Genetic Mutations in the LTR Region of SRLV Viruses in Capra ibex

The lentivirus (genus of the retroviruses family) can integrate a significant amount of viral cDNA into the DNA of the host cell and can efficiently infect dividing cells. They are able to spill over from their natural host species to induce new infections and pathologies among hosts of new species. This defines the crossing of species barrier that originates emergent viruses causing emergent diseases. The transmission of lentiviruses was observed between different species (domestic & wild). The small ruminant lentiviruses (SRLV) transmission is accompanied by genetic mutations in the genome of the virus. The study investigated the genetic mutations that accompany the infection and adaptation of SRLV to the new host. Genetic mutations were studied by amplifying and sequencing the Long Terminal Repeat (LTR) region.Blood samples were taken from Capra ibex living in the French Alps. Sera were tested using a commercially available ELISA. Peripheral blood mononuclear cells (PBMC) isolated on a Ficoll gradient were cultured in a macrophage differentiation medium to obtain monocyte-derived macrophage (MDM) monolayers for virus isolation. DNAs from non-cultured PBMC were used as templates for the PCR amplification of proviral DNA. PCR products (270 nt) were cloned and sequenced. Sequences were analysed using ClustalW.The alignments of the LTR fragment show three types of nucleotide mutations: replacement, addition, and deletion of nucleotide. Sequence analysis shows that the TATA box and the poly (A) site were highly conserved. The divergence of the LTR region between sequences obtained varied by 0.3 - 5.7 %. These differences were also shown by the phylogenetic tree. It can be seen that proviruses from the Capra ibex sequences are a closely related group, quite distinct from the reference sequence.

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CD4-Independent, CCR5-Dependent Simian Immunodeficiency Virus Infection and Chemotaxis of Human Cells

Journal of Virology ◽

10.1128/jvi.74.15.6720-6724.2000 ◽

2000 ◽

Vol 74 (15) ◽

pp. 6720-6724 ◽

Cited By ~ 7

Author(s):

Sujatha Iyengar ◽

David H. Schwartz ◽

Janice E. Clements ◽

James E. K. Hildreth

Keyword(s):

Simian Immunodeficiency Virus ◽

Mononuclear Cells ◽

Selective Advantage ◽

Human Pbmcs ◽

Macrophage Infection ◽

Independent Manner ◽

Peripheral Blood Mononuclear ◽

New Host ◽

Immunodeficiency Virus ◽

Hiv 1

ABSTRACT Most simian immunodeficiency virus (SIV), human immunodeficiency virus type 2 (HIV-2), and HIV-1 infection of host peripheral blood mononuclear cells (PBMCs) is CD4 dependent. In some cases, X4 HIV-1 chemotaxis is CD4 independent, and cross-species transmission might be facilitated by CD4-independent entry, which has been demonstrated for some SIV strains in CD4− non-T cells. As expected for CCR5-dependent virus, SIV required CD4 on rhesus and pigtail macaque PBMCs for infection and chemotaxis. However, SIV induced the chemotaxis of human PBMCs in a CD4-independent manner. Furthermore, in contrast to the results of studies using transfected human cell lines, SIV did not require CD4 binding to productively infect primary human PBMCs. CD4-independent lymphocyte and macrophage infection may facilitate cross-species transmission, while reacquisition of CD4 dependence may confer a selective advantage for the virus within new host species.

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Derivation of simian tropic HIV-1 infectious clone reveals virus adaptation to a new host

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1818059116 ◽

2019 ◽

Vol 116 (21) ◽

pp. 10504-10509 ◽

Cited By ~ 4

Author(s):

Fabian Schmidt ◽

Brandon F. Keele ◽

Gregory Q. Del Prete ◽

Dennis Voronin ◽

Christine M. Fennessey ◽

...

Keyword(s):

Mononuclear Cells ◽

Infectious Clone ◽

Host Protein ◽

Peripheral Blood Mononuclear ◽

New Host ◽

Species Specific ◽

Hiv 1 ◽

Pigtail Macaques

To replicate in a new host, lentiviruses must adapt to exploit required host factors and evade species-specific antiviral proteins. Understanding how host protein variation drives lentivirus adaptation allowed us to expand the host range of HIV-1 to pigtail macaques. We have previously derived a viral swarm (in the blood of infected animals) that can cause AIDS in this new host. To further exploit this reagent, we generated infectious molecular clones (IMCs) from the viral swarm. We identified clones with high replicative capacity in pigtail peripheral blood mononuclear cells (PBMC) in vitro and used in vivo replication to select an individual IMC, named stHIV-A19 (for simian tropic HIV-1 clone A19), which recapitulated the phenotype obtained with the viral swarm. Adaptation of HIV-1 in macaques led to the acquisition of amino acid changes in viral proteins, such as capsid (CA), that are rarely seen in HIV-1–infected humans. Using stHIV-A19, we show that these CA changes confer a partial resistance to the host cell inhibitor Mx2 from pigtail macaques, but that complete resistance is associated with a fitness defect. Adaptation of HIV-1 to a new host will lead to a more accurate animal model and a better understanding of virus–host interactions.

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BCL-2/JH rearrangements in circulating B cells of healthy blood donors and patients with nonmalignant diseases.

Journal of Clinical Oncology ◽

10.1200/jco.1996.14.4.1333 ◽

1996 ◽

Vol 14 (4) ◽

pp. 1333-1344 ◽

Cited By ~ 137

Author(s):

G Dölken ◽

G Illerhaus ◽

C Hirt ◽

R Mertelsmann

Keyword(s):

Sequence Analysis ◽

Nucleotide Sequence ◽

Follicular Lymphoma ◽

Peripheral Blood ◽

Blood Donors ◽

Mononuclear Cells ◽

Nucleotide Sequence Analysis ◽

Peripheral Blood Mononuclear ◽

Healthy Donors ◽

Follicular Lymphomas

PURPOSE To answer the question whether t(14;18)-positive cells can be detected by polymerase chain reaction (PCR) in the peripheral blood of healthy blood donors and patients with nonmalignant diseases. PATIENTS AND METHODS Peripheral-blood mononuclear cells (PBMNC) from healthy donors (n = 36) and patients with nonmalignant diseases (n = 21) were examined by two-step PCR for the detection of t(14;18)-positive cells with a breakpoint within the major breakpoint region (MBR). Approximate numbers of t(14;18)-positive cells were determined using limiting dilution assays, as well as the stochastic multiple-tube approach. RESULTS We were able to detect t(14;18)-positive cells in PBMNC of approximately 50% of healthy donors and patients with nonmalignant diseases if DNA amounts up to 10 microg were tested. Compared with 17 t(14;18)-positive patients being in complete remission after radiotherapy for low-stage malignant follicular lymphoma, the majority of 26 healthy donors were found to have significantly lower numbers of t(14;18)-positive cells circulating in the peripheral blood. In the case of six healthy donors, more than one t(14;18) DNA fragment based on size and nucleotide sequence analysis was detected. In one healthy individual, four different t(14;18)-positive cell clones were found in nine samples found over 5 years. CONCLUSION The occurrence of the t(14;18) translocation is not restricted to follicular lymphoma cells. In healthy donors, long-lived t(14;18)-positive cells can be detected by PCR if the sensitivity is high enough. Based on nucleotide sequence analysis, the t(14;18) DNA fragments detected in healthy donors cannot be distinguished from those found in follicular lymphomas.

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Human Immunodeficiency Virus Type 1 Intergroup (M/O) Recombination in Cameroon

Journal of Virology ◽

10.1128/jvi.73.8.6810-6820.1999 ◽

1999 ◽

Vol 73 (8) ◽

pp. 6810-6820 ◽

Cited By ~ 90

Author(s):

Jun Takehisa ◽

Léopold Zekeng ◽

Eiji Ido ◽

Yumi Yamaguchi-Kabata ◽

Innocent Mboudjeka ◽

...

Keyword(s):

Human Immunodeficiency Virus ◽

Human Immunodeficiency Virus Type ◽

Virus Type ◽

Mononuclear Cells ◽

Pcr Amplification ◽

Phylogenetic Position ◽

Peripheral Blood Mononuclear ◽

Immunodeficiency Virus ◽

Hiv 1

ABSTRACT Here we describe, for the first time, recombinants between two highly divergent major groups of human immunodeficiency virus type 1 (HIV-1), M and O, within a Cameroonian woman infected with three different HIV-1 strains, a group O virus, a subtype D virus, and a recently reported IBNG (A/G)-like recombinant virus. Using nested extra-long PCR amplification, we sequenced from the polregion to the env region including accessory genes of the viral genome obtained from the patient’s uncultured peripheral blood mononuclear cells and examined the phylogenetic position of each gene. Compared with sequential blood samples obtained in 1995 and 1996, there were multiple segmental exchanges between three HIV-1 strains (O, D, and IBNG) and all the recombinants appeared to be derived from a common M/O ancestor. Importantly, recombination between groups M and O occurred, even though the homology between these two groups is 69, 76, 68, and 55% in the gag, pol,vif-vpr, and env regions, respectively. Recombination between strains with such distant lineages may contribute substantially to generating new HIV-1 variants.

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Isolation of an Equine Foamy Virus and Sero-Epidemiology of the Viral Infection in Horses in Japan

Viruses ◽

10.3390/v11070613 ◽

2019 ◽

Vol 11 (7) ◽

pp. 613

Author(s):

Rikio Kirisawa ◽

Yuko Toishi ◽

Hiromitsu Hashimoto ◽

Nobuo Tsunoda

Keyword(s):

Mononuclear Cells ◽

Foamy Virus ◽

Epidemiological Survey ◽

Nucleotide Sequence Analysis ◽

Peripheral Blood Mononuclear ◽

Wobbler Syndrome ◽

Positive Rate ◽

Infected Cells ◽

First Time ◽

Indirect Immunofluorescent

An equine foamy virus (EFV) was isolated for the first time in Japan from peripheral blood mononuclear cells of a broodmare that showed wobbler syndrome after surgery for intestinal volvulus and the isolate was designated as EFVeca_LM. Complete nucleotide sequences of EFVeca_LM were determined. Nucleotide sequence analysis of the long terminal repeat (LTR) region, gag, pol, env, tas, and bel2 genes revealed that EFVeca_LM and the EFV reference strain had 97.2% to 99.1% identities. For a sero-epidemiological survey, indirect immunofluorescent antibody tests were carried out using EFVeca_LM-infected cells as an antigen against 166 sera of horses in five farms collected in 2001 to 2002 and 293 sera of horses in eight farms collected in 2014 to 2016 in Hokkaido, Japan. All of the farms had EFV antibody-positive horses, and average positive rates were 24.6% in sera obtained in 2001 to 2002 and 25.6% in sera obtained in 2014 to 2016 from broodmare farms. The positive rate in a stallion farm (Farm A) in 2002 was 10.7%, and the positive rates in two stallion farms, Farms A and B, in 2015 were 40.9% and 13.3%, respectively. The results suggested that EFV infection is maintained widely in horses in Japan.

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A higher throughput assay for quantification of melphalan-induced DNA damage in peripheral blood mononuclear cells

Scientific Reports ◽

10.1038/s41598-019-55161-3 ◽

2019 ◽

Vol 9 (1) ◽

Author(s):

Maia van Kan ◽

Kathryn E. Burns ◽

Peter Browett ◽

Nuala A. Helsby

Keyword(s):

Individual Differences ◽

Mononuclear Cells ◽

Ex Vivo ◽

Clinical Investigation ◽

Pcr Amplification ◽

Dna Adduct ◽

Good Precision ◽

Direct Pcr ◽

Peripheral Blood Mononuclear ◽

Healthy Donors

AbstractInter-individual differences in DNA adduct formation and repair influence the response to melphalan treatment, however, further clinical investigation of this variability requires a logistically feasible and reproducible bioassay. Our improved fluorescence-based QPCR-block assay is robust, has good precision, and improved throughput. It also incorporates direct PCR amplification from melphalan exposed PBMC using commercially available blood tubes and extraction kits to maximise the utility of this assay for future clinical studies. Using this assay we have demonstrated reproducible inter-individual differences in melphalan-induced QPCR-block across individual PBMC donors. As proof-of-principle we assessed nine healthy donors and found a 7.8 fold range in sensitivity following exposure of PBMC ex vivo. This likely reflects differences in melphalan transport into cells as well as differences in DNA adduct repair proficiency. This improved bioassay may be useful for assessment of these processes in patients about to receive melphalan treatment.

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