Presentation Title

The Expression of Membrane Transport Proteins of Vaginal Epithelial Cells between no Anti-HIV Drugs and 3 Anti-HIV Drugs Embedded in Intravaginal Rings

Faculty Mentor

Paul Webster, John Cortez

Start Date

17-11-2018 8:45 AM

End Date

17-11-2018 9:00 AM

Location

C158

Session

Oral 1

Type of Presentation

Oral Talk

Subject Area

biological_agricultural_sciences

Abstract

Expression of drug transporter proteins was studied on vaginal epithelial cells (VEC) collected from woman female volunteers who participated in a study of the effects of wearing intravaginal ring (IVR) as a prophylactic device against HIV infection. During the study, the IVR first contained no drug formulation, but later the volunteers were exposed to IVRs containing a 3-drug formulation of anti-HIV drugs. The 3 drugs, TDF, (Tenofovir Disoproxil Fumarate) FTC (Emtricitabine), and MVC (Maraviroc) are known drugs that fight the HIV virus, and also known to reduce damage to the immune system as well as prevent HIV in non-infected individuals. Samples were collected from pap-smear like techniques and analyzed before drug exposure, and 2 weeks after drug exposure. The hypothesis tested whether or not levels of membrane transport protein would be affected with the presence of the 3 anti-HIV drugs from the VEC. Depending on whether the drugs promote transport expression, either no change or change in the amount of transporters will be seen, specifically a decrease or increase.

In order to track the protein presented in the vaginal cells that were collected from volunteers from Texas, we used a technique known as immunolabeling. Immunolabeling is a method used to detect a specific antigen presented in a cell. The samples were fixed in agarose gel to ensure minimal sample loss during immunolabeling procedure. Antigens are usually known as proteins that have the ability to bind to an antibody. Labeling a cell requires a primary antibody coated on the sample. When the primary antibody is coated, the proteins and antibodies bind together. A secondary antibody is than coated to bind to the primary antibody, which fluoresces at a certain wavelength and given a color. The samples were dried on microscopic slides and analyzed the day after for transport protein expression.

Summary of research results to be presented

The main result found was that the three Anti-HIV drugs working together are able to increase the amount of the transport protein in the vaginal cells. Before drug exposure to cells and after the Intravaginal rings were inserted, a major difference in protein expression was shown, with a major effect on the amount of transport protein expression for every antibody. Each antibody represents a protein in the vaginal epithelial cells, so all proteins tested experienced a major effect on the amount present. Different transport proteins expressed in different amount and ways, however all transport proteins expressed much more when the drug was introduced. Future work, including using electron microscopy, is currently being done for quantification on the amount of protein expression for the samples tested.

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Nov 17th, 8:45 AM Nov 17th, 9:00 AM

The Expression of Membrane Transport Proteins of Vaginal Epithelial Cells between no Anti-HIV Drugs and 3 Anti-HIV Drugs Embedded in Intravaginal Rings

C158

Expression of drug transporter proteins was studied on vaginal epithelial cells (VEC) collected from woman female volunteers who participated in a study of the effects of wearing intravaginal ring (IVR) as a prophylactic device against HIV infection. During the study, the IVR first contained no drug formulation, but later the volunteers were exposed to IVRs containing a 3-drug formulation of anti-HIV drugs. The 3 drugs, TDF, (Tenofovir Disoproxil Fumarate) FTC (Emtricitabine), and MVC (Maraviroc) are known drugs that fight the HIV virus, and also known to reduce damage to the immune system as well as prevent HIV in non-infected individuals. Samples were collected from pap-smear like techniques and analyzed before drug exposure, and 2 weeks after drug exposure. The hypothesis tested whether or not levels of membrane transport protein would be affected with the presence of the 3 anti-HIV drugs from the VEC. Depending on whether the drugs promote transport expression, either no change or change in the amount of transporters will be seen, specifically a decrease or increase.

In order to track the protein presented in the vaginal cells that were collected from volunteers from Texas, we used a technique known as immunolabeling. Immunolabeling is a method used to detect a specific antigen presented in a cell. The samples were fixed in agarose gel to ensure minimal sample loss during immunolabeling procedure. Antigens are usually known as proteins that have the ability to bind to an antibody. Labeling a cell requires a primary antibody coated on the sample. When the primary antibody is coated, the proteins and antibodies bind together. A secondary antibody is than coated to bind to the primary antibody, which fluoresces at a certain wavelength and given a color. The samples were dried on microscopic slides and analyzed the day after for transport protein expression.