A vaccinia-based elispot assay for detection of CD8+T cells from HIV-1 infected children

被引：9

作者：

Buseyne, F

Catteau, A

Scott-Algara, D

Corre, B

Porrot, F

Rouzioux, C

Blanche, S

Rivière, Y

机构：

[1] Inst Pasteur, CNRS, URA 1930, Unite Postulante Immunopathol Virale, F-75015 Paris, France

[2] Inst Pasteur, Unite Biol Retrovirus, F-75015 Paris, France

[3] Univ Paris 05, CHU Necker Enfants Malad, EA 3620, Virol Lab, F-75015 Paris, France

[4] CHU Necker Enfants Malad, Fed Pediatrie, F-75015 Paris, France

来源：

JOURNAL OF IMMUNOLOGICAL METHODS | 2005年 / 298卷 / 1-2期

关键词：

perinatal HIV infection; cytotoxic T lymphocytes (CTL); elispot;

D O I：

10.1016/j.jim.2005.01.003

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

HIV-specific CD8+ T lymphocytes participate in the control of viral replication in infected patients. These responses are of low intensity in young infants and are decreased by antiretroviral therapy. In the present study, we report on a recombinant Vaccinia virus (rVV)-based Elispot assay for the detection of HIV-specific CD8+ T cells immediately after isolation of peripheral blood mononuclear cells (PBMC). The rVV-based assay was highly sensitive; 48 out of 50 children had a positive response against the rVV encoding HIV Env-Gag-Pol antigen. Interferon-gamma was produced by CD8+ T cells, and CD14+/15+ cells were the main cell subset presenting antigens expressed by rVV. We observed that the cell input per well had a critical influence on the sensitivity of the assay. Results from the ex vivo Elispot assay correlated poorly with those of the Cr-51 release assay performed after expansion of PBMC in vitro; thus, both assays gave information on different subsets and/or functions of the HIV-specific T cell response. (c) 2005 Elsevier B.V. All rights reserved.

引用

页码：105 / 118

页数：14