Further Characterization of HLA Typing Cell Lines at CCR-5 Receptor by a Gene Amplification and Detection Method
Abstract
The co-chemokine receptor 5 or CCR-5 is one of the major co-receptors
for the macrophage-tropic (M-tropic) strain of HIV. The frequency of the 32
base pair (bp) deletion of CCR-5 is elevated in individuals who have had
multiple exposures to HIV but remain uninfected. This is evidence suggests
that the CCR-5 can act as a natural resistance to the infection. CCR-5 reduces
the ability of an individual's cells to support an entry of the M-tropic virus,
because the defective receptors are severely truncated.
In this study, a typing method was designed specifically to detect the
mutation that occurs on CCR-5 sequences. A gene amplification and
detection method that involved polymerase chain reaction (PCR), restriction
enzyme digestion, and gel electrophroresis was utilized. In order to establish
this new amplification typing method, primers were assigned to yield a 73S-bp
fragment spanning the site of the mutation, PCR condition optimized to
increase the original of DNA product, and control Human Lymphocyte
Antigens (HLAj) typing cell lines were tested to validate testing parameters.
As a result, the functional data suggested that the designed reagents
and method were found to be another effective way for detecting the
mutation on cell lines, by defining the 32 bp deletion on the heterozygote
mutant alleles, which furthered the characterization of HLA typing cell lines
at the CCR-5 receptor. With this knowledge in understanding the
mechanism of the resistance to HIV infection, it may provide additional
information for inducing similar resistance in others and successfully
combating AIDS.