Jeffreys Presentation 2 05 08

Induction of Humoral and Cell-mediated Anti-Human Immunodeficiency Virus (HIV) Responses in HIV Sero-Negative Volunteers by immunization with Recombinant gp 160
By: Jeffrey Bruning

Background Info: Find a vaccine. Use gp-160, a glycoprotein of the enveloped virus that attaches to our cell and is the way into our cells.
-Have response to specific epitopes of HIV
-Inject gp-160 for response from T-cells.

Goals of the study:
-Safety for humans
-Evaluate nature of response
-Establish a dosage with the level of immune response

Seronegative- Absent from the blood, Not HIV positive, no antibodies against it.
Seropositive- have antibodies against it,
Blastogenic- to make more antibodies
1. Immunization- gene codes for the gp 160. Introduced into a virulent virus that effects butterflies. Butterflies were used to make sure they could obtain a lot of gp 160 from the butterflies in a short amt. of time.

2. 138 volunteers with 8 groups of 15.
2nd part- Blind study with KLH (control) was done. We have an innate response to KLH, and gp 160 was adhered to it to see how it would react with our body. See question 1

3. Immunoblot- video-
Used wells of HIV with gp 160 to see if it would adhere to it. Completed the ELISA test to make sure it stuck.

Figure 1- Western blot from HIV + person. Antibody is sticking to gp 160, but antibody also was sticking to 120, 88, and 41. These gp’s are very similar, same epitopes, and will react to same antigens.

Figure 2- There is a trend that injecting gp 160 it shoots up the antibodies of the person but does begin to decrease a bit.

Table 2- See diagram of board in notes

Table 3- If the person reacts to KLH, we want to make sure that when KLH is combinant with gp 160 the people are reacting to the gp 160 and not KLH.

Robsons questions
What is KLH? What kinds of cells do we need to present to T-cells? KLH will be presenting gp 160 to innate immune system which will have an immune response to it.

Figure 1- Farthest right (control) HIV with all of its proteins. What is being used as primary antibody of this system? Ans= blood serum of HIV + patient. The control on the right has all the gp’s and only the one with blots on them in our other graphs show which gp the antibody is sticking to.

Table 2 - What do you mean by %’s were higher? Higher % of rgp 160 binding than HIV + patients.

Diagram!!! How do they know the antibody is prevalent or effective? (neutralizing antibody= virus can’t attach to host cell to infect it) Blood serum from volunteer / by 4-—-into a test tube with HIV particles. Ya, refer to diagram in your own notes.

4 micrograms is too little in order for our body to produce antibody against gp 160.

War of attrition- HIV attacks T-cells which help differentiate B-cells to make antibodies against the HIV.

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