1. Within-host evolution of X4 HIV-1 in a rare transmission pair revealed by phylogenetic reconstruction of deep-sequence data
Anh Q. Le, Jeremy Taylor, Winnie Dong, Rosemary McCloskey, Conan Woods, Kanna Hayashi, M-J Milloy, P. Richard Harrigan, Art F.Y. Poon, Zabrina L. Brumme
- Thank you organizers and the title of my poster is shown here.

2. Putative case of X4 transmission from CCR5-wt/wt donor to CCR5-∆32 homozygous recipient
Envelope-V3
- Getting straight into it, through phylogenetic analysis of bulk env-V3 sequences from acute and chronically infected individuals of VIDUS
Retrospectively identified a putative transmission case, donor CCR5wt and recipient CCR5∆32 homozygous
Shown is a phylogenetic tree created using env-V3 sequences, red branches are X4 and blue are R5
Zoomed in window shows our putative transmission pair, exhibited lowest genetic distances of any two in the tree

3. Single X4 transmitted/founder virus
(TFV)
Next performed env-v3 deep sequences on all plasma and pbmc samples
Did anc phylogenetic recons using only plasma sequences
Shown is a representative time stamped phylogenetic ancestral reconstruction, donor sequences are shown in green and recipient in purple
All 10 reconstructions indicated that a single X4 virus was transmitted from donor to recipient indicated by the single line connecting the sequences and labeled
Reconstruction by Dr. Art Poon

4. Transmission via transfer of an HIV infected cell?
Donor plasma and PBMC 1 month prior to transmission
Tracking the frequency of the exact nucleotide sequence of the TFV in the donor plasma and PBMC compartment 1 month prior to transmission
We found that in the plasma shown on the left, this virus represented a minority variant, whereas in the pbmc compartment it was a majority variant representing 33.5% of all sequences
This data possibly suggests that transmission and infection in the recipient possibly occurred through transfer of an HIV infected cell rather than free virus spread
Plasma
PBMC

5. Differential coreceptor evolution in donor and recipient
41%
60%
59%
40%
Lastly we wanted to compare the evolution of X4 in two genetically distinct hosts.
On the left are phylogenetic trees constructed using plasma deep sequences from the donor and recipient, again in red are X4 and blue R5
On the right is a summary of the frequency of X4 vs R5 overtime.
For the donor you can see a gradual reversion from X4 to R5 whereas the recipient remains consistently X4, this trend is also consistent for the PBMC compartment.
18%
60%
82%
26%

6. Summary Single X4 T/F virus from the donor to recipient
Infection occurred by transfer of HIV infected cell
Differential coreceptor evolution
In summary anc phylo recon revealed that a single X4 virus was transmitted, a number that is lower than expected for IDU
Data also suggests that transmission possibly occurred via the transfer of an HIV infected cell
Overtime in the donor and recipient exhibited differential coreceptor evolution

7. Acknowledgements Dr. Zabrina Brumme Dr. Richard Harrigan Jeremy Taylor
Dr. Mark Brockman
Gursev Anmole
Tallie Kuang
Tristan Markle
Bemulu Baraki
Philip Mwimanzi
Aniqa Shahid
Natalie Kinloch
Dr. Richard Harrigan
Jeremy Taylor
Winnie Dong
VIDUS participants