8fe4fad2c58e967b46b6874ff523c2ca pmc_9503691.pdf 2e7dac857f5b0e7e3719c95846a0288052c0a528 pmc_9503691.pdf d15e8246019ff14da33b183f1ee46433a44136239496344ec66d88e26e9f2b85 pmc_9503691.pdf Title: Autopsy Study Defines Composition and Dynamics of the HIV-1 Reservoir after Allogeneic Hematopoietic Stem Cell Transplantation with CCR532/32 Donor Cells Subject: Allo-HSCT with CCR532/32 donor cells is the only curative HIV-1 intervention. We investigated the impact of allo-HSCT on the viral reservoir in PBMCs and post-mortem tissue in two patients. IciS-05 and IciS-11 both received a CCR532/32 allo-HSCT. Before allo-HSCT, ultrasensitive HIV-1 RNA quantification; HIV-1-DNA quantification; co-receptor tropism analysis; deep-sequencing and viral characterization in PBMCs and bone marrow; and post-allo-HSCT, ultrasensitive RNA and HIV-1-DNA quantification were performed. Proviral quantification, deep sequencing, and viral characterization were done in post-mortem tissue samples. Both patients harbored subtype B CCR5-tropic HIV-1 as determined genotypically and functionally by virus culture. Pre-allo-HSCT, HIV-1-DNA could be detected in both patients in bone marrow, PBMCs, and T-cell subsets. Chimerism correlated with detectable HIV-1-DNA LTR copies in cells and tissues. Post-mortem analysis of IciS-05 revealed proviral DNA in all tissue biopsies, but not in PBMCs. In patient IciS-11, who was transplanted twice, no HIV-1-DNA could be detected in PBMCs at the time of death, whereas HIV-1-DNA was detectable in the lymph node. In conclusion, shortly after CCR532/32, allo-HSCT HIV-1-DNA became undetectable in PBMCs. However, HIV-1-DNA variants identical to those present before transplantation persisted in post-mortem-obtained tissues, indicating that these tissues play an important role as viral reservoirs. Keywords: HIV-1; HIV persistence; reservoir; cure; tissue; CCR532; allo-HSCT Author: Laura E. P. Huyveneers, Anke Bruns, Arjen Stam, Pauline Ellerbroek, Dorien de Jong, Noémi A. Nagy, Stephanie B. H. Gumbs, Kiki Tesselaar, Kobus Bosman, Maria Salgado, Gero Hütter, Lodewijk A. A. Brosens, Mi Kwon, Jose Diez Martin, Jan T. M. van der Meer, Theun M. de Kort, Asier Sáez-Cirión, Julian Schulze zur Wiesch, Jaap Jan Boelens, Javier Martinez-Picado, Jürgen H. E. Kuball, Annemarie M. J. Wensing and Monique Nijhuis Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Sep 21 04:32:59 2022 CEST ModDate: Wed Sep 21 04:46:50 2022 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 12 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 537230 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- ZZTQUO+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 JUTHNT+VnURWPalladioL Type 1 Custom yes yes no 15 0 EJAQZK+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 GFHGWB+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 MPUDFL+PazoMath Type 1 Builtin yes yes no 28 0 MAKAMH+URWPalladioL-Ital Type 1 Custom yes yes no 32 0 PDXFNG+PazoMath-Bold Type 1 Builtin yes yes no 36 0 HOWKDX+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 54 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 76 0 VIWIOF+CMSY10 Type 1 Builtin yes yes no 86 0 INJKKM+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 102 0 INJKKN+PalatinoLinotype CID TrueType Identity-H yes yes yes 105 0 INJKKO+PalatinoLinotype TrueType WinAnsi yes yes no 111 0 INJKLA+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 114 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-09-13 02:29:24 CEST RepresentationInformation: pmc_9503691.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-08-05 02:20:12 CEST Size: 537230 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 267 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Children: Item: Title: Cases Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: Patient IciS-05 Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Patient IciS-11 Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Children: Item: Title: Patients and Patient Material Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: DNA Isolation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Patient and Donor CCR5 Genotype Determination Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Ultra-Sensitive Viral Reservoir Quantification Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Micro-Chimerism Assessment Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Ultra-Sensitive Plasma HIV-1 RNA Determination Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: HIV-1 Co-Receptor Prediction and Determination Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Children: Item: Title: Patient IciS-05 Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Patient IciS-11 Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Info: Title: Autopsy Study Defines Composition and Dynamics of the HIV-1 Reservoir after Allogeneic Hematopoietic Stem Cell Transplantation with CCR532/32 Donor Cells Author: Laura E. P. Huyveneers, Anke Bruns, Arjen Stam, Pauline Ellerbroek, Dorien de Jong, Noémi A. Nagy, Stephanie B. H. Gumbs, Kiki Tesselaar, Kobus Bosman, Maria Salgado, Gero Hütter, Lodewijk A. A. Brosens, Mi Kwon, Jose Diez Martin, Jan T. M. van der Meer, Theun M. de Kort, Asier Sáez-Cirión, Julian Schulze zur Wiesch, Jaap Jan Boelens, Javier Martinez-Picado, Jürgen H. E. Kuball, Annemarie M. J. Wensing and Monique Nijhuis Subject: Allo-HSCT with CCR532/32 donor cells is the only curative HIV-1 intervention. We investigated the impact of allo-HSCT on the viral reservoir in PBMCs and post-mortem tissue in two patients. IciS-05 and IciS-11 both received a CCR532/32 allo-HSCT. Before allo-HSCT, ultrasensitive HIV-1 RNA quantification; HIV-1-DNA quantification; co-receptor tropism analysis; deep-sequencing and viral characterization in PBMCs and bone marrow; and post-allo-HSCT, ultrasensitive RNA and HIV-1-DNA quantification were performed. Proviral quantification, deep sequencing, and viral characterization were done in post-mortem tissue samples. Both patients harbored subtype B CCR5-tropic HIV-1 as determined genotypically and functionally by virus culture. Pre-allo-HSCT, HIV-1-DNA could be detected in both patients in bone marrow, PBMCs, and T-cell subsets. Chimerism correlated with detectable HIV-1-DNA LTR copies in cells and tissues. Post-mortem analysis of IciS-05 revealed proviral DNA in all tissue biopsies, but not in PBMCs. In patient IciS-11, who was transplanted twice, no HIV-1-DNA could be detected in PBMCs at the time of death, whereas HIV-1-DNA was detectable in the lymph node. In conclusion, shortly after CCR532/32, allo-HSCT HIV-1-DNA became undetectable in PBMCs. However, HIV-1-DNA variants identical to those present before transplantation persisted in post-mortem-obtained tissues, indicating that these tissues play an important role as viral reservoirs. Keywords: HIV-1; HIV persistence; reservoir; cure; tissue; CCR532; allo-HSCT Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Wed Sep 21 04:32:59 CEST 2022 ModDate: Wed Sep 21 04:46:50 CEST 2022 ID: 0xc238e65792533849913e426a1c8ce515, 0xc238e65792533849913e426a1c8ce515 Filters: FilterPipeline: FlateDecode Fonts: Type1: Font: BaseFont: MAKAMH+URWPalladioL-Ital FontSubset: true FirstChar: 2 LastChar: 121 FontDescriptor: FontName: MAKAMH+URWPalladioL-Ital Flags: Symbolic FontBBox: -170, -305, 1010, 941 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: EJAQZK+URWPalladioL-Bold FontSubset: true FirstChar: 2 LastChar: 252 FontDescriptor: FontName: EJAQZK+URWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: PDXFNG+PazoMath-Bold FontSubset: true FirstChar: 68 LastChar: 68 FontDescriptor: FontName: PDXFNG+PazoMath-Bold Flags: Symbolic FontBBox: -10, -266, 829, 695 FontFile: true Font: BaseFont: VIWIOF+CMSY10 FontSubset: true FirstChar: 0 LastChar: 48 FontDescriptor: FontName: VIWIOF+CMSY10 Flags: Symbolic FontBBox: -29, -960, 1116, 775 FontFile: true Font: BaseFont: GFHGWB+URWPalladioL-Roma FontSubset: true FirstChar: 2 LastChar: 252 FontDescriptor: FontName: GFHGWB+URWPalladioL-Roma Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: ZZTQUO+VnURWPalladioL-Bold FontSubset: true FirstChar: 161 LastChar: 226 FontDescriptor: FontName: ZZTQUO+VnURWPalladioL-Bold Flags: Symbolic FontBBox: -152, -301, 1000, 935 FontFile: true EncodingDictionary: Differences: true Font: BaseFont: MPUDFL+PazoMath FontSubset: true FirstChar: 68 LastChar: 68 FontDescriptor: FontName: MPUDFL+PazoMath Flags: Symbolic FontBBox: -40, -283, 878, 946 FontFile: true Font: BaseFont: OVNEXW+EURM10 FontSubset: true FirstChar: 22 LastChar: 22 FontDescriptor: FontName: OVNEXW+EURM10 Flags: Symbolic FontBBox: -32, -243, 1060, 720 FontFile: true Font: BaseFont: JUTHNT+VnURWPalladioL FontSubset: true FirstChar: 160 LastChar: 230 FontDescriptor: FontName: JUTHNT+VnURWPalladioL Flags: Symbolic FontBBox: -166, -283, 1021, 943 FontFile: true EncodingDictionary: Differences: true Pages: Page: Sequence: 1 Annotations: Annotation: Subtype: Link Rect: 34, 311, 147, 320 Annotation: Subtype: Link Rect: 34, 164, 96, 186 Annotation: Subtype: Link Rect: 126, 90, 154, 99 Annotation: Subtype: Link Rect: 34, 78, 147, 87 Annotation: Subtype: Link Rect: 34, 757, 147, 793 Annotation: Subtype: Link Rect: 515, 757, 560, 793 Annotation: Subtype: Link Rect: 148, 630, 159, 643 Annotation: Subtype: Link Rect: 233, 630, 244, 643 Annotation: Subtype: Link Rect: 408, 630, 419, 643 Annotation: Subtype: Link Rect: 145, 618, 156, 631 Annotation: Subtype: Link Rect: 163, 605, 175, 618 Annotation: Subtype: Link Rect: 274, 605, 285, 618 Annotation: Subtype: Link Rect: 427, 605, 438, 618 Annotation: Subtype: Link Rect: 543, 605, 554, 618 Annotation: Subtype: Link Rect: 293, 592, 304, 606 Annotation: Subtype: Link Rect: 112, 17, 241, 28 Annotation: Subtype: Link Rect: 411, 17, 560, 28 Annotation: Subtype: Link Rect: 34, 444, 87, 463 Page: Sequence: 2 Annotations: Annotation: Subtype: Link Rect: 367, 645, 374, 654 ActionDest: 127 Annotation: Subtype: Link Rect: 494, 607, 501, 616 ActionDest: 127 Annotation: Subtype: Link Rect: 272, 507, 279, 516 ActionDest: 127 Annotation: Subtype: Link Rect: 282, 507, 289, 516 ActionDest: 127 Annotation: Subtype: Link Rect: 234, 481, 241, 491 ActionDest: 127 Annotation: Subtype: Link Rect: 241, 481, 248, 491 ActionDest: 127 Annotation: Subtype: Link Rect: 249, 482, 256, 491 ActionDest: 127 Annotation: Subtype: Link Rect: 256, 481, 263, 491 ActionDest: 127 Annotation: Subtype: Link Rect: 206, 431, 213, 440 ActionDest: 127 Annotation: Subtype: Link Rect: 216, 431, 228, 440 ActionDest: 127 Annotation: Subtype: Link Rect: 168, 366, 246, 378 Annotation: Subtype: Link Rect: 531, 252, 538, 264 ActionDest: 80 Annotation: Subtype: Link Rect: 327, 164, 334, 176 ActionDest: 80 Page: Sequence: 3 Annotations: Annotation: Subtype: Link Rect: 466, 64, 473, 76 ActionDest: 80 Page: Sequence: 4 Annotations: Annotation: Subtype: Link Rect: 444, 683, 451, 695 ActionDest: 80 Annotation: Subtype: Link Rect: 544, 93, 556, 102 ActionDest: 127 Page: Sequence: 5 Annotations: Annotation: Subtype: Link Rect: 249, 559, 261, 568 ActionDest: 127 Annotation: Subtype: Link Rect: 366, 333, 378, 342 ActionDest: 131 Annotation: Subtype: Link Rect: 371, 308, 383, 317 ActionDest: 131 Annotation: Subtype: Link Rect: 455, 308, 467, 317 ActionDest: 131 Annotation: Subtype: Link Rect: 537, 94, 544, 103 ActionDest: 127 Annotation: Subtype: Link Rect: 429, 69, 441, 78 ActionDest: 131 Page: Sequence: 6 Annotations: Annotation: Subtype: Link Rect: 479, 735, 491, 744 ActionDest: 131 Annotation: Subtype: Link Rect: 198, 581, 205, 593 ActionDest: 96 Annotation: Subtype: Link Rect: 367, 556, 373, 569 ActionDest: 93 Page: Sequence: 7 Annotations: Annotation: Subtype: Link Rect: 295, 506, 301, 518 ActionDest: 93 Annotation: Subtype: Link Rect: 248, 468, 255, 480 ActionDest: 96 Annotation: Subtype: Link Rect: 290, 468, 297, 480 ActionDest: 93 Annotation: Subtype: Link Rect: 305, 393, 311, 405 ActionDest: 93 Annotation: Subtype: Link Rect: 368, 305, 375, 317 ActionDest: 96 Annotation: Subtype: Link Rect: 457, 280, 464, 292 ActionDest: 118 Annotation: Subtype: Link Rect: 221, 242, 228, 254 ActionDest: 118 Annotation: Subtype: Link Rect: 371, 167, 378, 179 ActionDest: 118 Annotation: Subtype: Link Rect: 414, 167, 421, 179 ActionDest: 96 Page: Sequence: 8 Annotations: Annotation: Subtype: Link Rect: 260, 195, 267, 207 ActionDest: 118 Annotation: Subtype: Link Rect: 358, 135, 365, 144 ActionDest: 127 Annotation: Subtype: Link Rect: 368, 135, 375, 144 ActionDest: 127 Annotation: Subtype: Link Rect: 375, 135, 382, 144 ActionDest: 127 Annotation: Subtype: Link Rect: 385, 135, 392, 144 ActionDest: 127 Annotation: Subtype: Link Rect: 393, 135, 405, 144 ActionDest: 127 Annotation: Subtype: Link Rect: 405, 135, 417, 144 ActionDest: 131 Annotation: Subtype: Link Rect: 420, 135, 432, 144 ActionDest: 131 Page: Sequence: 9 Annotations: Annotation: Subtype: Link Rect: 219, 698, 231, 707 ActionDest: 131 Annotation: Subtype: Link Rect: 544, 622, 556, 631 ActionDest: 131 Annotation: Subtype: Link Rect: 377, 522, 389, 531 ActionDest: 127 Annotation: Subtype: Link Rect: 336, 397, 343, 406 ActionDest: 127 Annotation: Subtype: Link Rect: 360, 233, 371, 242 ActionDest: 131 Annotation: Subtype: Link Rect: 372, 233, 384, 242 ActionDest: 131 Annotation: Subtype: Link Rect: 366, 208, 378, 217 ActionDest: 127 Annotation: Subtype: Link Rect: 379, 208, 391, 217 ActionDest: 131 Annotation: Subtype: Link Rect: 391, 208, 403, 217 ActionDest: 131 Page: Sequence: 10 Annotations: Annotation: Subtype: Link Rect: 536, 659, 560, 670 Annotation: Subtype: Link Rect: 165, 647, 369, 658 Page: Sequence: 11 Annotations: Annotation: Subtype: Link Rect: 117, 496, 155, 506 Annotation: Subtype: Link Rect: 161, 496, 197, 506 Annotation: Subtype: Link Rect: 59, 462, 96, 472 Annotation: Subtype: Link Rect: 102, 462, 138, 472 Annotation: Subtype: Link Rect: 332, 426, 369, 437 Annotation: Subtype: Link Rect: 519, 403, 557, 414 Annotation: Subtype: Link Rect: 59, 393, 95, 403 Annotation: Subtype: Link Rect: 149, 324, 186, 334 Annotation: Subtype: Link Rect: 192, 324, 228, 334 Annotation: Subtype: Link Rect: 99, 255, 137, 265 Annotation: Subtype: Link Rect: 143, 255, 179, 265 Annotation: Subtype: Link Rect: 454, 231, 491, 242 Annotation: Subtype: Link Rect: 497, 231, 533, 242 Annotation: Subtype: Link Rect: 181, 196, 219, 207 Annotation: Subtype: Link Rect: 423, 162, 460, 173 Annotation: Subtype: Link Rect: 157, 128, 194, 138 Annotation: Subtype: Link Rect: 289, 93, 326, 104 Annotation: Subtype: Link Rect: 332, 93, 369, 104 Annotation: Subtype: Link Rect: 203, 59, 241, 69 Annotation: Subtype: Link Rect: 247, 59, 283, 69 Page: Sequence: 12 Annotations: Annotation: Subtype: Link Rect: 117, 722, 155, 733 Annotation: Subtype: Link Rect: 446, 687, 483, 698 Annotation: Subtype: Link Rect: 489, 687, 525, 698 Annotation: Subtype: Link Rect: 157, 665, 194, 675 Annotation: Subtype: Link Rect: 200, 665, 236, 675 Annotation: Subtype: Link Rect: 477, 641, 515, 652 Annotation: Subtype: Link Rect: 521, 641, 557, 652 Annotation: Subtype: Link Rect: 219, 618, 257, 629 Annotation: Subtype: Link Rect: 237, 583, 275, 594 Annotation: Subtype: Link Rect: 358, 549, 396, 560 Annotation: Subtype: Link Rect: 356, 514, 393, 525 Annotation: Subtype: Link Rect: 308, 480, 346, 491 Annotation: Subtype: Link Rect: 352, 480, 388, 491 Annotation: Subtype: Link Rect: 131, 435, 168, 445 Annotation: Subtype: Link Rect: 104, 400, 141, 410 Annotation: Subtype: Link Rect: 147, 400, 183, 410 Annotation: Subtype: Link Rect: 59, 366, 96, 376 Annotation: Subtype: Link Rect: 102, 366, 138, 376 Annotation: Subtype: Link Rect: 362, 330, 400, 341 Annotation: Subtype: Link Rect: 406, 330, 442, 341 Checksum: 5112ab14 Type: CRC32 Checksum: 8fe4fad2c58e967b46b6874ff523c2ca Type: MD5 Checksum: 2e7dac857f5b0e7e3719c95846a0288052c0a528 Type: SHA-1