{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,10]],"date-time":"2026-02-10T10:12:07Z","timestamp":1770718327837,"version":"3.49.0"},"reference-count":62,"publisher":"American Association for the Advancement of Science (AAAS)","issue":"5338","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Science"],"published-print":{"date-parts":[[1997,10,24]]},"abstract":"<jats:p>\n            CD8\n            <jats:sup>+<\/jats:sup>\n            T lymphocytes from individuals infected with human immunodeficiency virus\u2013type 1 (HIV-1) secrete a soluble activity that suppresses infection by HIV-1. A protein associated with this activity was purified from the culture supernatant of an immortalized CD8\n            <jats:sup>+<\/jats:sup>\n            T cell clone and identified as the \u03b2-chemokine macrophage-derived chemokine (MDC). MDC suppressed infection of CD8\n            <jats:sup>+<\/jats:sup>\n            cell\u2013depleted peripheral blood mononuclear cells by primary non\u2013syncytium-inducing and syncytium-inducing isolates of HIV-1 and the T cell line\u2013adapted isolate HIV-1\n            <jats:sub>IIIB<\/jats:sub>\n            . MDC was expressed in activated, but not resting, peripheral blood mononuclear cells and binds a receptor on activated primary T cells. These observations indicate that \u03b2-chemokines are responsible for a major proportion of HIV-1\u2013specific suppressor activity produced by primary T cells.\n          <\/jats:p>","DOI":"10.1126\/science.278.5338.695","type":"journal-article","created":{"date-parts":[[2002,7,27]],"date-time":"2002-07-27T09:35:26Z","timestamp":1027762526000},"page":"695-698","source":"Crossref","is-referenced-by-count":153,"title":["Inhibition of HIV-1 Infection by the \u03b2-Chemokine MDC"],"prefix":"10.1126","volume":"278","author":[{"given":"Ranajit","family":"Pal","sequence":"first","affiliation":[{"name":"R. Pal, P. D. Markham, M. Brown, Advanced BioScience Laboratories, 5510 Nicholson Lane, Kensington, MD 20895, USA."},{"name":"A. Garzino-Demo, R. C. Gallo, A. L. DeVico, Institute of Human Virology, University of Maryland at Baltimore, 725 West Lombard Street, Baltimore, MD 21201\u20131192, USA."},{"name":"J. Burns, Institute of Human Virology and Department of Microbiology and Immunology, University of Maryland School of Medicine, University of Maryland at Baltimore, 655 West Baltimore Street, Baltimore, MD 21201\u20131192, USA."}]},{"given":"Alfredo","family":"Garzino-Demo","sequence":"additional","affiliation":[{"name":"R. Pal, P. D. Markham, M. Brown, Advanced BioScience Laboratories, 5510 Nicholson Lane, Kensington, MD 20895, USA."},{"name":"A. Garzino-Demo, R. C. Gallo, A. L. DeVico, Institute of Human Virology, University of Maryland at Baltimore, 725 West Lombard Street, Baltimore, MD 21201\u20131192, USA."},{"name":"J. Burns, Institute of Human Virology and Department of Microbiology and Immunology, University of Maryland School of Medicine, University of Maryland at Baltimore, 655 West Baltimore Street, Baltimore, MD 21201\u20131192, USA."}]},{"given":"Phillip D.","family":"Markham","sequence":"additional","affiliation":[{"name":"R. Pal, P. D. Markham, M. Brown, Advanced BioScience Laboratories, 5510 Nicholson Lane, Kensington, MD 20895, USA."},{"name":"A. Garzino-Demo, R. C. Gallo, A. L. DeVico, Institute of Human Virology, University of Maryland at Baltimore, 725 West Lombard Street, Baltimore, MD 21201\u20131192, USA."},{"name":"J. Burns, Institute of Human Virology and Department of Microbiology and Immunology, University of Maryland School of Medicine, University of Maryland at Baltimore, 655 West Baltimore Street, Baltimore, MD 21201\u20131192, USA."}]},{"given":"Jennifer","family":"Burns","sequence":"additional","affiliation":[{"name":"R. Pal, P. D. Markham, M. Brown, Advanced BioScience Laboratories, 5510 Nicholson Lane, Kensington, MD 20895, USA."},{"name":"A. Garzino-Demo, R. C. Gallo, A. L. DeVico, Institute of Human Virology, University of Maryland at Baltimore, 725 West Lombard Street, Baltimore, MD 21201\u20131192, USA."},{"name":"J. Burns, Institute of Human Virology and Department of Microbiology and Immunology, University of Maryland School of Medicine, University of Maryland at Baltimore, 655 West Baltimore Street, Baltimore, MD 21201\u20131192, USA."}]},{"given":"Michelle","family":"Brown","sequence":"additional","affiliation":[{"name":"R. Pal, P. D. Markham, M. Brown, Advanced BioScience Laboratories, 5510 Nicholson Lane, Kensington, MD 20895, USA."},{"name":"A. Garzino-Demo, R. C. Gallo, A. L. DeVico, Institute of Human Virology, University of Maryland at Baltimore, 725 West Lombard Street, Baltimore, MD 21201\u20131192, USA."},{"name":"J. Burns, Institute of Human Virology and Department of Microbiology and Immunology, University of Maryland School of Medicine, University of Maryland at Baltimore, 655 West Baltimore Street, Baltimore, MD 21201\u20131192, USA."}]},{"given":"Robert C.","family":"Gallo","sequence":"additional","affiliation":[{"name":"R. Pal, P. D. Markham, M. Brown, Advanced BioScience Laboratories, 5510 Nicholson Lane, Kensington, MD 20895, USA."},{"name":"A. Garzino-Demo, R. C. Gallo, A. L. DeVico, Institute of Human Virology, University of Maryland at Baltimore, 725 West Lombard Street, Baltimore, MD 21201\u20131192, USA."},{"name":"J. Burns, Institute of Human Virology and Department of Microbiology and Immunology, University of Maryland School of Medicine, University of Maryland at Baltimore, 655 West Baltimore Street, Baltimore, MD 21201\u20131192, USA."}]},{"given":"Anthony L.","family":"DeVico","sequence":"additional","affiliation":[{"name":"R. Pal, P. D. Markham, M. Brown, Advanced BioScience Laboratories, 5510 Nicholson Lane, Kensington, MD 20895, USA."},{"name":"A. Garzino-Demo, R. C. Gallo, A. L. DeVico, Institute of Human Virology, University of Maryland at Baltimore, 725 West Lombard Street, Baltimore, MD 21201\u20131192, USA."},{"name":"J. Burns, Institute of Human Virology and Department of Microbiology and Immunology, University of Maryland School of Medicine, University of Maryland at Baltimore, 655 West Baltimore Street, Baltimore, MD 21201\u20131192, USA."}]}],"member":"221","reference":[{"key":"e_1_3_1_2_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.2431484"},{"key":"e_1_3_1_2_3","doi-asserted-by":"crossref","first-page":"2961","DOI":"10.4049\/jimmunol.144.8.2961","volume":"144","author":"Brinchman J. E.","year":"1990","unstructured":"Brinchman J. E., et al., J. Immunol. 144, 2961 (1990).","journal-title":"J. Immunol."},{"key":"e_1_3_1_3_2","doi-asserted-by":"crossref","first-page":"470","DOI":"10.1016\/0008-8749(89)90259-1","volume":"119","author":"Walker C. M.","year":"1989","unstructured":"Walker C. M., Moody D., Stites D. P., Levy J. A., Cell. Immunol. 119, 470 (1989);","journal-title":"Cell. Immunol."},{"key":"e_1_3_1_3_3","doi-asserted-by":"publisher","DOI":"10.1172\/JCI115153"},{"key":"e_1_3_1_3_4","doi-asserted-by":"crossref","first-page":"13125","DOI":"10.1073\/pnas.93.23.13125","volume":"93","author":"Blackbourn D. J.","year":"1996","unstructured":"Blackbourn D. J., et al., Proc. Natl. Acad. Sci. U.S.A. 93, 13125 (1996).","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_4_2","doi-asserted-by":"crossref","first-page":"1811","DOI":"10.1126\/science.270.5243.1811","volume":"270","author":"Cocchi F.","year":"1996","unstructured":"Cocchi F., et al., Science 270, 1811 (1996).","journal-title":"Science"},{"key":"e_1_3_1_5_2","doi-asserted-by":"crossref","first-page":"4476","DOI":"10.4049\/jimmunol.156.11.4476","volume":"156","author":"Barker T. D.","year":"1996","unstructured":"Barker T. D., Weissman D., Daucher J. A., Roche K., Fauci A., J. Immunol. 156, 4476 (1996);","journal-title":"J. Immunol."},{"key":"e_1_3_1_5_3","doi-asserted-by":"crossref","first-page":"1317","DOI":"10.1097\/00002030-199610000-00002","volume":"10","author":"Paliard X.","year":"1996","unstructured":"Paliard X., Lee A. Y., Walker C. M., AIDS 10, 1317 (1996);","journal-title":"AIDS"},{"key":"e_1_3_1_5_4","doi-asserted-by":"crossref","first-page":"14076","DOI":"10.1073\/pnas.93.24.14076","volume":"93","author":"Kinter A. L.","year":"1996","unstructured":"Kinter A. L., et al., Proc. Natl. Acad. Sci. U.S.A. 93, 14076 (1996);","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_5_5","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1089\/aid.1997.13.63","volume":"13","author":"Rubbert A.","year":"1997","unstructured":"Rubbert A., et al., AIDS Res. Hum. Retroviruses 13, 63 (1997).","journal-title":"AIDS Res. Hum. Retroviruses"},{"key":"e_1_3_1_6_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.93.26.15341"},{"key":"e_1_3_1_7_2","unstructured":"CD8 + T cell clones immortalized in vitro were prepared as previously described ["},{"key":"e_1_3_1_7_3","doi-asserted-by":"crossref","first-page":"413","DOI":"10.1002\/ijc.2910310404","volume":"31","author":"Markham P. D.","year":"1983","unstructured":"Markham P. D., et al., Int. J. Cancer 31, 413 (1983);","journal-title":"Int. J. Cancer"},{"key":"e_1_3_1_7_4","unstructured":"; ibid. 33 13 (1984)]. Briefly CD8 + T cells were isolated from HIV-1\u2013infected individuals by positive selection with magnetic beads coated with antibodies to CD8 (Dynal) and then activated with phytohemagglutinin (PHA) for 48 hours in complete medium (RPMI 1640 containing 15% fetal bovine serum 1% glutamine and 1% penicillin-streptomycin) supplemented with 10% IL-2. Cells were washed and exposed to HTLV-I by coculture at a ratio of 3:1 with an HTLV-I\u2013producing T cell line that had been subjected to irradiation with 80 Gy for 30 min to prevent cell proliferation. Cultures were maintained until clusters of immortalized cells were observed (usually after 2 to 4 weeks of exposure to HTLV-I). The individual clusters were then transferred into separate culture flasks. Rapidly proliferating cells were subjected to single-cell cloning by limiting dilution and then expanded in complete medium containing 10% IL-2. A CD8 + phenotype of the immortalized cell lines was verified by flow cytometry with antibodies to CD8 and to CD4 (Becton-Dickinson San Jose CA)."},{"key":"e_1_3_1_8_2","unstructured":"PBMCs from normal donors were activated with PHA for 48 hours and depleted of CD8 + T cells by negative selection with magnetic beads coated with antibodies to CD8 (Dynal). After culture for 18 hours in complete medium containing recombinant human IL-2 (rIL-2) (Gemini Biotech Woodland TX) at 16 ng\/ml the cells (1 \u00d7 10 6 ) were exposed to 250 TCID 50 (median tissue culture infectious dose) of the indicated HIV-1 isolates for 3 hours at 37\u00b0C. Cells were then washed and suspended in complete medium with rIL-2 and placed into 48-well plates (2 \u00d7 10 5 cells per well) with a 1:4 dilution of culture supernatant (or the dilution of chemokine indicated in other experiments) in a total volume of 250 \u03bcl. Control infections were performed in complete medium with rIL-2 alone. After 48 hours the medium in each well was replenished with 250 \u03bcl of fresh medium containing the same culture supernatant (or chemokine). The extent of infection was measured on either day 5 or 6 with an HIV-1 p24 ELISA (Organon Teknika Durham NC). The suppressive activity of each supernatant was calculated as the percentage of inhibition of HIV-1 p24 antigen production compared with controls. HIV-1 IIIB virus stock was prepared from chronically infected Molt3\u2013HIV-1 IIIB cell lines whereas the previously described primary NSI and SI isolates (8) ["},{"key":"e_1_3_1_8_3","doi-asserted-by":"crossref","first-page":"1772","DOI":"10.1128\/jvi.67.4.1772-1777.1993","volume":"67","author":"Connor R. I.","year":"1993","unstructured":"Connor R. I., et al., J. Virol. 67, 1772 (1993);","journal-title":"J. Virol."},{"key":"e_1_3_1_8_4","unstructured":"] were propagated in primary PBMCs. All isolates were titrated to determine TCID 50 in PHA-stimulated normal PBMCs."},{"key":"e_1_3_1_9_2","doi-asserted-by":"crossref","first-page":"621","DOI":"10.1084\/jem.185.4.621","volume":"185","author":"Connor R. I.","year":"1997","unstructured":"Connor R. I., et al., J. Exp. Med. 185, 621 (1997).","journal-title":"J. Exp. Med."},{"key":"e_1_3_1_10_2","doi-asserted-by":"crossref","first-page":"661","DOI":"10.1038\/381667a0","volume":"381","author":"Dragic T.","year":"1996","unstructured":"Dragic T., et al., Nature 381, 661 (1996);","journal-title":"Nature"},{"key":"e_1_3_1_10_3","doi-asserted-by":"publisher","DOI":"10.1126\/science.272.5270.1955"},{"key":"e_1_3_1_10_4","doi-asserted-by":"publisher","DOI":"10.1038\/381661a0"},{"key":"e_1_3_1_11_2","doi-asserted-by":"crossref","first-page":"15382","DOI":"10.1073\/pnas.93.26.15382","volume":"93","author":"Jansson M.","year":"1996","unstructured":"Jansson M., et al., Proc. Natl. Acad. Sci. U.S.A. 93, 15382 (1996).","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_12_2","unstructured":"F3B clone 19 cells were grown in complete medium containing rIL-2 (16 ng\/ml) at 37\u00b0C in a CO 2 incubator. After expanding the culture to 200 ml the cells were isolated by centrifugation and resuspended in RPMI medium containing HB101 (Irvine Scientific Santa Ana CA) and supplemented with rIL-2 (16 ng\/ml) 1% glutamine and 1% penicillin-streptomycin. The cells were grown to confluence and the medium was then harvested by centrifugation at 670 g for 10 min."},{"key":"e_1_3_1_13_2","unstructured":"Culture supernatant (1200 ml) from F3b clone 19 grown to high cell density in serum-free medium supplemented with rIL-2 (11) was subjected to centrifugation at 100 000 g for 60 min at 4\u00b0C and the resulting soluble fraction was applied to a 5-ml HiTrap heparin affinity FPLC (fast protein liquid chromatography) column (Pharmacia) that had been equilibrated with 10 mM tris-HCl (pH 7.6) containing 0.1 M NaCl (column buffer). The column was then washed extensively with column buffer after which the bound proteins were eluted with 10 mM tris-HCl (pH 7.6) containing 2.0 M NaCl at a flow rate of 1 ml\/min. The column eluate was adjusted to pH 2.0 by addition of trifluoroacetic acid (TFA) and subjected to reversed-phase HPLC on a PEEK C 18 column (Waters Instruments Millford MA) that had been equilibrated with H 2 O containing 0.1% TFA. Proteins bound to the column were eluted with a 5-min linear gradient of aqueous acetonitrile (0 to 35%) containing 0.1% TFA. After 10 min at 35% acetonitrile the column was further subjected to a 60-min linear gradient of 35 to 70% aqueous acetonitrile containing 0.1% TFA. The flow rate was maintained at 0.5 to 1 ml\/min. The resulting fractions were tested for suppressor activity in the infectivity assay with HIV-1 IIIB . Active fractions were pooled diluted twofold in H 2 O containing 0.1% TFA and reapplied to the column. Proteins were eluted with a 30-min linear gradient of aqueous acetonitrile (0 to 60%) containing 0.1% TFA at a flow rate of 0.5 to 1 ml\/min. The fractions obtained were assayed as above. Active fractions were pooled diluted with H 2 O containing 0.1% TFA and fractionated under the same conditions to obtain a single protein peak. The fraction corresponding to the peak and flanking fractions were tested in the infectivity assay to verify that suppressor activity cofractionated with the protein."},{"key":"e_1_3_1_14_2","doi-asserted-by":"crossref","first-page":"394","DOI":"10.1016\/S0960-9822(00)00088-9","volume":"4","author":"Witt D. P.","year":"1994","unstructured":"Witt D. P., Lander A. D., Curr. Biol. 4, 394 (1994);","journal-title":"Curr. Biol."},{"key":"e_1_3_1_14_3","doi-asserted-by":"crossref","first-page":"82","DOI":"10.1006\/meth.1996.0082","volume":"10","author":"Proost P.","year":"1996","unstructured":"Proost P., et al., Methods 10, 82 (1996).","journal-title":"Methods"},{"key":"e_1_3_1_15_2","unstructured":"A. L. DeVico and R. Pal unpublished data."},{"key":"e_1_3_1_16_2","unstructured":"Amino acid analysis of the purified protein was performed with a Beckman 6300 amino acid analyzer. Samples were hydrolyzed for 24 hours at 110\u00b0C in the presence of 6 M HCl and then reconstituted in loading buffer. Amino acids were analyzed by postcolumn derivatization with ninhydrin. NH 2 -terminal microsequencing was performed by automated Edman degradation with a Hewlett-Packard G1005A protein sequencing system which was operated with standard reagents solvents and programs (routine 3.0) supplied by the manufacturer. Abbreviations for the amino acid residues are A Ala; D Asp; E Glu; G Gly; L Leu; M Met; N Asn; P Pro; R Arg; S Ser; V Val; and Y Tyr."},{"key":"e_1_3_1_17_2","doi-asserted-by":"crossref","first-page":"1595","DOI":"10.1084\/jem.185.9.1595","volume":"185","author":"Godiska R.","year":"1997","unstructured":"Godiska R., et al., J. Exp. Med. 185, 1595 (1997).","journal-title":"J. Exp. Med."},{"key":"e_1_3_1_18_2","doi-asserted-by":"crossref","first-page":"9199","DOI":"10.1073\/pnas.85.23.9199","volume":"85","author":"Lindley I.","year":"1988","unstructured":"Lindley I., et al., Proc. Natl. Acad. Sci. U.S.A. 85, 9199 (1988);","journal-title":"Proc. Natl. Acad. Sci. U.S.A."},{"key":"e_1_3_1_18_3","doi-asserted-by":"crossref","first-page":"87","DOI":"10.1016\/0161-5890(89)90024-2","volume":"26","author":"Yoshimura T.","year":"1989","unstructured":"Yoshimura T., et al., Mol. Immunol. 26, 87 (1989);","journal-title":"Mol. Immunol."},{"key":"e_1_3_1_18_4","doi-asserted-by":"crossref","first-page":"337","DOI":"10.1111\/j.1432-1033.1989.tb14729.x","volume":"181","author":"VanDamme J.","year":"1989","unstructured":"VanDamme J., et al., Eur. J. Biochem. 181, 337 (1989);","journal-title":"Eur. J. Biochem."},{"key":"e_1_3_1_18_5","doi-asserted-by":"crossref","first-page":"3033","DOI":"10.4049\/jimmunol.145.9.3033","volume":"145","author":"Hebert C. A.","year":"1990","unstructured":"Hebert C. A., et al., J. Immunol. 145, 3033 (1990) .","journal-title":"J. Immunol."},{"key":"e_1_3_1_19_2","doi-asserted-by":"crossref","first-page":"449","DOI":"10.1084\/jem.171.2.449","volume":"171","author":"Walz A.","year":"1990","unstructured":"Walz A., Baggiolini M., J. Exp. Med. 171, 449 (1990);","journal-title":"J. Exp. Med."},{"key":"e_1_3_1_19_3","doi-asserted-by":"crossref","first-page":"2113","DOI":"10.1002\/eji.1830200933","volume":"20","author":"VanDamme J.","year":"1990","unstructured":"VanDamme J., et al., Eur. J. Immunol. 20, 2113 (1990);","journal-title":"Eur. J. Immunol."},{"key":"e_1_3_1_19_4","doi-asserted-by":"crossref","first-page":"23128","DOI":"10.1016\/S0021-9258(18)54472-0","volume":"266","author":"Clark-Lewis I.","year":"1991","unstructured":"Clark-Lewis I., Schumacher C., Baggiolini M., Moser B., J. Biol. Chem. 266, 23128 (1991).","journal-title":"J. Biol. Chem."},{"key":"e_1_3_1_20_2","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1002\/eji.1830230329","volume":"23","author":"Bischoff S. C.","year":"1993","unstructured":"Bischoff S. C., et al., Eur. J. Immunol. 23, 761 (1993);","journal-title":"Eur. J. Immunol."},{"key":"e_1_3_1_20_3","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1016\/S0065-2776(08)60509-X","volume":"55","author":"Baggiolini M.","year":"1994","unstructured":"Baggiolini M., Dewald B., Moser B., Adv. Immunol. 55, 97 (1994);","journal-title":"Adv. Immunol."},{"key":"e_1_3_1_20_4","doi-asserted-by":"crossref","first-page":"1727","DOI":"10.1126\/science.7569902","volume":"269","author":"Bacon K. B.","year":"1995","unstructured":"Bacon K. B., Premack B. A., Gardner P., Schall T. J., Science 269, 1727 (1995).","journal-title":"Science"},{"key":"e_1_3_1_21_2","unstructured":"Intracellular Ca 2+ was measured by flow cytometry according to a modification (J. Burns and G. Lewis Biotechniques in press) of previously described methods [R. Badolato et al. J. Immunol. 155 4004 (1995);"},{"key":"e_1_3_1_21_3","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1002\/(SICI)1097-0320(19960301)23:3<205::AID-CYTO4>3.0.CO;2-H","volume":"23","author":"Greimers R.","year":"1996","unstructured":"Greimers R., et al., Cytometry 23, 205 (1996);","journal-title":"Cytometry"},{"key":"e_1_3_1_21_4","unstructured":"]. Briefly unfractionated or CD8 + cell\u2013depleted PBMCs (1 \u00d7 10 6 cells\/ml) prepared as described (7) were cultured in the absence of IL-2 for 1 hour washed with RPMI 1640 (GIBCO BRL) containing 25 mM Hepes but no phenol red or sodium bicarbonate and resuspended in the same solution at a density of 2 \u00d7 10 7 cells\/ml. Cells (1 \u00d7 10 6 ) were then added to sample tubes loaded for 20 min at 37\u00b0C with 2 \u03bcM fluo-3 (Molecular Probes) reconstituted in a solution containing 20% Pluronic F-127 (Molecular Probes) and dimethyl sulfoxide and stained with 7-aminoactinomycin D (Molecular Probes) to discriminate dead cells ["},{"key":"e_1_3_1_21_5","doi-asserted-by":"crossref","first-page":"204","DOI":"10.1002\/cyto.990130216","volume":"13","author":"Schmid I.","year":"1992","unstructured":"Schmid I., et al., Cytometry 13, 204 (1992);","journal-title":"Cytometry"},{"key":"e_1_3_1_21_6","unstructured":"]. The samples were then washed once as before and resuspended in 1 ml of RPMI 1640 without phenol red and sodium bicarbonate. All samples were maintained at 20\u00b0C in the dark until 5 min before analysis at which time the sample tube was placed in a 37\u00b0C water bath. Cells were maintained at 37\u00b0C throughout data acquisition. Cells were stimulated by addition of test chemokine to a final concentration of 3 nM. Data were acquired with a FACSCalibur (Becton-Dickinson) flow cytometer with excitation at 488 nm. Cells were gated by forward- and side-scatter properties as well as by exclusion of 7-aminoactinomycin D fluorescence by use of emission above 650 nm in the FL-3 window. Calcium mobilization was determined by a two-parameter density plot of linear emissions collected at 530 nm in the FL-1 window for the gated cell population over time."},{"key":"e_1_3_1_22_2","unstructured":"PBMCs (American Red Cross) from a healthy donor were purified by centrifugation in Histopaque (Sigma) and harvested either immediately or after activation for 48 hours with PHA (5 \u03bcg\/ml) and rIL-2 (10 ng\/ml). The HUT 78 human T cell line was cultured in the presence (50 U\/ml) or absence of IL-2 (Boehringer Mannheim Germany). RNA was isolated by the RNAzol procedure (Tel-Test Friendswood TX) and 10 \u03bcg of total cellular RNA was separated by electrophoresis on a denaturing formaldehyde-agarose gel and then transferred to a nylon membrane. The membrane was subjected to hybridization with an MDC-specific probe and washed under stringent conditions as described ["},{"key":"e_1_3_1_22_3","doi-asserted-by":"crossref","first-page":"177","DOI":"10.1089\/hum.1995.6.2-177","volume":"6","author":"Garzino-Demo A.","year":"1995","unstructured":"Garzino-Demo A., Gallo R. C., Arya S. K., Hum. Gene Ther. 6, 177 (1995);","journal-title":"Hum. Gene Ther."},{"key":"e_1_3_1_22_4","unstructured":"]. The probe for Northern hybridizations was generated by reverse transcription and polymerase chain reaction with MDC-specific primers."},{"key":"e_1_3_1_23_2","unstructured":"A. Garzino-Demo A. DeVico R. Pal unpublished results."},{"key":"e_1_3_1_24_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.272.5263.872"},{"key":"e_1_3_1_24_3","doi-asserted-by":"publisher","DOI":"10.1016\/S0092-8674(00)81313-6"},{"key":"e_1_3_1_24_4","doi-asserted-by":"crossref","unstructured":"; B. J. Doranz et al. ibid. p. 1149; J. F. Berson et al. J. Virol. 70 6288 (1996).","DOI":"10.1128\/jvi.70.9.6288-6295.1996"},{"key":"e_1_3_1_25_2","doi-asserted-by":"crossref","first-page":"829","DOI":"10.1038\/382829a0","volume":"382","author":"Bluel C.","year":"1996","unstructured":"Bluel C., et al., Nature 382, 829 (1996);","journal-title":"Nature"},{"key":"e_1_3_1_25_3","unstructured":"; E. Oberlin et al. ibid. p. 833."},{"key":"e_1_3_1_26_2","first-page":"645","volume":"385","author":"He J.","year":"1997","unstructured":"He J., et al., ibid. 385, 645 (1997).","journal-title":"ibid."},{"key":"#cr-split#-e_1_3_1_27_2.1","doi-asserted-by":"crossref","unstructured":"L. Zhang Y. Huang T. He Y. Cao D. D. Ho ibid. 383 768 (1996)","DOI":"10.1038\/383768a0"},{"key":"#cr-split#-e_1_3_1_27_2.2","doi-asserted-by":"crossref","unstructured":"M. T. Dittmar et al. ibid. 385 495 (1997);","DOI":"10.1038\/385495a0"},{"key":"e_1_3_1_27_3","doi-asserted-by":"crossref","first-page":"8355","DOI":"10.1128\/jvi.70.12.8355-8360.1996","volume":"70","author":"Simmons G.","year":"1996","unstructured":"Simmons G., J. Virol. 70, 8355 (1996).","journal-title":"J. Virol."},{"key":"e_1_3_1_28_2","doi-asserted-by":"crossref","first-page":"1177","DOI":"10.1099\/0022-1317-68-4-1177","volume":"68","author":"Patterson S.","year":"1987","unstructured":"Patterson S., Knight S. C., J. Gen. Virol. 68, 1177 (1987);","journal-title":"J. Gen. Virol."},{"key":"e_1_3_1_28_3","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1038\/362359a0","volume":"362","author":"Embretson J.","year":"1993","unstructured":"Embretson J., et al., Nature 362, 359 (1993);","journal-title":"Nature"},{"key":"e_1_3_1_28_4","doi-asserted-by":"crossref","first-page":"2045","DOI":"10.1084\/jem.182.6.2045","volume":"182","author":"Pope M.","year":"1995","unstructured":"Pope M., et al., J. Exp. Med. 182, 2045 (1995).","journal-title":"J. Exp. Med."},{"key":"e_1_3_1_29_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.184.6.2433"},{"key":"e_1_3_1_30_2","unstructured":"We thank B. Lambe for technical assistance; A. Wiznia G. Lambert M. Rosenberg J. Dobroszycki and O. Cohen for clinical samples; N. Miller for helpful discussions and support for our search for new HIV-1\u2013suppressor chemokines; K. Swiderek for amino acid analysis and microsequencing; and R. Connor for primary virus isolates. R.P. M.B. and P.D.M. were supported in part by National Institute of Allergy and Infectious Diseases contract NO1-AI-55279 to Advanced BioScience Laboratories."}],"container-title":["Science"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.science.org\/doi\/pdf\/10.1126\/science.278.5338.695","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,1,13]],"date-time":"2024-01-13T04:39:06Z","timestamp":1705120746000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.science.org\/doi\/10.1126\/science.278.5338.695"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1997,10,24]]},"references-count":62,"journal-issue":{"issue":"5338","published-print":{"date-parts":[[1997,10,24]]}},"alternative-id":["10.1126\/science.278.5338.695"],"URL":"https:\/\/doi.org\/10.1126\/science.278.5338.695","relation":{},"ISSN":["0036-8075","1095-9203"],"issn-type":[{"value":"0036-8075","type":"print"},{"value":"1095-9203","type":"electronic"}],"subject":[],"published":{"date-parts":[[1997,10,24]]}}}