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Table 2

Binding Epitopes of IL-4R–specific mAbs

	ELISA^*					Flow cytometry^‡		Epitope dependent on^§	Neutralization of IL-4 function^¶
	C57BL/6	BALB/c	C57BL/6- C34R-T49I	C57BL/6- M168T	C57BL/6- T49I	Flow cytometry^‡
	C57BL/6	BALB/c	C57BL/6- C34R-T49I	C57BL/6- M168T	C57BL/6- T49I	C57BL/6	BALB/c
Mouse mAb
999-31	Yes	No	No	Yes	Yes	Yes	No	Cys34	No
999-258	Yes	No	Yes	No	Yes	No	No	Met168	No
999-461	Yes	No	No	Yes	Yes	Yes	No	Cys34	No
999-643	Yes	No	Yes	No	Yes	No	No	Met168	No
999-707	Yes	No	Yes	No	Yes	No	No	Met168	No
999-927	Yes	No	Yes	No	Yes	Yes	No	Met168	No
999-1243	Yes	No	Yes	No	Yes	No	No	Met168	No
999-1366	Yes	No	No	Yes	Yes	Yes	No	Cys34	No
999-1378	Yes	No	Yes	No	Yes	No	No	Met168	No
Rat mAb
1046-957-14	Yes	Yes	Yes	Yes	Yes	Yes	Yes	—	No
1046-1658-13	Yes	Yes	Yes	Yes	Yes	No	No	—	No
M1	Yes	Yes^‖	Yes^‖	Yes	Yes^‖	Yes	Yes	Thr49^‖	Yes

	ELISA^*					Flow cytometry^‡		Epitope dependent on^§	Neutralization of IL-4 function^¶
	C57BL/6	BALB/c	C57BL/6- C34R-T49I	C57BL/6- M168T	C57BL/6- T49I	Flow cytometry^‡
	C57BL/6	BALB/c	C57BL/6- C34R-T49I	C57BL/6- M168T	C57BL/6- T49I	C57BL/6	BALB/c
Mouse mAb
999-31	Yes	No	No	Yes	Yes	Yes	No	Cys34	No
999-258	Yes	No	Yes	No	Yes	No	No	Met168	No
999-461	Yes	No	No	Yes	Yes	Yes	No	Cys34	No
999-643	Yes	No	Yes	No	Yes	No	No	Met168	No
999-707	Yes	No	Yes	No	Yes	No	No	Met168	No
999-927	Yes	No	Yes	No	Yes	Yes	No	Met168	No
999-1243	Yes	No	Yes	No	Yes	No	No	Met168	No
999-1366	Yes	No	No	Yes	Yes	Yes	No	Cys34	No
999-1378	Yes	No	Yes	No	Yes	No	No	Met168	No
Rat mAb
1046-957-14	Yes	Yes	Yes	Yes	Yes	Yes	Yes	—	No
1046-1658-13	Yes	Yes	Yes	Yes	Yes	No	No	—	No
M1	Yes	Yes^‖	Yes^‖	Yes	Yes^‖	Yes	Yes	Thr49^‖	Yes

ELISA data obtained with the different sIL-4R constructs as designated in Fig. 3. Supernatants of transfected 293-EBNA cells were tested.

‡

Flow cytometry was performed with transfected TF-1 cells expressing IL-4R allotypes.

Amino acids of the C57BL/6 IL-4R located in the binding epitopes of the mouse mAbs as judged by the ELISA measurements.

The neutralizing function of the mAbs was detected by their capability to inhibit the IL-4–induced proliferation of HT-2 cells.

‖

Binding of mAb M1 was present but the OD values in the ELISA were three-fold reduced as compared to the detection with a polyclonal antiserum raised against sIL-4R.

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