parent polypeptide (e.g., Fc parent polypeptide) is a human wild-type sequence, such as the Fc region from human IgG1, IgG2, IgG3, or IgG4. In the context of IL-2 variants, the parent polypeptide is human IL-2, whose mature sequence is shown in Figure 1. The protein variant sequence herein will preferably have at least about 80% identity and most preferably at least about 90% identity, more preferably at least about 95%-98%-99% identity with the parent protein sequence. Variant protein may refer to the variant protein itself, a composition comprising the protein variant, or a DNA sequence encoding the protein variant.

[0124] As used herein, "Fc" or "Fc region" or "Fc domain" means a polypeptide comprising the constant region of an IgG antibody excluding the first constant region immunoglobulin domain and, in some cases, all or a portion of the hinge. For IgG, the Fc domain includes the immunoglobulin domains Cγ2 and Cγ3 (CH2 and CH3) and the hinge region between Cγ1 (CH1) and Cγ2 (CH2). In the context of IgG antibodies, each IgG isotype has three CH regions. Thus, in the context of IgG, the "CH" domains are as follows: "CH1" refers to positions 118-220 according to the EU index as in Kabat. "CH2" refers to positions 237-340 according to the EU index as in Kabat, and "CH3" refers to positions 341-447 according to the EU index as in Kabat. Unless otherwise indicated, the Fc domain of the present invention comprises a hinge that begins at position 216 (EU numbering) and ends at the C-terminus of the CH3 domain at position 447; this is referred to as "hinge-CH2-CH3" for IgG. In some cases, such as in the Fc fusions herein, the hinge serves as a domain linker as discussed herein. In some embodiments, as described more fully below, the Fc region is amino acid modified, for example, to alter binding to one or more FcγR receptors or FcRn receptors or to promote heterodimerization of the Fc domain.

[0125] Thus, as used herein, "Fc variant" or "variant Fc" refers to a protein comprising amino acid modifications in the Fc domain. The Fc variants of the present invention are defined according to the amino acid modifications that constitute them. Thus, for example, N434S or 434S is an Fc variant having a serine substitution at position 434 relative to the parent Fc polypeptide, wherein numbering is according to the EU index. Similarly, M428L / N434S defines an Fc variant having substitutions M428L and N434S relative to the parent Fc polypeptide. The identity of the WT amino acids may not be specified, in which case the variant is referred to as 428L / 434S. It should be noted that the order in which the substitutions are provided is arbitrary, that is, for example, 428L / 434S is an Fc variant identical to M428L / N434S, etc. For all positions discussed herein relating to antibodies, unless otherwise indicated, the amino acid position numbering is according to the EU index. The EU index, or EU index as in the Kabat or EU numbering scheme, refers to the EU antibody numbering (Edelman et al., 1969, Proc. Natl. Acad. Sci. USA 63:78-85, which is incorporated herein by reference in its entirety). Modifications may be additions, deletions, or substitutions. Substitutions may include naturally occurring amino acids, and in some cases, synthetic amino acids. Examples include U.S. Pat. No. 6,586,207; WO 98 / 48032; WO 03 / 073238; US2004-0214988A1; WO 05 / 35727A2; WO 05 / 74524A2; J.W. Chin et al. (2002), Journal of the American Chemical Society, 124:9026-9027; J.W. Chin & P.G. Schultz, (2002), ChemBioChem, 11:1135-1137; J.W. Chin et al. (2002), PICAS United States of America), 99: 11020-11024; and L. Wang & P. ​​G. Schultz, (2002), Chemistry, 1-10, all of which are incorporated by reference in their entirety.

[0126] "Interleukin-2" or "IL-2" herein refers to human IL-2 having the sequence shown in FIG1 .

[0127] By "IL-2 variant" or "variant IL-2" herein is meant a protein comprising amino acid modifications in the mature human IL-2 sequence shown in Figure 1. As described above, the IL-2 variants of the present invention are defined according to the amino acid modifications that constitute them using the numbering of the mature human form.

[0128] By "Fc fusion protein" or "immunoadhesin" herein is meant a protein comprising an Fc region, typically linked (optionally via a linker moiety, which may be a hinge region of an IgG (e.g., IgG1), as described herein) to a different protein, such as IL-2. Thus, an IL-2 Fc fusion protein is a protein comprising IL-2 (in this case, a variant IL-2) and an Fc domain as outlined herein (again, typically an Fc variant). These proteins typically have the structure IL-2-hinge-CH2-CH3. As will be understood in the art, the two Fc domains will self-assemble to provide the dimeric Fc fusion protein outlined herein.

[0129] As used herein, "position" refers to a position in the sequence of a protein. Positions can be numbered sequentially or according to an established format, such as the EU index for antibody numbering.

[0130] As used herein, "non-naturally occurring modification" means an amino acid modification that is not an isotype. For example, because none of the IgGs include a serine at position 434, the substitution 434S of IgG1, IgG2, IgG3, or IgG4 (or a hybrid thereof) is considered a non-naturally occurring modification.

[0131] As used herein, "amino acid" and "amino acid identity" mean one of the 20 naturally occurring amino acids encoded by DNA and RNA.

[0132] As used herein, "effector function" refers to the biochemical events that result in the interaction of the Fc region of an antibody with an Fc receptor or ligand. Effector functions include, but are not limited to, ADCC, ADCP, and CDC.

[0133] As used herein, "Fcγ receptor" or "FcγR (FcgammaR)" means any member of the family of proteins that bind to the Fc region of an IgG antibody and are encoded by an FcγR gene. In humans, this family includes, but is not limited to, FcγRI (CD64), which includes isoforms FcγRIa, FcγRIb, and FcγRIc; FcγRII (CD32), which includes isoforms FcγRIIa (including allotypes H131 and R131), FcγRIIb (including FcγRIIb-1 and FcγRIIb-2), and FcγRIIc; and FcγRIII (CD16), which includes isoforms FcγRIIIa (including allotypes V158 and F158) and FcγRIIIb (including allotypes FcγRIIb-NA1 and FcγRIIb-NA2) (Jefferis et al., 2002, Immunol Lett.). "Lett" 82:57-65, which is incorporated by reference in its entirety); and any undiscovered human FcγRs or FcγR isoforms or allotypes. FcγRs can be from any organism, including, but not limited to, humans, mice, rats, rabbits, and monkeys. Mouse FcγRs include, but are not limited to, FcγRI (CD64), FcγRII (CD32), FcγRIII (CD16), and FcγRIII-2 (CD16-2), as well as any undiscovered mouse FcγRs or FcγR isoforms or allotypes.

[0134] As used herein, "FcRn" or "neonatal Fc receptor" refers to a protein that binds the Fc region of an IgG antibody and is encoded at least in part by an FcRn gene.

[0135] As used herein, "parent polypeptide" means the starting polypeptide that is subsequently modified to generate a variant. A parent polypeptide can be a naturally occurring polypeptide or a variant or engineered version of a naturally occurring polypeptide. A parent polypeptide can refer to the polypeptide itself, a composition comprising the parent polypeptide, or an amino acid sequence encoding the parent polypeptide. Thus, as used herein, "parent IL-2" means the unmodified human IL-2 protein that is modified to generate a variant, and as used herein, "parent Fc" or "parent Fc domain" means the unmodified human IgG Fc domain that is modified to generate a variant Fc domain.

[0136] Herein, in the context of monomers of the heterodimeric Fc fusions of the present invention, "strandedness" means that, similar to the "matching" of two strands of DNA, heterodimerization variants are incorporated into each monomer to maintain the ability to "match" to form heterodimers. For example, if some pI variants are engineered as monomer A (e.g., to make the pI higher), then steric variants that can also be utilized as "charge pairs" do not interfere with the pI variants, such as charge variants that make the pI higher are placed on the same "strand" or "monomer" to maintain both functions. Similarly, for "skewed" variants that appear in pairs as more fully outlined below, one would expect that the pI will determine which strand or monomer will be incorporated into one of the pairs, such that the pI of the skewed variant is also used to maximize pI separation.

[0137] As used herein, "wild type" or "WT" refers to an amino acid sequence or nucleotide sequence found in nature, including allelic variations. A WT protein has an amino acid sequence or nucleotide sequence that has not been intentionally modified.

[0138] The proteins of the present invention are typically isolated or recombinant. When used to describe the various polypeptides disclosed herein, "isolated" means a polypeptide that has been identified and separated and / or recovered from the cell or cell culture in which it is expressed. Generally, an isolated polypeptide will be prepared by at least one purification step. "Recombinant" means that the antibody is produced in an exogenous host cell using recombinant nucleic acid technology.

[0139] "Percentage (%) of amino acid sequence identity" with respect to protein sequences is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in a particular (parent) sequence, after aligning the sequences and introducing gaps (if necessary) to achieve the maximum percentage of sequence identity and without considering any conservative substitutions as part of the sequence identity. Alignment for the purpose of determining percentage of amino acid sequence identity can be achieved in various ways within the art, for example, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. One skilled in the art can determine appropriate parameters for measuring alignment, including any algorithm required to achieve maximum alignment over the full length of the sequences being compared. A particular program is the ALIGN-2 program outlined in paragraphs

[0279] to

[0280] of U.S. Publication No. 20160244525, which is incorporated herein by reference.

[0140] The degree of identity between an amino acid sequence of the invention ("inventive sequence") and a parental amino acid sequence is calculated as the number of exact matches in the alignment of the two sequences divided by the length of the "inventive sequence" or the length of the parental sequence, whichever is shortest. The result is expressed as a percentage of identity.

[0141] In some embodiments, two or more amino acid sequences are at least 50%, 60%, 70%, 80% or 90% identical. In some embodiments, two or more amino acid sequences are at least 95%, 97%, 98%, 99% or even 100% identical.

[0142] " connexon " herein means the protein connexon for connecting two other protein domains (for example, variant IL-2 domain and variant Fc domain).In some cases, connexon is " domain connexon ", for being linked together any two domains as outlined herein.Although any suitable connexon can be used, many embodiments utilize glycine-serine polymer and allow to reorganize any peptide sequence connecting these two domains with the length and flexibility that are enough to allow each domain to keep its biological function, the glycine-serine polymer includes such as (GS) n, (GSGGS) n, (GGGGS) n and (GGGS) n, wherein n is at least one (and generally 3 to 4 to 5) integer.In some cases and when noting " chain type ", as outlined below, charged domain connexon can be used.In addition, the hinge domain of human IgG1 protein can also be domain connexon.

[0143] "Regulatory T cells" or "Tregs" herein refer to T cells that are CD3+ / CD4+ / CD8- / CD25+ / FOXP3+.

[0144] VII. IL-2 Fc Fusion Protein of the Present Invention

[0145] The present invention provides as herein shown and Figure 19 As will be appreciated by those skilled in the art, the fusion protein of the present invention is actually two different polypeptides that self-assemble into a homodimeric protein ( Figure 19 B) or heterodimeric protein ( Figure 19 A) The proteins of the present invention typically have three different domains: an Fc domain, one or more domain linkers, and an IL-2 domain.

[0146] A. IL-2 Domains of the Invention

[0147] The IL-2 Fc fusion proteins of the present invention comprise IL-2 domains that are variant human IL-2 domains. As discussed herein, these domains are engineered to comprise specific variants that increase activation of CD25+ T cells (such as Tregs) compared to CD25- T cell populations; and optionally further comprise amino acid substitutions designed to increase binding of IL-2 to IL-2Rα at pH 6, such that the IL-2 variant Fc fusion protein is recycled through the endocytic pathway rather than degraded.

[0148] 1. Expression variants

[0149] First, the IL-2 variants of the present invention also include the C125S variant that has previously been shown to increase the expression of human IL-2. Therefore, unless otherwise specified, all variants described herein include the C125S variant; in some cases, the C125A variant may also be used.

[0150] Additionally, in some cases, the IL-2 variants of the present invention comprise a T3A variant in which the O-glycosylation site has been removed to reduce complexity.

[0151] Additionally, the IL-2 variants of the present invention comprise additional mutations.

[0152] 2. Specific variants

[0153] Thus, the present invention provides IL-2 variant proteins with increased specificity for CD25+ T cells (including Tregs). This is generally achieved by increasing binding to IL-2Rα, or reducing binding to IL-2Rβ and IL-2Rγ, or binding to the IL-2Rβγ interface, or both.

[0154] In one embodiment, the variant IL-2 comprises a D20N amino acid substitution in addition to a C125S expression variant and thus has the amino acid group D20N / C125S. It should be noted that the D20N variant has previously been reported to result in loss of binding to both the high affinity receptor (IL-2Rαβ) and the intermediate affinity receptor (IL-2Rβ); see Collins et al., Proc. Natl. Acad. Sci. USA 85:7709-7713 (1988), which showed that "differential binding or activation between IL-2Rβγ or IL-2Rαβγ could not be achieved by substituting Asp at position 20," see U.S. Pat. No. 6,955,807 for a description of related art.

[0155] In one embodiment, the variant IL-2 includes a T3A amino acid variant in addition to a D20N amino acid substitution and a C125S expression variant, and thus has the amino acid set T3A / D20N / C125S.

[0156] In one embodiment, the variant IL-2 comprises, in addition to the T3A amino acid variant, the D20N amino acid substitution, and the C125S expression variant, a T37R amino acid variant and thus has the amino acid group T3A / D20N / T37R / C125S.

[0157] In one embodiment, the variant IL-2 comprises, in addition to the T3A amino acid variant, the D20N amino acid substitution, and the C125S expression variant, the N71K amino acid variant and thus has the amino acid set T3A / D20N / N71K / C125S.

[0158] In one embodiment, the variant IL-2 comprises the amino acid variant N29S / Y31H / K35R / T37A / K48E / N71R / N88D / I89V in addition to the C125S expression variant, and thus has the amino acid group N29S / Y31H / K35R / T37A / K48E / N71R / N88D / I89V / C125S.

[0159] a. Determination of Treg specificity

[0160] As known in the art, activation of STAT5 proteins (STAT5a and STAT5b) by phosphorylation is one of the early signaling events mediated by IL-2. Therefore, specificity can be assessed by observing STAT5 phosphorylation on different T cell populations using the constructs of the present invention.

[0161] Typically, as described in the Examples, STAT5 phosphorylation is determined using the methods outlined in the Examples. As described in the Examples, five different cell types are typically tested, including CD4+ / CD45RA+, CD4+ / CD45RA-, CD8+CD45RA+, CD8+CD45RA-, and Treg (CD3+ / CD4+ / CD8- / CD25+ / FOXP3+), to provide sampling of other T cell types (e.g., CD45RA is expressed on memory T cells but not on naive T cells).

[0162] Typically, the increase in activity is compared to human wild-type IL-2.

[0163] 3. pH variants

[0164] Additionally, the present invention provides IL-2 variant proteins with increased pH specificity, wherein binding is increased at pH 6 (endosomal pH).

[0165] In this embodiment, the IL-2 variant may have one or more amino acid substitutions selected from the group consisting of: R38A, R38D, R38E, R38F, R38G, R38H, R38I, R38K, R38L, R38M, R38N, R38P, R38Q, R38S, R38T, R38V, R38W, R38Y, T41A, T41D, T41E, T41F, T41G, T41 25, F42A, F42D, F42E, F42G, F42H, F42I, F42K, F42L, F42M, F42N, F42P, F42Q, F42R, F42S, F42T, F42V, F42W, and F42Y. In addition, these mutations can be combined with C125S.

[0166] In this embodiment, the IL-2 variant may have one or more amino acid substitutions selected from the group consisting of: R38Q / T41K, R38Q / 41Q, R38E / T41K, R38Q / T41R, R38N / T41Q, R38Q / T41V, R38N / T41V, R38Q / T41M, R38Q / T41S, R38Q / T41L, R38N / T41M, T41I / F42Y, T41E / F42Y, T41D / F42Y, T41M / F42Y, T41Q / F42Y, T41E / F42H, T41E / F42L, T41E / F42P, R38Q / F42Y, R38N / T41R, R38N / T41K , R38V / T41R, R38P / T41R, T41E / F42K, T41D / F42K, T41M / F42K, T41Q / F42K, R 38Q / F42K, T41I / F42K, R38N / F42K, T41H / F42K, R38Q / T41K / F42Y, R38Q / T41R / F42Y, R38Q / T41Q / F42Y, R38Q / T41V / F42Y, R38N / T41K / F42K, R38Q / T41H / F42K, R38Q / T41K / F42K, R38Q / T41Q / F42K, R38Q / T41V / F42K and R38Q / T41R / F42K. In addition, these mutations can be combined with C125S.

[0167] 4. Useful IL-2 variants

[0168] The present invention provides a number of particularly useful IL-2 variants having the desired activity both alone and when fused to an Fc domain, comprising both a wild-type Fc domain and a variant Fc domain as outlined herein. In addition, these IL-2 variants can be used in monovalent constructs (e.g., Figure 19 A) or a bivalent construct (e.g., Figure 19 B).

[0169] In one embodiment, the variant IL-2 domain has the amino acid substitution R38I / C125S and is used in a bivalent construct. For example, in this embodiment, the variant IL-2 domain can be fused to a wild-type Fc domain from IgG2 or IgG4. Alternatively, the variant IL-2 domain can be fused to a variant Fc domain, for example, a variant Fc structure containing an ablation variant and an FcRn variant.

[0170] In one embodiment, the variant IL-2 domain has the amino acid substitution R38I / C125S and is used in a monovalent construct.

[0171] In one embodiment, the variant IL-2 domain has the amino acid substitution R38L / C125S and is used in a monovalent construct.

[0172] In one embodiment, the variant IL-2 domain has the amino acid substitution R38L / C125S and is used in a bivalent construct.

[0173] In one embodiment, the variant IL-2 domain has the amino acid substitution D20N / C125S and is used in a bivalent construct.

[0174] In one embodiment, the variant IL-2 domain has the amino acid substitution D20N / C125S and is used in a monovalent construct.

[0175] In one embodiment, the variant IL-2 domain has the amino acid substitutions T3A / D20N / C125S and is used in a bivalent construct.

[0176] In one embodiment, the variant IL-2 domain has the amino acid substitutions T3A / D20N / C125S and is used in a monovalent construct.

[0177] In one embodiment, the variant IL-2 domain has the amino acid substitutions N29S / Y31H / K35R / T37A / K48E / N71R / N88D / I89V / C125S and is used in a bivalent construct.

[0178] In one embodiment, the variant IL-2 domain has the amino acid substitutions N29S / Y31H / K35R / T37A / K48E / N71R / N88D / I89V / C125S and is used in a monovalent construct.

[0179] In one embodiment, a variant IL-2 domain has the amino acid substitutions T3A / D20N / T37R / C125S and is used in a bivalent construct.

[0180] In one embodiment, the variant IL-2 domain has the amino acid substitutions T3A / D20N / T37R / C125S and is used in a monovalent construct.

[0181] In one embodiment, the variant IL-2 domain has the amino acid substitutions T3A / D20N / N71K / C125S and is used in a bivalent construct.

[0182] In one embodiment, the variant IL-2 domain has the amino acid substitutions T3A / D20N / N71K / C125S and is used in a monovalent construct.

[0183] In one embodiment, the variant IL-2 domain has the amino acid substitutions T3A / D20N / T37R / R38I / C125S and is used in a bivalent construct.

[0184] In one embodiment, the variant IL-2 domain has the amino acid substitutions T3A / D20N / T37R / R38I / C125S and is used in a monovalent construct.

[0185] In one embodiment, a variant IL-2 domain has the amino acid substitutions T3A / D20N / R38I / N71K / C125S and is used in a bivalent construct.

[0186] In one embodiment, the variant IL-2 domain has the amino acid substitutions T3A / D20N / R38I / N71K / C125S and is used in a monovalent construct.

[0187] Particularly preferred proteins include XENP14142, XENP14144, XENP23833, XENP25720, XENP26086, XENP26105, XENP26987, XENP27003, XENP26109, XENP26994, XENP26841, XENP27004, XENP26839, XENP26996, XENP26990, XENP27006, XENP26840, XENP27001, XENP26991, XENP27007, XENP27563, XENP26105, XENP27564, and XENP26109.

[0188] B. Fc Domains of the Invention

[0189] As discussed herein, the present invention provides Fc fusion proteins comprising two Fc domains, wherein at least one of the Fc domains comprises a fused 11-2 variant, optionally comprising a domain linker. Figure 19 As shown, the dimeric proteins of the present invention can be configured to have one IL-2 variant, sometimes referred to herein as "monovalent IL-2," such as Figure 19 As shown in A, one of the Fc domains is covalently linked to the IL-2 variant protein and the other is "empty" or "Fc-only". As discussed below, this embodiment relies on heterodimeric Fc domains. Alternatively, use Figure 19 B shows a "bivalent" IL-2 construct in which each Fc domain is fused to an IL-2 variant; these examples utilize homodimeric Fc domains, as discussed above.

[0190] In both embodiments, whether a homodimeric or heterodimeric Fc fusion construct is used, the Fc domain typically contains some specific amino acid variants for several functions.

[0191] 1. Fc variants with additional functions

[0192] In addition to pi amino acid variants, there are many useful Fc amino acid modifications that can be made for a variety of reasons, including but not limited to altered binding to one or more FcTR receptors, altered binding to FcRn receptors, etc.

[0193] Thus, the proteins of the present invention may comprise amino acid modifications, including heterodimerization variants as outlined herein, including pI variants and steric variants. Each variant group may be independently and optionally included or not included in any particular heterodimer protein.

[0194] (i) FcγR variants

[0195] Thus, there are a number of useful Fc substitutions that can be made to alter binding to one or more of the FcγR receptors. Substitutions that result in increased binding as well as decreased binding can be useful. For example, it is known that increased binding to FcγRIIIa results in increased ADCC (antibody-dependent cell-mediated cytotoxicity; a cell-mediated reaction in which nonspecific toxic cells expressing FcγRs recognize bound antibody on a target cell and subsequently cause lysis of the target cell). Similarly, decreased binding to FcγRIIb (an inhibitory receptor) can also be beneficial in some circumstances. Amino acid substitutions useful in the present invention include those described in USSN 11 / 124,620 (particularly Figure 41 ), amino acid substitutions listed in 11 / 174,287, 11 / 396,495, and 11 / 538,406, all of which are expressly incorporated herein by reference in their entirety and are particularly useful for the variants disclosed herein. Specific variants used include, but are not limited to, 236A, 239D, 239E, 332E, 332D, 239D / 332E, 267D, 267E, 328F, 267E / 328F, 236A / 332E, 239D / 332E / 330Y, 239D, 332E / 330L, 243A, 243L, 264A, 264V, and 299T.

[0196] In addition, as specifically disclosed in USSN 12 / 341,769, which is incorporated herein by reference in its entirety, there are additional Fc substitutions for increased binding to FcRn and increased serum half-life, including but not limited to: 434S, 434A, 428L, 308F, 259I, 428L / 434S, 428L / 434A, 259I / 308F, 436I / 428L, 436I or V / 434S, 436V / 428L, and 259I / 308F / 428L.

[0197] (ii) Ablation variant

[0198] Similarly, another category of functional variants is "FcγR ablation variants" or "Fc knockout (FcKO or KO) variants." In these embodiments, for some therapeutic applications, it is desirable to reduce or eliminate the normal binding of the Fc domain to one or more or all of the Fcγ receptors (e.g., FcγR1, FcγRIIa, FcγRIIb, FcγRIIIa, etc.) to avoid additional mechanisms of action. That is, for example, it is desirable to ablate FcγRIIIa binding to eliminate or significantly reduce ADCC activity, such that one of the Fc domains includes one or more Fcγ receptor ablation variants. Figure 4These ablation variants are depicted in , and each can independently and optionally be included or not, with preferred aspects utilizing an ablation variant selected from the group consisting of: G236R / L328R, E233P / L234V / L235A / G236del / S239K, E233P / L234V / L235A / G236del / S267K, E233P / L234V / L235A / G236del / S239K / A327G, E233P / L234V / L235A / G236del / S267K / A327G, and E233P / L234V / L235A / G236del. It should be noted that the ablation variants cited herein ablate FcγR binding but generally do not ablate FcRn binding.

[0199] 2. Homodimeric Fc domain

[0200] In some embodiments, the present invention provides as generally described in Figure 19 A bivalent homodimeric protein comprising a homodimeric Fc domain is depicted in B. In this example, each monomer is identical and typically comprises variant-IL-2-linker-Fc domain, wherein the linker is typically a hinge from IgG1.

[0201] In this embodiment, the Fc domain may have Figure 4 Ablation variant commonly shown. Figure 4 Suitable ablated variants that generally ablate binding to FcγRI, FcγRIIb, and FcγRIIIa are shown in Particularly useful in this IgG1 example is the ablated amino acid set of E233P / L234V / L235A / G236_ / S267K ("G236_" is a deletion as described herein).

[0202] In addition, as specifically disclosed in USSN 12 / 341,769, which is incorporated herein by reference in its entirety, there are additional Fc substitutions for increased binding to FcRn and increased serum half-life, including but not limited to: 434S, 434A, 428L, 308F, 259I, 428L / 434S, 428L / 434A, 259I / 308F, 436I / 428L, 436I or V / 434S, 436V / 428L, and 259I / 308F / 428L.

[0203] 3. Heterodimeric Fc domain

[0204] In addition to homodimeric, bivalent IL-2 fusion proteins, alternative embodiments utilize monovalent IL-2 fusion proteins in which one of the Fc domains is "empty" and the invention relies on heterodimeric variants to bring the two Fc domains together, such as Figure 19 B. These examples rely on the use of two different variant Fc sequences that will self-assemble to form heterodimeric Fc domains and heterodimeric Fc fusion proteins.

[0205] Heterodimeric protein constructs are based on the self-assembly properties of the two Fc domains of the heavy chain of an antibody (e.g., two "monomers" assembled into a "dimer"). Heterodimeric proteins are prepared by altering the amino acid sequence of each monomer, as discussed more fully below. Thus, the present invention generally relates to the generation of heterodimeric Fc fusion proteins that rely on amino acid variants in the constant region that differ on each chain to promote heterodimer formation and / or allow for easier purification of heterodimers compared to homodimers.

[0206] There are a variety of mechanisms that can be used to generate heterodimers of the present invention. In addition, as will be appreciated by those skilled in the art, these mechanisms can be combined to ensure high heterodimerization. Therefore, amino acid variants that result in heterodimer production are referred to as "heterodimerization variants." As discussed below, heterodimerization variants can include spatial variants (e.g., "knobs and holes" variants or "skewed" variants and "charge pairs" variants described below) and "pI variants," which allow purification of homodimers away from heterodimers. As generally described in WO2014 / 145806, which is hereby incorporated by reference in its entirety, and specifically as described below with respect to the discussion of "heterodimerization variants," useful mechanisms of heterodimerization include "knob and hole" ("KIH"; sometimes referred to herein as "skewed" variants (see discussion in WO2014 / 145806)), "electrostatic steering" or "charge pairs" as described in WO2014 / 145806, pi variants as described in WO2014 / 145806, and generally additional Fc variants as summarized in WO2014 / 145806 and below.

[0207] In the present invention, there are several basic mechanisms that can facilitate the purification of heterodimeric proteins; one mechanism relies on the use of pI variants, so that each monomer has a different pI, thereby allowing isoelectric purification of AA, AB, and BB dimeric proteins. In addition, the heterodimeric proteins of the present invention also allow for size-based separation. As further outlined below, it is also possible to "skew" the formation of heterodimers compared to homodimers. Therefore, the combination of steric heterodimerization variants with pI variants or charge pair variants is particularly useful in the present invention.

[0208] In general, embodiments particularly useful in the present invention rely on variant panels comprising skewed variants that encourage heterodimer formation over homodimer formation, combined with pi variants that increase the pi difference between the two monomers.

[0209] Additionally, as more fully outlined herein, pi variants can be contained within the constant domains and / or Fc domains of the monomer, or charged linkers, such as domain linkers, can be used.

[0210] In the present invention, where pI is used as a separation mechanism to allow purification of heterodimeric proteins, amino acid variants can be introduced into one or both monomeric polypeptides; that is, the pI of one of the monomers (referred to herein as "monomer A" for simplicity) can be engineered to be away from monomer B, or both monomer A and monomer B can be altered as the pI of monomer A increases and the pI of monomer B decreases. As discussed, the pI of either or both monomers can be altered by removing or adding charged residues (e.g., replacing a neutral amino acid with a positively charged amino acid residue or a negatively charged amino acid residue, e.g., glycine to glutamic acid), changing a charged residue from positively or negatively charged to the opposite charge (e.g., aspartic acid to lysine), or changing a charged residue to a neutral residue (e.g., loss of charge; lysine to serine). A variety of these variants are shown in the accompanying drawings.

[0211] Thus, this embodiment of the invention provides for generating a sufficient change in the pI of at least one of the monomers such that heterodimers can be separated from homodimers. As will be appreciated by those skilled in the art and as discussed further below, such separation can be performed by using a "wild-type" heavy chain constant region and a variant region that has been engineered to increase or decrease its pI (wt A- +B or wt A- -B) or to increase one region and decrease the other (A+ -B- or A- B+).

[0212] Thus, in general, components of some embodiments of the present invention are amino acid variants in the Fc domain that alter the isoelectric point (pI) of at least one, if not both, of the monomers of the dimeric protein by incorporating amino acid substitutions ("pI variants" or "pI substitutions") into one or both of the monomers. As shown herein, separation of a heterodimer from two homodimers can be accomplished when the pIs of the two monomers differ by as little as 0.1 pH units, with 0.2, 0.3, 0.4, and 0.5 or more pH units being useful in the present invention.

[0213] a. Heterodimerization variants

[0214] The present invention provides heterodimeric proteins comprising heterodimeric antibodies in various forms that utilize heterodimeric variants to allow heterodimer formation and / or purification away from homodimers. A plurality of heterodimerization variants are shown in FIG2 .

[0215] There are multiple suitable pairs of heterodimerization-skewed variants. These variants appear as "sets" of "pairs." That is, one set of the pair is incorporated into the first monomer and the other set of the pair is incorporated into the second monomer. It should be noted that these sets do not necessarily behave as "knob-into-hole" variants, where there is a one-to-one correspondence between residues on one monomer and residues on the other; that is, these pairs form an interface between the two monomers that encourages heterodimer formation and discourages homodimer formation, thereby allowing the percentage of heterodimers that spontaneously form under biological conditions to exceed 90%, rather than the expected 50% (25% homodimer A / A: 50% heterodimer A / B: 25% homodimer B / B).

[0216] b. Spatial variants

[0217] In some embodiments, heterodimer formation can be promoted by adding steric variants. That is, by varying the amino acids in each heavy chain, different heavy chains are more likely to associate to form a heterodimer structure rather than forming homodimers with the same Fc amino acid sequence. Suitable steric variants are included in the accompanying drawings.

[0218] A mechanism often referred to in the art as "knob and hole" refers to amino acid engineering that creates steric effects that favor heterodimer formation and disfavor homodimer formation and can also optionally be used; this is sometimes referred to as "knob and hole," as described in USSN 61 / 596,846, Ridgway et al., Protein Engineering 9(7):617 (1996); Atwell et al., J. Mol. Biol. 1997 270:26; U.S. Pat. No. 8,216,805, all of which are incorporated herein by reference in their entireties. The accompanying figures identify multiple "monomer A-monomer B" pairs that rely on "knob and hole." Additionally, as described in Merchant et al., Nature Biotech. 16:677 (1998), these "knob and hole" mutations can be combined with disulfide bonds to bias formation toward heterodimerization.

[0219] Another mechanism for generating heterodimers is sometimes referred to as "electrostatic steering," as described in Gunasekaran et al., J. Biol. Chem. 285(25): 19637 (2010), which is incorporated herein by reference in its entirety. This is sometimes referred to herein as a "charge pair." In this example, electrostatics were used to skew the formation toward heterodimerization. As will be appreciated by those skilled in the art, these charge pairs may also affect pI and thus purification, and therefore may also be considered pI variants in some cases. However, since these charge pairs are generated to force heterodimerization and are not used as a purification tool, they are classified as "steric variants." These charge pairs include, but are not limited to, D221E / P228E / L368E paired with D221R / P228R / K409R (eg, these are the "monomer counterparts") and C220E / P228E / 368E paired with C220R / E224R / P228R / K409R.

[0220] Additional monomer A and monomer B variants can be optionally and independently combined in any number with other variants, such as the pi variants outlined herein or those of US 2012 / 0149876. Figure 37 The other spatial variants shown, the figures and legends as well as the SEQ ID NOs of said document are expressly incorporated herein by reference.

[0221] In some embodiments, the steric variants outlined herein may optionally and independently be incorporated into one or two monomers with any pi variant (or other variant, such as an Fc variant, an FcRn variant, etc.), and may independently and optionally be included or not included in a protein of the invention.

[0222] A list of suitable skew variants is shown in Figure 2, which illustrates the specific use of some pairs in many embodiments. Pairs including, but not limited to, the following are particularly useful in some embodiments: S364K / E357Q:L368D / K370S; L368D / K370S:S364K; L368E / K370S:S364K; T411T / E360E / Q362E:D401K; L368D / K370S:S364K / E357L, K370S:S364K / E357Q and T366S / L368A / Y407V:T366W (optionally comprising a bridging disulfide T366S / L368A / Y407V / Y349C:T366W / S354C). In terms of nomenclature, "S364K / E357Q:L368D / K370S" means that one of the monomers has the double variant set S364K / E357Q and the other has the double variant set L368D / K370S; as above, these "strand-type" pairs depend on the starting pi.

[0223] c. pI (isoelectric point) of the heterodimer

[0224] In general, as will be appreciated by those skilled in the art, there are two general categories of pI variants: variants that increase the pI of a protein (alkaline changes) and variants that decrease the pI of a protein (acidic changes). As described herein, all combinations of these variants can be made: one monomer can be wild-type or a variant that does not exhibit a pI significantly different from wild-type, and another can be more basic or more acidic. Alternatively, individual monomers can be altered, one to be more basic and one to be more acidic.

[0225] Fig. 5 shows the preferred combination of pI variants. As outlined herein and shown in the accompanying drawings, these changes are shown relative to IgG1, but all isotypes and isotype hybrids can be changed in this way. In the case where the heavy chain constant domain is from IgG2-4, R133E and R133Q can also be used.

[0226] In one embodiment, a preferred combination of pI variants has one monomer (minus side) comprising the 295E / 384D / 418E / 421D variant (Q295E / N384D / Q418E / N421D when relative to human IgG1) and a second monomer (plus side) comprising a positively charged scFv linker comprising (GKPGS)4.

[0227] d. Isotype variants

[0228] Additionally, some embodiments of the present invention rely on the "import" of pi amino acids at specific positions from one IgG isotype to another, thereby reducing or eliminating the possibility of introducing unwanted immunogenicity into the variant. Several of these are shown in U.S. Publication No. 2014 / 0370013. Figure 21 , which is incorporated herein by reference. That is, for a variety of reasons, IgG1 is a commonly used isotype for therapeutic antibodies, comprising high effector functions. However, the heavy constant region of IgG1 has a higher pI than that of IgG2 (8.10 compared to 7.31). By introducing IgG2 at a specific position into the IgG1 main chain, the pI of the resulting monomer is reduced (or increased) and a longer serum half-life is additionally exhibited. For example, IgG1 has a glycine at position 137 (pI of 5.97), and IgG2 has a glutamic acid (pI of 3.22); importing glutamic acid will affect the pI of the resulting protein. As described below, multiple amino acid substitutions are generally required to significantly affect the pI of the variant antibody. However, it should be noted that, as discussed below, even changes in the IgG2 molecule allow for an increase in serum half-life.

[0229] In other embodiments, non-isotypic amino acid changes are made to lower the overall charge state of the resulting protein (e.g., by changing a higher pi amino acid to a lower pi amino acid) or to allow for structural adjustment for stabilization, etc., as described further below.

[0230] In addition, by engineering the pI of the heavy and light constant domains, significant changes can be seen in each monomer of the heterodimer. As discussed herein, having the pI of the two monomers differ by at least 0.5 can allow separation by ion exchange chromatography or isoelectric focusing or other methods sensitive to the isoelectric point.

[0231] e. Calculate pI

[0232] As outlined herein, the pi of each monomer may depend on the pi of the variant IL-2 domain and the Fc domain. Thus, in some embodiments, the pi of U.S. Publication No. 2014 / 0370013 is used. Figure 19 The chart in [ ] calculates the change in pI based on the variant heavy chain constant domain. As discussed herein, which monomer to engineer is typically determined by the inherent pI of the Fv and scaffold regions. Alternatively, the pI of each monomer can be compared.

[0233] f. Combination of heterodimer variants and Fc variants

[0234] As will be understood by those skilled in the art, for use in heterodimeric Fc domains, all of the listed heterodimerization variants (including skew variants and / or pI variants) can be optionally and independently combined in any manner, as long as the heterodimerization variants maintain their "chain type" or "monomer separation." In addition, all of these variants can be combined into any heterodimerization format.

[0235] In the case of pi variants, while particularly useful examples are shown in the figures, other combinations can be generated following the general rule of varying the pi difference between two monomers to facilitate purification.

[0236] Additionally, any of the heterodimerization variants, skew variants, and pi variants are independently and optionally combined with Fc ablation variants, Fc variants, and FcRn variants, as generally outlined herein.

[0237] In addition to the ablated variants, the Fc domain typically also comprises a C220S variant in which the light chain is eliminated because the light chain is not included herein and this cysteine ​​is used to form a disulfide with the light chain (e.g., because the Fc domain of the present invention comprises a hinge region starting at position 216 (EU numbering)).

[0238] Additionally, the Fc domain of the fusion protein of the present invention optionally may include half-life extending amino acid substitutions.

[0239] Recently, it has been proposed that antibodies with variable regions having lower isoelectric points may also have longer serum half-lives (Igawa et al., 2010 Protein Engineering Design and Selection (PEDS) 23(5):385-392, which is incorporated by reference in its entirety. However, the mechanism for this remains poorly understood. Moreover, variable regions vary from antibody to antibody. Constant region variants with reduced pI and extended half-life would provide a more modular approach to improving the pharmacokinetic properties of antibodies, as described herein.

[0240] VIII. Useful Constructs of the Invention

[0241] As outlined herein, the present invention provides a variety of useful monovalent and bivalent constructs.

[0242] A. Heterodimer Monovalent Constructs

[0243] In some embodiments, the 11-2-Fc fusion protein of the present invention is a heterodimeric monovalent construct, such as Figure 19 Heterodimer monovalent constructs depicted in Figures A and C. In this example, the variant IL-2 domain is typically fused to a variant human IgG1 Fc domain using a hinge (typically comprising a C220S variant) as a domain linker or using an additional linker attached to the hinge, while the other Fc domain (comprising the hinge) remains "empty."

[0244] In some embodiments, the variant IL-2 domain is attached to a "+" monomer side comprising (see Figure 5A ): variant human IgG1 Fc domain (comprising hinge with C220S variant), S364K / E357Q "skew variant" and ablated variant E233P / L234V / L235A / G236del / S267K, and the "empty Fc side" is variant human IgG1 Fc domain (comprising hinge with C220S variant), L368D / K370S "skew variant" and ablated variant E233P / L234V / L235A / G236del / S267K. In this embodiment, preferred constructs comprise a variant IL-2 domain having amino acid substitutions selected from the group consisting of: T3A / D20N / T37R, T3A / D20N / T37R / C125S, T3A / D20N / T37R / C125A, T3A / D20N / N71K, T3A / D20N / N71K / C125S, and T3A / D20N / N71K / C125A.

[0245] In some embodiments, the variant IL-2 domain is attached to a "+" monomer side comprising (see Figure 5A): variant human IgG1 Fc domain (comprising hinge with C220S variant), S364K / E357Q "skew variant" and ablated variant E233P / L234V / L235A / G236del / S267K, as well as FcRn variant M428L / N434S, and the "empty Fc side" is variant human IgG1 Fc domain (comprising hinge with C220S variant), L368D / K370S "skew variant" and ablated variant E233P / L234V / L235A / G236del / S267K, as well as FcRn variant M428L / N434S. In this embodiment, preferred constructs comprise a variant IL-2 domain having amino acid substitutions selected from the group consisting of: T3A / D20N / T37R, T3A / D20N / T37R / C125S, T3A / D20N / T37R / C125A, T3A / D20N / N71K, T3A / D20N / N71K / C125S, and T3A / D20N / N71K / C125A.

[0246] In one embodiment, "monomer 1" comprises a variant IL-2 domain comprising amino acid substitutions T3A / D20N / N71K / C125S (SEQ ID NO: 2 compared to wild-type IL-2) and a "monomer 1 Fc backbone" comprising SEQ ID NO: 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or 33 selected from those of Figure 6. In this embodiment, "monomer 2" comprises an "empty Fc" of a "monomer 2 Fc backbone" selected from SEQ ID NO: 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32 and 34.

[0247] In one embodiment, "monomer 1" comprises a variant IL-2 domain comprising amino acid substitutions T3A / D20N / N71K / C125S (SEQ ID NO: 2 compared to wild-type IL-2), and a "monomer 1 Fc backbone" comprising SEQ ID NO: 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, or 33, but with the FcRn variant M428L / N434S, selected from those of Figure 6. In this embodiment, "monomer 2" comprises an "empty Fc" selected from SEQ ID NO: 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, and 34, but with the FcRn variant M428L / N434S, "monomer 2 Fc backbone."

[0248] In one embodiment, "monomer 1" comprises a variant IL-2 domain comprising amino acid substitutions T3A / D20N / T37R / C125S (SEQ ID NO: 2 compared to wild-type IL-2), and a "monomer 1 Fc backbone" comprising SEQ ID NOs: 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, or 33 selected from those of Figure 6. In this embodiment, "monomer 2" comprises an "empty Fc" of a "monomer 2 Fc backbone" selected from SEQ ID NOs: 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, and 34.

[0249] In one embodiment, "monomer 1" comprises a variant IL-2 domain comprising the amino acid substitutions T3A / D20N / T37R / C125S (SEQ ID NO: 2 compared to wild-type IL-2) and an "Fc backbone" comprising SEQ ID NO: 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31 or 33 but with the FcRn variant M428L / N434S selected from those of Figure 6. In this embodiment, "monomer 2" comprises an "empty Fc" selected from SEQ ID NO: 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32 and 34 but with the FcRn variant M428L / N434S "monomer 2 Fc backbone".

[0250] In these embodiments, preferred constructs comprise XENP26105, XENP27563, XENP26109, and XENP27564.

[0251] B. Homodimeric Bivalent Constructs

[0252] In some embodiments, the 11-2-Fc fusion protein of the present invention is a homodimeric bivalent construct, such as Figure 19 Homodimeric bivalent constructs are depicted in Figures B and D. In this example, variant IL-2 domains are each fused to a variant human IgG1 Fc domain, typically using a hinge (typically comprising a C220S variant) as a domain linker or using an additional linker attached to the hinge.

[0253] In some embodiments, the variant IL-2 domain is linked to a variant human IgG1 Fc domain comprising a hinge with a C220S variant and ablated variants E233P / L234V / L235A / G236del / S267K. In this embodiment, the preferred construct comprises a variant IL-2 domain having amino acid substitutions selected from the group consisting of: T3A / D20N / T37R, T3A / D20N / T37R / C125S, T3A / D20N / T37R / C125A, T3A / D20N / N71K, T3A / D20N / N71K / C125S, and T3A / D20N / N71K / C125A.

[0254] In some embodiments, the variant IL-2 domain is linked to a variant human IgG1 Fc domain comprising a hinge having a C220S variant, an ablated variant E233P / L234V / L235A / G236del / S267K, and an FcRn variant M428L / N434S. In this embodiment, the preferred construct comprises a variant IL-2 domain having an amino acid substitution selected from the group consisting of: T3A / D20N / T37R, T3A / D20N / T37R / C125S, T3A / D20N / T37R / C125A, T3A / D20N / N71K, T3A / D20N / N71K / C125S, and T3A / D20N / N71K / C125A.

[0255] IX. Nucleic Acids of the Invention

[0256] The present invention further provides nucleic acid compositions encoding homodimeric bivalent IL-2-Fc fusion proteins and heterodimeric monovalent IL-2 fusion proteins.

[0257] As will be appreciated by those skilled in the art, the nucleic acid composition will depend on the format of the heterodimeric protein. Thus, for example, when the format requires two amino acid sequences, e.g., Figure 19 In the heterodimeric monovalent form of A, the two nucleic acid sequences can be incorporated into one or more expression vectors for expression.

[0258] Alternatively, when the homodimer bivalent formula is as Figure 19 As shown in B, a single nucleic acid construct and a single expression vector are used.

[0259] As is known in the art, nucleic acids encoding the components of the present invention can be incorporated into expression vectors as are known in the art and depending on the host cell used to produce the heterodimeric Fc fusion protein of the present invention. Typically, the nucleic acid is operably linked to any number of regulatory elements (promoter, replication origin, selectable marker, ribosome binding site, inducer, etc.). The expression vector can be an extrachromosomal vector or an integrating vector.

[0260] The nucleic acids and / or expression vectors of the invention are then transformed into any number of different types of host cells as are well known in the art, including mammalian cells, bacterial cells, yeast cells, insect cells, and / or fungal cells, with mammalian cells (e.g., CHO cells) being used in many embodiments.

[0261] In some embodiments, the nucleic acids encoding each monomer are each contained within a single expression vector, typically under different or identical promoter control conditions, if applicable depending on the format. In embodiments particularly useful in the present invention, each of the two or three nucleic acids is contained on a different expression vector.

[0262] The heterodimeric Fc fusion protein of the present invention is prepared by culturing a host cell containing one or more expression vectors as are well known in the art. Once produced, conventional fusion protein or antibody purification steps are performed, including ion exchange chromatography steps. As discussed herein, making the pI of the two monomers differ by at least 0.5 allows for separation by ion exchange chromatography or isoelectric focusing or other methods sensitive to the isoelectric point. That is, pI substitutions that alter the isoelectric point (pI) of each monomer result in each monomer having a different pI and the heterodimer also having a different pI, thereby promoting isoelectric purification of the heterodimer (e.g., anion exchange column, cation exchange column). These substitutions also help determine and monitor any contaminated homodimers after purification (e.g., IEF gel, cIEF, and analytical IEX columns).

[0263] X. Preparations

[0264] The formulations of the compositions used in accordance with the present invention are prepared by mixing the fusion protein having the desired degree of purity with optional pharmaceutically acceptable carriers, excipients, or stabilizers (as generally outlined in Remington's Pharmaceutical Sciences, 16th edition, Osol, A. ed.

[1980] ) for storage as lyophilized formulations or aqueous solutions.

[0265] XI. Treatment

[0266] The IL-2-Fc fusion protein compositions of the present invention can be used, for example, to treat autoimmune diseases by activating CD25+ cells of patients using the dimeric protein of the present invention.

[0267] XII. Examples

[0268] A. Example 1: Engineering IL-2 to Increase Half-Life

[0269] As discussed above, IL-2 is cleared very rapidly. This rapid clearance is partly dependent on the internalization of the IL-2:IL-2R complex. Following internalization into endosomes, IL-2, IL-2Rβ, and γc are degraded, while IL-2Rα is constitutively recycled to the cell surface. The IL-2 variant 2D1, which harbors the L18M / L19S mutations, has been shown to exhibit a longer half-life than wild-type IL-2. Fallon et al. reported that wild-type IL-2 has a lower affinity for IL-2Rα at endosomal pH, suggesting pH-dependent binding between IL-2 and IL-2Rα. The group further found that the extended half-life of 2D1 is due to recycling with IL-2Rα, as 2D1 has a higher affinity for IL-2Rα than for WT IL-2 at endosomal pH. Notably, residues L18 and L19 on IL-2 have been described to contact IL-2Rβ and γc. In contrast, we hypothesized that modifying residues at the IL-2:IL-2Rα interface would be more suitable to attenuate pH-dependent binding, increase recycling of IL-2 to IL-2Rα, and enhance half-life by prolonging its activity.

[0270] 1. Engineering IL-2 at the IL-2 / IL-2Rα interface to attenuate pH-dependent binding

[0271] By examining the crystal structure of the IL-2 and IL-2Rα interface (PDB code 2ERJ), a possible natural "pH switch" consisting of interactions between Arg38, Thr41 and / or Phe42 of IL-2 and His120 of IL-2Rα at the IL-2 / IL-2Rα interface was identified (see Figure 8). It is hypothesized that after internalization of the IL-2:IL-2Rαβγ complex, at low pH in the endosome, His20 will become protonated, allowing IL-2 to be released from IL-2Rα and subsequently subjecting the remaining IL-2:IL-2Rβγ complex to lysosomal degradation. IL-2Rα is known to recycle to the cell surface, and increasing the binding affinity of IL-2 to IL-2Rα at pH 6.0 by replacing one or more of these interacting residues can improve IL-2 recycling and extend half-life. Using computational predictions performed by protein design automation techniques (see, eg, WO 1998 / 047089 published October 22, 1998), variants were generated that saturated these three contact residues.

[0272] The plasmid encoding IL-2 was constructed by standard gene synthesis and then subcloned into the pTT5 expression vector. IL-2 comprises a C-terminal polyhistidine tag (8xHis) for purification and a C125S replacement for improved expression. Replacement as predicted above was introduced by standard mutagenesis techniques. Protein was produced by transient transfection in HEK293E cells and purified by Ni-NTA chromatography. The sequence of an illustrative variant is depicted in Figure 9, wherein the polyhistidine tag has been removed.

[0273] The affinity of IL-2 for IL-2Rα at pH 7.4 and pH 6.0 was determined using Biacore, a surface plasmon resonance (SPR)-based technology. The experimental steps of Biacore generally include the following: immobilization (capturing the ligand onto the sensor chip); association (flow of various concentrations of analyte on the sensor chip); and dissociation (flow buffer on the sensor chip) to determine the affinity of the test article. A reference flow using buffer alone is also included in the method for background correction during data processing. In this particular screening, human CD25 (IL-2Rα) was first captured on the sensor chip, and then various concentrations of IL-2 variants were flowed on the sensor chip. Separate experiments were performed using a buffer at pH 7.4 to simulate cell surface conditions, and a buffer at pH 6.0 to simulate endosomal conditions. Figures 10 and 11 depict the resulting dissociation constant (K D ), association rate (k a ) and dissociation rate (k dThe processed data were analyzed using a 1:1 binding model to obtain binding affinities and kinetic rate constants. Figure 12 The fold increase in the off-rate at pH 6.0 is depicted, whereas Figure 13 The fold increase in off-rate at pH 7.4 is depicted.

[0274] A number of variants with various substitutions at R38, T41, and F42 had off-rates that were worse or similar at pH 6.0 compared to WT IL-2; however, some variants with greatly improved off-rates were identified (i.e., IL-2 variants that were more likely to recycle with IL-2Rα). Favored variants included XENP14142 (R38I) and XENP14144 (R38L). Figure 14 Depicted are Biacore sensorgrams of XENP14135 (wild-type IL-2 with a C125S mutation) and XENP14142 (variant IL-2 with R38I and C125S). Similar dissociation curves of XENP14142 at pH 7.4 and pH 6.0 suggest that pH-dependent binding was successfully attenuated.

[0275] 2. Attenuation of pH-dependent binding in the context of other IL-2 variants

[0276] Next, R38L was combined with prior art IL-2 variants (mutants 2-4 with Q126T as described in WO 2009 / 061853 published on May 14, 2009) to investigate whether the off-rate was improved in the context of other IL-2 variants.

[0277] As above, substitutions were introduced by standard mutagenesis techniques. Protein was produced by transient transfection in HEK293E cells and purified by Ni-NTA chromatography. Figure 15 The sequences of the above described prior art variants with and without R38L are depicted in , wherein the polyhistidine tag is removed.

[0278] The affinity of IL-2 for IL-2Rα at pH 7.4 and pH 6.0 was determined using Biacore as described in Example 1A. Figure 16 The resulting dissociation constant (K D ), association rate (k a ) and dissociation rate (k d ) and the ratio of affinity at pH 7.4 to affinity at pH 6.0. The data show that the ratio of affinities increases with inclusion of the R38L substitution, indicating successful attenuation of pH-dependent binding.

[0279] B. Example 2: Engineering IL-2 Selectively for Tregs

[0280] IL-2Rβ and γc mediate proliferative signaling by IL-2 as part of the IL-2 intermediate affinity receptor complex (IL-2Rβγ) or as part of the IL-2 high affinity receptor complex (IL-2Rαβγ). CD25 confers high affinity binding to the IL-2Rαβγ complex for IL-2, but is otherwise deficient in its own signaling. Due to its high affinity binding to the IL-2Rαβγ complex, IL-2 favors Tregs that constitutively express IL-2Rα. Therefore, it is hypothesized that increasing the affinity of IL-2 for IL-2Rα can further skew the binding to favor the IL-2Rαβγ complex on Tregs. Alternatively, reducing the affinity of IL-2 for IL-2Rβ, γc, or IL-2Rβγ can skew the binding away from CD25-negative T cells and NK cells.

[0281] By examining the crystal structure of the interface between IL-2 and its receptor and by modeling using MOE software, we predicted residues that could be substituted to increase the affinity of IL-2 for IL-2Rα or decrease the affinity of IL-2 for IL-2Rβ, γc, and / or IL-2Rβγ.

[0282] The plasmid encoding IL-2 was constructed by standard gene synthesis and then subcloned into the pTT5 expression vector. IL-2 comprises a C-terminal polyhistidine tag (8xHis) for purification and a C125S replacement for improved expression. Replacement as predicted above was introduced by standard mutagenesis techniques. Protein was produced by transient transfection in HEK293E cells and purified by Ni-NTA chromatography. The sequence of an illustrative variant is depicted in Figure 17, wherein the polyhistidine tag has been removed.

[0283] The binding of IL-2 to its receptor components was determined using Octet, a method based on biolayer interferometry (BLI). The experimental steps of Octet generally include the following: immobilization (capturing the ligand to the biosensor); association (immersing the ligand-coated biosensor into wells containing serial dilutions of the analyte); and dissociation (returning the biosensor to wells containing buffer) to determine the affinity of the test article. Reference wells containing only buffer are also included in the method for background correction during data processing. Specifically, anti-human Fc (AHC) biosensors were used to capture bivalent CD25 (IL-2Rα)-Fc fusions, bivalent CD122 (IL-2Rβ)-Fc fusions, or heterodimeric CD122:CD132 (IL-2Rβγ)-Fc fusions and immersed in various concentrations of IL-2 variants. The BLI responses of the resulting IL-2 variants were normalized to that of XENP14135 (wild-type IL-2 with C125S) and are depicted in Figure 18. Notably, several substitutions at the IL-2:IL-2Rβ interface, such as D20N and N88D, drastically reduced or abolished IL-2 binding to IL-2Rβ.

[0284] C. Example 3: IL-2-FC fusion protein

[0285] To further address the short half-life of IL-2, IL-2 was generated as an Fc fusion (hereinafter referred to as IL-2-Fc fusion) with the aim of promoting production and promoting FcRn-mediated recycling of the complex and extending the half-life.

[0286] 1. Generation of IL-2-Fc Fusion

[0287] Plasmids encoding IL-2 are constructed by standard gene synthesis and then subcloned into a pTT5 expression vector containing an Fc fusion partner (e.g., a constant region as depicted in Figure 6). IL-2 may contain a C125S substitution for improved expression and a T3A substitution for removal of an O-glycosylation site. Figure 19 A cartoon schematic of an illustrative IL-2-Fc fusion format is depicted in . Selected substitutions were introduced by standard mutagenesis techniques.

[0288] Monovalent IL-2-Fc or "monovIL-2-Fc" format ( Figure 19A) comprises IL-2 fused to the N-terminus of the first heterodimeric Fc region (see, e.g., IL-2-Fc backbone 1 - monomer 2 in FIG6 ), while the other side of the molecule is an "Fc-only" or "empty Fc" heterodimeric Fc (see, e.g., IL-2-Fc backbone 1 - monomer 1 in FIG6 ). The sequence of an illustrative monovIL-2-Fc fusion is depicted in FIG20 . The bivalent IL-2-Fc or "bivIL-2-Fc" format ( Figure 19 B) comprises IL-2 fused to the N-terminus of a homodimeric Fc region (see, e.g., IL-2-Fc backbone 12 in FIG6 ). Figure 23 The sequence of an illustrative bivIL-2-Fc fusion is depicted in . The monovIL-2-Fc fusion and bivIL-2-Fc fusion can have variable length linkers located between the C-terminus of IL-2 and the N-terminus of the Fc region (for non-limiting examples of domain linkers that can be used in IL-2-Fc fusions, see Figure 7 , and for format images, see Figure 19 CD). The sequences of illustrative IL-2-Fc fusions with variable length linkers are depicted in FIG25 .

[0289] The protein was produced by transient transfection in HEK293E cells and purified by a two-step purification process involving protein A chromatography and anion exchange chromatography.

[0290] 2. Studies of prior art IL-2 variants engineered as monovIL-2-Fc fusions

[0291] To investigate the robustness and efficacy of the monovIL-2-Fc fusion format, several prior art IL-2 variants were generated in this format. These IL-2-Fc fusions included: XENP24637 (based on mutants described in WO 2012 / 107417 published on August 16, 2012), XENP24638 (based on mutants 2-4 as described in WO 2005 / 007121 published on January 27, 2005), XENP24639 (based on mutant M1 as described in WO 2005 / 007121 published on January 27, 2005), XENP24640 (based on mutants 2-4 with Q126T as described in WO 2009 / 061853 published on May 14, 2009), XENP24642 ​​(based on mutants 2-4 with Q126T as described in WO 2009 / 061853 published on November 25, 1999), and XENP24643 (based on mutants 2-4 with Q126T as described in WO 2009 / 061853 published on November 25, 1999). 1999 / 060128), XENP24728 (based on H9-RET as described in Mitra et al. 2015), and XENP24729 (based on H9-RETR as described in Mitra et al. 2015). Additional variants such as XENP24641, XENP24730, XENP24731, and XENP24732 are based on individual substitutions or combinations of substitutions described in the prior art. Figure 20 depicts the sequences.

[0292] a. Affinity screening of monovIL-2-Fc fusions

[0293] The affinity of the monovIL-2-Fc fusions described above for various IL-2 receptors was determined using Octet as generally described in Example 2. Specifically, to determine affinity for IL-2Rα, CD25 (IL-2Rα)-Fc fusion (R&D Systems, Minneapolis, MN) was loaded onto AR2G biosensors and immersed in various concentrations of IL-2-Fc fusion. To determine affinity for IL-2Rβ and IL-2RβT, bivalent CD122 (IL-2Rβ)-Fc-His fusion or heterodimeric CD122:CD132 (IL-2Rβγ)-Fc-His fusion was loaded onto HIS1K biosensors and immersed in various concentrations of IL-2-Fc fusion. Figure 21 The resulting dissociation constant (K D ), association rate (k a ) and dissociation rate (k d ).

[0294] b. Induction of STAT5 phosphorylation by monovIL-2-Fc fusion

[0295] After the cytokine binds to its receptor, the Janus kinase (JAK) associated with the receptor phosphorylates the STAT protein, which is then transported to the nucleus to regulate further downstream processes. Therefore, phosphorylation of STAT proteins (specifically, STAT5 comprising STAT5a and STAT5b) is one of the earliest signaling events triggered by the binding of IL-2 to high-affinity or intermediate-affinity IL-2 receptors (Lin and Leonard (2000); Wuest et al. (2008)). Therefore, the monovIL-2-Fc fusion described above was studied to induce CD8 + T cells and CD4 + The ability to phosphorylate STAT5 in various cell types including T cells and Tregs.

[0296] Fresh PBMCs were incubated with the indicated IL-2-Fc test articles at the indicated concentrations for 15 minutes. After incubation, PBMCs were stained with anti-CD3-BV510 (UCHT1), anti-CD4-BV605 (RPA-T4) and anti-CD8-Alexa700 (SK1) at room temperature for 30-45 minutes. The cells were washed and incubated with 90% methanol pre-cooled (-20°C) for 20-60 minutes. After methanol incubation, the cells were washed again and stained with anti-CD25-BV421 (M-A251), anti-CD45RA-PE (HI100), anti-FOXP3-Alexa488 (259D) and anti-pSTAT5-Alexa647 (pY687) to mark various cell populations and STAT5 phosphorylation. Figure 22 shows a graph depicting the induction of CD8 + T cells (CD3 + CD8 + CD25 - ), CD4 + T cells (CD3 + CD4 + CD25 - ) and Treg (CD3 + CD4 + CD25 + FOXP3 + ) data on STAT5 phosphorylation on Tregs. Notably, the IL-2-Fc fusions XENP24638 and XENP24642 ​​were potent activators of Tregs (as indicated by STAT5 phosphorylation), with the exception of CD8 + T cells and CD4 + T cell activation was minimal, consistent with the reported activity of IL-2 variants constituting IL-2-Fc fusions.

[0297] 3. Studies of prior art IL-2 variants engineered as bivIL-2-Fc fusions

[0298] To investigate the robustness and efficacy of the bivIL-2-Fc fusion format, a prior art IL-2 variant (described in WO 1999 / 060128 published on November 25, 1999) was generated in this format, as well as a control IL-2 (with a C125S substitution and a T3A substitution), the sequence of which is shown in FIG. Figure 23 As described in .

[0299] a. Induction of STAT5 phosphorylation by bivIL-2-Fc fusion

[0300] Fresh PBMCs were incubated with the indicated IL-2-Fc test articles at the indicated concentrations for 15 minutes. After incubation, PBMCs were stained with anti-CD3-BUV395 (UCHT1), anti-CD4-BV605 (RPA-T4) and anti-CD8-Alexa700 (SK1) at room temperature for 30-45 minutes. The cells were washed and incubated with 90% methanol pre-cooled (-20°C) for 20-60 minutes. After the methanol incubation, the cells were washed again and stained with anti-CD25-BV510 (M-A251), anti-CD45RA-PE (HI100), anti-FOXP3-Alexa488 (259D) and anti-pSTAT5-Alexa647 (pY694) to mark various cell populations and STAT5 phosphorylation. Figure 24 shows a depiction of the induction of CD8 + T cells, CD4 + Data on STAT5 phosphorylation on T cells and Tregs.

[0301] 4. Study of prior art IL-2 variants engineered as IL-2-Fc fusions with domain linkers

[0302] To investigate the effect of including a linker between IL-2 and the Fc region, prior art IL-2 variants (as described in WO 1999 / 060128, published on November 25, 1999, and WO 2012 / 107417, published on August 16, 2012) were generated as monovIL-2-Fc fusions or bivIL-2-Fc fusions with a Gly-Ser linker. Figure 25 depicts the sequences of these IL-2-Fc fusions.

[0303] a. Induction of STAT5 phosphorylation by IL-2-Fc fusion with a domain linker

[0304] Fresh PBMCs were incubated with the indicated IL-2-Fc test articles at the indicated concentrations for 15 minutes. After incubation, PBMCs were stained with anti-CD3-BUV395 (UCHT1), anti-CD4-BV605 (RPA-T4) and anti-CD8-Alexa700 (SK1) at room temperature for 30-45 minutes. The cells were washed and incubated with 90% methanol pre-cooled (-20°C) for 20-60 minutes. After the methanol incubation, the cells were washed again and stained with anti-CD25-BV510 (M-A251), anti-CD45RA-PE (HI100), anti-FOXP3-Alexa488 (259D) and anti-pSTAT5-Alexa647 (pY694) to mark various cell populations and STAT5 phosphorylation. Figure 26 shows a depiction of the induction of CD8 + T cells, CD4 + Data on STAT5 phosphorylation on T cells and Tregs.

[0305] D. Example 4: Engineering Variant IL-2-Fc Fusions with Increased Affinity for CD25 and Reduced Affinity for CD122

[0306] As discussed in Example 2, increasing the affinity of IL-2 for IL-2Rα can further skew binding in favor of the IL-2RαβT complex on Tregs, while reducing the affinity of IL-2 for IL-2Rβ, γc, or IL-2Rβγ can skew binding away from CD25-negative T cells and NK cells. Here, in the context of an IL-2-Fc fusion, substitutions that increase IL-2Rα binding are combined with substitutions that decrease IL-2Rβ binding to enhance Treg selectivity.

[0307] The plasmid encoding IL-2 was constructed by standard gene synthesis and then subcloned into the pTT5 expression vector containing an Fc fusion partner (e.g., constant region as described in Fig. 6). IL-2 comprises a C125S replacement for improving expression and a T3A replacement for removing the O-glycosylation site. Selected replacements as described in Example 2 were introduced by standard mutagenesis techniques. Protein was produced by transient transfection in HEK293E cells, and the protein was purified by the two-step purification process comprising protein A chromatography and anion exchange chromatography. The sequence of the illustrative IL-2-Fc fusions engineered to increase CD25 avidity and / or reduce CD122 avidity is depicted in Figure 27.

[0308] 1. Affinity screening of variant IL-2-Fc fusions engineered to increase CD25 affinity and decrease CD122 affinity

[0309] As generally described in Example 3B (a), the affinity of the variant IL-2-Fc fusions described above for various IL-2 receptors was determined using Octet. Specifically, to determine affinity for IL-2Rα, CD25 (IL-2Rα)-Fc fusion (R&D Systems, Minneapolis, MN) was loaded onto AR2G biosensors and immersed in various concentrations of IL-2-Fc fusion. To determine affinity for IL-2Rβ and IL-2Rβγ, bivalent CD122 (IL-2Rβ)-Fc-His fusion or heterodimeric CD122:CD132 (IL-2Rβγ)-Fc-His fusion was loaded onto HIS1K biosensors and immersed in various concentrations of IL-2-Fc fusion. Figure 33 The resulting dissociation constant (K D ), association rate (k a ) and dissociation rate (k d ).

[0310] 2. STAT5 phosphorylation by variant IL-2-Fc fusions engineered to increase CD25 affinity and decrease CD122 affinity on various cell populations

[0311] At indicated concentration, fresh PBMC and indicated IL-2-Fc test article are incubated for 15 minutes. After incubation, PBMC are dyed at room temperature with anti-CD3-BV396 (UCHT1), anti-CD4-BV605 (RPA-T4) and anti-CD8-Alexa700 (SK1), and continue for 30-45 minutes. Cells are washed, and the cells are incubated with 90% methanol pre-cooled (-20 ° C), and continue for 20-60 minutes. After methanol incubation, cells are washed again, and the cells are dyed with anti-CD25-BV421 (M-A251), anti-CD45RA-PE (HI100), anti-FOXP3-Alexa488 (259D) and anti-pSTAT5-Alexa647 (pY687), to mark various cell populations and STAT5 phosphorylation. The data depicting the STAT5 phosphorylation on inducing various cell populations are depicted in Figures 28-29.

[0312] The data showed that many of the illustrative variants were Treg-positive on CD25-negative T cells (CD8 + and CD4+ ) is a potent activator with minimal induction of STAT5 phosphorylation.

[0313] E. Example 5: Engineering Additional Variant IL-2-Fc Fusions

[0314] Additional IL-2-Fc fusions were engineered, incorporating features including valency, domain linker, pH switch, and Treg selectivity as described in the previous examples, and produced as described in Example 2. Illustrative sequences are depicted in Figure 30. Additionally, an Fc-IL-2 (V91K / C125A) fusion engineered to increase the ratio of Treg to non-regulatory T cells as described in WO 2014 / 153111 was generated as a comparator (referred to herein as XENP27193; its sequence is depicted in Figure 30). Figure 34 middle).

[0315] 1. Induction of STAT5 phosphorylation by an additional monovalent IL-2-Fc fusion

[0316] Fresh PBMCs were incubated with the indicated IL-2-Fc test articles at the indicated concentrations for 15 minutes at 37°C. After incubation, PBMCs were first stained with anti-CD3-BUV395 (UCHT1), anti-CD4-BV605 (RPA-T4), anti-CD8-AF700 (SK1), and anti-CD56-PE antibodies. After the first staining, cells were permeabilized using PerFix EXPOSE (Beckman Coulter, Indianapolis, Indiana). After permeabilization, cells were stained with anti-CD25-BV421 (M-A251), anti-CD45RA-BV510 (HI100), anti-FoxP3-AF488 (259D), and anti-pSTAT5-AF647 (47 / Stat5 (pY694)) antibodies. After the second staining, cells were analyzed by flow cytometry to investigate STAT5 phosphorylation on various lymphocyte populations. Figure 35 depicts data depicting pSTAT5 MFI on various lymphocyte populations, indicating signaling via the IL-2 receptor by IL-2-Fc fusions.

[0317] The data show that each of the variants induced STAT5 phosphorylation on Tregs. Notably, the variant IL-2-Fc fusions were significantly more phosphorylated relative to CD4 Tregs compared to both recombinant IL-2 and WT monovalent IL-2-Fc fusion (XENP24635). + Memory T cells (CD45RA-), CD8 +Memory T cells (CD45RA-), NK cells and γδ T cells preferentially induce Tregs. In contrast, the Treg selectivity of the prior art variant Fc-IL-2 fusion XENP27193 is relatively low.

[0318] 2. Bivalent IL-2-Fc fusions are more effective than their monovalent counterparts

[0319] Induction of STAT5 phosphorylation by various IL-2-Fc test articles was studied as described in Example 5A. Figures 36-42 The Treg and CD4 + Data for pSTAT5 MFI on memory T cells (CD45RA-), indicating signaling via the IL-2 receptor by IL-2-Fc fusion.

[0320] The data showed that for each IL-2 variant, the bivalent IL-2-Fc version was more effective in inducing STAT5 phosphorylation on Tregs than the corresponding monovalent IL-2-Fc version. Notably, engineering the domain linker between the IL-2 component and the Fc component (e.g., in XENP27002, XENP27003, XENP27004, XENP27005, XENP27006, and XENP27007) further improved the potency of the bivalent IL-2-Fc fusion. Notably, compared to other lymphocyte populations, such as CD4 + CD45RA - T cells, each of the bivalent IL-2-Fc fusion constructs (with and without a linker) retained selectivity for Tregs.

[0321] F. Example 6: Maximizing Exposure of IL-2-Fc Fusions

[0322] 1. Incorporation of Xtend Fc

[0323] The IL-2-Fc fusion described above was engineered with Xtend Fc (M428L / N434S) for enhanced binding to FcRn to further promote FcRn-mediated recycling of the fusion and subsequently extend circulating half-life. The sequence of an illustrative bivalent IL-2-Fc fusion with Xtend Fc is depicted in FIG43 , and the sequence of an illustrative monovalent IL-2-Fc fusion with Xtend Fc is depicted in FIG44 .

[0324] 2. Selecting IL-2-Fc Fusions with a Balance of Treg Selectivity and Potency

[0325] It was also reasoned that IL-2-Fc fusions with lower potency would reduce antigen silencing and therefore increase circulating half-life. Given that the data in Example 5B showed that bivalent IL-2-Fc fusions and IL-2-Fc fusions with domain linkers had enhanced potency, monovalent IL-2-Fc fusions lacking a domain linker were of particular interest. Therefore, to identify IL-2-Fc fusions with the best balance between selectivity and potency, the in vitro potency of monovalent IL-2 fusions (XENP26105 and XENP26109, comprising the D20N / T37R variant and the D20N / N71K variant, respectively) was compared with XENP24635 (a monovalent IL-2-Fc fusion with a C125S mutation), as well as XENP25908 and XENP27193 (comparator IL-2-Fc fusions indicated for the treatment of autoimmune diseases) in a STAT5 phosphorylation assay (as indicated by induction of STAT5 phosphorylation on various lymphocyte populations), the data of which are depicted in FIG45 .

[0326] It is noteworthy that the efficacy of both XENP26105 and XENP26109 is lower than that of XENP24635 and XENP27193 (EC50 is 1nM and 5nM to 0.02nM and 0.01nM respectively), but it is possible to achieve similar activity levels on Treg with higher doses while maintaining selectivity to Treg. Although the efficacy of XENP26105 is comparable to that of XENP25908 (EC50 is 1nM to 0.7nM respectively), data show that XENP26105 and XENP26109 can achieve activity levels far higher than XENP25908 on Treg. The observed efficacy and selectivity reduction of XENP26105 and XENP26109 indicate that it will be useful for selective and sustained Treg expansion in a clinical setting. Therefore, the potential of XENP27563 and XENP27564, Xtend Fc analogs of XENP26105 and XENP26109, was further investigated.

[0327] G. Example 7: In vitro characterization of XENP27563 and XENP27564

[0328] 1. Treg cultures treated with CD25-selective IL-2-Fc fusion showed higher CD25 expression

[0329] It has been previously reported that rapamycin promotes the expansion of CD4+CD25+FOXP3+ Tregs in vitro, and that the resulting expanded Tregs inhibit CD4+ and CD8+ T cell proliferation (see, e.g., Battaglia et al., (2006), “Rapamycin promotes expansion of functional CD4+CD25+FOXP3+ regulatory T cells of both healthy subjects and type 1 diabetic patients,” J Immunol. 177(12):8338-8347; and Strauss et al., (2007), “Selective survival of naturally occurring human CD4+CD25+Foxp3+ regulatory T cells cultured with rapamycin.” rapamycin), Journal of Immunology, 178(1)320-329).

[0330] Using EasySep TM Human CD4+ T cell enrichment kit (STEMCELL Technologies, Vancouver, Canada) was used to enrich CD4+ T cells from human PBMCs by negative selection. Dynabeads TM Human Treg expander (Thermo Fisher Scientific, Waltham, MA) was used to expand Tregs in RPMI1640 + 10% fetal bovine serum + 0.1 μg / ml rapamycin + 500 U / ml IL-2 for 1-4 days. Tregs were transferred to T75 flasks coated with 0.5 μg / ml anti-CD3 (OKT3, Biolegend, San Diego, CA) and cultured with RPMI1640 + 10% fetal bovine serum + 0.1 μg / ml rapamycin + 100 U / ml IL-2 + 0.5 μg / ml anti-CD28 mAb. Experiments were performed at least 8 days after the initial enrichment of CD4+ T cells from PBMCs. Tregs thus enriched and cultured are referred to below as rapamycin Tregs.

[0331] Rapamycin Tregs were further cultured with 0.5 μg / ml plate-bound anti-CD3 (OKT3) in RPMI1640 medium containing 10% FBS, 0.5 μg / ml anti-CD28 mAb, 100 ng / ml rapamycin, and 10 ng / ml recombinant IL-2 or 10 μg / ml XENP27564 (IL-2-Fc fusion with IL-2 (D20N / N71K / C125S) variant). After 14 days of culture, cells were incubated with anti-CD25-FITC (M-A251), anti-FoxP3-PE (PCH101), anti-CTLA-4-PE-Dazzle594 (L3D10), anti-PD-1-BB700 (EH12.1), anti-GITR-PE-Cy7 (108-17), anti-Ki67-Alexa647, anti-ICOS-Alexa700 (C398.4a), anti-TIGIT-BV421 (A15153G) Tregs were stained with anti-LAG-3 (11C3C65), anti-CCR4-BV605 (L291H4), anti-CD8-BV650 (SK1), anti-CD39-BV711 (A1), anti-TIM-3-BV785 (F38-2E2), anti-CD40BUV396 (SK3), anti-CD3-BUV496 (UCHT1), anti-CD45-BUV805 (HI30), anti-CD45RA-BUV737 (HI100), and Zombie NIR (APC-Cy7), and analyzed by flow cytometry. The data are depicted in Figure 4. Figures 46-47 The data showed that Tregs treated with CD25 selective XENP27564 showed higher CD25 expression. Figure 48 As depicted, XENP27564 showed effector Treg (CD45RA-FoxP3 mid-high )A greater expansion of the population.

[0332] 2.7B: Treg cultures treated with a CD25-selective IL-2-Fc fusion display enhanced suppressive function.

[0333] On day 15, rapamycin Tregs were further cultured with IL-2 or XENP27564 as described in Example 7A for suppressive function. 1×10 5 CFSE-labeled PBMCs were incubated with the indicated number of Tag-it Violet-labeled Tregs for 4 days, and the expansion of CD8+ and CD4+ responders was determined by CFSE dilution. Lymphocyte populations were stained as follows: anti-CD8-PerCp-By5.5 (SKI), anti-CD3-PE-Cy7 (OKT3), anti-CD127-APC (A019D5), anti-CD25-APC-Fire750 (M-A251), anti-CD45RO-Alexa700 (UCHL1), anti-CD16-BV605 (3G6), anti-CD56-BV605 (HCD56), anti-CD45RA-BV785 (HI100), anti-CD4-BUV395 (SK3) and ZombieAqua (BV510). It should be noted that, as indicated by the data depicted in Figure 49, Tregs amplified by Treg selective IL-2-Fc fusions can have enhanced suppressive function.

[0334] In addition, the expression of CD25 and CD127 on Tregs in the suppression assay was studied, and the data are depicted in Figures 50-51. Consistent with the data above, Tregs amplified by XENP27564 showed higher levels of CD25 expression. It is noteworthy that Tregs amplified by XENP27564 showed lower CD127 expression, which is a sign that has previously been found to be inversely correlated with the suppressive function of Tregs (Liu et al., (2006), "CD127 expression inversely correlates with FoxP3 and suppressive function of human CD4+ Treg cells", Journal of Experimental Medicine (J Exp Med.) 203 (7): 1701-1711). This may explain the observed enhanced suppressive function of Tregs amplified by XENP27564.

[0335] 3. Following activation with anti-CD3, a CD25-selective IL-2-Fc fusion exhibits Treg selectivity and suppressive effects over other lymphocyte populations.

[0336] In the inhibition assay, CFSE-labeled PBMCs and Tag-itViolet-labeled rapamycin Tregs were incubated with the indicated test articles at the indicated concentrations for 4 days with 100 ng / ml plate-bound anti-CD3 (OKT3). Lymphocyte populations were stained as follows: anti-CD8-PerCp-Cy5.5 (SK1), anti-CD3-PE-Cy7 (OKT3), anti-CD25-APC-Cy7 (M-A251), anti-CD45RO-Alexa700 (UCHL1), anti-CD16-BV605 (3G6), anti-CD56-BV605 (HCD56), anti-CD45RA-BV785 (HI100), anti-CD4-BUV395 (SK3), and Zombie Aqua (BV510). Figure 52 depicts the proliferation of various lymphocyte populations after treatment with the test article (as determined by CFSE or Tag-it Violet dilution; Zombie dye for exclusion of dead cells). The data show that the CD25-selective IL-2-Fc fusions XENP27563 and XENP27564 selectively expand Tregs compared to XENP24635 (IL-2-Fc with only C125S), recombinant IL-2, and recombinant IL-15. In fact, the data depicted in Figures 53 and 54 show that the CD25-selective IL-2-Fc fusions are significantly less effective in inducing CD8+ T cell and CD4+ T cell proliferation than XENP24635, recombinant IL-2, and recombinant IL-15.

[0337] In the proliferation assay, PBMCs were incubated with the indicated concentrations of the indicated test articles using plate-bound anti-CD3 mAb (OKT3). Lymphocyte populations were stained as follows: anti-FoxP3-PE (PCH101), anti-CD8-PerCP-Cy5.5 (SK1), anti-CD3-PE-Cy7 (OKT3), anti-Ki67-APC, anti-CD45RO-Alexa700 (UCHL1), anti-CD25-BV421 (M-A251), anti-CD16-BV605 (3G6), anti-CD56-BV605 (HCD56), anti-CD45RA-BV785 (H1100), anti-CD4-BUV396 (SK3), and Zombie NIR (APC-Cy7). The proliferation of various lymphocyte populations was determined based on the percentage expressing the proliferation marker Ki67, and the data are depicted in Figure 2. Figures 55-65 In agreement with the data described above based on the inhibition assay, Figures 55-61The proliferation assay data depicted in show that XENP27563 and XENP27564 (IL-2-Fc fusions engineered selective for CD25) are impaired in inducing proliferation of CD8+ T cells, CD8+CD45RA- T cells, CD8+CD45RA+ T cells, CD4+ T cells, CD4+CD45RA- T cells, CD4+CD45RA+ T cells, and NK cells; and Figures 63-65 The data depicted in show that XENP27563 and XENP27564 selectively expand Tregs compared to other lymphocyte populations.

[0338] H. Example 8: IL-2-Fc fusion drives selective and sustained Treg expansion in cynomolgus monkeys.

[0339] To investigate the clinical potential of XENP27563 and XENP27564, their activity was investigated in cynomolgus monkeys. Prior to dosing, the activity of the IL-2-Fc fusions on cynomolgus monkey lymphocytes was confirmed. Two assays were performed, described below.

[0340] In the first assay, human PBMCs were stimulated with various concentrations of XENP27563 or XENP27564 at 37°C for 15 minutes. PBMCs were then stained with anti-CD3-BUV395 (UCHT1), anti-CD4-BV605 (RPA-T4), anti-CD8-BV711 (RPA-T8), anti-CD25-BV421 (M-A251), anti-CD45RA-BV510 (HI100), and anti-CD56-PE. The cells were then permeabilized using PerFix EXPOSE (Beckman Coulter, Indianapolis, Indiana). After permeabilization, the cells were stained with anti-CD16-AF700 (DJ130C), anti-FoxP3-AF488 (259D), and pSTAT5 (pY694), and analyzed by flow cytometry for STAT5 phosphorylation on various lymphocyte populations, and the data are depicted in Figure 2. Figure 78 middle.

[0341] In the second assay, cynomolgus monkey PBMCs were stimulated with various concentrations of XENP27563 or XENP27564 at 37°C for 15 minutes. PBMCs were then stained with anti-CD3-BV421 (SP34), anti-CD4-BV785 (OKT4), anti-CD8-BUV395 (RPA-T8), anti-CD25-BV510 (M-A251), anti-CD45RA-APC / H7 (HI100), and anti-CD56-PE. The cells were then permeabilized using PerFixEXPOSE (Beckman Coulter, Indianapolis, Indiana). After permeabilization, the cells were stained with anti-CD16-AF700 (DJ130C), anti-FoxP3-AF488 (259D), and pSTAT5 (pY694), and analyzed by flow cytometry for STAT5 phosphorylation on various lymphocyte populations, and the data are depicted in Figure 2. Figure 79 middle.

[0342] The data showed that XENP27563 and XENP27564 were equally selective and effective against human and cynomolgus macaque Tregs.

[0343] 1. 8A: Comparison of PD and PK of XENP27563 and XENP27564

[0344] In the first study conducted on cynomolgus monkeys, animals (n=2) were intravenously administered a 3X dose of XENP27563 or a 3X dose of XENP27564 on days 0 and 15. Blood was collected over time to study the expansion of various lymphocyte populations and to study serum concentrations of the test article. Serum albumin concentration and blood pressure were also measured to study the tolerability of the test article.

[0345] Figure 66 depicts the expansion of various lymphocyte populations over time. The data show that both test articles were able to expand Tregs while maintaining close to baseline levels of CD8+CD45RA- T cells, CD4+CD45RA- T cells, and CD16+ NK cells. In addition, the data show that both test articles promoted similar pharmacological effects in monkeys. Figure 67 depicts the serum concentrations of the test articles. The data show that both test articles exhibited similar pharmacokinetic profiles, with XENP27564 having a half-life of 1.5 days. In summary, this confirms that the IL-2-Fc fusion engineered for CD25 selectivity and reduced potency provides selective and sustained Treg expansion.

[0346] Vascular leak syndrome is a hallmark toxicity associated with treatment with cytokines such as IL-2. One indicator of vascular leak is hypoalbuminemia, a decrease in serum albumin concentration. Therefore, changes in serum albumin concentration in animals were studied, and the data are presented in Figure 68 Notably, in one animal dosed with XENP27563, a sustained decrease in albumin was observed after both the first and second doses. In one animal dosed with XENP27564, a decrease in albumin was observed after the second dose, but concentrations quickly returned to baseline. These data suggest that XENP27564, despite its lower potency than XENP27563, can promote superior tolerability and therapeutic index.

[0347] Another index of vascular leakage is that blood pressure drops sharply.Therefore, the blood pressure of animal at day 0, day 1, day 3, day 5, day 9, day 16, day 18, day 20 and day 24 was recorded, and its data are depicted in Figure 69.It should be noted that, there is a decrease in blood pressure in the first monkey of administration XENP27563 at day 1 (day 1 after the 1st administration), and there is a decrease in blood pressure in the second monkey of administration XENP27563 at day 16 (day 1 after the 2nd administration), and in the monkey of administration XENP27564, the blood pressure observed in all days is all stable (the telemetry data of the 2nd monkey of administration XENP27564 is damaged).This further confirms that the IL-2-Fc fusions of lower effectiveness can promote the viewpoint of superior tolerability and therapeutic index.

[0348] Finally, eosinophil counts and basophil counts were also studied as additional indicators of tolerability, the data for which are depicted in Figure 76. In summary, the data demonstrate repeat dosing of XENP27564.

[0349] 2. 8B: XENP27564 dose-escalation study

[0350] In the first study in cynomolgus monkeys, animals (n=3) were intravenously administered 1X, 3X, or 10X doses of XENP27564. Blood was collected over time to study the expansion of various lymphocyte populations, as well as serum albumin and C-reactive protein (CRP) concentrations.

[0351] The expansion of various lymphocyte populations is depicted in Figures 70-71. Consistent with the data from the first cynomolgus monkey study, XENP27564 provides selective and sustained Treg expansion. In addition, the data show that the 1X and 3X doses promoted similar pharmacological effects (as indicated by Treg expansion) in monkeys. It is worth noting that the higher dose (10X dose) of XENP27564 did not enhance the pharmacokinetic effect. Consistent with the data depicted above, Figure 75 Pharmacokinetic effects in cynomolgus monkeys for up to several days were shown at all tested doses of XENP27564.

[0352] As in the first study, albumin reduction was investigated as an indicator of vascular leakage and resistance, the data of which are depicted in Figure 72 In addition, the serum concentration of CRP, an acute phase protein associated with inflammation, was studied as another indicator of tolerance, and the data are presented in Figure 73 Sodium concentration, chloride concentration, eosinophil count, and basophil count were also studied as additional indicators of tolerability (data for which are depicted in Figure 74). Notably, the data showed that higher doses of XENP27564 increased toxicity, as indicated by both a decrease in albumin and an increase in serum CRP concentration (as well as sodium concentration, chloride concentration, eosinophil count, and basophil count), while lower doses that still resulted in a significant increase in Tregs were more tolerable.

[0353] I. Example 9: IL-2-Fc fusion is also selective for Tregs in mice.

[0354] Splenocytes from B6 mice are incubated with IL-2-Fc fusions and recombinant human IL-2 for 15 minutes. After incubation, cells are stained with anti-CD4-PE (GK1.5), anti-CD25-BV605 (PC61) and anti-CD44-BV510 (IM7). PerFix EXPOSE (Beckman Coulter, Indianapolis, Indiana) is then used to permeabilize cells. After permeabilization, cells are stained with anti-CD3-AF700 (2C11), anti-CD8-AF488 (53-6.7), anti-FoxP3-eF450 (FJK-16S) and anti-pSTAT5 (pY694), and analyzed by flow cytometry for the STAT5 phosphorylation on various lymphocyte populations, and its data are depicted in Figure 77. The data indicate that the engineered IL-2-Fc fusion is also selective and potent for Tregs in mice, making it suitable for studying autoimmune diseases using preclinical mouse models. Sequence Listing <110> Xencor, Inc. <120> Engineered IL-2 Fc fusion <130> 067461-5217-WO <140> Submit with this application <141> 2018-11-27 <150> US 62 / 607,850 <151> 2017-12-19 <150> US 62 / 675,070 <151> 2018-05-22 <160> 308 <170> PatentIn version 3.5 <210> 1 <211> 153 <212> PRT <213> Artificial sequence <220> <223> Human IL-2 sequence <400> 1 Met Tyr Arg Met Gln Leu Leu Ser Cys Ile Ala Leu Ser Leu Ala Leu 1 5 10 15 Val Thr Asn Ser Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu 20 25 30 Gln Leu Glu His Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile 35 40 45 Asn Asn Tyr Lys Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe 50 55 60 Tyr Met Pro Lys Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu 65 70 75 80 Glu Glu Leu Lys Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys 85 90 95 Asn Phe His Leu Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile 100 105 110 Val Leu Glu Leu Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala 115 120 125 Asp Glu Thr Ala Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe 130 135 140 Cys Gln Ser Ile Ile Ser Thr Leu Thr 145 150 <210> 2 <211> 133 <212> PRT <213> Artificial Sequence <220> <223> Mature form of human IL-2 <400> 2 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Cys Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 3 <211> 272 <212> PRT <213> Artificial Sequence <220> <223> Human IL-2R (CD25) Sequence <400> 3 Met Asp Ser Tyr Leu Leu Met Trp Gly Leu Leu Thr Phe Ile Met Val 1 5 10 15 Pro Gly Cys Gln Ala Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro 20 25 30 His Ala Thr Phe Lys Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn 35 40 45 Cys Glu Cys Lys Arg Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr 50 55 60 Met Leu Cys Thr Gly Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys 65 70 75 80 Gln Cys Thr Ser Ser Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro 85 90 95 Gln Pro Glu Glu Gln Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro 100 105 110 Met Gln Pro Val Asp Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro 115 120 125 Pro Pro Trp Glu Asn Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val 130 135 140 Gly Gln Met Val Tyr Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His 145 150 155 160 Arg Gly Pro Ala Glu Ser Val Cys Lys Met Thr His Gly Lys Thr Arg 165 170 175 Trp Thr Gln Pro Gln Leu Ile Cys Thr Gly Glu Met Glu Thr Ser Gln 180 185 190 Phe Pro Gly Glu Glu Lys Pro Gln Ala Ser Pro Glu Gly Arg Pro Glu 195 200 205 Ser Glu Thr Ser Cys Leu Val Thr Thr Thr Asp Phe Gln Ile Gln Thr 210 215 220 Glu Met Ala Ala Thr Met Glu Thr Ser Ile Phe Thr Thr Glu Tyr Gln 225 230 235 240 Val Ala Val Ala Gly Cys Val Phe Leu Leu Ile Ser Val Leu Leu Leu 245 250 255 Ser Gly Leu Thr Trp Gln Arg Arg Gln Arg Lys Ser Arg Arg Thr Ile 260 265 270 <210> 4 <211> 219 3><212> PRT <213> Artificial sequence <220> <223> Human IL-2R (CD25), extracellular domain <400> 4 Glu Leu Cys Asp Asp Asp Pro Pro Glu Ile Pro His Ala Thr Phe Lys 1 5 10 15 Ala Met Ala Tyr Lys Glu Gly Thr Met Leu Asn Cys Glu Cys Lys Arg 20 25 30 Gly Phe Arg Arg Ile Lys Ser Gly Ser Leu Tyr Met Leu Cys Thr Gly 35 40 45 Asn Ser Ser His Ser Ser Trp Asp Asn Gln Cys Gln Cys Thr Ser Ser 50 55 60 Ala Thr Arg Asn Thr Thr Lys Gln Val Thr Pro Gln Pro Glu Glu Gln 65 70 75 80 Lys Glu Arg Lys Thr Thr Glu Met Gln Ser Pro Met Gln Pro Val Asp 85 90 95 Gln Ala Ser Leu Pro Gly His Cys Arg Glu Pro Pro Pro Trp Glu Asn 100 105 110 Glu Ala Thr Glu Arg Ile Tyr His Phe Val Val Gly Gln Met Val Tyr 115 120 125 Tyr Gln Cys Val Gln Gly Tyr Arg Ala Leu His Arg Gly Pro Ala Glu 130 135 140 Ser Val Cys Lys Met Thr His Gly Lys Thr Arg Trp Thr Gln Pro Gln 145 150 155 160 Leu Ile Cys Thr Gly Glu Met Glu Thr Ser Gln Phe Pro Gly Glu Glu 165 170 175 Lys Pro Gln Ala Ser Pro Glu Gly Arg Pro Glu Ser Glu Thr Ser Cys 180 185 190 Leu Val Thr Thr Thr Asp Phe Gln Ile Gln Thr Glu Met Ala Ala Thr 195 200 205 Met Glu Thr Ser Ile Phe Thr Thr Glu Tyr Gln 210 215 <210> 5 <211> 551 <212> PRT <213> Artificial Sequence <220> <223> Human IL-2R (CD122) Sequence <400> 5 Met Ala Ala Pro Ala Leu Ser Trp Arg Leu Pro Leu Leu Ile Leu Leu 1 5 10 15 Leu Pro Leu Ala Thr Ser Trp Ala Ser Ala Ala Val Asn Gly Thr Ser 20 25 30 Gln Phe Thr Cys Phe Tyr Asn Ser Arg Ala Asn Ile Ser Cys Val Trp 35 40 45 Ser Gln Asp Gly Ala Leu Gln Asp Thr Ser Cys Gln Val His Ala Trp 50 55 60 Pro Asp Arg Arg Arg Trp Asn Gln Thr Cys Glu Leu Leu Pro Val Ser 65 70 75 80 Gln Ala Ser Trp Ala Cys Asn Leu Ile Leu Gly Ala Pro Asp Ser Gln 85 90 95 Lys Leu Thr Thr Val Asp Ile Val Thr Leu Arg Val Leu Cys Arg Glu 100 105 110 Gly Val Arg Trp Arg Val Met Ala Ile Gln Asp Phe Lys Pro Phe Glu 115 120 125 Asn Leu Arg Leu Met Ala Pro Ile Ser Leu Gln Val Val His Val Glu 130 135 140 Thr His Arg Cys Asn Ile Ser Trp Glu Ile Ser Gln Ala Ser His Tyr 145 150 155 160 Phe Glu Arg His Leu Glu Phe Glu Ala Arg Thr Leu Ser Pro Gly His 165 170 175 Thr Trp Glu Glu Ala Pro Leu Leu Thr Leu Lys Gln Lys Gln Glu Trp 180 185 190 Ile Cys Leu Glu Thr Leu Thr Pro Asp Thr Gln Tyr Glu Phe Gln Val 195 200 205 Arg Val Lys Pro Leu Gln Gly Glu Phe Thr Thr Trp Ser Pro Trp Ser 210 215 220 Gln Pro Leu Ala Phe Arg Thr Lys Pro Ala Ala Leu Gly Lys Asp Thr 225 230 235 240 Ile Pro Trp Leu Gly His Leu Leu Val Gly Leu Ser Gly Ala Phe Gly 245 250 255 Phe Ile Ile Leu Val Tyr Leu Leu Ile Asn Cys Arg Asn Thr Gly Pro 260 265 270 Trp Leu Lys Lys Val Leu Lys Cys Asn Thr Pro Asp Pro Ser Lys Phe 275 280 285 Phe Ser Gln Leu Ser Ser Glu His Gly Gly Asp Val Gln Lys Trp Leu 290 295 300 Ser Ser Pro Phe Pro Ser Ser Ser Phe Ser Pro Gly Gly Leu Ala Pro 305 310 315 320 Glu Ile Ser Pro Leu Glu Val Leu Glu Arg Asp Lys Val Thr Gln Leu 325 330 335 Leu Leu Gln Gln Asp Lys Val Pro Glu Pro Ala Ser Leu Ser Ser Asn 340 345 350 His Ser Leu Thr Ser Cys Phe Thr Asn Gln Gly Tyr Phe Phe Phe His 355 360 365 Leu Pro Asp Ala Leu Glu Ile Glu Ala Cys Gln Val Tyr Phe Thr Tyr 370 375 380 Asp Pro Tyr Ser Glu Glu Asp Pro Asp Glu Gly Val Ala Gly Ala Pro 385 390 395 400 Thr Gly Ser Ser Pro Gln Pro Leu Gln Pro Leu Ser Gly Glu Asp Asp 405 410 415 Ala Tyr Cys Thr Phe Pro Ser Arg Asp Asp Leu Leu Leu Phe Ser Pro 420 425 430 Ser Leu Leu Gly Gly Pro Ser Pro Pro Ser Thr Ala Pro Gly Gly Ser 435 440 445 Gly Ala Gly Glu Glu Arg Met Pro Pro Ser Leu Gln Glu Arg Val Pro 450 455 460 Arg Asp Trp Asp Pro Gln Pro Leu Gly Pro Pro Thr Pro Gly Val Pro 465 470 475 480 Asp Leu Val Asp Phe Gln Pro Pro Pro Glu Leu Val Leu Arg Glu Ala 485 490 495 Gly Glu Glu Val Pro Asp Ala Gly Pro Arg Glu Gly Val Ser Phe Pro 500 505 510 Trp Ser Arg Pro Pro Gly Gln Gly Glu Phe Arg Ala Leu Asn Ala Arg 515 520 525 Leu Pro Leu Asn Thr Asp Ala Tyr Leu Ser Leu Gln Glu Leu Gln Gly 530 535 540 Gln Asp Pro Thr His Leu Val 545 550 <210> 6 <211> 214 <212> PRT <213> Artificial sequence <220> <223> Human IL-2R (CD122), extracellular domain <400> 6 Ala Val Asn Gly Thr Ser Gln Phe Thr Cys Phe Tyr Asn Ser Arg Ala 1 5 10 15 Asn Ile Ser Cys Val Trp Ser Gln Asp Gly Ala Leu Gln Asp Thr Ser 20 25 30 Cys Gln Val His Ala Trp Pro Asp Arg Arg Arg Trp Asn Gln Thr Cys 35 40 45 Glu Leu Leu Pro Val Ser Gln Ala Ser Trp Ala Cys Asn Leu Ile Leu 50 55 60 Gly Ala Pro Asp Ser Gln Lys Leu Thr Thr Val Asp Ile Val Thr Leu 65 70 75 80 Arg Val Leu Cys Arg Glu Gly Val Arg Trp Arg Val Met Ala Ile Gln 85 90 95 Asp Phe Lys Pro Phe Glu Asn Leu Arg Leu Met Ala Pro Ile Ser Leu 100 105 110 Gln Val Val His Val Glu Thr His Arg Cys Asn Ile Ser Trp Glu Ile 115 120 125 Ser Gln Ala Ser His Tyr Phe Glu Arg His Leu Glu Phe Glu Ala Arg 130 135 140 Thr Leu Ser Pro Gly His Thr Trp Glu Glu Ala Pro Leu Leu Thr Leu 145 150 155 160 Lys Gln Lys Gln Glu Trp Ile Cys Leu Glu Thr Leu Thr Pro Asp Thr 165 170 175 Gln Tyr Glu Phe Gln Val Arg Val Lys Pro Leu Gln Gly Glu Phe Thr 180 185 190 Thr Trp Ser Pro Trp Ser Gln Pro Leu Ala Phe Arg Thr Lys Pro Ala 195 200 205 Ala Leu Gly Lys Asp Thr 210 <210> 7 <211> 369 <212> PRT <213> Artificial Sequence <220> <223> Human common gamma chain (CD132) sequence <400> 7 Met Leu Lys Pro Ser Leu Pro Phe Thr Ser Leu Leu Phe Leu Gln Leu 1 5 10 15 Pro Leu Leu Gly Val Gly Leu Asn Thr Thr Ile Leu Thr Pro Asn Gly 20 25 30 Asn Glu Asp Thr Thr Ala Asp Phe Phe Leu Thr Thr Met Pro Thr Asp 35 40 45 Ser Leu Ser Val Ser Thr Leu Pro Leu Pro Glu Val Gln Cys Phe Val 50 55 60 Phe Asn Val Glu Tyr Met Asn Cys Thr Trp Asn Ser Ser Ser Glu Pro 65 70 75 80 Gln Pro Thr Asn Leu Thr Leu His Tyr Trp Tyr Lys Asn Ser Asp Asn 85 90 95 Asp Lys Val Gln Lys Cys Ser His Tyr Leu Phe Ser Glu Glu Ile Thr 100 105 110 Ser Gly Cys Gln Leu Gln Lys Lys Glu Ile His Leu Tyr Gln Thr Phe 115 120 125 Val Val Gln Leu Gln Asp Pro Arg Glu Pro Arg Arg Gln Ala Thr Gln 130 135 140 Met Leu Lys Leu Gln Asn Leu Val Ile Pro Trp Ala Pro Glu Asn Leu 145 150 155 160 Thr Leu His Lys Leu Ser Glu Ser Gln Leu Glu Leu Asn Trp Asn Asn 165 170 175 Arg Phe Leu Asn His Cys Leu Glu His Leu Val Gln Tyr Arg Thr Asp 180 185 190 Trp Asp His Ser Trp Thr Glu Gln Ser Val Asp Tyr Arg His Lys Phe 195 200 205 Ser Leu Pro Ser Val Asp Gly Gln Lys Arg Tyr Thr Phe Arg Val Arg 210 215 220 Ser Arg Phe Asn Pro Leu Cys Gly Ser Ala Gln His Trp Ser Glu Trp 225 230 235 240 Ser His Pro Ile His Trp Gly Ser Asn Thr Ser Lys Glu Asn Pro Phe 245 250 255 Leu Phe Ala Leu Glu Ala Val Val Ile Ser Val Gly Ser Met Gly Leu 260 265 270 Ile Ile Ser Leu Leu Cys Val Tyr Phe Trp Leu Glu Arg Thr Met Pro 275 280 285 Arg Ile Pro Thr Leu Lys Asn Leu Glu Asp Leu Val Thr Glu Tyr His 290 295 300 Gly Asn Phe Ser Ala Trp Ser Gly Val Ser Lys Gly Leu Ala Glu Ser 305 310 315 320 Leu Gln Pro Asp Tyr Ser Glu Arg Leu Cys Leu Val Ser Glu Ile Pro 325 330 335 Pro Lys Gly Gly Ala Leu Gly Glu Gly Pro Gly Ala Ser Pro Cys Asn 340 345 350 Gln His Ser Pro Tyr Trp Ala Pro Pro Cys Tyr Thr Leu Lys Pro Glu 355 360 365 Thr <210> 8 <211> 240 <212> PRT <213> Artificial sequence <220> <223> Human common gamma chain (CD132), extracellular domain <400> 8 Leu Asn Thr Thr Ile Leu Thr Pro Asn Gly Asn Glu Asp Thr Thr Ala 1 5 10 15 Asp Phe Phe Leu Thr Thr Met Pro Thr Asp Ser Leu Ser Val Ser Thr 20 25 30 Leu Pro Leu Pro Glu Val Gln Cys Phe Val Phe Asn Val Glu Tyr Met 35 40 45 Asn Cys Thr Trp Asn Ser Ser Ser Glu Pro Gln Pro Thr Asn Leu Thr 50 55 60 Leu His Tyr Trp Tyr Lys Asn Ser Asp Asn Asp Lys Val Gln Lys Cys 65 70 75 80 Ser His Tyr Leu Phe Ser Glu Glu Ile Thr Ser Gly Cys Gln Leu Gln 85 90 95 Lys Lys Glu Ile His Leu Tyr Gln Thr Phe Val Val Gln Leu Gln Asp 100 105 110 Pro Arg Glu Pro Arg Arg Gln Ala Thr Gln Met Leu Lys Leu Gln Asn 115 120 125 Leu Val Ile Pro Trp Ala Pro Glu Asn Leu Thr Leu His Lys Leu Ser 130 135 140 Glu Ser Gln Leu Glu Leu Asn Trp Asn Asn Arg Phe Leu Asn His Cys 145 150 155 160 Leu Glu His Leu Val Gln Tyr Arg Thr Asp Trp Asp His Ser Trp Thr 165 170 175 Glu Gln Ser Val Asp Tyr Arg His Lys Phe Ser Leu Pro Ser Val Asp 180 185 190 Gly Gln Lys Arg Tyr Thr Phe Arg Val Arg Ser Arg Phe Asn Pro Leu 195 200 205 Cys Gly Ser Ala Gln His Trp Ser Glu Trp Ser His Pro Ile His Trp 210 215 220 Gly Ser Asn Thr Ser Lys Glu Asn Pro Phe Leu Phe Ala Leu Glu Ala 225 230 235 240 <210> 9 <211> 231 <212> PRT <2​​​​​​​​​​​​​​​​​​​​​​​​​​​Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Gln Met Thr Lys 130 135 140 Asn Gln Val Lys Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 10 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc Monomer 2 <400> 10 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Glu Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Asp Val Ser Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asp Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Glu Gln Gly Asp Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 11 <211> 231 <212> PRT [[ID=3I]]<213> Artificial sequence <220> <223> IL-2-Fc monomer 1 <400> 11 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 2U 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135 140 Asn Gln Val Lys Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 12 <211> 231 <212> PRT <213> Artificial sequence <220> <223> IL-2-Fc monomer 2 <400> 12 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Glu Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Asp Val Ser Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asp Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Glu Gln Gly Asp Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 13 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc Monomer 1 <400> 13 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135 140 Asn Gln Val Lys Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 14 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc monomer 2 <400> 14 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp<000156⑧>50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Glu Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Glu Val Ser Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asp Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Glu Gln Gly Asp Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 15 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc Monomer 1 <400> 15 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110<00​​​ Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Lys Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 16 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc monomer 2 <400> 16 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Glu Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Glu 130 135 140 Asn Glu Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asp Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Glu Val Asp Lys Ser Arg Trp Glu Gln Gly Asp Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 17 <211> 231 <212> PRT <213> Artificial sequence <220> <223> IL-2-Fc monomer 1 <400> 17 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Gln Leu Thr Lys 130 135 140 Asn Gln Val Lys Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 18 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc monomer 2 <400> 18 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Glu Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Asp Val Ser Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asp Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Glu Gln Gly Asp Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 19 <211> 231 <212> PRT <213> Synthetic sequence <220> <223> IL-2-Fc monomer 1 <400> 19 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala[[ID=XXX]] 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Gln Met Thr Lys 130 135 140 Asn Gln Val Lys Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 20 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-₂-Fc monomer 2 <400> 20 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 [[ID=3,4]]Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Glu Tyr 65 70 75 80 Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp[[ID=]] 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Asp Val Ser Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asp Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Glu Gln Gly Asp Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 21 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc monomer 1 <400> 21 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Gln Met Thr Lys 130 135 140 Asn Gln Val Lys Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 22 <211> 231 <212> PRT <213> Artificial sequence <220> <223> IL-2-Fc monomer <400> 22 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Glu Tyr 65 70 75 80 Ser Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Asp Val Ser Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asp Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Glu Gln Gly Asp Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 23 <211> 229 <212> PRT <213> Artificial sequence <220> <223> IL-2-Fc monomer 1 <400> 23 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe 1 5 10 15 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 20 25 30 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 35 40 45 Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val 50 55 60 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser 65 70 75 80 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 85 90 95 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser 100 105 110 Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 115 120 125 Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Gln Met Thr Lys Asn Gln 130 135 140 Val Lys Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 145 150 155 160 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 165 170 175 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu 180 185 190 Thr Val Asp Lys Ser Arg Trp Gln Glu Gly Asn Val Phe Ser Cys Ser 195 200 205 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 210 215 220 Leu Ser Leu Gly Lys 225 <210> 24 <211> 229 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc Monomer 2 <400> 24 Glu Ser Lys Tyr Gly Pro Pro Cys Pro Pro Cys Pro Ala Pro Glu Phe 1 5 10 15 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 20 25 30 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 35 40 45 Ser Gln Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val 50 55 60 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Glu Phe Asn Ser 65 70 75 80 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 85 90 95 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ser 100 105 110 Ser Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 115 120 125 Gln Val Tyr Thr Leu Pro Pro Ser Gln Glu Glu Met Thr Lys Asn Gln 130 135 140 Val Ser Leu Thr Cys Asp Val Ser Gly Phe Tyr Pro Ser Asp Ile Ala 145 150 155 160 Val Glu Trp Glu Ser Asp Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 165 170 175 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Arg Leu 180 185 190 Thr Val Asp Lys Ser Arg Trp Glu Glu Gly Asp Val Phe Ser Cys Ser 195 200 205 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 210 215 220 Leu Ser Leu Gly Lys 225 <210> 25 <211> 228 <212> PRT <213> Artificial sequence <220> <223> IL-2-Fc monomer 1 <400> 25 Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val 1 5 10 15 Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 20 25 30 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 35 40 45 His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 50 55 60 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 65 70 75 �0 Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn 85 90 95 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro 100 105 110 Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln 115 120 125 Val Tyr Thr Leu Pro Pro Ser Arg Glu Gln Met Thr Lys Asn Gln Val 130 135 140 Lys Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 145 150 155 160 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 165 170 175 Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 180 185 190 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 195 200 205 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 210 215 220 Ser Pro Gly Lys 225 <210> 26 <211> 228 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc monomer 2 <400> 26 Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val 1 5 10 15 Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 20 25 30 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 35 40 45 His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 50 55 60 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Glu Phe Asn Ser Thr 65 70 75 80 Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn 85 90 95 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro 100 105 110 Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln 115 120 125 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 130 135 140 Ser Leu Thr Cys Asp Val Ser Gly Phe Tyr Pro Ser Asp Ile Ala Val 145 150 155 160 Glu Trp Glu Ser Asp Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 165 170 175 Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 180 185 190 Val Asp Lys Ser Arg Trp Glu Gln Gly Asp Val Phe Ser Cys Ser Val 195 200 205 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 210 215 220 Ser Pro Gly Lys 225 <210> 27 <211> 228 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc Monomer 1 <400> 27 Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val 1 5 10 15 Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 20 25 30 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Lys 35 40 45 His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 50 55 60 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr 65 70 75 80 Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn 85 90 95 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro 100 105 110 Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln 115 120 125 Val Tyr Thr Leu Pro Pro Ser Arg Glu Gln Met Thr Lys Asn Gln Val 130 135 140 Lys Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 145 150 155 160 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 165 170 175 Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 180 185 190 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 195 200 205 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 210 215 220 Ser Pro Gly Lys 225 <210> 28 <211> 228 <212> PRT <213> Artificial sequence <220> <223> IL-2-Fc monomer 2 <400> 28 Glu Arg Lys Cys Cys Val Glu Cys Pro Pro Cys Pro Ala Pro Pro Val 1 5 10 15 Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 20 25 30 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Lys 35 40 45 His Glu Asp Pro Glu Val Gln Phe Asn Trp Tyr Val Asp Gly Val Glu 50 55 60 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Glu Phe Asn Ser Thr 65 70 75 80 Phe Arg Val Val Ser Val Leu Thr Val Val His Gln Asp Trp Leu Asn 85 90 95 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Gly Leu Pro Ala Pro 100 105 110 Ile Glu Lys Thr Ile Ser Lys Thr Lys Gly Gln Pro Arg Glu Pro Gln 115 120 125 Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val 130 135 140 Ser Leu Thr Cys Asp Val Ser Gly Phe Tyr Pro Ser Asp Ile Ala Val 145 150 155 160 Glu Trp Glu Ser Asp Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 165 170 175 Pro Met Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 180 185 190 Val Asp Lys Ser Arg Trp Glu Gln Gly Asp Val Phe Ser Cys Ser Val 195 200 205 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 210 215 220 Ser Pro Gly Lys 225 <210> 29 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc monomer 1 <400> 29 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Gln Met Thr Lys 130 135 140 Asn Gln Val Lys Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 30 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc monomer 2 <400> 30 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Glu Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Asp Val Ser Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asp Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Glu Gln Gly Asp Val Phe Ser 195 200 205 Cys Ser Val Leu His Glu Ala Leu His Ser His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 31 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc monomer 1 <400> 31 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 32 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc Monomer 2 <400> 32 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 33 <211> 231 <212> PRT <213> Artificial sequence <220> <223> IL-2-Fc monomer 1 <400> 33 Glu Pro Lys Ser Ser Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Glu Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys 130 135 140 Asn Gln Val Ser Leu Thr Cys Asp Val Ser Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asp Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Glu Gln Gly Asp Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 34 <211> 231 <212> PRT <213> Artificial Sequence <220> <223> IL-2-Fc monomer 2 <400> 34 Glu Arg Lys Ser Ser Asp Lys Thr His Thr Cys Pro Arg Cys Pro Ala 1 5 10 15 Pro Pro Val Ala Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 20 25 30 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 35 40 45 Asp Val Lys His Glu Asp Pro Glu Val Lys Phe Lys Trp Tyr Val Asp 50 55 60 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 65 70 75 80 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 85 90 95 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 100 105 110 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 115 120 125 Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Glu Gln Met Thr Lys 130 135 140 Asn Gln Val Lys Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 145 150 155 160 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 165 170 175 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 180 185 190 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 195 200 205 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 210 215 220 Leu Ser Leu Ser Pro Gly Lys 225 230 <210> 35 <211> 5 <212> PRT <213> Artificial sequence <220> <223> Linker <400> 35 Gly Gly Gly Gly Ser 1 5 <210> 36 <211> 10 <212> PRT <213> Artificial sequence <220> <223> Linker <400> 36 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 <210> 37 <211> 15 <212> PRT <213> Artificial sequence <220> <223> Linker <400> 37 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 15 <210> 38 <211> 20 <212> PRT <213> Artificial sequence <220> <223> Linker <400> 38 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 1 5 10 15 Gly Gly Gly Ser 20 <210> 39 <211> 5 <212> PRT <213> Artificial Sequence <220> <223> Linker <400> 39 Gly Gly Gly Glu Ser 1 5 <210> 40 <211> 135 <212> PRT <213> Artificial Sequence <220> <223> XENP014135 hIL2_0.1 <400> 40 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys<00Gly Gly Gly Glu Ser35 40 45<0Gly Gly Gly Glu SerLys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 41 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP014136 hIL2_0.9 <400> 41 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Ala Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 42 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014137 hIL2_0.10 <400> 42 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Asp Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 43 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014138 hIL2_0.11 <400> 43 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Glu Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 44 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014139 hIL2_0.12 <400> 44 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Phe Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 45 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014140 hIL2_0.13 <400> 45 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gly Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 46 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014141 hIL2_0.14 <400> 46 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr His Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 47 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP014142 hIL2_0.15 <400> 47 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Ile Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 48 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014143 hIL2_0.16 <400> 48 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Lys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 49 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP014144 hIL2_0.17 <400> 49 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Leu Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 50 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014145 hIL2_0.18 <400> 50 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Met Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 51 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014146 hIL2_0.19 <400> 51 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Asn Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 52 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014147 hIL2_0.20 <400> 52 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Pro Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 53 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014148 hIL2_0.21 <400> 53 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 54 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014149 hIL2_0.22 <400> 54 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Ser Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 55 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014150 hIL2_0.23 <400> 55 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Thr Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 56 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014151 hIL2_0.24 <400> 56 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Val Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 57 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014152 hIL2_0.25 <400> 57 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Trp Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 58 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014153 hIL2_0.26 <400> 58 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Tyr Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 59 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014154 hIL2_0.27 <400> 59 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Ala Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 60 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014155 hIL2_0.28 <400> 60 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Asp Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 61 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014156 hIL2_0.29 <400> 61 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Glu Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 62 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014157 hIL2_0.30 <400> 62 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Phe Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 63 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014158 hIL2_0.31 <400> 63 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Gly Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 64 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014159 hIL2_0.32 <400> 64 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu His Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 65 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014160 hIL2_0.33 <400> 65 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Ile Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 66 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014161 hIL2_0.34 <400> 66 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Lys Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 67 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014162 hIL2_0.35 <400> 67 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Leu Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 68 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014163 hIL2_0.36 <400> 68 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Met Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 69 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014164 hIL2_0.37 <400> 69 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Asn Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 70 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014165 hIL2_0.38 <400> 70 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Pro Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 71 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014166 hIL2_0.39 <400> 71 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Gln Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 72 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014167 hIL2_0.40 <400> 72 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Arg Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 73 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014168 hIL2_0.41 <400> 73 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Ser Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 74 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014169 hIL2_0.42 <400> 74 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Val Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 75 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014170 hIL2_0.43 <400> 75 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Trp Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 76 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014171 hIL2_0.44 <400> 76 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Tyr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 77 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014172 hIL2_0.45 <400> 77 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Ala Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 78 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014173 hIL2_0.46 <400> 78 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Asp Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 79 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014174 hIL2_0.47 <400> 79 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Glu Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 80 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014175 hIL2_0.48 <400> 80 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Gly Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 81 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014176 hIL2_0.49 <400> 81 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr His Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 82 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014177 hIL2_0.50 <400> 82 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Ile Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 83 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014178 hIL2_0.51 <400> 83 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 84 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014179 hIL2_0.52 <400> 84 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Leu Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 85 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014180 hIL2_0.53 <400> 85 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Met Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 86 <211> 129 <212> PRT <213> artificial sequence <220> <223> XENP014181 hIL2_0.54 <400> 86 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Asn Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 authorities <210> 87 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014182 hIL2_0.55 <400> 87 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Pro Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 88 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014183 hIL2_0.56 <400> 88 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Gln Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 89 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014184 hIL2_0.57 <400> 89 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Arg Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 90 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014185 hIL2_0.58 <400> 90 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Ser Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 91 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014186 hIL2_0.59 <400> 91 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Thr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 92 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014187 hIL2_0.60 <400> 92 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Val Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 93 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014188 hIL2_0.61 <400> 93 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Trp Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 94 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014189 hIL2_0.62 <400> 94 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Tyr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 95 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014190 hIL2_0.63 <400> 95 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Lys Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 96 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014191 hIL2_0.64 <400> 96 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Gln Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 97 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014192 hIL2_0.65 <400> 97 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Glu Met Leu Lys Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 98 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014193 hIL2_0.66 <400> 98 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Arg Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 99 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014194 hIL2_0.67 <400> 99 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Asn Met Leu Gln Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 100 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014195 hIL2_0.68 <400> 100 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Val Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 101 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014196 hIL2_0.69 <400> 101 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Asn Met Leu Val Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 102 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014201 hIL2_0.70 <400> 102 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Met Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 103 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014202 hIL2_0.71 <400> 103 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Ser Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 104 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014203 hIL2_0.72 <400> 104 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Leu Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 105 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014204 hIL2_0.73 <400> 105 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Asn Met Leu Met Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 106 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014205 hIL2_0.74 <400> 106 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Ile Tyr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 107 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014206 hIL2_0.75 <400> 107 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Glu Tyr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 108 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014207 hIL2_0.76 <400> 108 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Asp Tyr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 109 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014208 hIL2_0.77 <400> 109 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Met Tyr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 110 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014209 hIL2_0.78 <400> 110 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Gln Tyr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 111 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014210 hIL2_0.79 <400> 111 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Glu His Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 112 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014211 hIL2_0.80 <400> 112 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Glu Leu Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 113 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014212 hIL2_0.81 <400> 113 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Glu Pro Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 114 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014213 hIL2_0.82 <400> 114 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Thr Tyr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 115 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014214 hIL2_0.83 <400> 115 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Asn Met Leu Arg Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 116 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014215 hIL2_0.84 <400> 116 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Asn Met Leu Lys Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 117 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014216 hIL2_0.85 <400> 117 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Val Met Leu Arg Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 118 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014217 hIL2_0.86 <400> 118 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Pro Met Leu Arg Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 119 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014218 hIL2_0.87 <400> 119 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Glu Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 120 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014219 hIL2_0.88 <400> 120 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Asp Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 121 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014220 hIL2_0.89 <400> 121 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Met Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 122 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014221 hIL2_0.90 <400> 122 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Gln Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 123 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014222 hIL2_0.91 <400> 123 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Thr Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 124 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014223 hIL2_0.92 <400> 124 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Ile Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 125 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014224 hIL2_0.93 <400> 125 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Asn Met Leu Thr Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 126 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014225 hIL2_0.94 <400> 126 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu His Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 127 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014226 hIL2_0.95 <400> 127 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Lys Tyr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 128 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014227 hIL2_0.96 <400> 128 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Arg Tyr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 129 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014228 hIL2_0.97 <400> 129 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Gln Tyr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 130 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014229 hIL2_0.98 <400> 130 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Val Tyr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 131 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014230 hIL2_0.99 <400> 131 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Asn Met Leu Lys Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 132 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014231 hIL2_0.100 <400> 132 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu His Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 133 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014232 hIL2_0.101 <400> 133 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Lys Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 134 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014233 hIL2_0.102 <400> 134 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Gln Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 135 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014234 hIL2_0.103 <400> 135 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Val Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 136 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014235 hIL2_0.104 <400> 136 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Gln Met Leu Arg Lys Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 137 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP014277 hIL2_0.6 <400> 137 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Ser Asn His Lys 20 25 30 Asn Pro Arg Leu Ala Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Glu 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Ala Leu Arg Leu Ala Pro Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asp Val Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Thr Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 138 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP014381 hIL2_0.202 <400> 138 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Ser Asn His Lys 20 25 30 Asn Pro Arg Leu Ala Leu Met Leu Thr Phe Lys Phe Tyr Met Pro Glu 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Ala Leu Arg Leu Ala Pro Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asp Val Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Thr Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 139 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP014135 hIL2_0.1 <400> 139 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 140 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP014142 hIL2_0.15 <400> 140 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Ile Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 141 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014143 hIL2_0.16 <400> 141 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Lys Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 142 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP014144 hIL2_0.17 <400> 142 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Leu Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 143 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014168 hIL2_0.41 <400> 143 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Ser Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 144 <211> 133 <212> PRT <213> artificial sequence <220> <223> XENP014189 hIL2_0.62 <400> 144 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Tyr Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr 130 <210> 145 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023816 hIL2_0.205 <400> 145 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Glu Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 146 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023817 hIL2_0.206 <400> 146 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Asp Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 147 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023820 hIL2_0.209 <400> 147 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Glu Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 148 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023824 hIL2_0.213 <400> 148 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Gln His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 149 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023827 hIL2_0.216 <400> 149 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu Tyr 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 150 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023830 hIL2_0.219 <400> 150 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Asp Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 151 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023833 hIL2_0.222 <400> 151 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asn Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 152 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023834 hIL2_0.223 <400> 152 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Glu Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 153 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023836 hIL2_0.225 <400> 153 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Arg Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 154 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023837 hIL2_0.226 <400> 154 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Ser Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 155 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023838 hIL2_0.227 <400> 155 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Arg Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 156 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023839 hIL2_0.228 <400> 156 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Tyr Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 157 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023840 hIL2_0.229 <400> 157 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Phe Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 158 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023842 hIL2_0.231 <400> 158 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Arg 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 159 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023843 hIL2_0.232 <400> 159 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Glu Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 160 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023846 hIL2_0.235 <400> 160 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Gln Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 161 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023848 hIL2_0.237 <400> 161 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Gln Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 162 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023849 hIL2_0.238 <400> 162 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Arg 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 163 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023851 hIL2_0.240 <400> 163 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Gln Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 164 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023852 hIL2_0.241 <400> 164 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Leu Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 165 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023853 hIL2_0.242 <400> 165 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Ile Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 166 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023855 hIL2_0.244 <400> 166 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Asp Pro Arg Asp Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 167 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023859 hIL2_0.248 <400> 167 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asn Leu Ile Ser Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 168 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023861 hIL2_0.250 <400> 168 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Thr Asn Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 169 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023862 hIL2_0.251 <400> 169 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asp Ile Asn Val Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 170 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023868 hIL2_0.257 <400> 170 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys 50 55 60 Pro Leu Glu Glu Val Leu Asn Leu Ala Gln Ser Lys Asn Phe His Leu 65 70 75 80 Arg Pro Arg Asp Leu Ile Ser Asn Ile Asn Leu Ile Val Leu Glu Leu 85 90 95 Lys Gly Ser Glu Thr Thr Phe Met Cys Glu Tyr Ala Asp Glu Thr Ala 100 105 110 Thr Ile Val Glu Phe Leu Asn Arg Trp Ile Thr Phe Ser Gln Ser Ile 115 120 125 Ile Ser Thr Leu Thr Gly Ser 130 135 <210> 171 <211> 135 <212> PRT <213> artificial sequence <220> <223> XENP023869 hIL2_0.258 <400> 171 Ala Pro Thr Ser Ser Ser Thr Lys Lys Thr Gln Leu Gln Leu Glu His 1 5 10 15 Leu Leu Leu Asp Leu Gln Met Ile Leu Asn Gly Ile Asn Asn Tyr Lys 20 25 30 Asn Pro Lys Leu Thr Arg Met Leu Thr Phe Lys Phe Tyr Met Pro Lys 35 40 45 Lys Ala Thr Glu Leu Lys His Leu Gln Cys Leu Glu Glu Glu Leu Lys...

Claims

1. A polypeptide composition for amplifying Tregs, which comprises a heterodimeric protein complex, wherein the heterodimeric protein complex consists of: - A first protein monomer, which consists of the amino acid sequence shown in SEQ ID NO: 297; and - A second protein monomer, which consists of the amino acid sequence shown in SEQ ID NO:

298.

2. A nucleic acid composition, which comprises: a) A first nucleic acid, which encodes the first protein monomer of claim 1; and b) A second nucleic acid, which encodes the second protein monomer of claim 1.

3. An expression vector composition, which comprises: a) A first expression vector, which comprises the first nucleic acid of claim 2; and b) A second expression vector, which comprises the second nucleic acid of claim 2.

4. A host cell, which comprises the expression vector composition according to claim 3.

5. A method for preparing a polypeptide composition, the method comprising: Culturing the host cell according to claim 4 under conditions for producing the composition and recovering the composition.

Images