Inhibition of t-cell engaging agent-related adverse effects

Abstract

The present invention relates to the inhibition of adverse effects related to T cell engaging agents, such as cytokine release syndrome. Specifically, the invention relates to the inhibition of such adverse effects using a tyrosine kinase inhibitor and a glucocorticoid.

Description

[0001] Case P39664-WO

[0002] Inhibition of T-cell engaging agent-related adverse effects

[0003] Field of the Invention

[0004] The present invention relates to the inhibition of adverse effects related to T cell engaging agents, such as cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS). Specifically, the invention relates to the inhibition of such side effects using the combination of a glucocorticoid (GC), such as dexamethasone and methylprednisolone, together with a tyrosine kinase inhibitor (TKI), such as an inhibitor of Janus kinase (JAK) and an inhibitor of mammalian target of rapamycin (mTOR).

[0005] Background

[0006] T cell engaging agents such as T cell bispecific antibodies (TCBs) or chimeric antigen receptor (CAR) expressing T cells (CAR-T cells) hold great promise as cancer immunotherapeutics. However, treatment with T cell engaging agent is sometimes associated with safety liabilities due to on-target on-tumor, on-target off-tumor cytotoxic activity and cytokine release. One of the most common adverse effects reported for T cell engaging agents is Cytokine Release Syndrome (CRS). This complex clinical syndrome is characterized by fever, hypotension and respiratory deficiency and associated with the release of pro-inflammatory cytokines such as IL-6, TNF-a, IFN-y, and IL-10 (see e.g. Shimabukuro-Vomhagen et al., J Immunother Cancer (2018) 6, 56). Approaches to mitigate these life-threatening toxicities are greatly needed.

[0007] Summary of the Invention

[0008] The present inventors have found that the combination of a GC and a TKI may be used to reduce TCB-induced cytokine release, even from pre-activated PBMCs, and thus to inhibit adverse effects of T cell engaging therapies. Surprisingly, the combination of these agents more strongly reduces cytokine release than either agent alone. It has also been found that combining a GC and a TKI at

[0009] LK / 29.10.2024 subclinical concentrations has comparable effects to GC at concentrations corresponding to clinical exposure, thus allowing to use these agents at lower concentrations and reducing the risk of unwanted side effects. Moreover, the combination of a GC and a JAK inhibitor and, in particular, the combination of a GC and an mTOR inhibitor has been found to potently reduce TCB-induced cytokine release while retaining TCB-mediated target cell killing, thus uncoupling TCB-dependent cytokine release and target cell killing and allowing such combinations to be used not only as a “safety switch” for mitigating adverse effects but more generally to reduce the undesirable adverse effects of T cell engaging therapies while preserving the desirable therapeutic effects. PBMCs exposed to a TCB, a GC and a JAK inhibitor or mTOR inhibitor also retain their capacity to kill tumor cells when restimulated with the TCB alone and then only secrete limited amounts of cytokines, demonstrating that the reduction of cytokine release is an enduring effect of the combination.

[0010] Embodiments provided by the present invention include the following:

[0011] El. A T cell engaging agent for use in the treatment of a disease in an individual, wherein said treatment comprises the administration of the following agents to the individual

[0012] (a) the T cell engaging agent,

[0013] (b) a TKI, and

[0014] (c) a GC.

[0015] E2. Use of a T cell engaging agent in the manufacture of a medicament for the treatment of a disease in an individual, wherein said treatment comprises the administration of the following agents to the individual

[0016] (a) the T cell engaging agent,

[0017] (b) TKI, and

[0018] (c) a GC.

[0019] E3. A method for treatment of a disease in an individual, wherein said method comprises the administration of the following agents to the individual

[0020] (a) a T cell engaging agent,

[0021] (b) a TKI, and

[0022] (c) a GC. E4. The T cell engaging agent for use, use or method according to any one of El to E3, wherein the administration of the TKI and the GC is for the inhibition of an adverse effect related to the administration of the T cell engaging agent.

[0023] E5. A TKI and a GC for use in the inhibition of an adverse effect related to the administration of a T cell engaging agent to an individual, wherein said inhibition comprises the administration of the TKI and the GC to the individual.

[0024] E6. Use of a TKI and a GC in the manufacture of a medicament for the inhibition of an adverse effect related to the administration of a T cell engaging agent, wherein said inhibition comprises the administration of the TKI and the GC to the individual.

[0025] E7. A method for inhibiting an adverse effect related to the administration of a T cell engaging agent to an individual, comprising the administration of a TKI and a GC to the individual.

[0026] E8. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E7, wherein the TKI is an inhibitor of JAK or mTOR.

[0027] E9. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E8, wherein the TKI is an mTOR inhibitor, optionally selected from the group consisting of sirolimus, temsirolimus and everolimus.

[0028] E10. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E8, wherein the TKI is a JAK inhibitor, optionally a JAK1 and / or JAK2 inhibitor, optionally selected from the group consisting of ruxolitinib, baricitinib, tofacitinib or fedratinib.

[0029] El l. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E10, wherein the GC is selected from dexamethasone and methylprednisolone.

[0030] El 2. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to El 1, wherein the TKI is an inhibitor of JAK or mTOR and the GC is dexamethasone.

[0031] El 3. The T cell engaging agent or TKI and GC for use, use or method according to any one of E4 to E12, wherein the adverse effect is one or more effect selected from the group consisting of

[0032] (i) cytokine secretion by immune cells, particularly by T cells;

[0033] (ii) cytokine release syndrome (CRS);

[0034] (iii) one or more of fever, hypotension and hypoxia; (iv) an elevated serum level of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL- 10, TNF-a, GM-CSF, MCP-1 and IL-1P;

[0035] (v) immune effector cell-associated neurotoxicity syndrome (ICANS);

[0036] (vi) one or more of mild tremor, confusion, disorientation, headache, attention deficits, agitation, seizures, cerebral oedema, hesitancy of speech, deterioration in handwriting, aphasia with expressive and / or receptive components, status epilepticus, fatal cerebral oedema, intracerebral haemorrhage and transient coma;

[0037] (vii) one or more of thrombocytopenia, an elevated serum level of one or more of lactate dehydrogenase, ferritin, IFN-y, IL-10, granzyme B, GM-CSF, MIP-la, TNF, IL-2, and an elevated level in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL- 10, IL-6 and granzyme B; and

[0038] (viii) T cell exhaustion.

[0039] El 4. The T cell engaging agent or TKI and GC for use, use or method according to any one of E4 to El 2, wherein the adverse effect is CRS and / or ICANS.

[0040] El 5. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E14, wherein administration of the TKI and GC does not cause inhibition of a desired effect related to the administration of the T cell engaging agent.

[0041] El 6. The T cell engaging agent or TKI and GC for use, use or method according to El 5, wherein the desired effect is one or more effect selected from the group consisting of

[0042] (i) activation of T cells;

[0043] (ii) cytotoxic activity of T cells; and

[0044] (iii) the therapeutic effect of the T cell engaging agent.

[0045] El 7. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E16, wherein the administration of the TKI and GC is for the inhibition of an adverse effect related to the administration of the T cell engaging agent, wherein the adverse effect is CRS and / or ICANS, wherein administration of the TKI and GC does not cause inhibition of a desired effect related to the administration of the T cell engaging agent, and wherein the desired effect is the therapeutic effect of the T cell engaging agent.

[0046] El 8. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E16, wherein the administration of the TKI and GC is for the inhibition of an adverse effect related to the administration of the T cell engaging agent, wherein the adverse effect is cytokine secretion by immune cells, particularly by T cells, wherein administration of the TKI and GC does not cause inhibition of a desired effect related to the administration of the T cell engaging agent, and wherein the desired effect is activation of T cells and / or cytotoxic activity of T cells.

[0047] El 9. The T cell engaging agent or TKI and GC for use, use or method according to any one of E4 to El 8, wherein administration of the TKI and GC is upon or after manifestation of the adverse effect.

[0048] E20. The T cell engaging agent or TKI and GC for use, use or method according to any one of E4 to El 8, wherein administration of the TKI and GC is before manifestation of the adverse effect.

[0049] E21. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E20, wherein administration of the inhibitor of the TKI and GC is one or more of

[0050] (i) before, concurrent to, or after the administration of the T cell engaging agent;

[0051] (ii) intermittently or continuously; and

[0052] (iii) oral or parenteral, particularly intravenous.

[0053] E22. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E21, wherein administration of the TKI and GC is associated with the first administration of the T cell engaging agent, and optionally is prior, concurrent or subsequent to the first administration of the T cell engaging agent.

[0054] E23. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E22, wherein the administration of the T cell engaging agent is one or more of

[0055] (i) at an effective dose;

[0056] (ii) parenteral, particularly intravenous; and

[0057] (iii) the first administration of the T cell engaging agent to the individual.

[0058] E24. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E23, wherein GC is dexamethasone, administered at a dose of below 0.5 mg and, optionally, wherein the individual is an adult.

[0059] E25. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E24, wherein the TKI is sirolimus and administered at a dose of below 0.5 mg or ruxolitinib and administered at a dose of below 5 mg and, optionally, wherein the individual is an adult. E26. The T cell engaging agent or TKI and GC for use, use or method according to any one of El to E25, wherein the T cell engaging agent is a T cell bispecific antibody or a CAR-T cell.

[0060] E27. The T cell engaging agent or TKI and GC for use, use or method according to E26, wherein the T cell bispecific antibody binds to CD3 and a target cell antigen.

[0061] E28. The T cell engaging agent or TKI and GC for use, use or method according to E26 or E27, wherein the T cell bispecific antibody comprises an antigen binding moiety that binds to CD3 and an antigen binding moiety that binds to a target cell antigen.

[0062] E29. The T cell engaging agent, inhibitor of JAK and / or mTOR signaling, use or method according to E27 or E38, wherein the target cell antigen is CD20.

[0063] E30. The T cell engaging agent, inhibitor of JAK and / or mTOR signaling, use or method according to E29, wherein the T cell bispecific antibody comprises

[0064] (i) a first antigen binding moiety that binds to CD3 and comprises a heavy chain variable region comprising the heavy chain CDR (HCDR) 1 of SEQ ID NO: 4, the HCDR2 of SEQ ID NO: 5, and the HCDR3 of SEQ ID NO: 6; and a light chain variable region comprising the light chain CDR (LCDR) 1 of SEQ ID NO: 7, the LCDR2 of SEQ ID NO: 8 and the LCDR3 of SEQ ID NO: 9; and

[0065] (ii) a second antigen binding moiety that binds to CD20 and comprises a heavy chain variable region comprising the heavy chain CDR (HCDR) 1 of SEQ ID NO: 12, the HCDR2 of SEQ ID NO: 13, and the HCDR3 of SEQ ID NO: 14; and a light chain variable region comprising the light chain CDR (LCDR) 1 of SEQ ID NO: 15, the LCDR2 of SEQ ID NO: 16 and the LCDR3 of SEQ ID NO: 17.

[0066] E31. The T cell engaging agent, inhibitor of JAK and / or mTOR signaling, use or method according to E29 or E30, wherein the T cell bispecific antibody comprises a third antigen binding moiety that binds to CD20 and / or an Fc domain composed of a first and a second subunit.

[0067] E32. The T cell engaging agent, inhibitor of JAK and / or mTOR signaling, use or method according to any one of E29 to E31, wherein the T cell bispecific antibody comprises

[0068] (i) a first antigen binding moiety that binds to CD3, comprising a heavy chain variable region comprising the heavy chain CDR (HCDR) 1 of SEQ ID NO: 4, the HCDR2 of SEQ ID NO: 5, and the HCDR3 of SEQ ID NO: 6; and a light chain variable region comprising the light chain CDR (LCDR) 1 of SEQ ID NO: 7, the LCDR2 of SEQ ID NO: 8 and the LCDR3 of SEQ ID NO: 9, wherein the first antigen binding moiety is a crossover Fab molecule wherein either the variable or the constant regions of the Fab light chain and the Fab heavy chain are exchanged;

[0069] (ii) a second and a third antigen binding moiety that bind to CD20, comprising a heavy chain variable region comprising the heavy chain CDR (HCDR) 1 of SEQ ID NO: 12, the HCDR2 of SEQ ID NO: 13, and the HCDR3 of SEQ ID NO: 14; and a light chain variable region comprising the light chain CDR (LCDR) 1 of SEQ ID NO: 15, the LCDR2 of SEQ ID NO: 16 and the LCDR3 of SEQ ID NO: 17, wherein the second and third antigen binding moiety are each a Fab molecule, particularly a conventional Fab molecule;

[0070] (iii) an Fc domain composed of a first and a second subunit, wherein the second antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the first antigen binding moiety, and the first antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first subunit of the Fc domain, and wherein the third antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the second subunit of the Fc domain.

[0071] E33. The T cell engaging agent, inhibitor of JAK and / or mTOR signaling, use or method according to any one of E29 to E32, wherein the first antigen binding moiety of the T cell bispecific antibody comprises a heavy chain variable region sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 10 and a light chain variable region sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 11, and / or the second and (where present) third antigen binding moiety of the T cell bispecific antibody comprise a heavy chain variable region sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 18 and a light chain variable region sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 19.

[0072] E34. The T cell engaging agent, inhibitor of JAK and / or mTOR signaling, use or method according to any one of E29 to E33, wherein the first antigen binding moiety of the T cell bispecific antibody is a crossover Fab molecule wherein the variable regions of the Fab light chain and the Fab heavy chain are exchanged, and wherein the second and (where present) third antigen binding moiety of the T cell bispecific antibody is a conventional Fab molecule wherein in the constant domain CL the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat) and the amino acid at position 123 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat) and in the constant domain CHI the amino acid at position 147 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index) and the amino acid at position 213 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index).

[0073] E35. The T cell engaging agent, inhibitor of JAK and / or mTOR signaling, use or method according to any one of E31 to E34, wherein the Fc domain of the T cell bispecific antibody comprises a modification promoting the association of the first and the second subunit of the Fc domain, and / or the Fc domain comprises one or more amino acid substitution that reduces binding to an Fc receptor and / or effector function.

[0074] E36. The T cell engaging agent, inhibitor of JAK and / or mTOR signaling, use or method according to any one of E26 to E35, wherein the T cell bispecific antibody is glofitamab.

[0075] E37. The T cell engaging agent, inhibitor of JAK and / or mTOR signaling, use or method according to any one of El to E36, wherein the disease (to be treated by the T cell engaging agent) is cancer, particularly a cancer expressing the target cell antigen of the T cell engaging agent.

[0076] E38. The T cell engaging agent, inhibitor of JAK and / or mTOR signaling, use or method according to E37, wherein the cancer is

[0077] (i) a CD20-expressing cancer,

[0078] (ii) a B-cell cancer, and / or

[0079] (ii) selected from the group consisting of Non-Hodgkin lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle-cell lymphoma (MCL) and marginal zone lymphoma (MZL).

[0080] The T cell engaging agent or TKI and GC for use, uses or methods described above and herein, may incorporate, singly or in combination, any of the features described in the following (unless the context dictates otherwise).

[0081] Terms are used herein as generally used in the art, unless otherwise defined herein. Brief description of the Drawings

[0082] Figure 1. Assay set-up. PBMCs were co-cultured with CTV-labeled Toledo tumor cells (Effector : Tumor cells ratio = 5: 1) with escalating doses of CD20-TCB (glofitamab) in the presence and absence of 10 nM dexamethasone, 50 nM JAK inhibitor (ruxolitinib), 10 mTOR inhibitor (sirolimus), 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib) or 10 nM dexamethasone + 10 nM mTOR inhibitor (sirolimus) for 15-18 hrs. At assay end point, tumor cell killing and immune cell phenotyping were performed by flow cytometry and cytokine levels were measured in the supernatant by Legendplex.

[0083] Figure 2. Effects of the combination of subclinical dexamethasone concentrations with subclinical JAK (ruxolitinib) and mTOR (sirolimus) inhibitors on tumor cell killing. At assay end point, the killing of CTV-labeled Toledo cells was measured by flow cytometry using a Live / dead NIR stain. A. Dose response killing curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0084] Figure 3. Effects of the combination of subclinical dexamethasone concentrations with subclinical JAK (ruxolitinib) and mTOR (sirolimus) inhibitors on CD8+ T cell activation. At assay end point, the expression of CD69 on CD8+ T cells was measured by flow cytometry. A. Dose response curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0085] Figure 4. Effects of the combination of subclinical dexamethasone concentrations with subclinical JAK (ruxolitinib) and mTOR (sirolimus) inhibitors on CD8+ T cell activation. At assay end point, the expression of CD25 on CD8+ T cells was measured by flow cytometry. A. Dose response curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0086] Figure 5. Effects of the combination of subclinical dexamethasone concentrations with subclinical JAK (ruxolitinib) and mTOR (sirolimus) inhibitors on CD4+ T cell activation. At assay end point, the expression of CD69 on CD4+ T cells was measured by flow cytometry. A. Dose response curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0087] Figure 6. Effects of the combination of subclinical dexamethasone concentrations with subclinical JAK (ruxolitinib) and mTOR (sirolimus) inhibitors on CD4+ T cell activation. At assay end point, the expression of CD25 on CD4+ T cells was measured by flow cytometry. A. Dose response curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM. Figure 7. Effects of the combination of subclinical dexamethasone concentrations with subclinical JAK (ruxolitinib) and mTOR (sirolimus) inhibitors on IL-2 release. At assay end point, the levels of IL-2 were measured in the supernatants by Legendplex. A. Dose response curves of a representative donor. B. AUC values of N= 4 PBMCs donors. Mean + / - SEM.

[0088] Figure 8. Effects of the combination of subclinical dexamethasone concentrations with subclinical JAK (ruxolitinib) and mTOR (sirolimus) inhibitors on TNF-a release. At assay end point, the levels of TNF-a were measured in the supernatants by Legendplex. A. Dose response curves of a representative donor. B. AUC values of N= 4 PBMCs donors. Mean + / - SEM.

[0089] Figure 9. Effects of the combination of subclinical dexamethasone concentrations with subclinical JAK (ruxolitinib) and mTOR (sirolimus) inhibitors on IFN-y release. At assay end point, the levels of IFN-y were measured in the supernatants by Legendplex. A. Dose response curves of a representative donor. B. AUC values of N= 4 PBMCs donors. Mean + / - SEM.

[0090] Figure 10. Effects of the combination of subclinical dexamethasone concentrations with subclinical JAK (ruxolitinib) and mTOR (sirolimus) inhibitors on IL-6 release. At assay end point, the levels of IL-6 were measured in the supernatants by Legendplex. A. Dose response curves of a representative donor. B. AUC values of N= 4 PBMCs donors. Mean + / - SEM.

[0091] Figure 11. Effects on tumor cell killing of the combination of dexamethasone and the JAK (ruxolitinib) inhibitor or the mTOR (sirolimus) inhibitor at subclinical concentrations in comparison to dexamethasone concentrations corresponding to Cmax and Cmin clinical values of a 20 mg dose administered intravenously to patients. At assay end point, the killing of CTV-labeled Toledo cells was measured by flow cytometry using a Live / dead NIR stain. A. Dose response killing curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0092] Figure 12. Effects on CD8+ T cell activation of the combination of dexamethasone and the JAK (ruxolitinib) inhibitor or the mTOR (sirolimus) inhibitor at subclinical concentrations in comparison to dexamethasone concentrations corresponding to Cmax and Cmin clinical values of a 20 mg dose administered intravenously to patients. At assay end point, the expression of CD69 on CD8+ T cells was measured by flow cytometry. A. Dose response curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0093] Figure 13. Effects on CD8+ T cell activation of the combination of dexamethasone and the JAK (ruxolitinib) inhibitor or the mTOR (sirolimus) inhibitor at subclinical concentrations in comparison to dexamethasone concentrations corresponding to Cmax and Cmin clinical values of a 20 mg dose administered intravenously to patients. At assay end point, the expression of CD25 on CD8+ T cells was measured by flow cytometry. A. Dose response curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0094] Figure 14. Effects on CD4+ T cell activation of the combination of dexamethasone and the JAK (ruxolitinib) inhibitor or the mTOR (sirolimus) inhibitor at subclinical concentrations in comparison to dexamethasone concentrations corresponding to Cmax and Cmin clinical values of a 20 mg dose administered intravenously to patients. At assay end point, the expression of CD69 on CD4+ T cells was measured by flow cytometry. A. Dose response curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0095] Figure 15. Effects on CD4+ T cell activation of the combination of dexamethasone and the JAK (ruxolitinib) inhibitor or the mTOR (sirolimus) inhibitor at subclinical concentrations in comparison to dexamethasone concentrations corresponding to Cmax and Cmin clinical values of a 20 mg dose administered intravenously to patients. At assay end point, the expression of CD25 on CD4+ T cells was measured by flow cytometry. A. Dose response curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0096] Figure 16. Effects on IL-2 release of the combination of dexamethasone and the JAK (ruxolitinib) inhibitor or the mTOR (sirolimus) inhibitor at subclinical concentrations in comparison to dexamethasone concentrations corresponding to Cmax and Cmin clinical values of a 20 mg dose administered intravenously to patients. At assay end point, the levels of IL-2 were measured in the supernatants by Legendplex. A. Dose response curves of a representative donor. B. AUC values of N= 4 PBMCs donors. Mean + / - SEM.

[0097] Figure 17. Effects on TNF-u release of the combination of dexamethasone and the JAK (ruxolitinib) inhibitor or the mTOR (sirolimus) inhibitor at subclinical concentrations in comparison to dexamethasone concentrations corresponding to Cmax and Cmin clinical values of a 20 mg dose administered intravenously to patients. At assay end point, the levels of TNF-u were measured in the supernatants by Legendplex. A. Dose response curves of a representative donor. B. AUC values of N= 4 PBMCs donors. Mean + / - SEM.

[0098] Figure 18. Effects on IFN-y release of the combination of dexamethasone and the JAK (ruxolitinib) inhibitor or the mTOR (sirolimus) inhibitor at subclinical concentrations in comparison to dexamethasone concentrations corresponding to Cmax and Cmin clinical values of a 20 mg dose administered intravenously to patients. At assay end point, the levels of IFN-y were measured in the supernatants by Legendplex. A. Dose response curves of a representative donor. B. AUC values of N= 4 PBMCs donors. Mean + / - SEM.

[0099] Figure 19. Effects on IL-6 release of the combination of dexamethasone and the JAK (ruxolitinib) inhibitor or the mTOR (sirolimus) inhibitor at subclinical concentrations in comparison to dexamethasone concentrations corresponding to Cmax and Cmin clinical values of a 20 mg dose administered intravenously to patients. At assay end point, the levels of IL-6 were measured in the supernatants by Legendplex. A. Dose response curves of a representative donor. B. AUC values of N= 4 PBMCs donors. Mean + / - SEM.

[0100] Figure 20. Pre-activation assay set-up to assess the effect of the mitigation strategies on preactivated PBMCs. PBMCs were co-cultured with Toledo tumor cells (Effector : Tumor cells ratio = 5: 1) with escalating doses of CD20-TCB (glofitamab) for 15 to 18 hrs as a preactivation step (in the absence of any mitigations). At 18 hrs, 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib), 10 nM dexamethasone + 10 nM mTOR inhibitor (sirolimus) or 305 nM dexamethasone (corresponding to Cmax concentrations after administering 20mg I V. in patients) were added to the assay supernatant to stop further cytokine release from pre-activated PBMCs. Tumor cell killing and immune cell phenotyping were performed by flow cytometry and cytokine levels were measured in the assay supernatant by Legendplex before (18 hrs) and after addition (40 hrs) of the different mitigations.

[0101] Figure 21. Effects of the different mitigation strategies on tumor cell killing by pre-activated PBMCs. At assay end point (15-18 hrs and 40 hrs), the killing of CTV-labeled Toledo cells was measured by flow cytometry using a Live / dead NIR stain. A. Dose response killing curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0102] Figure 22. Effects of the different mitigation strategies on T cell activation by pre-activated PBMCs. At assay end point (15-18 hrs and 40 hrs), the expression of CD69 on CD8+ T cells was measured by flow cytometry. A. Dose response curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0103] Figure 23. Effects of the different mitigation strategies on T cell activation by pre-activated

[0104] PBMCs. At assay end point (15-18 hrs and 40 hrs), the expression of CD25 on CD8+ T cells was measured by flow cytometry. A. Dose response curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0105] Figure 24. Effects of the different mitigation strategies on T cell activation by pre-activated PBMCs. At assay end point (15-18 hrs and 40 hrs), the expression of CD69 on CD4+ T cells was measured by flow cytometry. A. Dose response curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0106] Figure 25. Effects of the different mitigation strategies on T cell activation by pre-activated PBMCs. At assay end point (15-18 hrs and 40 hrs), the expression of CD25 on CD4+ T cells was measured by flow cytometry. A. Dose response curves of a representative donor. B. EC50 values of N = 4 PBMCs donors. Mean + / - SEM.

[0107] Figure 26. IL-6 release before and after addition of the mitigation strategies by pre-activated PBMCs. At assay end point, the levels of IL-6 were measured in the supernatants by Legendplex. Dose response curves of a representative donor.

[0108] Figure 27. Restimulation assay set-up. PBMCs were co-cultured with CTV-labeled Toledo tumor cells (Effector : Tumor cells ratio = 5: 1) with escalating doses of CD20-TCB (glofitamab) in the presence and absence of 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib) or 10 nM dexamethasone + 10 nM mTOR inhibitor (sirolimus) or 305 nM dexamethasone (corresponding to Cmax concentrations after administering 20 mg I V. in patients) for 72 hrs. At 72 hrs, activated PBMCs were re-stimulated with escalating doses of CD20-TCB (glofitamab) on fresh CFSE- labeled Toledo cells in the absence of any mitigations. Before the restimulation (72 hrs) and after the restimulation (90 hrs), tumor cell killing was measured by flow cytometry and cytokine levels were measured in the supernatant by Legendplex to assess the lasting effect of the different combinations at subclinical concentrations in contrast to the clinical doses of dexamethasone.

[0109] Figure 28. Tumor cell killing after the first (in the presence of mitigation strategies) and second stimulation (in the absence of mitigation strategies). At 40 hrs and 72 hrs respectively, the killing of CTV-labeled Toledo cells (used in the first stimulation) and CFSE-labeled Toledo cells (used in the second stimulation) was measured by flow cytometry using a Live / dead NIR stain. Dose response killing curves of one donor. Figure 29. Level of IL-6 after the first (in the presence of mitigation strategies) and second stimulation (in the absence of mitigation strategies). At 40 hrs and 72 hrs, the level of IL-6 was measured in the assay supernatant by Legendplex. Dose response curves of one donor.

[0110] Figure 30. Assay set-up. Cytokine-enriched supernatants from co-culture of PBMCs and Toledo cells with 100 pM CD20-TCB (glofitamab) in the presence and absence of 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib), 10 nM dexamethasone + 10 nM mTOR inhibitor (sirolimus) and 305 nM dexamethasone (corresponding to Cmax concentrations after administering 20mg I V. in patients) were incubated on HUVECs for 5 hrs to assess the effects of endothelial cell activation.

[0111] Figure 31. Effects of the combination of subclinical doses of dexamethasone and JAK (ruxolitinib) or mTOR (sirolimus) inhibitor on endothelial cell activation. A. ICAM and B. VCAM on HUVECs were measured by flow cytometry 5 hours after incubation with cytokine-enriched supernatants. Means of technical duplicates + / -SEM.

[0112] Figure 32. In vivo study design to evaluate the effects of dexamethasone combination with JAK (ruxolitinib) or mTOR (sirolimus) inhibitors on CD20-TCB-mediated cytokine release in WSU (DLBCL)-bearing humanized BRGS-47 mice treated with CD20-TCB.

[0113] Figure 33. Effects of dexamethasone combination with JAK (ruxolitinib) or mTOR (sirolimus) inhibitors on CD20-TCB-mediated IFN-y release in WSU (DLBCL)-bearing humanized BRGS- 47 mice treated with 0.15 mg / kg CD20-TCB. Cytokines were measured by Luminex in the serum collected 4 hrs and 24 hrs post CD20-TCB treatment. Mean of N=7 mices + / - SEM. Ordinary oneway ANOVA, Dunnett’s multiple comparisons with *, P <0.05; **, P<0.01; ***, P < 0.001; ****, PO.OOOL

[0114] Figure 34. Effects of dexamethasone combination with JAK (ruxolitinib) or mTOR (sirolimus) inhibitors on CD20-TCB-mediated TNF-a release in WSU (DLBCL)-bearing humanized BRGS- 47 mice treated with 0.15 mg / kg CD20-TCB. Cytokines were measured by Luminex in the serum collected 4 hrs and 24 hrs post CD20-TCB treatment. Mean of N=7 mices + / - SEM. Ordinary oneway ANOVA, Dunnett’s multiple comparisons with *, P <0.05; **, P<0.01; ***, P < 0.001; ****, PO.OOOl.

[0115] Figure 35. Effects of dexamethasone combination with JAK (ruxolitinib) or mTOR (sirolimus) inhibitors on CD20-TCB-mediated IL-2 release in WSU (DLBCL)-bearing humanized BRGS-47 mice treated with 0.15 mg / kg CD20-TCB. Cytokines were measured by Luminex in the serum collected 4 hrs and 24 hrs post CD20-TCB treatment. Mean of N=7 mices + / - SEM. Ordinary oneway ANOVA, Dunnett’s multiple comparisons with *, P <0.05; **, P<0.01; ***, P < 0.001; ****, PO.OOOL

[0116] Figure 36. Effects of dexamethasone combination with JAK (ruxolitinib) or mTOR (sirolimus) inhibitors on CD20-TCB-mediated IL-6 release in WSU (DLBCL)-bearing humanized BRGS-47 mice treated with 0.15 mg / kg CD20-TCB. Cytokines were measured by Luminex in the serum collected 4 hrs and 24 hrs post CD20-TCB treatment. Mean of N=7 mices + / - SEM. Ordinary oneway ANOVA, Dunnett’s multiple comparisons with *, P <0.05; **, P<0.01; ***, P < 0.001; ****, PO.OOOl.

[0117] Figure 37. Effects of dexamethasone combination with JAK (ruxolitinib) or mTOR (sirolimus) inhibitors on CD20-TCB-mediated MCP-1 (CCL2) release in WSU (DLBCL)-bearing humanized BRGS-47 mice treated with 0.15 mg / kg CD20-TCB. Cytokines were measured by Luminex in the serum collected 4 hrs and 24 hrs post CD20-TCB treatment. Mean of N=7 mices + / - SEM. Ordinary one-way ANOVA, Dunnett’s multiple comparisons with *, P <0.05; **, P<0.01; ***, P < 0.001; ****, P<0.0001.

[0118] Detailed description of the Invention

[0119] Pharmaceutical agents

[0120] In some aspects, the TKI is an inhibitor of JAK or mTOR. In some aspects, the TKI is an inhibitor of mTOR. In some aspects, the TKI is a small molecule TKI.

[0121] “TKI” stands for tyrosine kinase inhibitor. A TKI is a compound that binds to one or more tyrosine kinase and inhibits its signaling, e.g., through competing with ATP and / or the substrate for binding to the kinase or by causing a conformational change that affects activity of the kinase. Inhibition in this context need not be complete or 100%. Instead, inhibition includes partially inhibiting the signaling of a tyrosine kinase, e.g., by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 95%.

[0122] “mTOR” stands for mammalian target of rapamycin (also known as FK506-binding protein 12- rapamycin complex-associated protein 1 (FRAP1)), and is a serine / threonine-specific protein kinase that belongs to the family of phosphatidylinositol-3 kinase (PI3K) related kinases. It serves as core component of two distinct protein complexes, mTOR complex 1 (TORC1) and mTOR complex 2 (TORC2), which regulate different cellular processes. Human mTOR is described, e.g., in UniProt entry P42345 (version 218). An mTOR inhibitor is a compound that binds to mTOR and inhibits mTOR signaling. Inhibition in this context need not be complete or 100%. Instead, inhibition includes partially inhibiting the signaling of an mTOR, e.g., by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 95%. The most established inhibitors of mTOR are rapamycin (also known as sirolimus) and the so-called rapalogs, which are derivatives of rapamycin, including temsirolimus, everolimus and ridaforolimus. A second generation of mTOR inhibitors are ATP-competitive mTOR inhibitors, designed to compete with ATP in the catalytic site of mTOR.

[0123] Exemplary mTOR inhibitors that might be useful in the present invention are provided in Table 1 below.

[0124] Table 1. mTOR inhibitors.

[0125] In some aspects, the inhibitor mTOR is a small molecule mTOR inhibitor. In some aspects, the mTOR inhibitor is a derivative of rapamycin (also known as a rapalog).

[0126] In some aspects, the mTOR inhibitor is selected from the group consisting of sirolimus, temsirolimus, everolimus and ridaforolimus, particularly the group consisting of sirolimus, temsirolimus and everolimus.

[0127] In specific aspects, the mTOR inhibitor is sirolimus. In further specific aspects, the mTOR inhibitor is temsirolimus. In yet further specific aspects, the mTOR inhibitor is everolimus.

[0128] In some aspects, the TKI is a JAK inhibitor, e.g., a small molecule JAK inhibitor. “JAK” stands for Janus kinase and refers to a family of intracellular, non-receptor tyrosine kinases that transduce cytokine-mediated signals via the JAK / STAT pathway. JAKs possess two nearidentical phosphate-transferring domains, one exhibiting the kinase activity, and the other one negatively regulating the kinase activity of the first. The four JAK family members are JAK1, JAK2, JAK3 and TYK2 (tyrosine kinase 2). In particular aspects herein, JAK is JAK1 and / or JAK2 (JAK1 / 2). Human JAK1 and JAK2 are described, e.g., in UniProt entries P23458 (version 221) and P60674 (version 224), respectively. A JAK inhibitor (also sometimes referred to as a jakinib) is a compound that binds to and inhibits the signaling of one or more JAKs (JAK1, JAK2, JAK3, TYK2). Inhibition in this context need not be complete or 100%. Instead, inhibition includes partially inhibiting the signaling of a JAK, e.g., by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 95%.

[0129] Exemplary JAK inhibitors that might be useful in the present invention are provided in Table 2 below.

[0130] Table 2. JAK inhibitors.

[0131]

[0132]

[0133] In some aspects, the JAK inhibitor is a small molecule JAK inhibitor. In some aspects, the JAK inhibitor is a JAK1 and / or JAK2 (JAK1 / 2) inhibitor. In some aspects, the JAK inhibitor is selected from the group consisting of ruxolitinib, baricitinib, momelotinib, upadacitinib, filgotinib, abrocitinib, itacitinib, solcitinib, oclacitinib, fedratinib, gandotinib, lestaurtinib and pacritinib.

[0134] In particular aspects, the JAK inhibitor is a JAK1 and JAK2 inhibitor. In specific such aspects, the JAK inhibitor is selected from the group consisting of ruxolitinib, baricitinib and momelotinib.

[0135] In some aspects, the JAK inhibitor is a JAK1 inhibitor. In specific such aspects, the JAK inhibitor is selected from the group consisting of upadacitinib, filgotinib, abrocitinib, itacitinib, solcitinib and oclacitinib.

[0136] In some aspects, the JAK inhibitor is a JAK2 inhibitor. In specific such aspects, the JAK inhibitor is selected from the group consisting of fedratinib, gandotinib, lestaurtinib and pacritinib. In a particular such aspect, the JAK inhibitor is fedratinib.

[0137] In some aspects, the JAK inhibitor is a pan-JAK inhibitor. In specfic such aspects, the JAK inhibitor is tofacitinib or peficitinib, particularly tofacitinib. In particular aspects, the JAK inhibitor is ruxolitinib. In further particular aspects, the JAK inhibitor is baricitinib. In some aspects, the JAK inhibitor is tofacitinib. In some aspects, the JAK inhibitor is fedratinib.

[0138] In particular aspects, the TKI is selected from the group consisting of sirolimus, temsirolimus, everolimus and ruxolitinib. In further particular aspects, the TKI is selected from the group consisting of sirolimus, temsirolimus, everolimus, ruxolitinib and baricitinib. In further particular aspects, the TKI is selected from sirolimus and ruxolitinib.

[0139] “GC” stands for glucocorticoid. Glucocorticoids are steroid hormones that bind to the glucocorticoid receptor. They can exert, e.g., anti-inflammatory, anti-allergic and immune- suppressive effects and can be categorized into natural and synthetic types.

[0140] In some aspects, the GC is selected from the group consisting of cortisol, cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, deflazacort, fludrocortisone acetate, deoxycorticosterone acetate, fluprednisolone, fludrocortisone, meprednisone, hydrocortisone, aldosterone, paramethasone and beclomethasone. In some aspects, the GC is a synthetic glucocorticoid. In some aspects, the GC is dexamethasone or methylprednisolone. In some aspects, the GC is dexamethasone. In some aspects, the GC is dexamethasone and the TKI is an mTOR inhibitor.

[0141] Inhibition of effects of the T cell

[0142] In some aspects, administration of the TKI and GC causes inhibition of an effect of the T cell engaging agent. In some aspects, administration of the TKI and GC does not cause inhibition of another effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes inhibition of a first effect of the T cell engaging agent but does not cause inhibition of a second effect of the T cell engaging agent. In some of these aspects, said inhibition is a complete inhibition. In some of these aspects, said inhibition is a partial inhibition.

[0143] In some aspects, administration of the TKI and GC causes inhibition of a first effect of the T cell engaging agent and inhibition of a second effect of the T cell engaging agent, wherein said inhibition of the first effect is stronger than said inhibition of the second effect. In some aspects, administration of the TKI and GC causes inhibition of a first effect of the T cell engaging agent and inhibition of a second effect of the T cell engaging agent, wherein said inhibition of the first effect is a complete inhibition and said inhibition of the second effect is a partial inhibition. “Effect” of a T cell engaging agent refers to responses in an individual’s body caused by the T cell engaging agent. Such effect may include cellular response(s) of immune cells, such as T cells, particularly CD4+ and / or CD8+ T cells, such as proliferation, differentiation, activation, cytokine secretion, cytotoxic effector molecule release, cytotoxic activity, and expression of activation markers, and / or effects on target cells, particularly target cells (e.g. tumor cells) expressing the target cell antigen of the T cell engaging agent, such as lysis of target cells. Such effect may also, more generally, relate to affecting symptoms, such as fever, hypotension and hypoxia, and syndromes, such as CRS and ICANS, caused by the T cell engaging agent.

[0144] The “inhibition” of an effect, as referred to herein, may refer to the complete or the partial inhibition of the effect. For instance, partial inhibition may refer to the inhibition of such effect by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90% or at least 95%. In some aspects, partial inhibition refers to the inhibition of such effect by less than 100%, less than 95%, less than 90%, less than 80%, less than 70%, less than 60% or less than 50%. In some aspects, complete inhibition refers to the inhibition of such effect by at least 70%, at least 80%, at least 90%, at least 95% or 100%. In some aspects, the complete inhibition refers to an inhibition that is substantially complete, e.g., where the effect is no longer clinically meaningful. In some aspects, complete inhibition refers to 100% inhibition of the effect. In some aspects, inhibition refers to a clinically meaningful and / or statistically significant inhibition.

[0145] In some aspects, administration of the TKI and GC causes inhibition of an adverse effect related to the administration of the T cell engaging agent. In some aspects, administration of the TKI and GC does not cause inhibition of a desired effect related to the administration of the T cell engaging agent. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of an adverse effect related to the administration of the T cell engaging agent. In some aspects, administration of the TKI and GC does not cause a clinically meaningful and / or statistically significant inhibition of a desired effect related to the administration of the T cell engaging agent.

[0146] In some aspects, administration of the TKI and GC causes inhibition of an adverse effect related to the administration of the T cell engaging agent but does not cause inhibition of a desired effect related to the administration of the T cell engaging agent. In some aspects, administration of the TKI and GC causes inhibition of an adverse effect related to the administration of the T cell engaging agent and inhibition of a desired effect related to the administration of the T cell engaging agent, wherein said inhibition of the adverse effect is stronger than said inhibition of the desired effect. In some aspects, administration of the TKI and GC causes inhibition of an adverse effect related to the administration of the T cell engaging agent and inhibition of a desired effect related to the administration of the T cell engaging agent, wherein said inhibition of the adverse effect is a complete inhibition and said inhibition of the beneficial effect is a partial inhibition. In some aspects, administration of the TKI and GC causes inhibition of an adverse effect related to the administration of the T cell engaging agent and inhibition of a desired effect related to the administration of the T cell engaging agent, wherein said inhibition of the adverse effect is a clinically meaningful and / or statistically significant inhibition and said inhibition of the beneficial effect is not a clinically meaningful and / or statistically significant inhibition.

[0147] A “desired effect” is a beneficial and desired effect resulting from medication in the treatment of an individual, herein particularly with a T cell engaging agent, e.g., a therapeutic and / or prophylactic effect, such as, e.g., killing of tumor cells, reduction or retardation of tumor growth, reduction of tumor volume, reduction or prevention of tumor metastasis, increase of progression- free or overall survival, alleviation of disease symptoms, and the like.

[0148] An “adverse effect”, which is sometimes also denoted as “side effect” or “adverse event” (especially in clinical studies), is a harmful and undesired effect resulting from medication in the treatment of an individual, herein particularly with a T cell engaging agent.

[0149] The “inhibition” of an adverse effect may relate to both the prevention and / or the mitigation of the adverse effect. In this context, the term “prevention” refers to the complete or partial prevention or delay of an adverse effect in an individual that does not yet experience or display the adverse effect, i.e. before manifestation of the adverse effect, and the term “mitigation” refers to the complete or partial mitigation or alleviation of an adverse effect in an individual that already experiences or displays the adverse effect, i.e. upon or after manifestation of the adverse effect.

[0150] According to the invention, the adverse effect is related to the administration of the T cell engaging agent. In some aspects, the adverse effect is related to the first administration of the T cell engaging agent. In some aspects, the adverse effect occurs upon the first administration of the T cell engaging agent. In some aspects, the adverse effect occurs predominantly or only upon the first administration of the T cell engaging agent. In some aspects, the adverse effect occurs within 12 hours, 24 hours, 36 hours, 48 hours, 72 hours or 96 hours of the administration, particularly the first administration, of the T cell engaging agent. In some aspects, in particular wherein only a single administration of the T cell engaging is made (in the course of the treatment with the T cell engaging agent), the adverse effect occurs within 3 days, 4 days, 5 days, 6 days, 7 days, 10 days, 14 days or 21 days of the administration of the T cell engaging agent.

[0151] In some aspects, said adverse effect is cytokine secretion by immune cells, particularly T cells. Immune cells may include various immune cell types, such as T cells, macrophages, monocytes, NK cells, neutrophils and dendritic cells. In some aspects, said T cells are CD8+ T cells or CD4+ cells. In some aspects, said cytokine is one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-1 .

[0152] In some aspects, said adverse effect is cytokine release syndrome (CRS).

[0153] “Cytokine release syndrome” (abbreviated as “CRS”) refers to an increase in the levels of cytokines, such as TNF-a, IFN-y, IL-6, IL- 10 and others, in the blood of a subject during or shortly after (e.g. within 1 day of) administration of a therapeutic agent (e.g., a T cell engaging agent), resulting in adverse symptoms. CRS is an adverse reaction to a therapeutic agent and timely related to administration of the therapeutic agent. It typically occurs during or shortly after an administration of the therapeutic agent, i.e. typically within 24 hours after administration (typically infusion), predominantly at the first administration. In some instances, e.g. after the administration of CAR-T cells, CRS can also occur only later, e.g. several days after administration upon expansion of the CAR-T cells. The incidence and severity typically decrease with subsequent administrations. Symptoms may range from symptomatic discomfort to fatal events, and may include fever, chills, dizziness, hypertension, hypotension, hypoxia, dyspnea, restlessness, sweating, flushing, skin rash, tachycardia, tachypnoea, headache, tumour pain, nausea, vomiting and / or organ failure. CRS may be graded, e.g., based on the ASTCT (American Society for Transplantation and Cellular Therapy) grading system (DW Lee et al., Biol Blood Marrow Transplant. 2019 Apr;25(4):625-638).

[0154] In some aspects, said adverse effect is one or more of fever, hypotension and hypoxia.

[0155] In some aspects, said adverse effect is an elevated serum level of one of more cytokine. For instance, the serum level may be elevated as compared to the serum level in a healthy individual, and / or the serum level in an individual (including the same individual) without administration of the T cell engaging agent. In some aspects, said one or more cytokine is selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-10. In some aspects, said adverse effect is immune effector cell-associated neurotoxicity syndrome (ICANS).

[0156] “Immune effector cell-associated neurotoxicity syndrome” (abbreviated as “ICANS”) refers to a neurological condition that can occur as a side effect after administration of a therapeutic agent (e.g., a T cell engaging agent). Symptoms vary in severity and may include mild tremor, confusion, disorientation, headache, attention deficits, agitation, seizures, cerebral oedema, hesitancy of speech, deterioration in handwriting, aphasia with expressive and / or receptive components, status epilepticus, fatal cerebral oedema, intracerebral haemorrhage and transient coma. ICANS may be graded, e.g., based on the ASTCT (American Society for Transplantation and Cellular Therapy) grading system (DW Lee et al., Biol Blood Marrow Transplant. 2019 Apr;25(4):625-638).

[0157] In some aspects, said adverse effect comprises one or more of thrombocytopenia, an elevated serum level, e.g., as compared to the serum level in a healthy individual, and / or the serum level in an individual (including the same individual) without administration of the T cell engaging agent, of one or more of lactate dehydrogenase, ferritin, IFN-y, IL-10, granzyme B, GM-CSF, MIP-la, TNF, IL-2, and an elevated level, e.g., as compared to the serum level in a healthy individual, and / or the serum level in an individual (including the same individual) without administration of the T cell engaging agent, in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL-10, IL-6 and granzyme B.

[0158] In some aspects, said adverse effect is T cell exhaustion.

[0159] “T cell exhaustion” refers to a state of T cell dysfunction, e.g., arising during chronic infections or cancer. It is characterized by a progressive loss of effector functions, such as cytokine production and cytotoxic activity, along with sustained expression of inhibitory receptors like PD-1, CTLA- 4, TIM-3, and LAG-3. In some aspects, the inhibition of T cell exhaustion refers to one or more of the following in the T cells: increased production and / or secretion of cytokines, in particular, one or more cytokine of TNF-a, IFN-y, and IL-2, decreased expression of inhibitory receptors, in particular one or more inhibitory receptor of PD-1, CTLA-4, TIM-3, and LAG-3, increased cytotoxicity, increased proliferation, and increased memory cell formation. In each case, the respective markers are increased or decreased in comparison, e.g., to a state where no treatment occurred with the TKI and GC combination.

[0160] In some aspects, said desired effect is the activation of T cells. “Activation of T cells” or “T cell activation” as used herein refers to one or more cellular response of a T lymphocyte, particularly a CD4+ or CD8+ T cell, selected from: proliferation, differentiation, cytotoxic effector molecule release, cytotoxic activity, and expression of activation markers. Suitable assays to measure T cell activation are known in the art and described herein. In particular aspects, T cell activation is the expression of activation markers, particularly expression of CD25 and / or CD69, e.g., as measured by flow cytometry. In particular aspects, T cell activation is determined by measuring expression of CD25 and / or CD69 on the T cell, e.g. by flow cytometry.

[0161] In some aspects, said desired effect is the cytotoxic activity of T cells.

[0162] “Cytotoxic activity” of a T cell refers to the induction of lysis (i.e. killing) of target cells by a T lymphocyte, particularly a CD4+ or CD8+ T cell. Cytotoxic activity typically involves degranulation of the T lymphocyte, associated with the release of cytotoxic effector molecules such as granzyme B and / or perforin from the T lymphocyte.

[0163] In some aspects, said desired effect is the therapeutic effect of the T cell engaging agent.

[0164] The “therapeutic effect” of a T cell engaging agent refers to beneficial or desired results upon the treatment of an individual with the T cell engaging agent, including preventing occurrence or recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, in case of a tumor preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, remission or improved prognosis, or other beneficial results.

[0165] In some aspects, administration of the TKI and GC causes inhibition of cytokine secretion by immune cells, particularly by T cells. In some aspects, administration of the TKI and GC causes inhibition of cytokine secretion by immune cells, particularly by T cells, but does not cause inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of cytokine secretion by immune cells, particularly by T cells, but does not cause a clinically meaningful and / or statistically significant inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC inhibits cytokine secretion by immune cells, particularly by T cells, more strongly than the activation and / or the cytotoxic activity of T cells. For instance, cytokine secretion may be reduced by 50%, whereas activation and / or cytotoxic activity of the T cells is reduced by only 20%. In some aspects, administration of the TKI and GC causes inhibition of cytokine secretion by immune cells, particularly by T cells, and inhibition of the activation and / or the cytotoxic activity of T cells, wherein said inhibition of cytokine secretion is a complete inhibition and said inhibition of activation and / or cytotoxic activity of T cells is a partial inhibition.

[0166] In some aspects, administration of the TKI and GC causes inhibition of cytokine secretion by immune cells, particularly by T cells, and inhibition of the activation and / or the cytotoxic activity of T cells.

[0167] In some aspects, administration of the TKI and GC causes inhibition of cytokine secretion by immune cells, particularly by T cells, but does not cause inhibition of the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of cytokine secretion by immune cells, particularly by T cells, but does not cause a clinically meaningful and / or statistically significant inhibition of the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC inhibits cytokine secretion by immune cells, particularly by T cells, more strongly than the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes inhibition of cytokine secretion by immune cells, particularly by T cells, and inhibition of the therapeutic effect of the T cell engaging agent, wherein said inhibition of cytokine secretion is a complete inhibition and said inhibition of the therapeutic effect is a partial inhibition.

[0168] In some aspects, administration of the TKI and GC causes inhibition of cytokine secretion by immune cells, particularly by T cells, and inhibition of the therapeutic effect of the T cell engaging agent.

[0169] In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level, i.e. reduction of the serum level, of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-ip. In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level, i.e. reduction of the serum level, of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-ip, but does not cause inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of the elevated serum level of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-ip, but does not cause a clinically meaningful and / or statistically significant inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC inhibits the elevated serum level of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-ip, more strongly than the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-ip, and inhibition of the activation and / or the cytotoxic activity of T cells, wherein said inhibition of the elevated serum level of one or more cytokine is a complete inhibition and said inhibition of activation and / or cytotoxic activity of T cells is a partial inhibition.

[0170] In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-ip, and inhibition of the activation and / or the cytotoxic activity of T cells.

[0171] In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level, i.e. reduction of the serum level, of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-ip, but does not cause inhibition of the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of the elevated serum level of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-ip, but does not cause a clinically meaningful and / or statistically significant inhibition of the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC inhibits the elevated serum level of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-ip, more strongly than the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-ip, and inhibition of the therapeutic effect of the T cell engaging agent, wherein said inhibition of the elevated serum level of one of more cytokine is a complete inhibition and said inhibition of the therapeutic effect is a partial inhibition. In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL- 10, TNF-a, GM-CSF, MCP-1 and IL-ip, and inhibition of the therapeutic effect of the T cell engaging agent.

[0172] In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level, i.e. reduction of the serum level, of one of more of lactate dehydrogenase, ferritin, IFN-y, IL-10, granzyme B, GM-CSF, MIP-la, TNF, IL-2 and / or it causes inhibition of the elevated level, i.e. reduction of the level, in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SI 00b, glial fibrillary acidic protein, protein, IFN-y, IL- 10, IL-6 and granzyme B. In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level, i.e. reduction of the serum level, of one of more of lactate dehydrogenase, ferritin, IFN-y, IL- 10, granzyme B, GM-CSF, MIP-la, TNF, IL-2 and / or it causes inhibition of the elevated level, i.e. reduction of the level, in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL-10, IL-6 and granzyme B, but does not cause inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of the elevated serum level of one of more of lactate dehydrogenase, ferritin, IFN-y, IL- 10, granzyme B, GM-CSF, MIP-la, TNF, IL-2 and / or it causes a clinically meaningful and / or statistically significant inhibition of the elevated level in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL- 10, IL-6 and granzyme B, but does not cause a clinically meaningful and / or statistically significant inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC inhibits the elevated serum level of one of more of lactate dehydrogenase, ferritin, IFN-y, IL-10, granzyme B, GM-CSF, MIP-la, TNF, IL-2 and / or the elevated level in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL-10, IL-6 and granzyme B, more strongly than the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level of one of more of lactate dehydrogenase, ferritin, IFN-y, IL- 10, granzyme B, GM-CSF, MIP-la, TNF, IL-2 and / or it causes inhibition of the elevated level in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL-10, IL-6 and granzyme B, and inhibition of the activation and / or the cytotoxic activity of T cells, wherein said inhibition of the elevated level in the serum and / or in the cerebrospinal fluid is a complete inhibition and said inhibition of activation and / or cytotoxic activity of T cells is a partial inhibition.

[0173] In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level of one of more of lactate dehydrogenase, ferritin, IFN-y, IL-10, granzyme B, GM-CSF, MIP-la, TNF, IL-2 and / or it causes inhibition of the elevated level in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL- 10, IL-6 and granzyme B, and further causes inhibition of the activation and / or the cytotoxic activity of T cells.

[0174] In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level, i.e. reduction of the serum level, of one of more of lactate dehydrogenase, ferritin, IFN-y, IL-10, granzyme B, GM-CSF, MIP-la, TNF, IL-2 and / or it causes inhibition of the elevated level, i.e. reduction of the level, in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL-10, IL-6 and granzyme B, but does not cause inhibition of the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of the elevated serum level of one of more of lactate dehydrogenase, ferritin, IFN-y, IL- 10, granzyme B, GM-CSF, MIP-la, TNF, IL-2 and / or it causes a clinically meaningful and / or statistically significant inhibition of the elevated level in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL- 10, IL-6 and granzyme B, but does not cause a clinically meaningful and / or statistically significant inhibition of the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC inhibits the elevated serum level of one of more of lactate dehydrogenase, ferritin, IFN-y, IL-10, granzyme B, GM-CSF, MIP-la, TNF, IL-2 and / or the elevated level in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL- 10, IL-6 and granzyme B, more strongly than the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level of one of more of lactate dehydrogenase, ferritin, IFN-y, IL- 10, granzyme B, GM-CSF, MIP-la, TNF, IL-2 and / or it causes inhibition of the elevated level in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL-10, IL-6 and granzyme B, and inhibition of the therapeutic effect of the T cell engaging agent, wherein said inhibition of the elevated level in the serum and / or in the cerebrospinal fluid is a complete inhibition and said inhibition of the therapeutic effect is a partial inhibition.

[0175] In some aspects, administration of the TKI and GC causes inhibition of the elevated serum level of one of more of lactate dehydrogenase, ferritin, JFN-y, IL-10, granzyme B, GM-CSF, MIP-la, TNF, IL-2 and / or it causes inhibition of the elevated level in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL- 10, IL-6 and granzyme B, and further causes inhibition of the therapeutic effect of the T cell engaging agent.

[0176] In some aspects, administration of the TKI and GC causes inhibition of T cell exhaustion. In some aspects, administration of the TKI and GC causes inhibition of T cell exhaustion, but does not cause inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of T cell exhaustion, but does not cause a clinically meaningful and / or statistically significant inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC inhibits T cell exhaustion more strongly than the activation and / or cytotoxic activity of T cells. In some aspects, administration of the TKI and GC causes inhibition of T cell exhaustion and inhibition of the activation and / or the cytotoxic activity of T cells, wherein said inhibition of T cell exhaustion is a complete inhibition and said inhibition of activation and / or cytotoxic activity of T cells is a partial inhibition.

[0177] In some aspects, administration of the TKI and GC causes inhibition of T cell exhaustion and inhibition of the activation and / or the cytotoxic activity of T cells.

[0178] In some aspects, administration of the TKI and GC causes inhibition of T cell exhaustion, but does not cause inhibition of the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of T cell exhaustion, but does not cause a clinically meaningful and / or statistically significant inhibition of the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC inhibits T cell exhaustion more strongly than the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes inhibition of T cell exhaustion and inhibition of the therapeutic effect of the T cell engaging agent, wherein said inhibition of T cell exhaustion is a complete inhibition and said inhibition of the therapeutic effect of the T cell engaging agent is a partial inhibition. In some aspects, administration of the TKI and GC causes inhibition of T cell exhaustion and inhibition of the therapeutic effect of the T cell engaging agent.

[0179] In some aspects, administration of the TKI and GC causes inhibition of CRS. In some aspects, administration of the TKI and GC causes inhibition of CRS, but does not cause inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of CRS, but does not cause a clinically meaningful and / or statistically significant inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC inhibits CRS more strongly than the activation and / or cytotoxic activity of T cells. In some aspects, administration of the TKI and GC causes inhibition of CRS and inhibition of the activation and / or the cytotoxic activity of T cells, wherein said inhibition of CRS is a complete inhibition and said inhibition of activation and / or cytotoxic activity of T cells is a partial inhibition.

[0180] In some aspects, administration of the TKI and GC causes inhibition of CRS and inhibition of the activation and / or the cytotoxic activity of T cells.

[0181] In some aspects, administration of the TKI and GC causes inhibition of CRS, but does not cause inhibition of the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of CRS, but does not cause a clinically meaningful and / or statistically significant inhibition of the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC inhibits CRS more strongly than the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes inhibition of CRS and inhibition of the therapeutic effect of the T cell engaging agent, wherein said inhibition of CRS is a complete inhibition and said inhibition of the therapeutic effect of the T cell engaging agent is a partial inhibition.

[0182] In some aspects, administration of the TKI and GC causes inhibition of CRS and inhibition of the therapeutic effect of the T cell engaging agent.

[0183] In some aspects, administration of the TKI and GC causes inhibition of ICANS. In some aspects, administration of the TKI and GC causes inhibition of ICANS, but does not cause inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of ICANS, but does not cause a clinically meaningful and / or statistically significant inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC inhibits ICANS more strongly than the activation and / or cytotoxic activity of T cells. In some aspects, administration of the TKI and GC causes inhibition of ICANS and inhibition of the activation and / or the cytotoxic activity of T cells, wherein said inhibition of ICANS is a complete inhibition and said inhibition of activation and / or cytotoxic activity of T cells is a partial inhibition.

[0184] In some aspects, administration of the TKI and GC causes inhibition of ICANS and inhibition of the activation and / or the cytotoxic activity of T cells.

[0185] In some aspects, administration of the TKI and GC causes inhibition of ICANS, but does not cause inhibition of the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes a clinically meaningful and / or statistically significant inhibition of ICANS, but does not cause a clinically meaningful and / or statistically significant inhibition of the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC inhibits ICANS more strongly than the therapeutic effect of the T cell engaging agent. In some aspects, administration of the TKI and GC causes inhibition of ICANS and inhibition of the therapeutic effect of the T cell engaging agent, wherein said inhibition of ICANS is a complete inhibition and said inhibition of the therapeutic effect of the T cell engaging agent is a partial inhibition.

[0186] In some aspects, administration of the TKI and GC causes inhibition of ICANS and inhibition of the therapeutic effect of the T cell engaging agent.

[0187] In some aspects, administration of the TKI and GC does not cause inhibition of the activation and / or the cytotoxic activity of T cells. In some aspects, administration of the TKI and GC does not cause inhibition of the therapeutic effect of the T cell engaging agent.

[0188] Administration regimens

[0189] In some aspects, the inhibition of the adverse effect, e.g., the reduction of cytokine secretion or the serum level of one or more cytokine, is sustained after the TKI and GC has not been administered for a given amount of time. In some aspects, said amount of time is about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 12 hours, 16 hours, 20 hours, 24 hours, 36 hours, 48 hours, 72 hours, or 96 hours. In some aspects, said inhibition is sustained after a subsequent administration of the T cell engaging agent. Particularly, said inhibition may be sustained even after administration of the TKI and GC is stopped / no further administration of the TKI and GC is made. In some aspects, the TKI and GC are administered initially around the time that the T cell engaging agent is initially administered and the ensuing inhibition of the adverse effect is sustained beyond a subsequent administration of the T cell engaging agent that occurs without further administration of the TKI and GC.

[0190] In some aspects, administration of the TKI and GC is upon and / or after manifestation, in particular upon and / or after the clinical manifestation, of the adverse effect. In some aspects, administration of the TKI and GC is in response to the manifestation, in particular the clinical manifestation, of the adverse effect. Said administration may be, for example, within about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 12 hours, 16 hours, 20 hours or 24 hours after manifestation of the adverse effect, e.g., the occurrence of clinical symptoms of the adverse effect, such as fever.

[0191] In some aspects, administration of the TKI and GC is before the administration of the T cell engaging agent. In some aspects, administration of the inhibitor of the TKI and GC is concurrent with the administration of the T cell engaging agent. In some aspects, administration of the TKI and GC is after the administration of the T cell engaging agent. Where administration of the TKI and GC is before or after the administration of the T cell engaging agent, such administration of the TKI and GC may be, for example, within about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 12 hours, 16 hours, 20 hours or 24 hours before or after, respectively, the administration of the T cell engaging agent. In some aspects, administration of the TKI and GC is before manifestation, in particular before the clinical manifestation, of the adverse effect. Administration of the TKI and GC may be intermittently or continuously. In some aspects, administration of the TKI and GC is oral. In some aspects, administration of the TKI and GC is parenteral, particularly intravenous.

[0192] In some aspects, administration of the TKI and GC is at a dose sufficient to cause inhibition of an activity of the T cell engaging agent, in particular an adverse effect of the T cell engaging agent. In some aspects, administration of the TKI and GC is at a dose insufficient to cause inhibition of another activity of the T cell engaging agent, in particular a desired effect of the T cell engaging agent. In some aspects, administration of the TKI and GC is at a dose sufficient to cause inhibition of a first activity of the T cell engaging agent but insufficient to cause inhibition of a second activity of the T cell engaging agent. In some aspects, administration of the TKI and GC is at a dose sufficient to cause inhibition of an adverse effect of the T cell engaging agent but insufficient to cause inhibition of a desired effect of the T cell engaging agent. In some of these aspects, said inhibitions are complete inhibitions. In some of these aspects, said inhibitions are clinically meaningful and / or statistically significant.

[0193] In some aspects, administration of the inhibitor of the TKI and GC is at a dose sufficient to cause inhibition of cytokine secretion by immune cells, particularly T cells. In some of these aspects, said inhibition is a clinically meaningful and / or statistically significant inhibition.

[0194] In some aspects, administration of the inhibitor of the TKI and GC is at a dose sufficient to cause inhibition of the elevated serum level, i.e. to reduce the serum level, of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF- a, GM-CSF, MCP-1 and IL-ip. In some of these aspects, said inhibition is a clinically meaningful and / or statistically significant inhibition.

[0195] In some aspects, administration of the inhibitor of the TKI and GC is at a dose sufficient to cause inhibition of the elevated serum level, i.e. to reduce the serum level, of one of more of lactate dehydrogenase, ferritin, IFN-y, IL-10, granzyme B, GM-CSF, MIP-la, TNF, IL-2. In some of these aspects, said inhibition is a clinically meaningful and / or statistically significant inhibition.

[0196] In some aspects, administration of the inhibitor of the TKI and GC is at a dose sufficient to cause inhibition of the elevated level, i.e. to reduce the level, in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL- 10, IL-6 and granzyme B. In some of these aspects, said inhibition is a clinically meaningful and / or statistically significant inhibition.

[0197] In some aspects, administration of the TKI and GC is at a dose insufficient to cause inhibition of the activation of T cells. In some of these aspects, said inhibition is a clinically meaningful and / or statistically significant inhibition.

[0198] In some aspects, administration of the TKI and GC is at a dose insufficient to cause inhibition of the cytotoxic activity of T cells. In some of these aspects, said inhibition is a clinically meaningful and / or statistically significant inhibition.

[0199] In some aspects, administration of the TKI and GC is at a dose sufficient to causes inhibition of cytokine secretion by T cells, but insufficient to cause inhibition of the activation and / or the cytotoxic activity of T cells. In some of these aspects, said inhibitions are clinically meaningful and / or statistically significant. In some aspects, administration of the TKI and GC is at a dose sufficient to causes inhibition of the elevated serum level, i.e. reduces the serum level, of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP- 1 and IL-ip, but insufficient to cause inhibition of the activation and / or the cytotoxic activity of T cells. In some of these aspects, said inhibitions are clinically meaningful and / or statistically significant.

[0200] In some aspects, administration of the TKI and GC is at a dose sufficient to causes inhibition of the elevated serum level, i.e. reduces the serum level, of one of more of lactate dehydrogenase, ferritin, IFN-y, IL-10, granzyme B, GM-CSF, MIP-la, TNF, IL-2, but insufficient to cause inhibition of the activation and / or the cytotoxic activity of T cells. In some of these aspects, said inhibitions are clinically meaningful and / or statistically significant.

[0201] In some aspects, administration of the TKI and GC is at a dose sufficient to causes inhibition of the elevated level, i.e. to reduce the level, in the cerebrospinal fluid of one or more of white blood cells, T cells, CD 14+ cells, SI 00b, glial fibrillary acidic protein, protein, IFN-y, IL- 10, IL-6 and granzyme B, but insufficient to cause inhibition of the activation and / or the cytotoxic activity of T cells. In some of these aspects, said inhibitions are clinically meaningful and / or statistically significant.

[0202] In some aspects, administration of the TKI and GC is at an effective dose.

[0203] An “effective amount” or “effective dose” of an agent, e.g., the TKI, GC or T cell engaging agent, refers to an amount effective, at dosages and for periods of time necessary, to achieve the desired therapeutic and / or prophylactic result.

[0204] In some aspects, the administration of the TKI and GC is at a dose equaling a dose strength available for the TKI and GC. Typically, several dose strengths (i.e. dosage forms such as tablets or capsules with a specific amount of active ingredient) are available for a given TKI and GC. For example, if the TKI is everolimus, it may be administered at a dose of 2.5 mg, 5 mg, 7.5 mg or 10 mg (administration preferably being oral administration). For example, if the TKI is sirolimus, it may be administered at a dose of 0.5 mg, 1 mg or 2 mg (administration preferably being oral administration). For example, if the TKI is ruxolitinib, it may be administered at a dose of 5 mg, 10 mg, 15 mg, 20 mg or 25 mg (administration preferably being oral administration). For example, if the TKI is temsirolimus, it may be administered for example at a dose of 12.5 mg or 25 mg (administration preferably being intravenous administration, particularly using a solution of 25mg / ml active ingredient). For example, if the GC is dexamethasone, it may be administered for example at a dose of 0.5 mg, 0.75 mg, 1 mg, 2 mg, 4 mg, 6 mg, 9 mg, 10 mg, 12 mg, 20 mg or 30 mg (administration preferably being oral, intravenous, subcutaneous or intramuscular administration). For example, if the GC is methylprednisolone, it may be administered for example at a dose of 2 mg, 4 mg, 8 mg, 12 mg, 16 mg, 20 mg, 24 mg, 28 mg, 32 mg, 36 mg, 40 mg, 44 mg or 48 mg (administration preferably being oral, intravenous or intramuscular administration).

[0205] In some aspects, the administration of the TKI and GC is at a lower dose than currently approved or recommended for these drugs for a given indication. For example, if the TKI is everolimus, it may be administered at a dose of below 2.5 mg, below 5 mg, below 7.5 mg or below 10 mg (administration preferably being oral administration). For example, if the TKI is sirolimus, it may be administered at a dose of below 0.5 mg, below 1 mg or below 2 mg (administration preferably being oral administration). For example, if the TKI is ruxolitinib, it may be administered at a dose of below 5 mg, below 10 mg, below 15 mg, below 20 mg or below 25 mg (administration preferably being oral administration). For example, if the TKI is temsirolimus, it may be administered for example at a dose of below 12.5 mg or below 25 mg (administration preferably being intravenous administration, particularly using a solution of 25mg / ml active ingredient). For example, if the GC is dexamethasone, it may be administered for example at a dose of below 0.5 mg, below 0.75 mg, below 1 mg, below 2 mg, below 4 mg, below 6 mg, below 9 mg, below 10 mg, below 12 mg, below 20 mg or below 30 mg (administration preferably being oral, intravenous, subcutaneous or intramuscular administration). For example, if the GC is methylprednisolone, it may be administered for example at a dose of below 2 mg, below 4 mg, below 8 mg, below 12 mg, below 16 mg, below 20 mg, below 24 mg, below 28 mg, below 32 mg, below 36 mg, below 40 mg, below 44 mg or below 48 mg (administration preferably being oral, intravenous or intramuscular administration).

[0206] In some aspects, any of the above doses are for treating an adult.

[0207] In some aspects, administration of the TKI and / or GC is daily. In some aspects, administration of the TKI and / or GC is once daily. In some aspects, administration of the TKI and / or GC is once daily at a dose as mentioned hereinabove. In some aspects, administration of the TKI and / or GC is for the period of time during which the adverse effect persists (i.e. administration of the TKI and / or GC is from manifestation of the adverse effect until reduction or disappearance of the adverse effect). In some aspects, administration of the TKI and / or GC is stopped after the adverse effect is inhibited. In some aspects, administration of the TKI and / or GC is stopped after reduction or disappearance of the adverse effect, e.g., after its clinically meaningful and / or statistically significant reduction. In some aspects, administration of the TKI and / or GC is once, twice, three times, four times, five times, six times, seven times, eight times, nine times or ten times in the course of the treatment of the individual with the T cell engaging agent. In some aspects, administration of the TKI and / or GC is for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days. In some aspects, administration of the TKI and / or GC is once daily for 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days or 10 days. The administration of the TKI and / or GC is generally associated with the administration of the T cell engaging agent. In some aspects, administration of the TKI and GC is associated with the first administration of the T cell engaging agent. Said first administration is particularly the first administration of the T cell engaging agent in the course of the treatment of the individual with the T cell engaging agent. In some aspects, administration of the TKI and GC is concurrent with the first administration of the T cell engaging agent. In some aspects, administration of the TKI and GC is prior to the first administration of the T cell engaging agent. In some aspects, administration of the TKI and GC is subsequent to the first administration of the T cell engaging agent. In some aspects, administration of the TKI and GC is subsequent to the first administration of the T cell engaging agent and prior to a second administration of the T cell engaging agent. Where administration of the TKI and GC is prior or subsequent to the (first) administration of the T cell engaging agent, such administration of the TKI and GC may be, for example, within about 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 12 hours, 16 hours, 20 hours, 24 hours, 48 hours or 72 hours before or after, respectively, the administration of the T cell engaging agent.

[0208] In some aspects, the administration of the T cell engaging agent is for a longer period of time than the administration of the TKI and GC. In some aspects, the administration of the T cell engaging agent continues after the administration of the TKI and GC is stopped. In some aspects, the administration of the T cell engaging agent is a single administration or a repeated administration. In the course of the treatment of the individual with the T cell engaging agent, the T cell engaging agent may be administered once or several times. For example, treatment of the individual with the T cell engaging agent may comprise multiple treatment cycles which each comprise one or more administrations of the T cell engaging agent. In some aspects, the administration of the T cell engaging agent comprises a first and a second administration. For use in the present invention, the T cell engaging agent would be formulated, dosed, and administered in a fashion consistent with good medical practice. Factors for consideration in this context include the particular disorder being treated, the particular mammal being treated, the clinical condition of the individual patient, the cause of the disorder, the site of delivery of the agent, the method of administration, the scheduling of administration, and other factors known to medical practitioners.

[0209] In some aspects, the administration of the T cell engaging agent is at an effective dose. For systemic administration, an effective dose can be estimated initially from in vitro assays, such as cell culture assays. A dose can then be formulated in animal models to achieve a circulating concentration range that includes the IC50 as determined in cell culture. Such information can be used to more accurately determine useful doses in humans. Initial dosages can also be estimated from in vivo data, e.g., animal models, using techniques that are well known in the art. Dosage amount and interval may be adjusted individually to provide plasma levels of the T cell engaging agent which are sufficient to maintain therapeutic effect. Usual patient dosages for administration by injection range from about 0.1 to 50 mg / kg / day, typically from about 0.5 to 1 mg / kg / day. Therapeutically effective plasma levels may be achieved by administering multiple doses each day. Levels in plasma may be measured, for example, by HPLC.

[0210] An effective amount of the T cell engaging agent may be administered for disease treatment. The appropriate route of administration and dosage of the T cell engaging agent may be determined based on the type of disease to be treated, the type of the T cell engaging agent, the severity and course of the disease, the clinical condition of the individual, the individual’s clinical history and response to the treatment, and the discretion of the attending physician. Dosing can be done by any suitable route, e.g. by injections, such as intravenous or subcutaneous injections, depending in part on whether the administration is brief or chronic. Various dosing schedules including but not limited to single or multiple administrations over various time-points, bolus administration, and pulse infusion are contemplated herein.

[0211] The T cell engaging agent, the TKI and the GC can be administered by any suitable route and may be administered by the same route of administration or by different routes of administration. In some aspects, the administration of the T cell engaging agent is parenteral, particularly intravenous. In some aspects, the administration of the T cell engaging agent is the first administration of the T cell engaging agent to the individual, particularly the first administration of the T cell engaging agent in the course of the treatment of the individual with the T cell engaging agent.

[0212] In some aspects, administration of the T cell engaging agent induces (i.e. causes or increases) the adverse and / or desired effect. In some aspects, administration of the T cell engaging agent induces the activation of T cells. In some aspects, administration of the T cell engaging agent induces cytotoxic activity of T cells. In some aspects, administration of the T cell engaging agent induces cytokine secretion by immune cells, particularly by T cells. In some aspects, cytokine is one or more cytokine selected from the group consisting of IL-2, IL-6, IFN-y, IL- 10, TNF-a and GM- CSF. In some aspects, said T cells are CD8+ T cells or CD4+ T cells. In some aspects, administration of the T cell engaging agent induces CRS. In some aspects, administration of the T cell engaging agent induces ICANS. In some aspects, administration of the T cell engaging agent induces one or more of thrombocytopenia, an elevated serum level of one or more of lactate dehydrogenase, ferritin, IFN-y, IL-10, granzyme B, GM-CSF, MIP-la, TNF, IL-2, and an elevated level in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL- 10, IL-6 and granzyme B. In some aspects, administration of the T cell engaging agent induces T cell exhaustion.

[0213] In some aspects, administration of the T cell engaging agent results in activation of T cells, particularly cytotoxic T cells, particularly at the site of the cancer (e.g. within a solid tumor cancer). Said activation may comprise proliferation of T cells, differentiation of T cells, cytokine secretion by T cells, cytotoxic effector molecule release from T cells, cytotoxic activity of T cells, and expression of activation markers by T cells. In some aspects, the administration of the T cell engaging agent results in an increase of T cell, particularly cytotoxic T cell, numbers at the site of the cancer (e.g. within a solid tumor cancer).

[0214] By “T cell engaging agent” is meant an immunotherapeutic agent that exerts its effect through the activity of T cells, particularly cytotoxic T cells. Such activity of T cells may include cellular response(s) of T cells, particularly CD4+ and / or CD8+ T cells, such as proliferation, differentiation, expression of activation markers, cytokine secretion, cytotoxic effector molecule release and / or cytotoxic activity. T cell engaging agents as contemplated herein typically comprise an antigen binding moiety that enables their binding to a target cell antigen on a target cell such as a tumor cell. Such T cell engaging agents exert effects on their target cell, such as lysis of the target cell, through the activity of T cells. Exemplary T cell engaging agents include T cell bispecific antibodies, chimeric antigen receptor (CAR) expressing T cells (CAR-T cells), and T cell receptor (TCR)-based approaches such as ImmTACs (“Immune mobilising monoclonal T-cell receptors Against Cancer”; bispecific fusion proteins of an engineered TCR and an antibody fragment, capable of binding to a T cell and a target cell) or TCR-modified T cells featuring engineered T cell receptors capable of binding to a specific antigenic determinant on a target cell (TCR-T cells).

[0215] In particular aspects of the present invention, the T cell engaging agent is a T cell bispecific antibody.

[0216] In other aspects, the T cell engaging agent is a CAR-T cell. In some aspects, the CAR-T cell is a universal CAR-T cell. By “universal” CAR-T cell is meant a CAR-T cell that binds to a target cell antigen through an adaptor molecule, such as an antibody, that binds to the target cell antigen. A universal CAR-T cell expresses a CAR comprising an antigen binding moiety that binds to the adaptor molecule, and the adaptor molecule binds to the target cell antigen. Through different adaptor molecules (binding to different target cell antigens), a universal CAR-T cell can bind to different target cell antigens, without the need for expression of a different CAR for each target cell antigen. The adaptor molecule is a molecule that (i) can be bound by the CAR, and (ii) can bind to a target cell antigen, such as, for example, an antibody that binds to the target cell antigen and comprises an Fc region that can be bound by the CAR. In some aspects, the CAR-T cell expresses a CAR comprising an antigen binding moiety that binds to an antibody Fc region, particularly an IgG Fc region, more particularly an IgGi Fc region, and particularly a human Fc region. In some aspects, the CAR-T expresses a CAR comprising an antigen binding moiety that binds to an IgG Fc region, particularly a human IgGi Fc region, comprising the amino acid substitution P329G (Kabat EU index numbering). In some of these aspects, the antigen binding moiety is a scFv. In other aspects, the CAR-T expresses a CAR comprising an antigen binding moiety that binds to a wild-type Fc region, particularly a wild-type human IgGi Fc region. In some of these aspects, the antigen binding moiety is CD16 or an Fc-binding fragment thereof (for example, the extracellular domain of CD 16).

[0217] In some aspects, the T cell engaging agent is an ImmTAC. In some aspects, the T cell engaging agent is a TCR-T cell.

[0218] In the following, the T cell bispecific antibody that may be used in the present invention is described. By “T cell bispecific antibody” is meant an antibody that is able to bind, including simultaneously bind, to a T cell (typically via an antigenic determinant expressed on the T cell, such as CD3) and to a target cell (typically via an antigenic determinant expressed on the target cell, such CD20).

[0219] In some aspects, the T cell bispecific antibody is capable of simultaneous binding to the antigenic determinant on the T cell (i.e. a first antigen such as CD3) and the antigenic determinant on the target cell (i.e. a second antigen such as CD20). In some aspects, the T cell bispecific antibody is capable of crosslinking the T cell and the target cell by simultaneous binding to CD3 and a target cell antigen. In some aspects, such simultaneous binding results in lysis of the target cell, particularly a target cell antigen (e.g. CD20)-expressing tumor cell. In some aspects, such simultaneous binding results in activation of the T cell. In some aspects, such simultaneous binding results in a cellular response of the T cell, selected from the group of: proliferation, differentiation, cytokine secretion, cytotoxic effector molecule release, cytotoxic activity, and expression of activation markers. In some aspects, binding of the T cell bispecific antibody to CD3 without simultaneous binding to the target cell antigen does not result in T cell activation. In some aspects, the T cell bispecific antibody is capable of re-directing cytotoxic activity of a T cell to a target cell. In some aspects, said re-direction is independent of MHC-mediated peptide antigen presentation by the target cell and and / or specificity of the T cell.

[0220] The term “bispecific” means that the antibody is able to bind to at least two distinct antigenic determinants. Typically, a bispecific antibody comprises two antigen binding sites, each of which is specific for a different antigenic determinant. In certain aspects, the bispecific antibody is capable of simultaneously binding two antigenic determinants, particularly two antigenic determinants expressed on two distinct cells. In certain aspects, the bispecific antibody may also bind to more than two antigens, e.g., three, four or five antigens, which may be distinct or not.

[0221] As used herein, the term “antigenic determinant” is synonymous with “antigen” and “epitope”, and refers to a site (e.g., a contiguous stretch of amino acids or a conformational configuration made up of different regions of non-contiguous amino acids) on a biological molecule, such as a polypeptide or nucleic acid, to which an antigen binding moiety binds, forming an antigen binding moiety-antigen complex. Useful antigenic determinants can be found, for example, on the surfaces of tumor cells, on the surfaces of virus-infected cells, on the surfaces of other diseased cells, on the surface of immune cells, free in blood serum, and / or in the extracellular matrix (ECM). As used herein, the term “antigen binding moiety” refers to a polypeptide molecule that binds, including specifically binds, to an antigenic determinant. In some aspects, an antigen binding moiety is able to direct the entity to which it is attached (e.g. a second antigen binding moiety) to a target site, for example to a specific type of tumor cell bearing the antigenic determinant. In further aspects, an antigen binding moiety is able to activate signaling through its target antigen, for example a T cell receptor complex antigen. Antigen binding moieties include antibodies and fragments thereof as further defined herein. Particular antigen binding moieties include an antigen binding domain of an antibody, comprising an antibody heavy chain variable region and an antibody light chain variable region. In certain aspects, the antigen binding moieties may comprise antibody constant regions as further defined herein and known in the art. Useful heavy chain constant regions include any of the five isotypes: a, 5, a, y, or p. Useful light chain constant regions include any of the two isotypes: K and X.

[0222] By “specific binding” is meant that the binding is selective for the antigen and can be discriminated from unwanted or non-specific interactions. The term “bind” or “binding” herein generally refers to “specific binding”. The ability of an antigen binding moiety to bind to a specific antigenic determinant can be measured either through an enzyme-linked immunosorbent assay (ELISA) or other techniques familiar to one of skill in the art, e.g. surface plasmon resonance (SPR) technique (analyzed e.g. on a BIAcore instrument) (Liljeblad et al., Glyco J 17, 323-329 (2000)), and traditional binding assays (Heeley, Endocr Res 28, 217-229 (2002)). In some aspects, the extent of binding of an antigen binding moiety to an unrelated protein is less than about 10% of the binding of the antigen binding moiety to the antigen as measured, e.g., by SPR. In certain aspects, an antigen binding moiety that binds to the antigen, or an antibody comprising that antigen binding moiety, has a dissociation constant (KD) of < 1 pM, < 100 nM, < 10 nM, < 1 nM, < 0.1 nM, < 0.01 nM, or < 0.001 nM (e.g. 10'8M or less, e.g. from 10'8M to 10'13M, e.g., from 10'9M to 10'13M).

[0223] “Affinity” refers to the strength of the sum total of non-covalent interactions between a single binding site of a molecule (e.g., a receptor) and its binding partner (e.g., a ligand). Unless indicated otherwise, as used herein, “binding affinity” refers to intrinsic binding affinity which reflects a 1 : 1 interaction between members of a binding pair (e.g., an antigen binding moiety and an antigen, or a receptor and its ligand). The affinity of a molecule X for its partner Y can generally be represented by the dissociation constant (KD), which is the ratio of dissociation and association rate constants (koff and kon, respectively). Thus, equivalent affinities may comprise different rate constants, as long as the ratio of the rate constants remains the same. Affinity can be measured by well established methods known in the art, including those described herein. A particular method for measuring affinity is Surface Plasmon Resonance (SPR).

[0224] “CD3” refers to any native CD3 from any vertebrate source, including mammals such as primates (e.g. humans), non-human primates (e.g. cynomolgus monkeys) and rodents (e.g. mice and rats), unless otherwise indicated. The term encompasses “full-length,” unprocessed CD3 as well as any form of CD3 that results from processing in the cell. The term also encompasses naturally occurring variants of CD3, e.g., splice variants or allelic variants. In some aspects, CD3 is human CD3, particularly the epsilon subunit of human CD3 (CD3s). The amino acid sequence of human CD3s is shown in UniProt accession no. P07766 (version 144), or NCBI RefSeq NP_000724.1. See also SEQ ID NO: 1. The amino acid sequence of cynomolgus [Macaca fascicularis] CD3s is shown in NCBI GenBank no. BAB71849.1. See also SEQ ID NO: 2.

[0225] A “target cell antigen” as used herein refers to an antigenic determinant presented on the surface of a target cell, for example a cell in a tumor such as a cancer cell or a cell of the tumor stroma (in that case a “tumor cell antigen”). Preferably, the target cell antigen is not CD3, and / or is expressed on a different cell than CD3. In some aspects, the target cell antigen is CD20, particularly human CD20.

[0226] As used herein, the terms “first”, “second” or “third” with respect to antigen binding moieties etc., are used for convenience of distinguishing when there is more than one of each type of moiety. Use of these terms is not intended to confer a specific order or orientation of the bispecific antibody unless explicitly so stated.

[0227] The term “valent” as used herein denotes the presence of a specified number of antigen binding sites in an antibody. As such, the term “monovalent binding to an antigen” denotes the presence of one (and not more than one) antigen binding site specific for the antigen in the antibody.

[0228] The term “antibody” herein is used in the broadest sense and encompasses various antibody structures, including but not limited to monoclonal antibodies, polyclonal antibodies, multispecific antibodies (e.g. bispecific antibodies), and antibody fragments so long as they exhibit the desired antigen-binding activity.

[0229] The terms “full length antibody,” “intact antibody,” and “whole antibody” are used herein interchangeably to refer to an antibody having a structure substantially similar to a native antibody structure. An “antibody fragment” refers to a molecule other than an intact antibody that comprises a portion of an intact antibody and is suitable for binding to an antigen. Examples of antibody fragments include but are not limited to Fv, Fab, Fab', Fab’-SH, F(ab')2, diabodies, linear antibodies, singlechain antibody molecules (e.g. scFv), and single-domain antibodies. For a review of certain antibody fragments, see Hudson et al., Nat Med 9, 129-134 (2003). For a review of scFv fragments, see e.g. Pliickthun, in The Pharmacology of Monoclonal Antibodies, vol. 113, Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994); see also WO 93 / 16185; and U.S. Patent Nos. 5,571,894 and 5,587,458. For discussion of Fab and F(ab')2 fragments comprising salvage receptor binding epitope residues and having increased in vivo half-life, see U.S. Patent No. 5,869,046. Diabodies are antibody fragments with two antigen-binding sites that may be bivalent or bispecific. See, for example, EP 404,097; WO 1993 / 01161; Hudson et al., Nat Med 9, 129-134 (2003); and Hollinger et al., Proc Natl Acad Sci USA 90, 6444-6448 (1993). Triabodies and tetrabodies are also described in Hudson et al., Nat Med 9, 129-134 (2003). Single-domain antibodies are antibody fragments comprising all or a portion of the heavy chain variable domain or all or a portion of the light chain variable domain of an antibody. In certain aspects, a singledomain antibody is a human single-domain antibody (Domantis, Inc., Waltham, MA; see e.g. U.S. Patent No. 6,248,516 Bl). Antibody fragments can be made by various techniques, including but not limited to proteolytic digestion of an intact antibody as well as production by recombinant host cells (e.g. E. coli or phage), as described herein.

[0230] The term “variable region” or “variable domain” refers to the domain of an antibody heavy or light chain that is involved in binding the antibody to antigen. The variable domains of the heavy chain and light chain (VH and VL, respectively) of a native antibody generally have similar structures, with each domain comprising four conserved framework regions (FRs) and three hypervariable regions (HVRs). See, e.g., Kindt et al., Kuby Immunology, 6thed., W.H. Freeman and Co., page 91 (2007). A single VH or VL domain may be sufficient to confer antigen-binding specificity. As used herein in connection with variable region sequences, "Kabat numbering" refers to the numbering system set forth by Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991).

[0231] As used herein, the amino acid positions of all constant regions and domains of the heavy and light chain are numbered according to the Kabat numbering system described in Kabat, et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991), referred to as “numbering according to Kabat” or “Kabat numbering” herein. Specifically the Kabat numbering system (see pages 647-660 of Kabat, et al., Sequences of Proteins of Immunological Interest, 5th ed., Public Health Service, National Institutes of Health, Bethesda, MD (1991)) is used for the light chain constant domain CL of kappa and lambda isotype and the Kabat EU index numbering system (see pages 661-723) is used for the heavy chain constant domains (CHI, Hinge, CH2 and CH3), which is herein further clarified by referring to “numbering according to Kabat EU index” in this case.

[0232] The term “hypervariable region” or “HVR”, as used herein, refers to each of the regions of an antibody variable domain which are hypervariable in sequence and which determine antigen binding specificity, for example “complementarity determining regions” (“CDRs”). Generally, antibodies comprise six CDRs; three in the VH (HCDR1, HCDR2, HCDR3), and three in the VL (LCDR1, LCDR2, LCDR3). Exemplary CDRs herein include:

[0233] (a) hypervariable loops occurring at amino acid residues 26-32 (LI), 50-52 (L2), 91-96 (L3), 26-32 (Hl), 53-55 (H2), and 96-101 (H3) (Chothia and Lesk, J. Mol. Biol. 196:901-917 (1987));

[0234] (b) CDRs occurring at amino acid residues 24-34 (LI), 50-56 (L2), 89-97 (L3), 31-35b (Hl), 50-65 (H2), and 95-102 (H3) (Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991)); and

[0235] (c) antigen contacts occurring at amino acid residues 27c-36 (LI), 46-55 (L2), 89-96 (L3), 30-35b (Hl), 47-58 (H2), and 93-101 (H3) (MacCallum et al. J. Mol. Biol. 262: 732-745 (1996)).

[0236] Unless otherwise indicated, the CDRs are determined according to Kabat et al., supra. One of skill in the art will understand that the CDR designations can also be determined according to Chothia, supra, McCallum, supra, or any other scientifically accepted nomenclature system.

[0237] “Framework” or “FR” refers to variable domain residues other than hypervariable region (HVR) residues. The FR of a variable domain generally consists of four FR domains: FR1, FR2, FR3, and FR4. Accordingly, the HVR and FR sequences generally appear in the following order in VH (or VL) : FR1 -H 1 (L 1 )-FR2-H2(L2)-FR3 -H3 (L3 )-FR4.

[0238] The “class” of an antibody or immunoglobulin refers to the type of constant domain or constant region possessed by its heavy chain. There are five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM, and several of these may be further divided into subclasses (isotypes), e.g., IgGi, IgG?, IgGs, IgG4, IgAi, and IgA2. The heavy chain constant domains that correspond to the different classes of immunoglobulins are called a, 5, a, y, and p, respectively.

[0239] A “Fab molecule” refers to a protein consisting of the VH and CHI domain of the heavy chain (the “Fab heavy chain”) and the VL and CL domain of the light chain (the “Fab light chain”) of an immunoglobulin.

[0240] By a “crossover” Fab molecule (also termed “Crossfab”) is meant a Fab molecule wherein the variable domains or the constant domains of the Fab heavy and light chain are exchanged (i.e. replaced by each other), i.e. the crossover Fab molecule comprises a peptide chain composed of the light chain variable domain VL and the heavy chain constant domain 1 CHI (VL-CH1, in N- to C-terminal direction), and a peptide chain composed of the heavy chain variable domain VH and the light chain constant domain CL (VH-CL, in N- to C-terminal direction). For clarity, in a crossover Fab molecule wherein the variable domains of the Fab light chain and the Fab heavy chain are exchanged, the peptide chain comprising the heavy chain constant domain 1 CHI is referred to herein as the “heavy chain” of the (crossover) Fab molecule. Conversely, in a crossover Fab molecule wherein the constant domains of the Fab light chain and the Fab heavy chain are exchanged, the peptide chain comprising the heavy chain variable domain VH is referred to herein as the “heavy chain” of the (crossover) Fab molecule.

[0241] In contrast thereto, by a “conventional” Fab molecule is meant a Fab molecule in its natural format, i.e. comprising a heavy chain composed of the heavy chain variable and constant domains (VH- CH1, in N- to C-terminal direction), and a light chain composed of the light chain variable and constant domains (VL-CL, in N- to C-terminal direction).

[0242] The term “immunoglobulin molecule” refers to a protein having the structure of a naturally occurring antibody. For example, immunoglobulins of the IgG class are heterotetrameric glycoproteins of about 150,000 daltons, composed of two light chains and two heavy chains that are disulfide-bonded. From N- to C-terminus, each heavy chain has a variable domain (VH), also called a variable heavy domain or a heavy chain variable region, followed by three constant domains (CHI, CH2, and CH3), also called a heavy chain constant region. Similarly, from N- to C-terminus, each light chain has a variable domain (VL), also called a variable light domain or a light chain variable region, followed by a constant light (CL) domain, also called a light chain constant region. The heavy chain of an immunoglobulin may be assigned to one of five types, called a (IgA), 5 (IgD), a (IgE), y (IgG), or p (IgM), some of which may be further divided into subtypes, e.g. 71 (IgGi), 72 (IgG?), 73 (IgGs), 74 (IgG4), ai (IgAi) and a? (IgA2). The light chain of an immunoglobulin may be assigned to one of two types, called kappa (K) and lambda (X), based on the amino acid sequence of its constant domain. An immunoglobulin essentially consists of two Fab molecules and an Fc domain, linked via the immunoglobulin hinge region.

[0243] The term “Fc domain” or “Fc region” herein is used to define a C-terminal region of an immunoglobulin heavy chain that contains at least a portion of the constant region. The term includes native sequence Fc regions and variant Fc regions. Although the boundaries of the Fc region of an IgG heavy chain might vary slightly, the human IgG heavy chain Fc region is usually defined to extend from Cys226, or from Pro230, to the carboxyl-terminus of the heavy chain. However, antibodies produced by host cells may undergo post-translational cleavage of one or more, particularly one or two, amino acids from the C-terminus of the heavy chain. Therefore an antibody produced by a host cell by expression of a specific nucleic acid molecule encoding a full- length heavy chain may include the full-length heavy chain, or it may include a cleaved variant of the full-length heavy chain. This may be the case where the final two C-terminal amino acids of the heavy chain are glycine (G446) and lysine (K447, numbering according to Kabat EU index). Therefore, the C-terminal lysine (Lys447), or the C-terminal glycine (Gly446) and lysine (K447), of the Fc region may or may not be present. Unless otherwise specified herein, numbering of amino acid residues in the Fc region or constant region is according to the EU numbering system, also called the EU index, as described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD, 1991 (see also above). A “subunit” of an Fc domain as used herein refers to one of the two polypeptides forming the dimeric Fc domain, i.e. a polypeptide comprising C-terminal constant regions of an immunoglobulin heavy chain, capable of stable self-association. For example, a subunit of an IgG Fc domain comprises an IgG CH2 and an IgG CH3 constant domain.

[0244] A “modification promoting the association of the first and the second subunit of the Fc domain” is a manipulation of the peptide backbone or the post-translational modifications of an Fc domain subunit that reduces or prevents the association of a polypeptide comprising the Fc domain subunit with an identical polypeptide to form a homodimer. A modification promoting association as used herein particularly includes separate modifications made to each of the two Fc domain subunits desired to associate (i.e. the first and the second subunit of the Fc domain), wherein the modifications are complementary to each other so as to promote association of the two Fc domain subunits. For example, a modification promoting association may alter the structure or charge of one or both of the Fc domain subunits so as to make their association sterically or electrostatically favorable, respectively. Thus, (hetero)dimerization occurs between a polypeptide comprising the first Fc domain subunit and a polypeptide comprising the second Fc domain subunit, which might be non-identical in the sense that further components fused to each of the subunits (e.g. antigen binding moieties) are not the same. In some aspects the modification promoting association comprises an amino acid mutation in the Fc domain, specifically an amino acid substitution. In particular aspects, the modification promoting association comprises a separate amino acid mutation, specifically an amino acid substitution, in each of the two subunits of the Fc domain.

[0245] The term “effector functions” refers to those biological activities attributable to the Fc region of an antibody, which vary with the antibody isotype. Examples of antibody effector functions include: Clq binding and complement dependent cytotoxicity (CDC), Fc receptor binding, antibody-dependent cell-mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), cytokine secretion, immune complex-mediated antigen uptake by antigen presenting cells, down regulation of cell surface receptors (e.g. B cell receptor), and B cell activation.

[0246] “Percent (%) amino acid sequence identity” with respect to a reference polypeptide sequence is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, Clustal W, Megalign (DNASTAR) software or the FASTA program package. Those skilled in the art can determine appropriate parameters for aligning sequences, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared. For purposes herein, however, % amino acid sequence identity values are generated using the ggsearch program of the FASTA package version 36.3.8c or later with a BLOSUM50 comparison matrix. The FASTA program package was authored by W. R. Pearson and D. J. Lipman (1988), “Improved Tools for Biological Sequence Analysis”, PNAS 85:2444-2448; W. R. Pearson (1996) “Effective protein sequence comparison” Meth. Enzymol. 266:227- 258; and Pearson et. al. (1997) Genomics 46:24-36, and is publicly available from http: / / fasta.bioch.virginia.edu / fasta_www2 / fasta_down.shtml. Alternatively, a public server accessible at http: / / fasta.bioch.virginia.edu / fasta_www2 / index.cgi can be used to compare the sequences, using the ggsearch (global protein: protein) program and default options (BLOSUM50; open: -10; ext: -2; Ktup = 2) to ensure a global, rather than local, alignment is performed. Percent amino acid identity is given in the output alignment header.

[0247] An “activating Fc receptor” is an Fc receptor that following engagement by an Fc domain of an antibody elicits signaling events that stimulate the receptor-bearing cell to perform effector functions. Human activating Fc receptors include FcyRIIIa (CD16a), FcyRI (CD64), FcyRIIa (CD32), and FcaRI (CD89).

[0248] “Reduced binding”, for example reduced binding to an Fc receptor, refers to a decrease in affinity for the respective interaction, as measured for example by SPR. For clarity, the term includes also reduction of the affinity to zero (or below the detection limit of the analytic method), i.e. complete abolishment of the interaction. Conversely, “increased binding” refers to an increase in binding affinity for the respective interaction.

[0249] By “fused” is meant that the components (e.g. a Fab molecule and an Fc domain subunit) are linked by peptide bonds, either directly or via one or more peptide linkers.

[0250] In particular aspects, the T cell bispecific antibody binds to CD3 and a target cell antigen. Accordingly, in some aspects, the T cell bispecific antibody comprises an antigen binding moiety that binds to CD3 and an antigen binding moiety that binds to a target cell antigen.

[0251] In some aspects, the first and / or the second antigen binding moiety is a Fab molecule. In some aspects, the first antigen binding moiety is a crossover Fab molecule wherein either the variable or the constant regions of the Fab light chain and the Fab heavy chain are exchanged. In such aspects, the second antigen binding moiety preferably is a conventional Fab molecule.

[0252] In some aspects wherein the first and the second antigen binding moiety of the T cell bispecific antibody are both Fab molecules, and in one of the antigen binding moi eties (particularly the first antigen binding moiety) the variable domains VL and VH of the Fab light chain and the Fab heavy chain are replaced by each other, i) in the constant domain CL of the first antigen binding moiety the amino acid at position 124 is substituted by a positively charged amino acid (numbering according to Kabat), and wherein in the constant domain CHI of the first antigen binding moiety the amino acid at position 147 or the amino acid at position 213 is substituted by a negatively charged amino acid (numbering according to Kabat EU index); or ii) in the constant domain CL of the second antigen binding moiety the amino acid at position 124 is substituted by a positively charged amino acid (numbering according to Kabat), and wherein in the constant domain CHI of the second antigen binding moiety the amino acid at position 147 or the amino acid at position 213 is substituted by a negatively charged amino acid (numbering according to Kabat EU index).

[0253] The T cell bispecific antibody does not comprise both modifications mentioned under i) and ii). The constant domains CL and CHI of the antigen binding moiety having the VH / VL exchange are not replaced by each other (i.e. remain unexchanged).

[0254] In more specific aspects, i) in the constant domain CL of the first antigen binding moiety the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat), and in the constant domain CHI of the first antigen binding moiety the amino acid at position 147 or the amino acid at position 213 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index); or ii) in the constant domain CL of the second antigen binding moiety the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat), and in the constant domain CHI of the second antigen binding moiety the amino acid at position 147 or the amino acid at position 213 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index).

[0255] In some aspects, in the constant domain CL of the second antigen binding moiety the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat), and in the constant domain CHI of the second antigen binding moiety the amino acid at position 147 or the amino acid at position 213 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index).

[0256] In further aspects, in the constant domain CL of the second antigen binding moiety the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat), and in the constant domain CHI of the second antigen binding moiety the amino acid at position 147 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index).

[0257] In preferred aspects, in the constant domain CL of the second antigen binding moiety the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat) and the amino acid at position 123 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat), and in the constant domain CHI of the second antigen binding moiety the amino acid at position 147 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index) and the amino acid at position 213 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index).

[0258] In some aspects, in the constant domain CL of the second antigen binding moiety the amino acid at position 124 is substituted by lysine (K) (numbering according to Kabat) and the amino acid at position 123 is substituted by lysine (K) (numbering according to Kabat), and in the constant domain CHI of the second antigen binding moiety the amino acid at position 147 is substituted by glutamic acid (E) (numbering according to Kabat EU index) and the amino acid at position 213 is substituted by glutamic acid (E) (numbering according to Kabat EU index).

[0259] In some aspects, in the constant domain CL of the second antigen binding moiety the amino acid at position 124 is substituted by lysine (K) (numbering according to Kabat) and the amino acid at position 123 is substituted by arginine (R) (numbering according to Kabat), and in the constant domain CHI of the second antigen binding moiety the amino acid at position 147 is substituted by glutamic acid (E) (numbering according to Kabat EU index) and the amino acid at position 213 is substituted by glutamic acid (E) (numbering according to Kabat EU index).

[0260] In particular aspects, if amino acid substitutions according to the above aspects are made in the constant domain CL and the constant domain CHI of the second antigen binding moiety, the constant domain CL of the second antigen binding moiety is of kappa isotype.

[0261] In some aspects, the first and the second antigen binding moiety are fused to each other, optionally via a peptide linker.

[0262] In some aspects, the first and the second antigen binding moiety are each a Fab molecule and either (i) the second antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N- terminus of the Fab heavy chain of the first antigen binding moiety, or (ii) the first antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second antigen binding moiety.

[0263] In some aspects, the T cell bispecific antibody provides monovalent binding to CD3.

[0264] In particular aspects, the T cell bispecific antibody comprises a single antigen binding moiety that binds to CD3, and two antigen binding moi eties that bind to the target cell antigen. Thus, in some aspects, the T cell bispecific antibody comprises a third antigen binding moiety, particularly a Fab molecule, more particularly a conventional Fab molecule, that binds to the target antigen. The third antigen binding moiety may incorporate, singly or in combination, all of the features described herein in relation to the second antigen binding moiety (e.g. the CDR sequences, variable region sequences, and / or amino acid substitutions in the constant regions). In some aspects, the third antigen moiety is identical to the first antigen binding moiety (e.g. is also a conventional Fab molecule and comprises the same amino acid sequences).

[0265] In particular aspects, the T cell bispecific antibody further comprises an Fc domain composed of a first and a second subunit. In some aspects, the Fc domain is an IgG Fc domain. In particular aspects, the Fc domain is an IgGi Fc domain. In other aspects, the Fc domain is an IgG4 Fc domain. In more specific aspects, the Fc domain is an IgG4 Fc domain comprising an amino acid substitution at position S228 (Kabat EU index numbering), particularly the amino acid substitution S228P. This amino acid substitution reduces in vivo Fab arm exchange of IgG4 antibodies (see Stubenrauch et al., Drug Metabolism and Disposition 38, 84-91 (2010)). In further particular aspects, the Fc domain is a human Fc domain. In particularly preferred aspects, the Fc domain is a human IgGi Fc domain. An exemplary sequence of a human IgGi Fc region is given in SEQ ID NO: 3.

[0266] In some aspects wherein the first, the second and, where present, the third antigen binding moiety are each a Fab molecule, (a) either (i) the second antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the first antigen binding moiety and the first antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N- terminus of the first subunit of the Fc domain, or (ii) the first antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the second antigen binding moiety and the second antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first subunit of the Fc domain; and (b) the third antigen binding moiety, where present, is fused at the C-terminus of the Fab heavy chain to the N-terminus of the second subunit of the Fc domain.

[0267] In some aspects, the T cell bispecific antibody essentially consists of the first, the second and the third antigen binding moiety (particularly Fab molecule), the Fc domain composed of a first and a second subunit, and optionally one or more peptide linkers.

[0268] The components of the T cell bispecific antibody may be fused to each other directly or, preferably, via one or more suitable peptide linkers. Where fusion of a Fab molecule is to the N-terminus of a subunit of the Fc domain, it is typically via an immunoglobulin hinge region.

[0269] The antigen binding moieties may be fused to the Fc domain or to each other directly or through a peptide linker, comprising one or more amino acids, typically about 2-20 amino acids. Peptide linkers are known in the art and are described herein. Suitable, non-immunogenic peptide linkers include, for example, (G4S)n, (SG4)n, (G4S)n, G4(SG4)nor (G4S)nGs peptide linkers, “n” is generally an integer from 1 to 10, typically from 2 to 4. In some aspects, said peptide linker has a length of at least 5 amino acids, in some aspects a length of 5 to 100, in further aspects of 10 to 50 amino acids. In some aspects said peptide linker is (GxS)nor (GxS)nGmwith G=glycine, S=serine, and (x=3, n= 3, 4, 5 or 6, and m=0, 1, 2 or 3) or (x=4, n=l, 2, 3, 4 or 5 and m= 0, 1, 2, 3, 4 or 5), in some aspects x=4 and n=2 or 3, in further aspects x=4 and n=2, in yet further aspects x=4, n=l and m=5. In some aspects, said peptide linker is (648)2. In other aspects, said peptide linker is G4SG5. Additionally, linkers may comprise (a portion of) an immunoglobulin hinge region. Particularly where a Fab molecule is fused to the N-terminus of an Fc domain subunit, it may be fused via an immunoglobulin hinge region or a portion thereof, with or without an additional peptide linker.

[0270] In particular aspects, the Fc domain comprises a modification promoting the association of the first and the second subunit of the Fc domain. The site of most extensive protein-protein interaction between the two subunits of a human IgGFc domain is in the CH3 domain. Thus, in some aspects, said modification is in the CH3 domain of the Fc domain.

[0271] In specific aspects, said modification promoting the association of the first and the second subunit of the Fc domain is a so-called “knob-into-hole” modification, comprising a “knob” modification in one of the two subunits of the Fc domain and a “hole” modification in the other one of the two subunits of the Fc domain. The knob-into-hole technology is described e.g. in US 5,731,168; US 7,695,936; Ridgway et al., Prot Eng 9, 617-621 (1996) and Carter, J Immunol Meth 248, 7-15 (2001). Generally, the method involves introducing a protuberance (“knob”) at the interface of a first polypeptide and a corresponding cavity (“hole”) in the interface of a second polypeptide, such that the protuberance can be positioned in the cavity so as to promote heterodimer formation and hinder homodimer formation. Protuberances are constructed by replacing small amino acid side chains from the interface of the first polypeptide with larger side chains (e.g. tyrosine or tryptophan). Compensatory cavities of identical or similar size to the protuberances are created in the interface of the second polypeptide by replacing large amino acid side chains with smaller ones (e.g. alanine or threonine).

[0272] Accordingly, in some aspects, an amino acid residue in the CH3 domain of the first subunit of the Fc domain is replaced with an amino acid residue having a larger side chain volume, thereby generating a protuberance within the CH3 domain of the first subunit which is positionable in a cavity within the CH3 domain of the second subunit, and an amino acid residue in the CH3 domain of the second subunit of the Fc domain is replaced with an amino acid residue having a smaller side chain volume, thereby generating a cavity within the CH3 domain of the second subunit within which the protuberance within the CH3 domain of the first subunit is positionable. Preferably said amino acid residue having a larger side chain volume is selected from the group consisting of arginine (R), phenylalanine (F), tyrosine (Y), and tryptophan (W). Preferably said amino acid residue having a smaller side chain volume is selected from the group consisting of alanine (A), serine (S), threonine (T), and valine (V). The protuberance and cavity can be made by altering the nucleic acid encoding the polypeptides, e.g. by site-specific mutagenesis, or by peptide synthesis.

[0273] In specific such aspects, in the first subunit of the Fc domain the threonine residue at position 366 is replaced with a tryptophan residue (T366W), and in the second subunit of the Fc domain the tyrosine residue at position 407 is replaced with a valine residue (Y407V) and optionally the threonine residue at position 366 is replaced with a serine residue (T366S) and the leucine residue at position 368 is replaced with an alanine residue (L368A) (numbering according to Kabat EU index). In further aspects, in the first subunit of the Fc domain additionally the serine residue at position 354 is replaced with a cysteine residue (S354C) or the glutamic acid residue at position 356 is replaced with a cysteine residue (E356C) (particularly the serine residue at position 354 is replaced with a cysteine residue), and in the second subunit of the Fc domain additionally the tyrosine residue at position 349 is replaced by a cysteine residue (Y349C) (numbering according to Kabat EU index). In preferred aspects, the first subunit of the Fc domain comprises the amino acid substitutions S354C and T366W, and the second subunit of the Fc domain comprises the amino acid substitutions Y349C, T366S, L368A and Y407V (numbering according to Kabat EU index).

[0274] In some aspects, the Fc domain comprises one or more amino acid substitution that reduces binding to an Fc receptor and / or effector function.

[0275] In particular aspects, the Fc receptor is an Fey receptor. In some aspects, the Fc receptor is a human Fc receptor. In some aspects, the Fc receptor is an activating Fc receptor. In specific aspects, the Fc receptor is an activating human Fey receptor, more specifically human FcyRIIIa, FcyRI or FcyRIIa, most specifically human FcyRIIIa. In some aspects, the effector function is one or more selected from the group of complement dependent cytotoxicity (CDC), antibody-dependent cell- mediated cytotoxicity (ADCC), antibody-dependent cellular phagocytosis (ADCP), and cytokine secretion. In particular aspects, the effector function is ADCC.

[0276] Typically, the same one or more amino acid substitution is present in each of the two subunits of the Fc domain. In some aspects, the one or more amino acid substitution reduces the binding affinity of the Fc domain to an Fc receptor. In some aspects, the one or more amino acid substitution reduces the binding affinity of the Fc domain to an Fc receptor by at least 2-fold, at least 5-fold, or at least 10-fold.

[0277] In some aspects, the Fc domain comprises an amino acid substitution at a position selected from the group of E233, L234, L235, N297, P331 and P329 (numberings according to Kabat EU index). In more specific aspects, the Fc domain comprises an amino acid substitution at a position selected from the group of L234, L235 and P329 (numberings according to Kabat EU index). In some aspects, the Fc domain comprises the amino acid substitutions L234A and L235A (numberings according to Kabat EU index). In some such aspects, the Fc domain is an IgGi Fc domain, particularly a human IgGi Fc domain. In some aspects, the Fc domain comprises an amino acid substitution at position P329. In more specific aspects, the amino acid substitution is P329A or P329G, particularly P329G (numberings according to Kabat EU index). In some aspects, the Fc domain comprises an amino acid substitution at position P329 and a further amino acid substitution at a position selected from E233, L234, L235, N297 and P331 (numberings according to Kabat EU index). In more specific aspects, the further amino acid substitution is E233P, L234A, L235A, L235E, N297A, N297D or P331S. In particular aspects, the Fc domain comprises amino acid substitutions at positions P329, L234 and L235 (numberings according to Kabat EU index). In more particular aspects, the Fc domain comprises the amino acid mutations L234A, L235A and P329G (“P329G LALA”, “PGLALA” or “LALAPG”). Specifically, in preferred aspects, each subunit of the Fc domain comprises the amino acid substitutions L234A, L235A and P329G (Kabat EU index numbering), i.e. in each of the first and the second subunit of the Fc domain the leucine residue at position 234 is replaced with an alanine residue (L234A), the leucine residue at position 235 is replaced with an alanine residue (L235A) and the proline residue at position 329 is replaced by a glycine residue (P329G) (numbering according to Kabat EU index). In some such aspects, the Fc domain is an IgGi Fc domain, particularly a human IgGi Fc domain.

[0278] In some aspects, the target cell antigen of the T cell bispecific antibody is CD20.

[0279] “CD20”, also known as “B-lymphocyte antigen Bl”, refers to any native CD20 from any vertebrate source, including mammals such as primates (e.g. humans), non-human primates (e.g. cynomolgus monkeys) and rodents (e.g. mice and rats), unless otherwise indicated. The term encompasses “full-length,” unprocessed CD20 as well as any form of CD20 that results from processing in the cell. The term also encompasses naturally occurring variants of CD20, e.g., splice variants or allelic variants. In some aspects, CD20 is human CD20. Human CD20 is described in UniProt accession no. Pl 1836 (entry version 200), and an amino acid sequence of human CD20 is also shown in SEQ ID NO: 24.

[0280] Useful T cell bispecific antibodies for the present invention that bind to CD20 are described e.g. in PCT publication no. WO 2016 / 020309.

[0281] In some aspects, the T cell bispecific antibody comprises a first antigen binding moiety that binds to CD3, and a second antigen binding moiety that binds to CD20.

[0282] In some aspects, the first antigen binding moiety comprises a heavy chain variable region comprising the heavy chain CDR (HCDR) 1 of SEQ ID NO: 4, the HCDR2 of SEQ ID NO: 5, and the HCDR3 of SEQ ID NO: 6; and a light chain variable region comprising the light chain CDR (LCDR) 1 of SEQ ID NO: 7, the LCDR2 of SEQ ID NO: 8 and the LCDR3 of SEQ ID NO: 9.

[0283] In some aspects, the second antigen binding moiety comprises a heavy chain variable region comprising the heavy chain CDR (HCDR) 1 of SEQ ID NO: 12, the HCDR2 of SEQ ID NO: 13, and the HCDR3 of SEQ ID NO: 14; and a light chain variable region comprising the light chain CDR (LCDR) 1 of SEQ ID NO: 15, the LCDR2 of SEQ ID NO: 16 and the LCDR3 of SEQ ID NO: 17. In some aspects, the T cell bispecific antibody comprises

[0284] (i) a first antigen binding moiety that binds to CD3 and comprises a heavy chain variable region comprising the heavy chain CDR (HCDR) 1 of SEQ ID NO: 4, the HCDR2 of SEQ ID NO: 5, and the HCDR3 of SEQ ID NO: 6; and a light chain variable region comprising the light chain CDR (LCDR) 1 of SEQ ID NO: 7, the LCDR2 of SEQ ID NO: 8 and the LCDR3 of SEQ ID NO: 9; and

[0285] (ii) a second antigen binding moiety that binds to CD20 and comprises a heavy chain variable region comprising the heavy chain CDR (HCDR) 1 of SEQ ID NO: 12, the HCDR2 of SEQ ID NO: 13, and the HCDR3 of SEQ ID NO: 14; and a light chain variable region comprising the light chain CDR (LCDR) 1 of SEQ ID NO: 15, the LCDR2 of SEQ ID NO: 16 and the LCDR3 of SEQ ID NO: 17.

[0286] In some aspects, the first antigen binding moiety comprises a heavy chain variable region sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 10 and a light chain variable region sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 11. In some aspects, the first antigen binding moiety comprises the heavy chain variable region sequence of SEQ ID NO: 10 and the light chain variable region sequence of SEQ ID NO: 11.

[0287] In some aspects, the second antigen binding moiety comprises a heavy chain variable region sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 18 and a light chain variable region sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 19. In some aspects, the second antigen binding moiety comprises the heavy chain variable region sequence of SEQ ID NO: 18 and the light chain variable region sequence of SEQ ID NO: 19.

[0288] In some aspects, the T cell bispecific antibody comprises a third antigen binding moiety that binds to CD20 and / or an Fc domain composed of a first and a second subunit, as described herein.

[0289] In preferred aspects, the T cell bispecific antibody comprises

[0290] (i) a first antigen binding moiety that binds to CD3, comprising a heavy chain variable region comprising the heavy chain CDR (HCDR) 1 of SEQ ID NO: 4, the HCDR2 of SEQ ID NO: 5, and the HCDR3 of SEQ ID NO: 6; and a light chain variable region comprising the light chain CDR (LCDR) 1 of SEQ ID NO: 7, the LCDR2 of SEQ ID NO: 8 and the LCDR3 of SEQ ID NO: 9, wherein the first antigen binding moiety is a crossover Fab molecule wherein either the variable or the constant regions, particularly the variable regions, of the Fab light chain and the Fab heavy chain are exchanged;

[0291] (ii) a second and a third antigen binding moiety that bind to CD20, comprising a heavy chain variable region comprising the heavy chain CDR (HCDR) 1 of SEQ ID NO: 12, the HCDR2 of SEQ ID NO: 13, and the HCDR3 of SEQ ID NO: 14; and a light chain variable region comprising the light chain CDR (LCDR) 1 of SEQ ID NO: 15, the LCDR2 of SEQ ID NO: 16 and the LCDR3 of SEQ ID NO: 17, wherein the second and third antigen binding moiety are each a Fab molecule, particularly a conventional Fab molecule;

[0292] (iii) an Fc domain composed of a first and a second subunit, wherein the second antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the Fab heavy chain of the first antigen binding moiety, and the first antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the first subunit of the Fc domain, and wherein the third antigen binding moiety is fused at the C-terminus of the Fab heavy chain to the N-terminus of the second subunit of the Fc domain.

[0293] In some aspects, the first antigen binding moiety of the T cell bispecific antibody (that binds to CD20 and CD3) is a crossover Fab molecule wherein the variable regions of the Fab light chain and the Fab heavy chain are exchanged, and wherein the second and (where present) third antigen binding moiety of the T cell bispecific antibody is a conventional Fab molecule wherein in the constant domain CL the amino acid at position 124 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat) and the amino acid at position 123 is substituted independently by lysine (K), arginine (R) or histidine (H) (numbering according to Kabat) and in the constant domain CHI the amino acid at position 147 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index) and the amino acid at position 213 is substituted independently by glutamic acid (E), or aspartic acid (D) (numbering according to Kabat EU index).

[0294] Particularly, in the above aspects, in the constant domain CL of the second and the third Fab molecule under (ii) the amino acid at position 124 may be substituted by lysine (K) (numbering according to Kabat) and the amino acid at position 123 may be substituted by lysine (K) or arginine (R), particularly by arginine (R) (numbering according to Kabat), and in the constant domain CHI of the second and the third Fab molecule under (ii) the amino acid at position 147 may be substituted by glutamic acid (E) (numbering according to Kabat EU index) and the amino acid at position 213 may be substituted by glutamic acid (E) (numbering according to Kabat EU index). In some aspects, the first antigen binding moiety of the T cell bispecific antibody (that binds to CD20 and CD3) comprises a heavy chain variable region sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 10 and a light chain variable region sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 11. In some aspects, the first antigen binding moiety comprises the heavy chain variable region sequence of SEQ ID NO: 10 and the light chain variable region sequence of SEQ ID NO: 11.

[0295] In some aspects, the second and (where present) third antigen binding moiety of the T cell bispecific antibody (that binds to CD20 and CD3) comprise a heavy chain variable region sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 18 and a light chain variable region sequence that is at least about 95%, 96%, 97%, 98%, 99% or 100% identical to the amino acid sequence of SEQ ID NO: 19. In some aspects, the second and (where present) third antigen binding moiety comprise the heavy chain variable region of SEQ ID NO: 18 and the light chain variable region of SEQ ID NO: 19.

[0296] The Fc domain according to the above aspects may incorporate, singly or in combination, all of the features described hereinabove in relation to Fc domains.

[0297] In some aspects, the Fc domain of the T cell bispecific antibody (that binds to CD20 and CD3) comprises a modification promoting the association of the first and the second subunit of the Fc domain, and / or the Fc domain comprises one or more amino acid substitution that reduces binding to an Fc receptor and / or effector function.

[0298] In some aspects, the antigen binding moieties and the Fc region are fused to each other by peptide linkers, particularly by peptide linkers as in SEQ ID NO: 21 and SEQ ID NO: 23.

[0299] In some aspects, the T cell bispecific antibody (that binds to CD20 and CD3) comprises a polypeptide (particularly two polypeptides) comprising a sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 20, a polypeptide comprising a sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 21, a polypeptide comprising a sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 22, and a polypeptide comprising a sequence that is at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical to the sequence of SEQ ID NO: 23. In some aspects, the T cell bispecific antibody (that binds to CD20 and CD3) comprises a polypeptide (particularly two polypeptides) comprising the sequence of SEQ ID NO: 20, a polypeptide comprising the sequence of SEQ ID NO: 21, a polypeptide comprising the sequence of SEQ ID NO: 22, and a polypeptide comprising the sequence of SEQ ID NO: 23.

[0300] In preferred aspects, the T cell bispecific antibody is glofitamab (WHO Drug Information (International Nonproprietary Names for Pharmaceutical Substances), Recommended INN: List 83, 2020, vol. 34, no. 1, p. 39).

[0301] In some aspects, the disease (to be treated by the T cell engaging agent) is cancer.

[0302] As used herein, “treatment” (and grammatical variations thereof such as “treat” or “treating”) refers to clinical intervention in an attempt to alter the natural course of a disease in the individual being treated, and can be performed either for prophylaxis or during the course of clinical pathology. Desirable effects of treatment include, but are not limited to, preventing occurrence or recurrence of disease, alleviation of symptoms, diminishment of any direct or indirect pathological consequences of the disease, preventing metastasis, decreasing the rate of disease progression, amelioration or palliation of the disease state, and remission or improved prognosis.

[0303] The term “cancer” refers to the physiological condition in mammals that is typically characterized by unregulated cell proliferation. Examples of cancer include but are not limited to, carcinoma, lymphoma, blastoma, sarcoma and leukemia. More non-limiting examples of cancers include haematological cancer such as leukemia, bladder cancer, brain cancer, head and neck cancer, pancreatic cancer, biliary cancer, thyroid cancer, lung cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, endometrial cancer, esophageal cancer, colon cancer, colorectal cancer, rectal cancer, gastric cancer, prostate cancer, skin cancer, squamous cell carcinoma, sarcoma, bone cancer, and kidney cancer. Other cell proliferation disorders include, but are not limited to neoplasms located in the: abdomen, bone, breast, digestive system, liver, pancreas, peritoneum, endocrine glands (adrenal, parathyroid, pituitary, testicles, ovary, thymus, thyroid), eye, head and neck, nervous system (central and peripheral), lymphatic system, pelvic, skin, soft tissue, spleen, thoracic region, and urogenital system. Also included are pre-cancerous conditions or lesions and cancer metastases.

[0304] In some aspects, the cancer is a cancer expressing the target cell antigen of the T cell engaging agent (e.g., the T cell bispecific antibody). In some aspects, the cancer is a target cell antigen-expressing cancer. By “target cell antigenpositive cancer” or “target cell antigen-expressing cancer” is meant a cancer characterized by expression or overexpression of the target cell antigen on cancer cells. The expression of the target cell antigen may be determined for example by an immunohistochemistry (IHC) or flow cytometric assay. In some aspects, the cancer expresses the target cell antigen. In some aspects, the cancer expresses the target cell antigen in at least 20%, preferably at least 50% or at least 80% of tumor cells as determined by immunohistochemistry (IHC) using an antibody specific for the target cell antigen.

[0305] In some aspects, the cancer is colon cancer, lung cancer, ovarian cancer, gastric cancer, bladder cancer, pancreatic cancer, endometrial cancer, breast cancer, kidney cancer, esophageal cancer, prostate cancer, or other cancers described herein.

[0306] In particular aspects, the cancer is a cancer selected from the group consisting of colorectal cancer, lung cancer, pancreatic cancer, breast cancer, and gastric cancer. In preferred aspects, the cancer is colorectal cancer (CRC). In some aspects, the colorectal cancer is metastatic colorectal cancer (mCRC). In some aspects, the colorectal cancer is microsatellite-stable (MSS) colorectal cancer. In some aspects, the colorectal cancer is microsatellite-stable metastatic colorectal cancer (MSS mCRC).

[0307] In some aspects, the cancer is a CD20-expressing cancer (in particular in aspects, wherein the target cell antigen of the T cell engaging agent, e.g. T cell bispecific antibody, is CD20). By “CD20-positive cancer” or “CD20-expressing cancer” is meant a cancer characterized by expression or overexpression of CD20 in cancer cells. The expression of CD20 may be determined for example by quantitative real-time PCR (measuring CD20 mRNA levels), flow cytometry, immunohistochemistry (IHC) or western blot assays. In some aspects, the cancer expresses CD20. In some aspects, the cancer expresses CD20 in at least 20%, preferably at least 50% or at least 80% of tumor cells as determined by immunohistochemistry (IHC) using an antibody specific for CD20.

[0308] In some aspects, the cancer is a B-cell cancer, particularly a CD20-positive B-cell cancer. In some aspects, the cancer is selected from the group consisting of Non-Hodgkin lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle-cell lymphoma (MCL), marginal zone lymphoma (MZL), Multiple myeloma (MM) or Hodgkin lymphoma (HL). In particular aspects, the cancer is selected from the group consisting of Non-Hodgkin lymphoma (NHL), acute lymphocytic leukemia (ALL), chronic lymphocytic leukemia (CLL), diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), mantle-cell lymphoma (MCL) and marginal zone lymphoma (MZL). In more particular aspects, the cancer is NHL, particularly relapsed / refractory (r / r) NHL. In some aspects, the cancer is DLBCL. In some aspects, the cancer is FL. In some aspects, the cancer is MCL. In some aspects, the cancer is MZL.

[0309] In some aspects, the cancer is a cancer selected from the group consisting of lung cancer, head and neck cancer, bladder cancer, esophageal cancer, skin cancer, gastric cancer and ovarian cancer.

[0310] In some aspects, the cancer is treatable by the T cell engaging agent. In some aspects, the T cell engaging agent is indicated for the treatment of the cancer.

[0311] In some aspects, the cancer is a solid tumor cancer. By a “solid tumor cancer” is meant a malignancy that forms a discrete tumor mass (including also tumor metastasis) located at specific location in the patient’s body, such as sarcomas or carcinomas (as opposed to e.g. blood cancers such as leukemia, which generally do not form solid tumors). Non-limiting examples of solid tumor cancers include bladder cancer, brain cancer, head and neck cancer, pancreatic cancer, lung cancer, breast cancer, ovarian cancer, uterine cancer, cervical cancer, endometrial cancer, esophageal cancer, colon cancer, colorectal cancer, rectal cancer, gastric cancer, prostate cancer, skin cancer, squamous cell carcinoma, bone cancer, liver cancer and kidney cancer. Other solid tumor cancers that are contemplated in the context of the present invention include, but are not limited to neoplasms located in the: abdomen, bone, breast, digestive system, liver, pancreas, peritoneum, endocrine glands (adrenal, parathyroid, pituitary, testicles, ovary, thymus, thyroid), eye, head and neck, nervous system (central and peripheral), lymphatic system, pelvic, skin, soft tissue, muscles, spleen, thoracic region, and urogenital system. Also included are pre-cancerous conditions or lesions and cancer metastases.

[0312] An “individual” or “subject” herein is a mammal. Mammals include, but are not limited to, domesticated animals (e.g. cows, sheep, cats, dogs, and horses), primates (e.g. humans and nonhuman primates such as monkeys), rabbits, and rodents (e.g. mice and rats). In certain aspects, the individual or subject is a human. In some aspects, the individual has a disease, particularly a disease treatable or to be treated by the T cell engaging agent. In some aspects, the individual has cancer, particularly a cancer treatable or to be treated by the T cell engaging agent. In particular, an individual herein is any single human subject eligible for treatment who is experiencing or has experienced one or more signs, symptoms, or other indicators of cancer. In some aspects, the individual has cancer or has been diagnosed with cancer, in particular any of the cancers described hereinabove. In some aspects, the individual has locally advanced or metastatic cancer or has been diagnosed with locally advanced or metastatic cancer. The individual may have been previously treated with a T cell engaging agent (e.g. a T cell bispecific antibody) or another drug, or not so treated. In particular aspects, the patient has not been previously treated with a T cell engaging agent (e.g. a T cell bispecific antibody). The patient may have been treated with a therapy comprising one or more drugs other than T cell engaging agent (e.g. other than a T cell bispecific antibody) before the T cell engaging agent therapy is commenced.

[0313] In some aspects, the individual has an elevated serum level of one of more cytokine. In some aspects, said elevated serum level is related to the administration of the T cell engaging agent to the individual. Said elevated serum level is in particular as compared to the serum level in a healthy individual, and / or the serum level in an individual (including the same individual) without administration of the T cell engaging agent (i.e. in such case the serum level is elevated as compared to the serum level without administration of the T cell engaging agent). In some aspects, said one or more cytokine is selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-ip. In some aspects, the individual has CRS and / or shows one or more symptoms of fever, hypotension and hypoxia.

[0314] In some aspects, the individual has ICANS and / or shows one or more symptoms of mild tremor, confusion, disorientation, headache, attention deficits, agitation, seizures, cerebral oedema, hesitancy of speech, deterioration in handwriting, aphasia with expressive and / or receptive components, status epilepticus, fatal cerebral oedema, intracerebral haemorrhage and transient coma.

[0315] In some aspects, the individual has one or more of thrombocytopenia, an elevated serum level of one or more of lactate dehydrogenase, ferritin, IFN-y, IL- 10, granzyme B, GM-CSF, MIP-la, TNF, IL-2, and an elevated level in the cerebrospinal fluid of one or more of white blood cells, T cells, CD 14+ cells, SI 00b, glial fibrillary acidic protein, protein, IFN-y, IL- 10, IL-6 and granzyme B. In some aspects, said state is related to the administration of the T cell engaging agent to the individual. Said state is in particular as compared to the respective state in a healthy individual, and / or the respective state in an individual (including the same individual) without administration of the T cell engaging agent (i.e. in such case the respective state is elevated as compared to the state without administration of the T cell engaging agent). A cytokine according to any of the aspects of the invention may be one or more cytokine selected from the group consisting of interleukin (IL)-6, interferon (IFN)-y, IL-10, tumor necrosis factor (TNF)-a, granulocyte-macrophage colony-stimulating factor (GM-CSF), monocyte chemoattractant protein (MCP)-l, IL-ip, IL-8, IL-4 and IL-2. In some aspects, the cytokine is one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a, GM-CSF, MCP-1 and IL-ip. In some aspects, the cytokine is one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10, TNF-a and GM-CSF. In some aspects, the cytokine is one or more cytokine selected from the group consisting of IL-6, IFN-y, IL-10 and TNF-a. In some aspects, the cytokine is one or more cytokine selected from the group consisting of IL-6, IFN-y and IL- 10. In some aspects, the cytokine is IL-6. In some aspects, the cytokine is IFN-y. In some aspects, the cytokine is IL-10. In some aspects, the cytokine is TNF-a. In some aspects, the cytokine is GM-CSF. In some aspects, the cytokine is MCP-1. In some aspects, the cytokine is IL- ip. In some aspects, the cytokine is IL-8. In some aspects, the cytokine is IL-4. In some aspects, the cytokine is IL-2.

[0316] Preferably, a T cell according to any of the aspects of the invention is a cytotoxic T cell. In some aspects, the T cell is a CD4+or a CD8+T cell. In some aspects, the T cell is a CD8+T cell. In some aspects, the T cell is a CD4+T cell.

[0317] In some aspects, the treatment with or administration of the T cell engaging agent may result in a response in the individual. In some aspects, the response may be a complete response. In some aspects, the response may be a sustained response after cessation of the treatment. In some aspects, the response may be a complete response that is sustained after cessation of the treatment. In other aspects, the response may be a partial response. In some aspects, the response may be a partial response that is sustained after cessation of the treatment. In some aspects, the treatment with or administration of the T cell engaging agent and the TKI and GC may improve the response as compared to treatment with or administration of the T cell engaging agent alone (i.e. without the TKI and GC). In some aspects, the treatment or administration of the T cell engaging agent and the TKI and GC may increase response rates in a patient population, as compared to a corresponding patient population treated with the T cell engaging agent alone (i.e. without the TKI and GC).

[0318] The T cell engaging agent may be used alone or together with other agents in a therapy. For instance, a T cell engaging agent may be co-administered with at least one additional therapeutic agent. In certain aspects, an additional therapeutic agent is an anti-cancer agent, e.g. a chemotherapeutic agent, an inhibitor of tumor cell proliferation, or an activator of tumor cell apoptosis.

[0319] The TKI and GC may be used alone or together with one or more other agents for the inhibition of an adverse effect related to the administration of the T cell engaging agent.

[0320] Amino Acid Sequences

[0321] Examples

[0322] The following are examples of methods and compositions of the invention. It is understood that various other aspects may be practiced, given the general description provided above.

[0323] Example 1. Effect of the TKI and GC combination on TCB-induced cytokine release, T cell activation and target cell killing.

[0324] To assess whether a TKI and a GC can synergize to mitigate TCB-induced cytokine release and how this combination affects T cell cytotoxicity, we conducted killing assays using peripheral blood mononuclear cells (PBMCs), CTV-labeled Toledo tumor cells and escalating doses of

[0325] CD20-TCB (glofitamab) in media supplemented with 10 nM dexamethasone, 50 nM JAK inhibitor (ruxolitinib), 10 mTOR inhibitor (sirolimus), 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib) or 10 nM dexamethasone + 10 nM mTOR inhibitor (sirolimus) for 15-18 hrs (Figure 1).

[0326] At assay endpoint (15-18 hrs), cells were stained using a live dead NIR stain to assess the effects of the combination of dexamethasone with JAK (ruxolitinib) / mTOR (sirolimus) inhibitor on CD20-TCB-mediated CTV-labeled Toledo cell killing by exclusion of NIR positive dead cells using flow cytometry. The single treatment of 10 nM dexamethasone, 50 nM JAK inhibitor (ruxolitinib) and 10 nM mTOR inhibitor (sirolimus) did not interfere with Toledo cell killing by CD20-TCB, as indicated by the dose response curve (Figure 2A) and the EC50 values of N=4 donors (Figure 2B). The combination of 10 nM dexamethasone with 50 nM ruxolitinib or 10 nM sirolimus minimally interfered with Toledo cell killing by CD20-TCB, as indicated by the dose response curve (Figure 2A) and the increase in EC50 values of N=4 donors (Figure 2B).

[0327] Expression of CD69 (early T cell activation marker) and CD25 (late T cell activation marker) was also measured on CD8+ and CD4+ T cells by flow cytometry as a readout for T cell activation. The single treatment of 10 nM dexamethasone and 10 nM mTOR inhibitor (sirolimus) or the combination of both did not prevent the expression of the early activation marker CD69 on either CD8+ or CD4+ T cells (Figure 3 and 5) while the single treatment of 50 nM JAK inhibitor (ruxolitinib) and the combination with 10 nM dexamethasone partially prevented it. In addition, the expression of the late activation marker CD25 on both CD4+ and CD8+ T cells was most impacted by the single treatment of 50 nM JAK inhibitor (ruxolitinib) and the combination with 10 nM dexamethasone (Figure 4 and 6).

[0328] Lastly the levels of cytokines were measured by Legendplex in the assay supernatants to determine the impact of the different mitigation strategies on CD20-TCB-mediated cytokine release. In the presence of the single treatment of 10 nM dexamethasone, 10 nM mTOR inhibitor (sirolimus) or 10 nM JAK inhibitor (ruxolitinib), the release of IL-2 (Figure 7), TNF-a (Figure 8), IFN-y (Figure 9) and IL-6 (Figure 10) was suboptimally decreased. When combined with 10 nM dexamethasone, treatment with 10 nM mTOR inhibitor (sirolimus) or 10 nM JAK inhibitor (ruxolitinib) induced a stronger cytokine reduction, highlighting the synergistic effect of both dexamethasone and mTOR (sirolimus) or JAK inhibitor (ruxolitinib) in reducing IL-2 (Figure 7), TNF-a (Figure 8), IFN-y (Figure 9) and IL-6 (Figure 10). Due to the blocking of the IL-2R signaling, treatment with the JAK inhibitor (ruxolitinib) led to an accumulation of IL-2 (Figure 7) in the supernatant. When combined with dexamethasone, the effects on IL-2 are mainly driven by the latter.

[0329] It was thus demonstrated that dexamethasone synergizes with both the JAK inhibitor (ruxolitinib) and the mTOR inhibitor (sirolimus) to mitigate TCB-induced cytokine release while minimally interfering with T cell activation and tumor cell killing, suggesting that it could be used as a CRS mitigation approach.

[0330] Example 2. Comparison of TKI and dexamethasone combination at subclinical concentrations to clinical dexamethasone concentrations.

[0331] To compare the effect of the JAK or mTOR inhibitor and dexamethasone combination at subclinical concentrations to the effect of dexamethasone concentrations corresponding to clinical exposure, we conducted killing assays using peripheral blood mononuclear cells (PBMCs), CTV- labeled Toledo tumor cells and escalating doses of CD20-TCB (glofitamab) in media supplemented with 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib), 10 nM dexamethasone + 10 nM mTOR inhibitor (sirolimus), 305 nM dexamethasone (corresponding to Cmax concentrations after administering 20mg I.V. in patients) and 153 nM (corresponding to Cmin concentrations after administering 20mg I.V. in patients) for 15-18 hrs (Figure 1). In this assay, the Cmin and Cmax concentrations of dexamethasone were used as a minimum and maximum gate for the effects of the clinical exposure of dexamethasone.

[0332] At assay endpoint (15-18 hrs), cells were stained using a live dead NIR stain to assess the effects of the combination of the JAK (ruxolitinib) or mTOR (sirolimus) inhibitor with dexamethasone to the effect of clinical dexamethasone concentrations on CD20-TCB-mediated killing of CTV- labeled Toledo by exclusion of NIR positive dead cells using flow cytometry. 10 nM dexamethasone + 10 nM mTOR inhibitor (sirolimus) and 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib) retained Toledo cell killing comparably to Cmin dexamethasone concentration (153nM) and better than Cmax dexamethasone concentration (305nM) as indicated by the dose response curve and EC50 values of N=4 donors (Figure 11A & B).

[0333] Expression of CD69 (early T cell activation marker) and CD25 (late T cell activation marker) was also measured on CD8+ and CD4+ T cells by flow cytometry as a readout for T cell activation. 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib) minimally reduced CD69 and CD25 expression on CD4+ and CD8+ T cells compared to Cmax concentrations of dexamethasone while 10 nM dexamethasone + 50 nM mTOR inhibitor (sirolimus) retained T cell activation, comparably to Cmin / Cmax dexamethasone concentration (Figure 12-15).

[0334] Lastly, the levels of IL-2, TNF-a, IFN-y and IL-6 were measured in the assay supernatant by Legendplex. The combination of 10 nM dexamethasone + 10 nM mTOR inhibitor (sirolimus) reduced the levels of CD20-TCB -induced IL-2 (Figure 16), TNF-a (Figure 17), IFN-y (Figure 18) and IL-6 (Figure 19) release comparably to Cmin / Cmax dexamethasone concentrations. The combination of 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib) reduced the levels of CD20-TCB-induced TNF-a (Figure 17), IFN-y (Figure 18) and IL-6 (Figure 19) release, comparably to Cmin / Cmax dexamethasone concentrations. Due to the blocking of the IL-2R signaling, the combination of dexamethasone with the JAK inhibitor (ruxolitinib) led to a smaller inhibition of IL-2 release (Figure 16).

[0335] It was thus demonstrated that subclinical amounts of dexamethasone combined with subclinical amounts of ruxolitinib or sirolimus are comparable in effect to current clinical amounts of dexamethasone.

[0336] Example 3. Effect of the TKI and dexamethasone combination when mimicking CRS treatment in vitro.

[0337] To assess whether the JAK or mTOR inhibitor combination with dexamethasone is not only suitable for preventing TCB-induced cytokine release but also for treating a cytokine release that had already been induced, pre-activated PBMCs were treated with dexamethasone alone or in combination with TKI (JAK and mTOR inhibitors). In this assay, PBMCs were pre-activated on CTV-labeled Toledo tumor cells with CD20-TCB (glofitamab) in the absence of any mitigations for 15-18 hrs. At 15-18 hrs, the assay medium was supplemented with 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib), 10 nM dexamethasone + 10 nM mTOR inhibitor (sirolimus), 305 nM dexamethasone (corresponding to Cmax concentrations after administering 20mg I V. in patients) to assess the effects of the different treatments in further reducing CD20-TCB-mediated cytokine release while retaining further tumor cell killing and T cell activation (Figure 20).

[0338] Before (15-18 hrs) and after (40 hrs) the addition of Cmax dexamethasone concentrations, 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib), 10 nM dexamethasone + 10 nM mTOR inhibitor (sirolimus), cells were stained using a live dead NIR stain to assess CD20-TCB-mediated killing of CTV-labeled Toledo by exclusion of NIR positive dead cells using flow cytometry. Similarly to the Cmax dexamethasone concentration, the combination of dexamethasone and mTOR (sirolimus) / JAK (ruxolitinib) inhibitors did not prevent further tumor cell killing between 15-18 hrs and 40 hrs (Figure 21).

[0339] Expression of CD69 (early T cell activation marker) and CD25 (late T cell activation marker) was also measured on CD8+ and CD4+ T cells by flow cytometry as a readout for T cell activation. As shown by the levels of CD25 and CD69 levels on CD4+ and CD8+ T cells at 15-18 hrs (in absence of mitigations) and 40 hrs (in presence of mitigations), the combination of dexamethasone and mTOR (sirolimus) / JAK (ruxolitinib) inhibitor did not interfere with further T cell activation by pre-activated PBMCs, similarly to the Cmax dexamethasone concentrations (Figure 22-24). Of note, the expression of CD69 (early activation marker) is peaking at 15-18 hrs and then down- regulated from 15-18 hrs to 40 hrs while the expression of CD25 (late activation marker) is upregulated on CD4+ and CD8+ T cells from 15-18 hrs to 40 hrs.

[0340] Lastly, the level of IL-6 (monocyte-derived cytokine) was measured in the assay supernatant by Legendplex (Figure 26). As indicated by the IL-6 levels measured at 15 hrs (absence of mitigations) and the levels measured at 40 hrs (presence of mitigations), the combination of dexamethasone with mTOR (sirolimus) or JAK (ruxolitinib) inhibitors stopped further release of IL-6 by pre-activated PBMCs, similarly to Cmax dexamethasone concentrations.

[0341] It was thus demonstrated that the combination of dexamethasone and the JAK inhibitor (ruxolitinib) or the mTOR inhibitor (sirolimus) is suitable to reduce cytokine secretion from pre-activated PBMCs while retaining T cell activation and tumor cell killing to the same extent as clinical concentrations of dexamethasone.

[0342] Example 4. Duration of the effect of the TKI and GC combination.

[0343] To assess if the combination of dexamethasone and JAK / mTOR inhibitor has a lasting effect beyond the initial stimulation, PBMCs were first stimulated on CTV-labeled Toledo tumor cells with escalating doses of CD20-TCB (glofitamab) in the presence of 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib), 10 nM dexamethasone + 10 nM mTOR inhibitor (sirolimus) or 305 nM dexamethasone (corresponding to Cmax concentrations after administering 20mg I.V. in patients) for 72 hrs after which they were restimulated on fresh CFSE-labeled Toledo cells in the absence of any mitigations for 18 hrs.

[0344] After the first (72 hrs) and second (90 hrs) stimulation, cells were stained using a live dead NIR stain to assess CD20-TCB-mediated killing of CTV- (used in the first stimulation) and CFSE- labeled (used in the second stimulation) Toledo cells by exclusion of NIR positive dead cells using flow cytometry. As shown by the killing of CTV- and CFSE-labeled Toledo cells, the treatment with dexamethasone and JAK (ruxolitinib) / mTOR (sirolimus) inhibitor during the first stimulation did not prevent further Toledo cell killing when removed during the second stimulation, similarly to Cmax dexamethasone concentrations (Figure 28).

[0345] The levels of IL-6 were measured in the assay supernatant by Legendplex after the first (72 hrs) and second (90 hrs) stimulation (Figure 29). When present during the first stimulation, the combination of dexamethasone and JAK (ruxolitinib) / mTOR (sirolimus) inhibitor greatly reduced the levels of CD20-TCB-mediated IL-6 release, similarly to Cmax dexamethasone concentrations (Figure 29). During the second stimulation in the absence of treatment with dexamethasone and JAK (ruxolitinib) / mTOR (sirolimus) inhibitors, the levels of IL-6 remained low in the assay supernatant similarly to Cmax dexamethasone concentration, showing the lasting effects of these mitigations in reducing IL-6 release over different treatment cycle (Figure 29).

[0346] It was thus demonstrated that the combination of dexamethasone and the JAK inhibitor (ruxolitinib) or the mTOR inhibitor (sirolimus) has a lasting effect on reducing cytokine release beyond the first TCB stimulation, when PBMCs are restimulated with the TCB in the absence of the combination. Moreover, there is still no negative impact on tumor cell killing when PBMCs are restimulated with the TCB in the absence of the combination.

[0347] Example 5. Effect of the TKI and GC combination on endothelial cell activation.

[0348] To assess whether the combination of dexamethasone and JAK / mTOR inhibitor can prevent endothelial cell activation to the same extent as current clinical amounts of dexamethasone, human umbilical vein endothelial cells (HUVEC) were incubated with cytokine-enriched supernatants from co-cultures of PBMCs and Toledo cells with 100 pM CD20-TCB (glofitamab) in the presence and absence of 10 nM dexamethasone + 50 nM JAK inhibitor (ruxolitinib), 10 nM dexamethasone + 10 nM mTOR inhibitor (sirolimus) and 305 nM dexamethasone (corresponding to Cmax concentrations after administering 20mg I V. in patients) (Figure 30). Media from the same cocultures in the absence of any treatment was included as a negative control and recombinant TNF- u was used as a positive control for HUVEC activation.

[0349] After a 5 hrs incubation, ICAM and VCAM expression were measured on HUVEC by flow cytometry to assess their activation state in the presence of cytokine-enriched supernatants. The combination of dexamethasone with mTOR (sirolimus) or JAK (ruxolitinib) inhibitor induced a stronger prevention of ICAM and VCAM upregulation HUVEC than the single treatment of JAK (ruxolitinib) or mTOR (sirolimus) inhibitor, which was comparable to the effects of Cmax dexamethasone concentrations (Figure 31).

[0350] It was thus demonstrated that subclinical amounts of dexamethasone combined with JAK (ruxolitinib) or mTOR (sirolimus) inhibitor can prevent endothelial cell activation to the same extent as the current clinical amounts of dexamethasone. By preventing endothelial cell activation, the combination of dexamethasone with JAK or mTOR inhibitor could prevent the risk of ICANs in addition to preventing CRS

[0351] Example 6. Effect of the TKI and GC combination on cytokine release induced by CD20- TCB in a humanized diffuse large B cell lymphoma model in humanized NSG mice.

[0352] To assess whether a JAK or mTOR inhibitor can synergize with dexamethasone to further reduce cytokine release in vivo, WSU-bearing BRG47-S humanized mice were treated with CD20-TCB (glofitamab) as a monotherapy or in combination with different schedules of dexamethasone and combinations of dexamethasone and a JAK or mTOR inhibitor. There were six different treatment groups (Group A-E; Figure 32):

[0353] Group A was administered vehicle only on day 14 and all other groups, i.e. Groups B-E, were administered 0.15 mg / kg CD20-TCB on day 14. In addition, Group C was administered 4 mg / kg dexamethasone one hour prior to administration of CD20-TCB on day 14, Group D was administered 4 mg / kg dexamethasone 24 hours before, i.e. on day 13, one hour before, i.e. on day 14, and 24 hours after, i.e. on day 15, administration of CD20-TCB on day 14, Group E was administered 60 mg / kg ruxolitinib 24 hours before, i.e. on day 13, one hour before, i.e. on day 14, and 24 hours after, i.e. on day 15, administration of CD20-TCB on day 14, on top of 4 mg / kg dexamethasone administered one hour before treatment with CD20-TCB on day 14, and Group F was administered 2 mg / kg sirolimus 24 hours before, i.e. on day 13, and 24 hours after, i.e. on day 15, administration of CD20-TCB on day 14, on top of 4 mg / kg dexamethasone administered one hour before treatment with CD20-TCB on day 14.

[0354] Cytokine levels were measured in the peripheral blood 4 hrs and 24 hrs post treatment with CD20- TCB to assess the effects of the different mitigation strategies on cytokine release. 4 hrs post treatment with CD20-TCB, all mitigations reduced the levels of IFN-y (Figure 33), TNF-a (Figure 34), IL-6 (Figure 36), MCP-1 (Figure 37) and partially reduced the levels of IL-2 (Figure 35). At 24 hrs, one single pre-treatment with dexamethasone (group C) was not sufficient to reduce the levels of the aforementioned cytokines. When the JAK inhibitor (ruxolitinib) or the mTOR (sirolimus) were added on top of the pretreatment with dexamethasone (group E and F), this resulted in a complete reduction of IFN-y (Figure 33), IL-6 (Figure 36), MCP-1 (Figure 37) and a stronger reduction of TNF-a (Figure 34), comparable to giving two pre-treatment and one posttreatment dose of dexamethasone (group D). For IL-2 (Figure 35), only the combination of the mTOR inhibitor (sirolimus) with dexamethasone resulted in a complete reduction at 24 hrs, comparable to giving two pre-treatment and one post-treatment dose of dexamethasone. Due to the blocking of the IL-2R signaling, the combination of dexamethasone and the JAK inhibitor (ruxolitinib) could have led to an accumulation of IL-2 (Figure 35).

[0355] This demonstrates that dexamethasone can synergize with JAK or mTOR inhibitors to further reduce TCB-mediated cytokine release in a diffuse large B cell lymphoma model in humanized NSG mice, thus also mitigating cytokine release in vivo.

[0356] * * *

[0357] Although the foregoing invention has been described in some detail by way of illustration and example for purposes of clarity of understanding, the descriptions and examples should not be construed as limiting the scope of the invention. The disclosures of all patent and scientific literature cited herein are expressly incorporated in their entirety by reference.

Claims

-n-Claims1. A T cell engaging agent for use in the treatment of a disease in an individual, wherein said treatment comprises the administration of the following agents to the individual(a) the T cell engaging agent,(b) a tyrosine kinase inhibitor (TKI), and(c) a glucocorticoid (GC).

2. The T cell engaging agent for use according to claim 1, wherein the administration of the TKI and the GC is for the inhibition of an adverse effect related to the administration of the T cell engaging agent.

3. A TKI and a GC for use in the inhibition of an adverse effect related to the administration of a T cell engaging agent to an individual, wherein said inhibition comprises the administration of the TKI and GC to the individual.

4. The T cell engaging agent or TKI and GC for use according to any one of claims 1 to 3, wherein the TKI is an inhibitor of JAK or mTOR.

5. The T cell engaging agent or TKI and GC for use according to any one of claims 1 to 4, wherein the TKI is an mTOR inhibitor, optionally selected from the group consisting of sirolimus, temsirolimus and everolimus.

6. The T cell engaging agent or TKI and GC for use according to any one of claims 1 to 4, wherein the TKI is a JAK inhibitor, optionally a JAK1 and / or JAK2 inhibitor, optionally selected from the group consisting of ruxolitinib, baricitinib, tofacitinib or fedratinib.

7. The T cell engaging agent or TKI and GC for use according to any one of claims 1 to 6, wherein the GC is selected from dexamethasone and methylprednisolone.

8. The T cell engaging agent or TKI and GC for use according to any one of claims 2 to 7, wherein the adverse effect is one or more effect selected from the group consisting of(i) cytokine secretion by immune cells, particularly by T cells;(ii) cytokine release syndrome (CRS);(iii) one or more of fever, hypotension and hypoxia;(iv) an elevated serum level of one of more cytokine, particularly one or more cytokine selected from the group consisting of IL-6, IFN-y, IL- 10, TNF-a, GM-CSF, MCP-1 and IL-1P;-78-(v) immune effector cell-associated neurotoxicity syndrome (ICANS);(vi) one or more of mild tremor, confusion, disorientation, headache, attention deficits, agitation, seizures, cerebral oedema, hesitancy of speech, deterioration in handwriting, aphasia with expressive and / or receptive components, status epilepticus, fatal cerebral oedema, intracerebral haemorrhage and transient coma;(vii) one or more of thrombocytopenia, an elevated serum level of one or more of lactate dehydrogenase, ferritin, IFN-y, IL-10, granzyme B, GM-CSF, MIP-la, TNF, IL-2, and an elevated level in the cerebrospinal fluid of one or more of white blood cells, T cells, CD14+ cells, SlOOb, glial fibrillary acidic protein, protein, IFN-y, IL- 10, IL-6 and granzyme B; and(viii) T cell exhaustion.

9. The T cell engaging agent or TKI and GC for use according to any one of claims 1 to 8, wherein administration of the TKI and GC does not cause inhibition of a desired effect related to the administration of the T cell engaging agent.

10. The T cell engaging agent or TKI and GC for use according to claim 9, wherein the desired effect is one or more effect selected from the group consisting of(i) activation of T cells;(ii) cytotoxic activity of T cells; and(iii) the therapeutic effect of the T cell engaging agent.

11. The T cell engaging agent or TKI and GC for use according to any one of claims 1 to 10, wherein the administration of the TKI and GC is for the inhibition of an adverse effect related to the administration of the T cell engaging agent, wherein the adverse effect is CRS and / or ICANS, wherein administration of the TKI and GC does not cause inhibition of a desired effect related to the administration of the T cell engaging agent, and wherein the desired effect is the therapeutic effect of the T cell engaging agent.

12. The T cell engaging agent or TKI and GC for use according to any one of claims 2 to 11, wherein administration of the TKI and GC is upon or after manifestation of the adverse effect.

13. The T cell engaging agent or TKI and GC for use according to any one of claims 2 to 11, wherein administration of the TKI and GC is before manifestation of the adverse effect.

14. The T cell engaging agent or TKI and GC for use according to any one of claims 1 to 13, wherein administration of the TKI and GC is associated with the first administration of the T cell-79- engaging agent, and optionally is prior, concurrent or subsequent to the first administration of the T cell engaging agent.

15. The T cell engaging agent or TKI and GC for use according to any one of claims 1 to 14, wherein the T cell engaging agent is a T cell bispecific antibody or a CAR-T cell.

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