Experiment Update #1
Today, our contract lab in New York grafted peripheral blood monocytes from an HLA(B57) volunteer into C57BL/6 NOG mice (mice inter-bread to have no native immune system). They will ship these mice in a few days to another contract lab in California where they will acclimatize for between one and two weeks prior to injection with the KF11 epitope loaded in our microspheres.
Published data suggests that the HIV epitope KF11 is a good “match” to B57. This means that the immune system of an HLAB57 positive person should be able to develop an immune response to KF11.
By injecting human blood cells into mice with no native immune system, we have created a way to vaccinate human blood in a living organism without actually giving the vaccine to a human subject. We expect, when we administer our vaccine to the mice, that the human immune system dendrocytes will gobble up our microspheres, process the epitope delivered with the sphere, and “present” the epitope to a killer T cell. We can later check to see if this event took place with an immuno-assay.
Here are our next steps: 14 days after injection, we will process splenic blood from the mice. We will test for evidence of an immune response to KF11 using immuno-assay plates.
This will be the first time we have tested our vaccine loaded with a human epitope in human blood looking for quantitative evidence of immune response.
If we see an immune response, we will separate CD8 from CD4 cells, infect the CD4 cells with HIV and mix the cell populations together in an in-vitro culture and look for evidence of HIV inhibition.
A positive result would help confirm that the immune response we documented with the immuno-assay is robust enough to produce a cellular “kill” event in tissue culture.
Image #1 Forced air circulation hood loaded with supplies for blood sample preparation.
Image #2 Tissue culture (inverted) microscope.