188 BuHetfn of Experimental Biology and Medicine, Vol. 115, N o. 2, February, 1993 Effect of Synthetic Fragments of HIV Protein Immunodominant Sites on Human Neutrophil Oxygen Metabolism V. M. Zemskov,V. M. Drukh, S. M. Subbotin,S. M. Andreev, M. V. Sidorova,M. G. Vafma,D. E. Tsvetkov,and A. A. Vorob'ev UDC616.112.91-08 Translated from Byulleten' EksperunentaI'noi Biologii i NIeditsiny, Vol. 115, No~ 2, pp. 185- 187, February, 1993 Original article submitted July 7, 1992 Key Words: neutrophils; HJV; peptides; oxygen metabolism The important role played by neutrophilic dysfunc- tion in AIDS pathogenesis is now considered proven [1]. The purpose of our study was to reveal a possi- ble influence of HIV glycoprotein gp 120 and gp 41 fragments on spontaneous and zymosan-induced che- miluminescence (CL) of neutrophils, which reflects the intensity of a "breathing burst". In addition, the results will help predict the in vivo side effects of the tested peptides when used as vaccine components. MATERIALS AND METHODS Blood samples were colleGted from healthy donors and from patients with various chronic allergic dis- orders. The neutrophils were isolated by isotonic lysis followed by differential centrifugation [2]; all the procedures were carried out at 4~ The studied peptides (Table 1) were synthesized by the solid-phase method and used in aqueous solutions. Zymosan (Reakhim, USSR) was opsonized by a mix- ture of blood sera from several donors. Luminol (3• xl04M) in Hanks solution (0.5 ml per cuvette) was Laboratory of Nonspecific Immunity and Laboratory of Peptides, Institute of Immunology, Russian Ministry of Health; Microbiology Department of the I. M. Sechenov Medical Academy, Moscow TABLE 1. Peptides Used in the Study No. of peptide 337 480 24 411 477 22t 551 Correlation with natural fragment of HIV glycoprotein HIV strain BRU BRU MN BRU BRU BRU BRU qlycoprotein gpl20 gpl20 gp120 gp120 gp120 gp 41 gp 41 amino acid fragment 252-272 307-329 307-329 423-450 495-516 573-593 841-861 used as a CL stimulant. The neutrophil concentration was 1 million/ml; CL was determined at 37~ The investigation was carried out on an L-1251 lurninometer (LKB, Finland) in the following vari- ants: 1) the cells were added to the lurninometer cuvette already containing peptide in the required concentration in Hanks solution; 2) the same as 1), but the cells were preincubated at 37~ for 20 rain with subsequent cooling to 4~ to "prime" them; 3) the neutrophils were incubated for 20 rain with the peptide in phosphate-buffered normal saline at 4~ washed, and then added to the luminometer cuvette with Hanks solution and luminol; 4) the same as 3), but the cells were preheated at 37~ (see variant 2); 0007 -- 4888/93/0002 --018a$12.50 1993Plenum Publishing Corporation