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Unformatted text preview: Discussion Questions for Liu et al. 1997 Mar. 3 rd 2009 1. Handling was previously shown to reduce stress hormone responses. What does handling do to the hypothalamus to induce this change? 2. What did the mothers of handled pups do that was different from the nonhandled pups? 3. There were three big differences in how the HPA axis responded to restraint stress in the high and low LG-ABN (maternal behavior) groups. What were they? of cells after exposure to the peptide. 14. T. G. Senkevich et al ., Science 273 , 813 (1996); M. R. MacDonald, X.-Y Li, H. W. Virgin IV, J. Virol. 71 , 1671 (1997). 15. R. Horuk, Trends Pharmacol. Sci. 15 , 159 (1994); P. M. Murphy, Annu. Rev. Immunol. 12 , 593 (1994); C. A. Power and T. N. C. Wells, Trends Pharmacol. Sci. 17 , 209 (1996). 16. G. M. Clore, E. A. Apella, M. Yamada, K. Matsu- shima, A. M. Gronenborn, J. Biol. Chem. 264 , 18907 (1989); N. J. Skelton, F. Aspiras, J. Ogez, T. J. Schall, Biochemistry 34 , 5329 (1995); C. Chung, R. M. Cooke, A. E. I. Proudfoot, T. N. C. Wells, ibid ., p. 9307. 17. M. Lusti-Narasimhan et al ., J. Biol. Chem. 271 , 3148 (1996). 18. E. Cesarman et al ., J. Virol. 70 , 8218 (1996); the HHV-8–encoded chemokine receptor ORF74 is de- posited in GenBank (accession number U24275). 19. L. Arvanitakis, E. Geras-Raaka, A. Varma, M. C. Ger- shengorn, E. Cesarman, Nature 385 , 347 (1997). 20. Y. Feng, C. C. Broder, P. E. Kennedy, E. A. Berger, Science 272 , 872 (1996); T. Drajic et al ., Nature 381 , 667 (1996); H. Deng et al ., ibid ., p. 661. 21. J. He et al ., Nature 385 , 645 (1997). 22. Actual binding of vMIP-II to CCR3 was not deter- mined because of the lack of success in establishing a valid binding assay with radioactively labeled eotaxin or MCP-3. However, the observation that pretreatment with vMIP-II could inhibit the calcium mobilizing effect of MCP-3 in cells expressing CCR3 indicates that vMIP-II does bind to CCR3 (Fig. 3A). 23. U87/CD4 cells expressing CCR3, CCR5, and CXCR4 were kindly provided by Dan Littman. Pe- ripheral blood mononuclear cells (PBMCs) were pre- pared from buffy coat white blood cells from blood banks. After separation by density-gradient centrifu- gation on Ficoll-Paque, cells were cultured and stim- ulated with PHA. After 2 days IL-2 was added and after 3 to 5 days cells were infected. The HIV-1 strains have been described ( 25, 26 ). Virus stocks were prepared in PBMC cultures stimulated with PHA and IL-2. Although the same tissue culture ID 50 (1900 for PBMC) of virus was used to challenge the cells, replication and infection were more efficient on the U87/CD4 cells expressing CXCR4 than on those expressing CCR3 or CCR5. However, lowering the amount of virus failed to substantially increase the inhibition by chemokines shown in Fig. 4 for the CXCR4-expressing cells. For determination of che- mokine inhibition of infectivity, cells were seeded into 96-well dishes at 4 10 3 cells per well. On the following day chemokines were added and incubat- ed for 30 min at 37°C before virus was added and...
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