Name: humanized nog mice for intravaginal hiv exposure and treatment of hiv infection
Uploaded: 2020-01-31T15:00:00
Description: Watch this Scientific Journal Video about Humanized NOG Mice for Intravaginal HIV Exposure and Treatment of HIV Infection at JoVE.com

The authors would like to thank the Biomedicine Animal Facility staff at Aarhus University, particularly Ms. Jani K&#230;r for colony maintenance efforts and for tracking mouse weights. The authors would like to thank Professor Florian Klein for developing standard-of-care cART and for guidance. The following reagent was obtained through the NIH AIDS Reagent Program, Division of AIDS, NIAID, NIH: pRHPA.c/2635 (cat# 11744) from Dr. John Kappes and Dr. Christina Ochsenbauer.

All cord blood samples were obtained in strict accordance with locally approved protocols, including informed consent of anonymous donation by the parents. All animal experiments were approved and performed in strict accordance with Danish national regulations under the license 2017-15-0201-01312.
CAUTION: Handle HIV exposed mice and blood with extreme caution. Decontaminate all surfaces and liquids that have been in contact with HIV with a confirmed HIV disinfectant (Table of Material...

<ol>
	<li>Skelton, J. K., Ortega-Prieto, A. M., Dorner, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Hitchhiker's+guide+to+humanized+mice:+new+pathways+to+studying+viral+infections.">A Hitchhiker's guide to humanized mice: new pathways to studying viral infections.</a> Immunology. 154 (1), 50-61 (2018).</li><li>Denton, P. W., Krisko, J. F., Powell, D. A., Mathias, M., Kwak, Y. 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M., Connick, E., Akkina, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mucosal+transmission+of+R5+and+X4+tropic+HIV-1+via+vaginal+and+rectal+routes+in+humanized+Rag2+-/-+&#947;c+-/-+(RAG-hu)+mice.">Mucosal transmission of R5 and X4 tropic HIV-1 via vaginal and rectal routes in humanized Rag2 -/- &#947;c -/- (RAG-hu) mice.</a> Virology. 373 (2), 342-351 (2008).</li><li>Veselinovic, M., Charlins, P., Akkina, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+HIV-1+Mucosal+Transmission+and+Prevention+in+Humanized+Mice.">Modeling HIV-1 Mucosal Transmission and Prevention in Humanized Mice.</a> Methods Mol Biol. , 203-220 (2016).</li><li>Neff, C. 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V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Long-term+engraftment+of+nonobese+diabetic/severe+combined+immunodeficient+mice+with+human+CD34++cells+transduced+by+a+self-inactivating+human+immunodeficiency+virus+type+1+vector.">Long-term engraftment of nonobese diabetic/severe combined immunodeficient mice with human CD34+ cells transduced by a self-inactivating human immunodeficiency virus type 1 vector.</a> Human Gene Therapy. 12 (9), 1079-1089 (2001).</li><li>Leth, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HIV-1+transcriptional+activity+during+frequent+longitudinal+sampling+in+aviremic+patients+on+antiretroviral+therapy.">HIV-1 transcriptional activity during frequent longitudinal sampling in aviremic patients on antiretroviral therapy.</a> AIDS. 30 (5), 713-721 (2016).</li><li>Halper-Stromberg, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Broadly+neutralizing+antibodies+and+viral+inducers+decrease+rebound+from+HIV-1+latent+reservoirs+in+humanized+mice.">Broadly neutralizing antibodies and viral inducers decrease rebound from HIV-1 latent reservoirs in humanized mice.</a> Cell. 158 (5), 989-999 (2014).</li><li>Rothenberger, M. 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L., Sauzade, M., Brouzes, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DPCR:+A+technology+review.">DPCR: A technology review.</a> Sensors (Switzerland). 18 (4), (2018).</li><li>Denton, P. 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The severely immunocompromised mouse strain NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac (NOG) is extremely well suited for transplantation of human cells and tissues. Both innate and adaptive immune pathways in these mice are compromised. NOG and NSG mice harbor a Prkdcscid mutation that results in defective T and B cell function. Furthermore, these mice lack a functional interleukin-2 receptor &#947;-chain (common gamma chain, IL2rg) which is indispensable in ...

Human immunodeficiency virus (HIV) is a chronic infection with more than 37 million infected individuals worldwide1. Combination antiviral therapy (cART) is a life-saving therapy, but a cure is still warranted. Thus, there is a need for animal models that mirror the human immune system and its responses in order to facilitate continued research in HIV. Multiple types of humanized mice that are capable of supporting cell and tissue engraftment have been developed by transplanting human cells into severely immunodeficient mice2. Such humanized mice are susceptible to HIV infection and provide an important alternative to nonhuman primate simian immunodeficiency virus models, as they are cheaper and simpler to use than nonhuman primates. Humanized mice have facilitated research in HIV viral transmission, pathogenesis, prevention, and treatment3,4,5,6,7,8,9,10,11.
We present a flexible humanized model system for HIV research developed by transplanting cord blood derived human stem cells into mice of the NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac (NOG) background. Besides being of non-fetal origin, the practical bioengineering of these mice is less technically demanding compared to the microsurgical procedures involved in the transplant of the blood-liver-thymus (BLT) construct.
We show how to establish HIV infection through intravaginal transmission and how to monitor the plasma viral load with a sensitive droplet digital PCR (ddPCR)-based setup. Subsequently, we describe the establishment of standard cART given as part of the daily mouse diet. The aim of these combined methods is to reduce stress to the animals and facilitate large-scale experiments where time spent handling each animal is limited12.
In humans, a CCR5&#916;32/wt or CCR5&#916;32/ &#916;32 genotype causes reduced susceptibility to HIV infection with transmitter/founder viruses13, and some precautions must be taken when bioengineering humanized mice with stem cells for the purpose of HIV studies. This is especially true in our region because naturally occurring variants in the CCR5 gene, particularly &#916;32 deletions, are more prevalent in Scandinavian and Baltic native populations compared to rest of the world14,15. Thus, our protocol includes an easy, high-throughput assay for screening donor hematopoietic stem cells for CCR5 variants prior to transplantation.
For the intravaginal exposure we chose the transmitter/founder R5 virus RHPA4259, isolated from a woman in an early stage of infection who was infected intravaginally16. We exposed the mice to a viral dose that was sufficient to yield successful transmission in the majority of mice, but below a 100% transmission rate. Choosing such a dose enables a sufficient dynamic range in transmission rate such that antiviral effects of a drug candidate can result in protected animals in HIV prevention experiments and decreased viral load for treatment studies.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Blue pad</td>
			<td>VWR</td>
			<td>56616-031</td>
			<td>Should be sterilized prior to use</td>
		</tr>
		<tr>
			<td>Bovine serum albumin (BSA)</td>
			<td>Sigma</td>
			<td>A8022</td>
			<td></td>
		</tr>
		<tr>
			<td>CD19 (clone sj25c1) PE-Cy7</td>
			<td>BD Bioscience</td>
			<td>557835</td>
			<td></td>
		</tr>
		<tr>
			<td>CD3 (clone OKT3) FITC</td>
			<td>Biolegend</td>
			<td>317306</td>
			<td></td>
		</tr>
		<tr>
			<td>CD3 (clone SK7) BUV395</td>
			<td>BD Bioscience</td>
			<td>564001</td>
			<td></td>
		</tr>
		<tr>
			<td>CD34 (clone AC136) FITC</td>
			<td>Miltenyi</td>
			<td>130-113-740</td>
			<td></td>
		</tr>
		<tr>
			<td>CD4 (clone SK3) BUV 496</td>
			<td>BD Bioscience</td>
			<td>564652/51</td>
			<td></td>
		</tr>
		<tr>
			<td>CD45 (clone 2D1) APC</td>
			<td>Biolegend</td>
			<td>368511/12</td>
			<td></td>
		</tr>
		<tr>
			<td>CD8 (clone RPA-T8) BV421</td>
			<td>BD Bioscience</td>
			<td>562428</td>
			<td></td>
		</tr>
		<tr>
			<td>ddPCR Supermix for probes (no dUTP)</td>
			<td>Bio-Rad</td>
			<td>1863025</td>
			<td></td>
		</tr>
		<tr>
			<td>DMSO</td>
			<td>Merck</td>
			<td>10,02,95,21,000</td>
			<td></td>
		</tr>
		<tr>
			<td>DNAse</td>
			<td>Sigma</td>
			<td>D4263</td>
			<td>For suspension buffer</td>
		</tr>
		<tr>
			<td>dNTP mix</td>
			<td>Life Technologies</td>
			<td>R0192</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbeccos phosphate-buffered saline (PBS)</td>
			<td>Biowest</td>
			<td>L0615-500</td>
			<td></td>
		</tr>
		<tr>
			<td>EasySep Human Cord Blood CD34 Positive Selection Kit II</td>
			<td>Stemcell</td>
			<td>17896</td>
			<td></td>
		</tr>
		<tr>
			<td>EDTA</td>
			<td>Invitrogen</td>
			<td>15575-038</td>
			<td></td>
		</tr>
		<tr>
			<td>FACS Lysing solution 10X</td>
			<td>BD</td>
			<td>349202</td>
			<td>Dilute 1:10 in dH20 immediately before use</td>
		</tr>
		<tr>
			<td>FACS tubes (Falcon 5 mL round-botton)</td>
			<td>Falcon</td>
			<td>352052</td>
			<td></td>
		</tr>
		<tr>
			<td>Fc Receptor blocking solution (Human Trustain FcX)</td>
			<td>Biolegend</td>
			<td>422302</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>Sigma</td>
			<td>F8192-500</td>
			<td></td>
		</tr>
		<tr>
			<td>Ficoll-Paque PLUS</td>
			<td>GE Healthcare</td>
			<td>17144002</td>
			<td></td>
		</tr>
		<tr>
			<td>Flowjo v.10</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Gauze</td>
			<td>Mesoft</td>
			<td>157300</td>
			<td>Should be sterilized prior to use</td>
		</tr>
		<tr>
			<td>Heating lamp</td>
			<td></td>
			<td></td>
			<td>Custom made</td>
		</tr>
		<tr>
			<td>Hemacytometer (B&#252;rker-T&#252;rk)</td>
			<td>VWR</td>
			<td>DOWC1597418</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane gas</td>
			<td>Orion Pharma</td>
			<td>9658</td>
			<td></td>
		</tr>
		<tr>
			<td>LSR Fortessa X20 flow cytometer</td>
			<td>BD</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Microcentrifuge tubes, PCR-PT approved</td>
			<td>Sarstedt</td>
			<td>72692405</td>
			<td></td>
		</tr>
		<tr>
			<td>Mouse cART food</td>
			<td>ssniff Spezialdi&#228;ten GmbH</td>
			<td>Custom made product</td>
			<td></td>
		</tr>
		<tr>
			<td>Mouse restrainer</td>
			<td></td>
			<td>Custom made product</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle, Microlance 3, 30G &#189;&#34;</td>
			<td>BD</td>
			<td>304000</td>
			<td></td>
		</tr>
		<tr>
			<td>NOG mice NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac</td>
			<td>Taconic</td>
			<td>NOG-F</td>
			<td></td>
		</tr>
		<tr>
			<td>Nuclease-free water</td>
			<td>VWR chemicals</td>
			<td>436912C</td>
			<td></td>
		</tr>
		<tr>
			<td>Nucleospin 96 Virus DNA and RNA isolation kit</td>
			<td>Macherey-Nagel</td>
			<td>740691</td>
			<td></td>
		</tr>
		<tr>
			<td>PCR-approved microcentrifuge tubes</td>
			<td>Sarstedt</td>
			<td>72.692.405</td>
			<td></td>
		</tr>
		<tr>
			<td>Penicillin-Streptomycin solution 100X</td>
			<td>Biowest</td>
			<td>L0022-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Phusion Hot Start II DNA polymerase</td>
			<td>Life Technologies</td>
			<td>F549S</td>
			<td></td>
		</tr>
		<tr>
			<td>Pipette tips, sterile, ART 20P Barrier</td>
			<td>ThermoScientific</td>
			<td>2149P</td>
			<td></td>
		</tr>
		<tr>
			<td>Proteinase K</td>
			<td>NEB</td>
			<td>100005398</td>
			<td></td>
		</tr>
		<tr>
			<td>QuantaSoft software</td>
			<td>Bio-Rad</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>QX100 Droplet Generator</td>
			<td>Bio-Rad</td>
			<td>1886-3008</td>
			<td></td>
		</tr>
		<tr>
			<td>QX100 Droplet Reader</td>
			<td>Bio-Rad</td>
			<td>186-3003</td>
			<td></td>
		</tr>
		<tr>
			<td>RBC lysis solution</td>
			<td>Biolegend</td>
			<td>420301</td>
			<td></td>
		</tr>
		<tr>
			<td>RNase-free DNAse size F + reaction buffer</td>
			<td>Macherey-Nagel</td>
			<td>740963</td>
			<td></td>
		</tr>
		<tr>
			<td>RNAseOUT Recombinant Ribonuclease inhibitor</td>
			<td>ThermoScientific</td>
			<td>10777-019</td>
			<td></td>
		</tr>
		<tr>
			<td>RPMI</td>
			<td>Biowest</td>
			<td>L0501-500</td>
			<td>Dissolve in H20</td>
		</tr>
		<tr>
			<td>Softject 1 mL syringe</td>
			<td>Henke Sass Wolf</td>
			<td>5010-200V0</td>
			<td></td>
		</tr>
		<tr>
			<td>Superscript III Reverse Transcriptase</td>
			<td>ThermoFisher Scientific</td>
			<td>18080044</td>
			<td></td>
		</tr>
		<tr>
			<td>Thermoshaker</td>
			<td>VWR</td>
			<td>89370-910</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypane blue</td>
			<td>Sigma</td>
			<td>T8154</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrapure 0.5 EDTA, pH 8.0</td>
			<td>ThermoFisher Scientific</td>
			<td>15575-020</td>
			<td></td>
		</tr>
		<tr>
			<td>Virkon S (virus disinfectant)</td>
			<td>Dupont</td>
			<td>7511</td>
			<td></td>
		</tr>
	</tbody>
</table>

humanized nog mice for intravaginal hiv exposure and treatment of hiv infection

Humanized mice provide a sophisticated platform to study human immunodeficiency virus (HIV) virology and to test antiviral drugs. This protocol describes the establishment of a human immune system in adult NOG mice. Here, we explain all the practical steps from isolation of umbilical cord blood derived human CD34+ cells and their subsequent intravenous transplantation into the mice, to the manipulation of the model through HIV infection, combination antiretroviral therapy (cART), and blood sampling. Approximately 75,000 hCD34+ cells are injected intravenously into the mice and the level of human chimerism, also known as humanization, in the peripheral blood is estimated longitudinally for months by flow cytometry. A total of 75,000 hCD34+ cells yields 20%&#8211;50% human CD45+ cells in the peripheral blood. The mice are susceptible to intravaginal infection with HIV and blood can be sampled once weekly for analysis, and twice monthly for extended periods. This protocol describes an assay for quantification of plasma viral load using droplet digital PCR (ddPCR). We show how the mice can be effectively treated with a standard-of-care cART regimen in the diet. The delivery of cART in the form of regular mouse chow is a significant refinement of the experimental model. This model can be used for preclinical analysis of both systemic and topical pre-exposure prophylaxis compounds as well as for testing of novel treatments and HIV cure strategies.

The gating strategy for the analysis of stem cell purity is depicted in Figure 1. Figure 1A&#8211;C shows the purified CD34+ population and Figure 1D&#8211;F the CD34- flow-through used to illustrate that the minimal amount of the CD34+ population is lost in the isolation process. The purity of isolated CD34+ stem cells was between 85%&#8211;95% with less than 1% T-cell contamination. <strong class=...

Watch this Scientific Journal Video about Humanized NOG Mice for Intravaginal HIV Exposure and Treatment of HIV Infection at JoVE.com

Humanized NOG Mice for Intravaginal HIV Exposure and Treatment of HIV Infection

We have developed a protocol for the generation and evaluation of a humanized and human immunodeficiency virus-infected NOG mouse model based on stem cell transplant, intravaginal human immunodeficiency virus exposure, and droplet digital PCR RNA quantification.

Humanized mice provide a sophisticated platform to study human immunodeficiency virus (HIV) virology and to test antiviral drugs. This protocol describes ...

Establishing this humanized mouse model can aid in preclinical HIV research studies. For example, both different viral strains and novel HIV interventions can be evaluated. These protocols can be used to evaluate stem cell donor CCR5 variant status and to efficiently infect humanized mice with HIV and to quantify mouse plasma viral loads.The RNA quantification protocol can be easily adapted to quantify any viral RNA sequence of choice that is detectable in plasma samples. For genetic screening of cord blood samples for CCR5 delta 32 variants, incubate 1.25 microliters of non-pelleted flowthrough with 11.25 microliters of PCR mix containing 200 micromolar dNTP mix, 0.01 unit per microliter of high-fidelity DNA polymerase, and forward and reverse primers as detailed in the table. Adjust the volume with nuclease-free water to approximately 12.5 microliters for each PCR reaction and amplify the genomic fragments with the PCR cycling program as indicated in the table.At the end of the reaction, separate the PCR products on a 2%agarose gel. PCR products from the wild type alleles and the delta 32 alleles yield PCR fragments of 196 base pairs and 164 base pairs respectively, making the bands easily distinguishable by gel electrophoresis. For intravaginal HIV exposure, place a heating lamp focused on the center of the workspace where the mouse will be located and place sterile 20 microliter pipette tips in an appropriate pipette into the hood.Place a container with liquid disinfectant into the hood for immediate inactivation of any materials and liquids that have been in contact with the virus. Place the anesthetized mouse onto a sterile blue pad in the supine position under the lamp with the tail of the mouse facing the back of the hood and confirm the lack of response to pedal reflex in the mouse. Grasp the animal at the base of the tail.Gently lifting the mouse by the tail base, support the animal in an upside down upright position. Use a sterile pipe tip to stimulate the genital area with gentle stroking up toward the anus to induce emptying of the rectum and to relieve pressure on the vagina. To expose the vaginal opening, carefully wrap the mouse tail across the fingers, such that the vulva naturally opens.Using a new pipette tip, place the tip at the level of the vaginal opening and atraumatically release 20 microliters of the virus into the vagina, allowing gravity to pull the viral suspension into the vagina. When all the virus has been delivered, retain the mouse in this position for five minutes to avoid gravity-induced leakage of the virus. Then, carefully return the mouse to its home cage in the supine position and monitor until full recovery.To isolate RNA from thawed mouse plasma samples, use a virus RNA isolation kit according to the manufacturer's instructions. Add the sample that binds to the column and add an on-column DNase treatment step to ensure the removal of all DNA within the plasma sample. Then store the RNA samples at 80 degrees Celsius for at least one hour.To synthesize cDNA, use reverse transcriptase according to standard protocols, including 0.5 microliters of an RNase inhibitor within the cDNA reaction to avoid RNA degradation, then perform cDNA synthesis as detailed in the table and store the cDNA samples at 80 degrees Celsius for at least one hour. To prepare samples for ddPCR, mix 11 microliters of ddPCR probe reaction mixture containing polymerase and dNTPs with 250 nanomolar of minor groove binding probe and 900 nanomolar of each of the forward and reverse primers, as indicated in the table. Add three microliters of each cDNA sample and adjust the total PCR volumes to 22 microliters with nuclease-free water.Use droplet generation oil for probes on a droplet generator according to the manufacturer's protocol to emulsify the PCR mixes. Then run the PCR program as outlined in the table. For cART treatment after confirmed HIV infection, feed the mice with pellets prepared by an external vendor containing a standard cART regimen, as indicated in the table.Use a red-colored cART diet to easily distinguish it from ordinary mouse chow and use a control chow diet without cART in a standard brown color. For initiation of the treatment, place the cART-containing chow diet into sterile cages before transferring the mice from the old cage to the new cage. Then monitor the weights of the mice and check the consumption of cART-containing chow daily to ensure that the mice are adjusting to the change.Here, the flow cytometry gating strategy for analysis of the stem cell purity is depicted, with the purity of the isolated CD34 positive stem cells typically ranging between 85 to 95%with a less than 1%T cell contamination. PCR analysis demonstrates CCR5 variant status of the purified donor stem cells, making CCR5 delta 32 heterozygous donors easily identifiable. The frequency of the mutant genotype in a group of 19 donors was around 15.8%in agreement with larger epidemiological studies of Scandinavian populations.Three to five months after the adoptive transfer of 7.5 times 10 to the 4th human CD45 positive cells into recipient mice, 20 to 50%of the white blood cells recovered from recipient animal peripheral blood samples are human CD45 positive cells and the mice will have developed human B and T cells. 64%of mice become infected with HIV following vaginal exposure as demonstrated. Four weeks after switching to a 40 day diet containing standard cART, the viral loads in HIV positive mice decrease below the detection limit.After cessation of the diet, the virus rebounds, mirroring clinical data. Importantly, mice on cART tolerate the change in diet well. It is crucial to remember that these animals are immunodeficient and therefore vulnerable to the environment.Adhering to aseptic techniques will increase the overall animal health and protocol success. After establishment of the humanized mouse cohort for HIV infection, the protocol may be adapted for viral RNA quantification in tissues or for testing novel interventions in different viral strains. The flexibility and accessibility of humanized mice for HIV infection studies have enabled critical progress in HIV virology, immunology, and treatment research.Before attempting protocols involving infectious HIV, be sure to obtain approval from local work environment officials and to adhere to approved decontamination procedures.

Research

JoVE Journal

Immunology and Infection

Visualizing Cell-to-cell Transfer of HIV using Fluorescent Clones of HIV and Live Confocal Microscopy

By fusing the green fluorescent protein to their favorite proteins, biologists now have the ability to study living complex cellular processes using ...

Genotypic Inference of HIV-1 Tropism Using Population-based Sequencing of V3

Background: Prior to receiving a drug from CCR5-antagonist class in HIV therapy, a patient must undergo an HIV tropism test to confirm that his or her ...

Prediction of HIV-1 Coreceptor Usage (Tropism) by Sequence Analysis using a Genotypic Approach

Prediction of HIV-1 Coreceptor Usage Tropism by Sequence Analysis using a Genotypic Approach

Maraviroc (MVC) is the first licensed antiretroviral drug from the class of coreceptor antagonists. It binds to the host coreceptor CCR5, which is used by ...

In vitro Uncoating of HIV-1 Cores

In vitro Uncoating of HIV-1 Cores

The genome of the retroviruses is encased in a capsid surrounded by a lipid envelope. For lentiviruses, such as HIV-1, the conical capsid shell is ...

Rapid Screening of HIV Reverse Transcriptase and Integrase Inhibitors

Although a number of anti HIV drugs have been approved, there are still problems with toxicity and drug resistance. This demonstrates a need to identify ...

An In Vitro Model for Measuring Immune Responses to Malaria in the Context of HIV Co-infection

An In Vitro Model for Measuring Immune Responses to Malaria in the Context of HIV Co-infection

Malaria and HIV co-infection is a growing health priority. However, most research on malaria or HIV currently focuses on each infection individually. ...

Measurement of In Vitro Integration Activity of HIV-1 Preintegration Complexes

Measurement of In Vitro Integration Activity of HIV-1 Preintegration Complexes

HIV-1 envelope proteins engage cognate receptors on the target cell surface, which leads to viral-cell membrane fusion followed by the release of the ...

SILAC Based Proteomic Characterization of Exosomes from HIV-1 Infected Cells

Proteomics is the large-scale analysis of proteins. Proteomic techniques, such as liquid chromatography tandem mass spectroscopy (LC-MS/MS), can ...

Single-cell Quantitation of mRNA and Surface Protein Expression in Simian Immunodeficiency Virus-infected CD4+ T Cells Isolated from Rhesus macaques

Single-cell Quantitation of mRNA and Surface Protein Expression in Simian Immunodeficiency Virus-infected CD4+ T Cells Isolated from Rhesus macaques

Single-cell analysis is an important tool for dissecting heterogeneous populations of cells. The identification and isolation of rare cells can be ...

Humanized NOD/SCID/IL2r&#947;null (hu-NSG) Mouse Model for HIV Replication and Latency Studies

Ethical regulations and technical challenges for research in human pathology, immunology, and therapeutic development have placed small animal models in ...