Lysozyme and RNases as anti-HIV components in β-core preparations of human chorionic gonadotropin

Lysozyme and RNases as anti-HIV components in β-core preparations of human chorionic gonadotropin

^*Department of Biochemistry, New York University School of Medicine, New York, NY 10016; ^‡Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114; ^§Harvard Community Health Plan and Harvard Medical School, Boston, MA 02114; ^¶Laboratory of Biochemical Physiology, National Cancer Institute-Frederick Cancer Research and Development Center, Frederick, MD 21701; and ^**Contraception and Reproductive Health Branch, and ^‡‡Endocrinology and Reproduction Research Branch, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892

Communicated by Bernhard Witkop, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD (received for review December 16, 1998)

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Figure 1
(A) HPLC separation of hCG β-core fragment, human urinary AVL, and AVR from a crude β-core preparations. The elution profile of a representative reverse-phase liquid chromatogram of 2.5 mg run is shown. Details are described in Methods. (B) Characterization of AVL and AVR. SDS/PAGE was carried out on crude β-core (Sephadex G100 gel chromatography fractionated), fractions P1-P7 of C18 reverse-phase HPLC, pure β-core, AVL, AVR, lysozyme, and RNase. Each lane contains about 5–10 μg of sample. The presence (+) or absence (−) of anti-HIV, lysozyme, and RNase activities are shown under the SDS/PAGE pattern. Proteins smaller than 11 kDa were not separated in this SDS/PAGE system.
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Figure 2
AVR and AVL are active in HIV-1 inhibition. Dose–response anti-HIV activities of AVL, AVR, pure β-core, chicken egg white lysozyme, and bovine pancreatic RNase were assayed by viral production (p24 ELISA) in (A) ACH2 lymphocytes and (B) U1 monocytes. Each data point is the mean of triplicate. Identical dose–response anti-HIV activity was obtained for human milk lysozyme and human neutrophil lysozyme (data not shown).
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Figure 3
RNase activity of AVR. (A) RNase activity of AVR was assayed by degradation of total RNA isolated from HIV-infected ACH2 lymphocytes. AVR demonstrated potent RNase activity comparable to that of bovine pancreatic RNase A, whereas pure β-core and AVL showed no RNase activity. (B) Dose-dependent RNase activity of AVR.