Anti-Mouse IgG2b monoclonal antibody, clone TB75g

Background: Allergen-specific immunotherapy (AIT) is the only causal treatment for allergy. However, success rates vary depending on the type of allergy and disease background of the patient. Hence, strategies targeting an increased therapeutic efficacy are urgently needed. Here, the effects of blockade of IL-4 and IL-13 signaling on different phases of AIT were addressed. Methods: The impact of the recombinantly produced IL-4 and IL-13 antagonist IL-4 mutein (IL-4M) on allergic sensitization and AIT outcome in experimental allergic asthma were analyzed in a murine model. The effects of IL-4M administration were assessed prior/during sensitization, immediately after AIT under allergen challenge, and two weeks post-treatment. Results: Intervention with IL-4M prior/during sensitization led to strong induction of IgG1, IgG2a, IgG2b, and IgG3, decrease of specific and total IgE, as well as of IL-5 in serum. Similar effects on the serum immunoglobulin levels were observed immediately after IL4M-supplemented AIT during allergen challenge. Additionally, IL4M markedly suppressed type-2 cytokine secretion of splenocytes beyond the effect of AIT alone. These effects were equaled to those of AIT alone two weeks post-treatment. Intriguingly, here, IL-4M induced a sustained decrease of Th2-biased Tregs (ST2(+) FOXP3(+)GATA3(intermediate)). Conclusions: IL-4 and IL-13 blockade during experimental AIT demonstrates beneficial effects on immunological key parameters such as immunoglobulin and cytokine secretion immediately after AIT. Although two weeks later these effects were dropped to those of AIT alone, the number of potentially disease-triggering Th2-biased Tregs was further significantly decreased by IL-4M treatment. Hence, IL-4/IL13-targeting therapies prime the immune memory in therapy success-favoring manner.

beta(2)-Microglobulin (beta 2M), the light chain of the class I major histocompatibilty complex (MHC-I), is a promising tumor target for monoclonal antibodies (mAbs) in cancer immunotherapy. Several reports indicate that chelation of cell-associated beta 2M by specific mouse mAbs promotes tumor cell destruction by inducing apoptosis or other cytotoxic signaling pathways. Human mAbs employed in cancer therapy are usually IgG1, which mediates cell-killing by effector mechanisms including complement dependent cytotoxicity (CDC). The analogous mouse IgG2a and IgG2b isotypes are similarly effective in activating complement. Therefore, we examined the complement-activating properties of anti-beta 2M mouse mAbs 1B749 (IgG2a) and HB28 (IgG2b) when either mAb was bound to tumor cell lines or normal cells; we compared these beta 2M-specific mAbs with mouse mAb W6/32 (IgG2a), specific for human leukocyte antigens in the MHC-I heavy chain. All three mAbs bind to most human cell lines and normal cells in approximately equal amounts, consistent with a 1:1 stoichiometry for the HLA heavy chain in association with beta 2M. The three mAbs promote rapid C3b deposition and substantial CDC of human cell lines, and mAbs 1B749 and W6/32 have robust cytotoxic activity on reaction with normal mononuclear cells and platelets. Curiously, mAb HB28 induces modest C3b deposition and little CDC of normal cells, and its weaker complement-fixing activity was confirmed by ELISA. Based on these findings, we suggest that human IgG mAbs that target beta 2M for cancer immunotherapy be selected or engineered so as not to activate complement, thus eliminating the potential adverse effects of complement-mediated lysis of normal cells. (C) 2013 Elsevier Ltd. All rights reserved.