American Society of Hematology

Figure 2.

$Figure 2. Among human leukocytes, arginase I is specifically expressed in PMNs. (A) Arginase activity is found in the PMN fraction of human peripheral blood leukocytes. Leukocytes of healthy human blood donors were separated into PBMCs (□) and PMNs (▪). Mean arginase activity of 31 PMN preparations was 1644 ± 423 mU/mg protein and of 25 PBMC preparations 26 ± 52 mU/mg protein (P < .0001 for comparison between PMNs and PBMCs; 19 of 25 PBMC preparations showed no arginase activity). (B) Arginase I is specifically expressed in FACS-purified human PMNs. PBMCs and PMNs were FACS-purified according to forward/side-scatter characteristics. The highly pure populations (purity > 99.5%) were analyzed by immunoblotting. Results shown are representative of 3 independent experiments. (C) Absence of arginase I protein and arginase activity in PMNs of a patient with arginase I deficiency. Purified PMNs of an 18-year-old man with arginase I-deficiency (ARGI-/-) and a control healthy blood donor (ARGI+/+) were lysed and analyzed by immunoblotting. Two experiments yielded identical results. In panels B and C, the results of parallel determinations of arginase activities (in mU/mg protein) in aliquots of the same cell lysates are noted above the respective lanes of the arginase blots. To control for equal protein loading, Western blots for ERK1/2 were done in parallel.$

Among human leukocytes, arginase I is specifically expressed in PMNs. (A) Arginase activity is found in the PMN fraction of human peripheral blood leukocytes. Leukocytes of healthy human blood donors were separated into PBMCs (□) and PMNs (▪). Mean arginase activity of 31 PMN preparations was 1644 ± 423 mU/mg protein and of 25 PBMC preparations 26 ± 52 mU/mg protein (P < .0001 for comparison between PMNs and PBMCs; 19 of 25 PBMC preparations showed no arginase activity). (B) Arginase I is specifically expressed in FACS-purified human PMNs. PBMCs and PMNs were FACS-purified according to forward/side-scatter characteristics. The highly pure populations (purity > 99.5%) were analyzed by immunoblotting. Results shown are representative of 3 independent experiments. (C) Absence of arginase I protein and arginase activity in PMNs of a patient with arginase I deficiency. Purified PMNs of an 18-year-old man with arginase I-deficiency (ARGI^-/-) and a control healthy blood donor (ARGI^+/+) were lysed and analyzed by immunoblotting. Two experiments yielded identical results. In panels B and C, the results of parallel determinations of arginase activities (in mU/mg protein) in aliquots of the same cell lysates are noted above the respective lanes of the arginase blots. To control for equal protein loading, Western blots for ERK1/2 were done in parallel.

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