Use of a replication-defective vector to track cells initially infected by SIV in vivo

Infected mononuclear cells rapidly appear in the draining lymph node after intradermal inoculation of rhesus monkeys

Yichuan Wang, Steven S. Kim, Ding Lu, Juan You Xue, Steven Joye, Hung Fan, Chris J Miller

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

A better understanding of the mechanisms of HIV dissemination, a key step in pathogenesis, would be possible if the cellular pathways of viral dissemination could be followed in simian immunodeficiency virus (SIV)- inoculated monkeys or HIV-infected people. In an initial attempt to follow this process using a traceable virus infection, we inoculated rhesus monkeys intradermally (ID) or directly into lymph nodes with a replication- defective SIV-based vector expressing the enhanced jellyfish green fluorescent protein (EGFP), V1EGFP. EGFP expression was detected in mononuclear cells isolated from the sites of inoculation (skin and lymph node) at 5 and 16 hr after inoculation and then cultured in vitro for 6 days to allow maximum EGFP expression. Similarly, EGFP-expressing, SIV-infected cells could be detected at 16 hr postinfection in the lymph nodes that drained the sites of ID inoculation. Since V1EGFP is a replication-defective vector, the EGFP-expressing cells are the initial target cells infected by the virions in the original inoculum. The results of flow cytometric analysis were confirmed by a nested PCR assay to detect SIV DNA and hence infection of cells and reverse transcription. These experiments indicate that 16 hr after ID inoculation newly infected cells either remain in the skin at the site of inoculation or have migrated to the draining lymph node. The results in this SIV vector model probably reflect the short time (less than 16 hr) required for HIV to move from a site of epithelial penetration to the lymphoid tissues via lymphatic vessels.

Original languageEnglish (US)
Pages (from-to)1298-1305
Number of pages8
JournalAIDS Research and Human Retroviruses
Volume20
Issue number12
StatePublished - Dec 2004

Fingerprint

Simian Immunodeficiency Virus
Macaca mulatta
Lymph Nodes
HIV
Defective Viruses
Skin
Lymphatic Vessels
Lymphoid Tissue
Virus Diseases
Virion
Reverse Transcription
Haplorhini
enhanced green fluorescent protein
Polymerase Chain Reaction
DNA
Infection

ASJC Scopus subject areas

  • Immunology
  • Virology

Cite this

Use of a replication-defective vector to track cells initially infected by SIV in vivo : Infected mononuclear cells rapidly appear in the draining lymph node after intradermal inoculation of rhesus monkeys. / Wang, Yichuan; Kim, Steven S.; Lu, Ding; Xue, Juan You; Joye, Steven; Fan, Hung; Miller, Chris J.

In: AIDS Research and Human Retroviruses, Vol. 20, No. 12, 12.2004, p. 1298-1305.

Research output: Contribution to journalArticle

@article{42e3de5ee6234fe691cd70b315627ce4,
title = "Use of a replication-defective vector to track cells initially infected by SIV in vivo: Infected mononuclear cells rapidly appear in the draining lymph node after intradermal inoculation of rhesus monkeys",
abstract = "A better understanding of the mechanisms of HIV dissemination, a key step in pathogenesis, would be possible if the cellular pathways of viral dissemination could be followed in simian immunodeficiency virus (SIV)- inoculated monkeys or HIV-infected people. In an initial attempt to follow this process using a traceable virus infection, we inoculated rhesus monkeys intradermally (ID) or directly into lymph nodes with a replication- defective SIV-based vector expressing the enhanced jellyfish green fluorescent protein (EGFP), V1EGFP. EGFP expression was detected in mononuclear cells isolated from the sites of inoculation (skin and lymph node) at 5 and 16 hr after inoculation and then cultured in vitro for 6 days to allow maximum EGFP expression. Similarly, EGFP-expressing, SIV-infected cells could be detected at 16 hr postinfection in the lymph nodes that drained the sites of ID inoculation. Since V1EGFP is a replication-defective vector, the EGFP-expressing cells are the initial target cells infected by the virions in the original inoculum. The results of flow cytometric analysis were confirmed by a nested PCR assay to detect SIV DNA and hence infection of cells and reverse transcription. These experiments indicate that 16 hr after ID inoculation newly infected cells either remain in the skin at the site of inoculation or have migrated to the draining lymph node. The results in this SIV vector model probably reflect the short time (less than 16 hr) required for HIV to move from a site of epithelial penetration to the lymphoid tissues via lymphatic vessels.",
author = "Yichuan Wang and Kim, {Steven S.} and Ding Lu and Xue, {Juan You} and Steven Joye and Hung Fan and Miller, {Chris J}",
year = "2004",
month = "12",
language = "English (US)",
volume = "20",
pages = "1298--1305",
journal = "AIDS Research and Human Retroviruses",
issn = "0889-2229",
publisher = "Mary Ann Liebert Inc.",
number = "12",

}

TY - JOUR

T1 - Use of a replication-defective vector to track cells initially infected by SIV in vivo

T2 - Infected mononuclear cells rapidly appear in the draining lymph node after intradermal inoculation of rhesus monkeys

AU - Wang, Yichuan

AU - Kim, Steven S.

AU - Lu, Ding

AU - Xue, Juan You

AU - Joye, Steven

AU - Fan, Hung

AU - Miller, Chris J

PY - 2004/12

Y1 - 2004/12

N2 - A better understanding of the mechanisms of HIV dissemination, a key step in pathogenesis, would be possible if the cellular pathways of viral dissemination could be followed in simian immunodeficiency virus (SIV)- inoculated monkeys or HIV-infected people. In an initial attempt to follow this process using a traceable virus infection, we inoculated rhesus monkeys intradermally (ID) or directly into lymph nodes with a replication- defective SIV-based vector expressing the enhanced jellyfish green fluorescent protein (EGFP), V1EGFP. EGFP expression was detected in mononuclear cells isolated from the sites of inoculation (skin and lymph node) at 5 and 16 hr after inoculation and then cultured in vitro for 6 days to allow maximum EGFP expression. Similarly, EGFP-expressing, SIV-infected cells could be detected at 16 hr postinfection in the lymph nodes that drained the sites of ID inoculation. Since V1EGFP is a replication-defective vector, the EGFP-expressing cells are the initial target cells infected by the virions in the original inoculum. The results of flow cytometric analysis were confirmed by a nested PCR assay to detect SIV DNA and hence infection of cells and reverse transcription. These experiments indicate that 16 hr after ID inoculation newly infected cells either remain in the skin at the site of inoculation or have migrated to the draining lymph node. The results in this SIV vector model probably reflect the short time (less than 16 hr) required for HIV to move from a site of epithelial penetration to the lymphoid tissues via lymphatic vessels.

AB - A better understanding of the mechanisms of HIV dissemination, a key step in pathogenesis, would be possible if the cellular pathways of viral dissemination could be followed in simian immunodeficiency virus (SIV)- inoculated monkeys or HIV-infected people. In an initial attempt to follow this process using a traceable virus infection, we inoculated rhesus monkeys intradermally (ID) or directly into lymph nodes with a replication- defective SIV-based vector expressing the enhanced jellyfish green fluorescent protein (EGFP), V1EGFP. EGFP expression was detected in mononuclear cells isolated from the sites of inoculation (skin and lymph node) at 5 and 16 hr after inoculation and then cultured in vitro for 6 days to allow maximum EGFP expression. Similarly, EGFP-expressing, SIV-infected cells could be detected at 16 hr postinfection in the lymph nodes that drained the sites of ID inoculation. Since V1EGFP is a replication-defective vector, the EGFP-expressing cells are the initial target cells infected by the virions in the original inoculum. The results of flow cytometric analysis were confirmed by a nested PCR assay to detect SIV DNA and hence infection of cells and reverse transcription. These experiments indicate that 16 hr after ID inoculation newly infected cells either remain in the skin at the site of inoculation or have migrated to the draining lymph node. The results in this SIV vector model probably reflect the short time (less than 16 hr) required for HIV to move from a site of epithelial penetration to the lymphoid tissues via lymphatic vessels.

UR - http://www.scopus.com/inward/record.url?scp=11244338256&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=11244338256&partnerID=8YFLogxK

M3 - Article

VL - 20

SP - 1298

EP - 1305

JO - AIDS Research and Human Retroviruses

JF - AIDS Research and Human Retroviruses

SN - 0889-2229

IS - 12

ER -