CROI 2016 Abstract eBook

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Poster Abstracts

ART (since 06/2014 ABC/3TC/DTG) and the viral load remained undetectable in plasma and liquor. He had a 2 nd relapse of AML in 06/2013 but re-entered molecular remission after a total of 8 courses of 5-azacytidine and 4 donor lymphocyte infusions. Concerning HIV, all collected samples were negative for proviral DNA by conventional and digital droplet PCR* in two different labs, namely PBMCs (06/2014, 01/2015* and 02/2015), rectal biopsy (04/2015) and bone marrow (08/2015*). Western blots from 06/2014 and 02/2015 showed incomplete patterns with fading bands. Conclusions: Like in the Berlin patient, all tests from the Duesseldorf patient so far suggest that HIV may have been eradicated and that he may be the second individual cured from HIV by allogeneic CCR5-d32 HSCT. Further investigations will be performed before considering the discontinuation of ART. 365 HIV-1 Viral Rebound Following Allogeneic Hematopoietic Stem-Cell Transplantation Kersten K. Koelsch 1 ; John J. Zaunders 2 ; Angie Pinto 1 ; Kazuo Suzuki 1 ; Sarah C. Sasson 3 ; Bonnie Hiener 4 ; Sarah Palmer 4 ; SamT. Milliken 2 ; Anthony Kelleher 5 ; David Cooper 6 ; for the Sydney HIV SCT Study Group 1 Kirby Inst, Univ of New South Wales, Sydney, Australia; 2 St Vincent’s Hosp, Darlinghurst, Australia; 3 St Vincent’s Hosp, Sydney, Australia; 4 Cntr for Virus Rsr, Westmead Millennium Inst, Westmead, Australia; 5 Univ of New South Wales, Sydney, Australia; 6 Kirby Inst, Sydney, Australia Background: Allogeneic stem cell transplantation (SCT) has been shown to lead to significant changes to the HIV-1 reservoir and the HIV-1 immune response. However, these changes may not result in viral control upon cessation of antiretroviral therapy (ART). Methods: We studied one patient who had undergone allogeneic SCT with reduced intensity conditioning (RIC) for treatment of non-Hodgkin’s lymphoma (NHL). HIV-1 antigens and antibodies (Ag/Ab) were measured by 4 th generation chemiluminescence microparticle immunoassay (CMIA), Western blot (WB) and HIV-1 Ab avidity assays. HIV-1 specific CD4+ T cell responses were measured by CD25/CD134 upregulation. HIV-1 RNA levels in plasma were measured by two separate real-time PCR assays with 20 and single copy/ml sensitivity; HIV-1 DNA levels were assessed in peripheral blood mononuclear cells (PBMCs) and in isolated CD4+ T cells by PCR using three different primer sets. Viral genotypic sequence analysis was performed by sequencing of the pol RT, PR and INT regions. The presence of the CCR5Δ32 mutation was tested by PCR. Results: The patient received an HLA matched, allogeneic SCT in 2010 for NHL. ART was continued throughout and following the procedure. Post-transplant he experienced systemic grade 2 graft-versus-host disease (GVHD). He was heterozygous for the CCR5Δ32 mutation post-transplant, had no detectable HIV-1 RNA in plasma by either real-time PCR assay, and no detectable HIV-1 DNA by PCR in PBMCs or CD4+ T cells. CD4+ T cell responses to HIV-1 antigen were absent. Ag/Abs to HIV-1 were detectable at very low levels by CMIA, WB and Ab avidity assays. However, the patient experienced viral rebound 4 ½ years following the SCT with high levels of HIV-1 pVL (>10,000,000 copies), seroconversion by CMIA and WB and increasing HIV-1 Ab avidity. HIV-1 DNA levels were readily detectable by real-time PCR following the viral rebound. The patient reported strict treatment adherence but ART drug levels were profoundly reduced at the time of viral rebound, possibly attributable to poor drug absorption. There was no new ART drug-resistance or superinfection based on genotypic sequence analyses. Conclusions: This case describes HIV-1 viral rebound in a patient with profoundly reduced HIV-1 Ag/Ab and DNA levels following SCT, underlining the poor predictive value for viral rebound of these markers. This is highly relevant for the interpretation of these measures in the context of interventions targeting the HIV reservoir. 366 A Tale of Two Stem-Cell Transplantations in HIV+ Patients: Clues to Eradicate HIV Maria Salgado 1 ; Mi Kwon 2 ; Monique Nijhuis 3 ; Jan van Lunzen 4 ; Julià Blanco 5 ; Julian Schulze zurWiesch 4 ; Gero Hutter 6 ; Anne M.Wensing 3 ; Jose Luis Diez 2 ; Javier Martinez-Picado 1 ; for the EpiStem Consortium 1 IrsiCaixa Inst for AIDS Rsr, Badalona, Spain; 2 Hosp General Universitario Gregorio Marañón, Madrid, Spain; 3 Univ Med Cntr Utrecht, Utrecht, Netherlands; 4 Univ Med Cntr Hamburg- Eppendorf, Hamburg, Germany; 5 Univ de Vic, Barcelona, Spain; 6 Cellex, Dresden, Germany Background: To date, Berlin patient’s case provides the only evidence of long-term HIV-1 control after allogeneic stem cell transplantation (SCT), potentially due to the CCR5Δ32/ Δ32 donor genotype. The case of the two Boston patients, without resulting in HIV cure, showed that the SCT procedure itself involves a tremendous reduction of the viral reservoir. The reason why this happened is unraveled yet. Herein we analyzed two patients from the European cohort EpiStem receiving different types of SCT Methods: Patient 1 (Pt1) and Patient 3 (Pt3) were transplanted in 2012 and 2013 in Hospital Gregorio Marañón due to a Burkitt NHL and NK-NHL respectively. Analysis of the HIV-1 reservoir was performed 29 (Pt1) and 20 (Pt3) months post-transplant using qVOA, ddPCR, and semiquantitative PCR in CD4 T cells from peripheral blood and ileum, and CD3+ cells from bone marrow (BM). SCA was performed in plasma and CSF. Chimerismwas performed by short tandem repeat PCR (STR-PCR). Antibody titers were determined by ELISA. Activation markers (CD38 and HLA-DR) were determined in CD4 and CD8 T-cells Results: Pt 1 received a myeloablative single cord blood SCT, supported with third party HLA-mismatched CD34+ cells (haplo-cord SCT). Pt 3 underwent reduced intensity conditioning SCT with peripheral blood progenitor cells (PBPC) of an HLA-matched sibling. Both patients were kept on cART. Donors were wt for CCR5. Pt 3 reached full chimera 1 month after SCT, while Pt 1 still harbors 0.2% of host cell in BM and 0.1% in PB. Pt 3 developed chronic GvH disease but not Pt 1. CD4 T cells from both patients were susceptible to in vitro R5 and X4-tropic HIV infection. IUPM in Pt 1 were 0.034 but undetectable (IUPM< 0.006) in Pt3. Total DNA in peripheral CD4 were 25 copies/10 6 cells in Pt1, with 5 HIV RNA copies/ml in plasma. On the contrary, we were unable to detect virus in peripheral CD4, ileum CD4, BM, plasma or CSF in Pt3. Although both patients had a reduction in HIV-specific antibody titers, Pt 3 had lower levels. CD4 and CD8 activation levels were lower in Pt 3 than Pt 1 Conclusions: Within the EpiStem cohort, we compared two different SCT procedures, with different outcome regarding the viral reservoir. Allogeneic SCT with PBPC of an HLA-matched sibling donor, with a short engraftment time and a chronic GvH disease resulted in a more drastic reduction of the latent reservoir down to undetectable levels. We hypothesize that the “graft versus HIV-1 reservoir effect” contributes to facilitate the clearance of the viral reservoir 367 Ex Vivo Determination of Stem-Cell Transplantation Graft-Versus-HIV Reservoir Effects Louise Hogan 1 ; Kristen S. Hobbs 1 ; Emily Hanhauser 1 ; Christine D. Palmer 2 ; Marisol Romero-Tejeda 2 ;Yvonne P. Robles 3 ; Stephanie Jost 2 ; Daniel Kuritzkes 4 ; Jerome Ritz 5 ;Timothy J. Henrich 1 1 Univ of California San Francisco, San Francisco, CA, USA; 2 Ragon Inst of MGH, MIT, and Harvard, Cambridge, MA, USA; 3 Brigham and Women’s Hosp, Harvard Med Sch, Boston, MA, USA; 4 Harvard Med Sch, Boston, MA, USA; 5 Dana-Farber Cancer Inst, Boston, MA, USA Background: Allogeneic hematopoietic stem cell transplantation (HSCT) is one of the few strategies that leads to substantial decreases in HIV-1 reservoir size. Graft-versus-host (GVH) responses are likely responsible for ongoing clearance of residual recipient cells capable of harboring HIV. Beneficial GVH responses, such as those that permit donor cells to clear tumor or residual host hematopoietic cells, may be mediated largely by the innate immune system. To investigate the role of NK cells and other lymphocytes in reactivating and eliminating latent HIV following HSCT, we designed a novel ex vivo assay to determine the activity of HLA-matched, post-HSCT donor effector cells on latently infected, pre-HSCT host CD4 T cells. Methods: We adapted a latency model to enable infection of high numbers of CD4 T-cells from individuals with hematopoietic malignancies prior to HSCT with an iGFP-gag HIV viral strain. The infected pre-HSCT CD4 T cells were then co-incubated with PBMC obtained from the same individuals 9-12 months after HSCT. The PBMC were obtained following engraftment and development of full donor cell chimerism ( i.e. post-HSCT cells are of donor origin despite being recovered from the transplant recipient). We then determined lymphocyte activation, proliferation, viral reactivation and death over a two week period using flow cytometric analyses. Results: We included samples from a total of 30 HIV-negative individuals who received either full myeloablative or reduced intensity conditioning HSCT. Up to 95% pre-HSCT CD4+ T cells were infected with iGFP-HIV-1, with subsequent resting resulting in large numbers of latently infected cells. Flow cytometry was performed 0-13 days following lymphocyte mixing and co-culture. Of note, higher levels of non-proliferating HIV reactivated cells were found in the autogeneic setting compared to that of the allogeneic samples. Conversely, higher levels of proliferating HIV infected cells were seen in the allogeneic samples, peaking at day 7. While expression of activation markers increased on NK,

Poster Abstracts

139

CROI 2016

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