Primary intestinal epithelial cells selectively transfer R5 HIV-1 to CCR5+ cells

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Abstract

The upper gastrointestinal tract is a principal route of HIV-1 entry in vertical transmission and after oral–genital contact. The phenotype of the newly acquired virus is predominantly R5 (CCR5-tropic) and not X4 (CXCR4-tropic), although both R5 and X4 viruses are frequently inoculated onto the mucosa. Here we show that primary intestinal (jejunal) epithelial cells express galactosylceramide, an alternative primary receptor for HIV-1, and CCR5 but not CXCR4. Moreover, we show that intestinal epithelial cells transfer R5, but not X4, viruses to CCR5+ indicator cells, which can efficiently replicate and amplify virus expression. Transfer was remarkably efficient and was not inhibited by the fusion blocker T-20, but was substantially reduced by colchicine and low (4 °C) temperature, suggesting endocytotic uptake and microtubule-dependent transcytosis of HIV-1. Our finding that CCR5+ intestinal epithelial cells select and transfer exclusively R5 viruses indicates a mechanism for the selective transmission of R5 HIV-1 in primary infection acquired through the upper gastrointestinal tract.

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Figure 1: Characteristics of primary IECs isolated by neutral protease from jejunal mucosa.
Figure 2: IECs express GalCer and CCR5, but not CXCR4.
Figure 3: IECs do not express SDF-1.
Figure 4: IECs preferentially transfer R5 virus to indicator cells.
Figure 5: Direct relationship between the duration of IEC–indicator-cell contact and HIV-1 production.
Figure 6: Sequence of gp41 and V3 regions of indicator-cell output virus was identical to input BaL but not IIIB HIV-1.

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Acknowledgements

This study was supported NIH grants and contracts DK-47322, DE-72621, AI-41530, HD-41361, AI-35467, CA-73470, AI-28147, DK34151; the Central AIDS Virus Core of the Birmingham Center for AIDS Research (P30-AI-27767); and the Research Service of the Department of Veterans Affairs.

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Correspondence to Phillip D. Smith.

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