Abstract:Described in this unit are basic protocols frequently used in the research of human immunodeficiency viruses (HIVs). Provided are methods for propagating and quantifying HIV, as well as recommendations for long-term storage. Background information about these methods is also provided and includes their advantages, disadvantages, and troubleshooting.
“…Sixty to 72 hr after transfection, SNs were harvested and syringe-filtered through a 0.45-lm filter before being aliquoted and stored at -80°C. Infectious HIV was generated and titered as described 24 with the following modifications: 10-cm dishes were plated with 2 · 10 6 293T cells the day before transfection. Twenty-five micrograms of cDNA encoding an infectious molecular clone of HIV-1 was used for transfection.…”
{The first three authors contributed equally to this manuscript.Despite nearly three decades of research, a safe and effective vaccine against human immunodeficiency virus type 1 (HIV-1) has yet to be achieved. More recently, the discovery of highly potent anti-gp160 broadly neutralizing antibodies (bNAbs) has garnered renewed interest in using antibody-based prophylactic and therapeutic approaches. Here, we encoded bNAbs in first-generation adenoviral (ADV) vectors, which have the distinctive features of a large coding capacity and ease of propagation. A single intramuscular injection of ADV-vectorized bNAbs in humanized mice generated high serum levels of bNAbs that provided protection against multiple repeated challenges with a high dose of HIV-1, prevented depletion of peripheral CD4 + T cells, and reduced plasma viral loads to below detection limits. Our results suggest that ADV vectors may be a viable option for the prophylactic and perhaps therapeutic use of bNAbs in humans.
INTRODUCTIONSince its emergence more than three decades ago, human immunodeficiency virus type 1 (HIV-1) remains a pandemic, with more than 60 million infected individuals to date and more than 32 million acquired immunodeficiency syndrome (AIDS)-related deaths.1,2 Despite intense research efforts, a safe and effective vaccine remains elusive. At present, highly active antiretroviral therapy (HAART) constitutes the mainstay of treatment and has resulted in HIV-infected individuals with plasma viral RNA loads (VLs) below the limits of detection, increased peripheral CD4 + T cell counts, and decreased patient morbidity and mortality. Despite the improved quality of life, HAART has a number of limitations including high cost, drug toxicity and interactions, emergence of virus resistance, and the need for indefinite treatment, necessitating alternative therapeutic approaches.
“…Sixty to 72 hr after transfection, SNs were harvested and syringe-filtered through a 0.45-lm filter before being aliquoted and stored at -80°C. Infectious HIV was generated and titered as described 24 with the following modifications: 10-cm dishes were plated with 2 · 10 6 293T cells the day before transfection. Twenty-five micrograms of cDNA encoding an infectious molecular clone of HIV-1 was used for transfection.…”
{The first three authors contributed equally to this manuscript.Despite nearly three decades of research, a safe and effective vaccine against human immunodeficiency virus type 1 (HIV-1) has yet to be achieved. More recently, the discovery of highly potent anti-gp160 broadly neutralizing antibodies (bNAbs) has garnered renewed interest in using antibody-based prophylactic and therapeutic approaches. Here, we encoded bNAbs in first-generation adenoviral (ADV) vectors, which have the distinctive features of a large coding capacity and ease of propagation. A single intramuscular injection of ADV-vectorized bNAbs in humanized mice generated high serum levels of bNAbs that provided protection against multiple repeated challenges with a high dose of HIV-1, prevented depletion of peripheral CD4 + T cells, and reduced plasma viral loads to below detection limits. Our results suggest that ADV vectors may be a viable option for the prophylactic and perhaps therapeutic use of bNAbs in humans.
INTRODUCTIONSince its emergence more than three decades ago, human immunodeficiency virus type 1 (HIV-1) remains a pandemic, with more than 60 million infected individuals to date and more than 32 million acquired immunodeficiency syndrome (AIDS)-related deaths.1,2 Despite intense research efforts, a safe and effective vaccine remains elusive. At present, highly active antiretroviral therapy (HAART) constitutes the mainstay of treatment and has resulted in HIV-infected individuals with plasma viral RNA loads (VLs) below the limits of detection, increased peripheral CD4 + T cell counts, and decreased patient morbidity and mortality. Despite the improved quality of life, HAART has a number of limitations including high cost, drug toxicity and interactions, emergence of virus resistance, and the need for indefinite treatment, necessitating alternative therapeutic approaches.
The current COVID‐19 pandemic caused by SARS‐Cov‐2 is responsible for more than 6 million deaths globally. The development of broad‐spectrum and cost‐effective antivirals is urgently needed. Medicinal plants are renowned as a complementary approach in which antiviral natural products have been established as safe and effective drugs. Here, we report that the percolation extract of Spatholobus suberectus Dunn (SSP) is a broad‐spectrum viral entry inhibitor against SARS‐CoV‐1/2 and other enveloped viruses. The viral inhibitory activities of the SSP were evaluated by using pseudotyped SARS‐CoV‐1 and 2, HIV‐1ADA and HXB2, and H5N1. SSP effectively inhibited viral entry and with EC50 values ranging from 3.6 to 5.1 μg/ml. Pre‐treatment of pseudovirus or target cells with SSP showed consistent inhibitory activities with the respective EC50 value of 2.3 or 2.1 μg/ml. SSP blocked both SARS‐CoV‐2 spike glycoprotein and the host ACE2 receptor. In vivo studies indicated that there was no abnormal toxicity and behavior in long‐term SSP treatment. Based on these findings, we concluded that SSP has the potential to be developed as a drug candidate for preventing and treating COVID‐19 and other emerging enveloped viruses.
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