实验动物技术

Mononuclear phagocytes (MP: monocytes, dendritic cells, and tissue macrophages) are host cells for the human immunodeficiency viruses types 1 and 2. MPs are both the first lines of defense and vehicles for viral dissemination in the infected human host. Viral infection of MP can affect the dise ...

Glial inflammation, principally involving astrocytes and microglia, underlies the pathogenesis of a broad range of neurodegenerative disorders, including, most notably, human immunodeficiency virus (HIV-1)-associated dementia. Indeed, for the latter, disease mechan ...

Semen and vaginal fluids transmit HIV infection. The virus is present as cell-free particles and as infected cells. Isolation of infectious virus from both genital tract fluids poses unique problems. Vaginal fluids are heavily contaminated with normal bacterial flora, and seminal pla ...

This chapter describes a standardized microculture technique adapted from a peripheral blood mononuclear cells (PBMC) microculture assay to isolate and quantify HIV from lymph nodes. This quantitative lymph node microculture estimates the number of infectious cells per milli ...

Human T-cell leukemia virus type 2 (HTLV-2) was first isolated from leukemia patients, but has been found to be endemic among asymptomatic groups worldwide, including certain American Indian tribes (1). The virus infection is associated with a low incidence of disease among infected subje ...

Infectious variants of human immunodeficiency virus (HIV) can be isolated from peripheral blood mononuclear cells (PBMC) of infected individuals by propagating the virus in co-cultures with healthy donor PBMC. Standardized culture protocols have been designed specifically f ...

With the discovery that changes in the biological properties of HIV-1 correlate with the progression to disease, it became more and more important to develop assays to distinguish between the viral phenotypes. In this chapter, it is described how the biological phenotype of HIV-1 with regard to ...

An assay has been established for quantitative evaluation of lentivirus coreceptor use with the help of GHOST(3) cells. GHOST(3) cells were derived from the human osteosarcoma cell line, HOS, and have been engineered to stably express CD4 and one or another of the chemokine receptors CCR3, CCR5, C ...

In addition to CD4, HIV-1 uses chemokine receptors for entry in their target cells. The most important chemokine receptors in this respect are β-chemokine receptor 5 (CCR5) and α-chemokine receptor 4 (CXCR4). Coreceptor usage is an important feature of the biological phenotype of HIV-1 varia ...

Monocytes play an important, yet only partly understood, role in HIV-1 pathogenesis. Two main subsets of peripheral blood monocytes have been described; the major subset of monocytes are phenotypically characterized as being CD14hi/CD16−, and a minor subset (5–15% of total monocytes in he ...

Differential phenotypes or properties of HIV-1 gene products in primary virus isolates are difficult to assess due to interference by the high degree of sequence variation across the entire genome. Thus, chimeric viruses provide a powerful tool to study the function of single gene products ...

The fitness of human immunodeficiency virus type-1 (HIV-1) appears to be an important piece of the puzzle in understanding the global HIV-1 epidemic and disease progression in infected individuals. We have developed a dual infection/competition assay followed by a sensitive heterodu ...

The development of mixtures of highly processive and high-fidelity thermostable DNA polymerases has enabled the routine recovery of DNA sequences in excess of 25 kb generated by polymerase chain reaction. This powerful tool has been instrumental in the ability to recover virtually ful ...

The genomes of human immunodeficiency virus type 2 (HIV-2), like those of HIV-1, are not only extremely variable but are also highly recombinogenic (1–5). Determination of subtypes based on partial genomes cannot predict the subtype classification of other regions of the genome owing to the fr ...

In retrovirus research, the generation of an infectious molecular clone is a landmark event, opening up new avenues of research using the cloned virus. A full-length proviral plasmid clone of the human T-cell leukemia virus (HTLV) makes possible reproducible viral genetic studies. Howev ...

The molecular engineering of recombinant plasmid DNA clones containing the full-length and replication-competent feline foamy (retro)virus (FFV) proviral genome is described. The methods used to combine subgenomic FFV DNA fragments can be applied to other retrovirus genomes, r ...

Molecular characterization of proteolytic processing of the human spumaretrovirus (HSRV) Gag proteins and the precise determination of cleavage sites was performed. For in vitro processing of recombinant HSRV Gag proteins, a recombinant enzymatically active HSRV protease w ...

HIV-infected monocyte/macrophage-derived cells are believed to play a major role in the spread of HIV through the body. Not only are fresh monocytes and more differentiated macrophages relatively insensitive to the cytopathic effect of HIV, but once infected they can efficiently infe ...

To illustrate the methods employed in gene expression profiling using cDNA microarrays, infection of CD4+ T cell lines with HIV-1LAI is used to identify expression changes relevant to in vitro HIV-1 infection. Cell lines are infected at a high multiplicity of infection to ensure a population of ...

DC-SIGN and DC-SIGNR efficiently bind HIV-1 and other viral as well as nonviral pathogens and assist either cis or trans infection. Both are type II transmembrane proteins that consist of an N-terminal cytoplasmic domain, a repeat region consisting of seven 23-amino-acid tandem repeats, and a C ...