SUPPLEMENTARY INFORMATION
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- Σίβύλ Μάγκας
- 6 χρόνια πριν
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1 SUPPLEMENTARY INFORMATION doi:.8/nture7 Supplementry Tble. Serum neutrlizing ctivity of selected donors. Clde A Clde B Clde C CRF_AE Donor presumed clde Score 94UG 9BR JRCSF MGRM-C6 9IN95 9TH #6 CRF_AG #84 A or D #7 A < #9 C Supplementry Tble : Antibody sequence chrcteristics Hevy Chin Sequences Puttive V- CDR length Somtic muttions Somtic muttions Muttion frequency Muttion frequency Nme Donor gene gene (mino cids) b CDR sequence (mino cids) (nucleotides) c (mino cids) c (nucleotides) c (mino cids) c PGT TLHGRRIYGIVAFNEWFTYFYMDV % % 7 IGHV4-59* IGHJ6* 4 Insertion/Deletions (number of mino cids/position) PGT TKHGRRIYGVVAFKEWFTYFYMDV 68 8 % 4 % PGT ALHGKRIYGIVALGELFTYFYMDV 79 5 % 7 % PGT5 FDGEVLVYNHWPKPAWVDL 78 6 % 7 % +6 in CDR PGT6 FDGEVLVYHDWPKPAWVDL 67 7 % % +6 in CDR PGT7 FGGEVLVYRDWPKPAWVDL 58 5 % % +6 in CDR PGT8 6 IGHV4-9*7 IGHJ5* 9 FGGEVLRYTDWPKPAWVDL % 8 % +6 in CDR PGT SGGDILYYYEWQKPHWFSP 8 4 % 4 % PGT SGGDILYYIEWQKPHWFYP 84 4 % % PGT5 HRHHDVFMLVPIAGWFDV % 9 % +5 in CDR PGT6 9 IGHV4-9*7 IGHJ5* 8 HKYHDIFRVVPVAGWFDP 6 5 % 5 % +5 in CDR/+ in CDR PGT7 HKYHDIVMVVPIAGWFDP 77 8 % 9 % +5 in CDR PGT4 GSKHRLRDYVLYDDYGLINYQEWNDYLEFLDV 5 9 % % or + in CDR d PGT4 GSKHRLRDYVLYDDYGLINYQEWNDYLEFLDV 5 % % or + in CDR d PGT4 84 IGHV-8* IGHJ6* / GSKHRLRDYVLYDDYGLINYQEWNDYLEFLDV 5 9 % % or + in CDR d PGT44 GSKHRLRDYVLYDDYGLINQQEWNDYLEFLDV 59 4 % 4 % or + in CDR d PGT45 GSKHRLRDYFLYNEYGPNYEEWGDYLATLDV % 9 % - or in CDR d Puttive J- Light Chin Sequences Puttive V- CDR length Somtic Somtic Muttion Muttion Insertion/Deletions muttions muttions frequency frequency (number of mino Nme Donor gene gene (mino cids) b CDR sequence (mino cids) (nucleotides) c (mino cids) c (nucleotides) c (mino cids) c cids/position) PGT HIWDSRVPTKWV 56 8 % % -7 in FR/+ in FR PGT 7 IGLV-* IGLJ* HIWDSRRPTNWV % 6 % -7 in FR/+ in FR PGT HIYDARGGTNWV % 8 % -7 in FR/+ in FR PGT5 GSLVGNWDVI 46 5 % % -5 in CDR PGT6 SSLVGNWDVI % % -5 in CDR PGT7 IGLJ* or SSLVGNWDVI 7 9 % % -5 in CDR 6 IGLV-8* PGT8 IGLJ* GSLVGNWDVI % 4 % -5 in CDR PGT SSLFGRWDVV 9 % 9 % PGT SSLSGRWDIV 44 5 % % PGT5 QQYEEWPRT % 8 % PGT6 9 IGKV-5* IGKJ* 9 QQYEEWPRT 9 % % PGT7 QQYEEWPRT 5 9 % 8 % PGT4 MQGLNRPWT 46 4 % % PGT4 MQGLNRPWT 46 4 % % IGKV-8* or PGT4 84 IGKJ* 9 IGKVD-8* MQGLNRPWT % % PGT44 MQGLNRPWT 44 5 % % PGT45 MQGLHSPWT % 4 % ) Puttive V- or J-gene of the common germline ncestor of ech clonlly relted ntibody cluster. b) CDR lenghts ccording to the Kbt definiton. c) Somtic muttions were counted over the whole vrible region s nucleotides or mino cids differing from puttive germline sequence. For ech cluster of clonlly relted ntibodies, germline sequence ws composed of the puttive V-gene, consensus junction, the puttive J-gene, nd consensus insertion where present. Sequences derived from ' cloning primers were excluded from the frequency clcultions. d) Either n insertion in PGT4 to 44 or deletion in PGT45. e) Insertion of two mino cids in J-region due to mispriming in cloning PCR.
2 RESEARCH SUPPLEMENTARY INFORMATION Supplementry Tble : Medin ICs nd percent viruses neutrlized A) Medin IC 5 (μg/ml) ginst viruses neutrlized with n IC 5 < 5 μg/ml Clde n PGT PGT PGT PGT5 PGT6 PGT7 PGT8 PGT PGT PGT5 PGT6 PGT7 PGT4 PGT4 PGT4 PGT44 PGT45 VRC PGV4 b G 4E A B C D F n 9..6 G AE n n n..77 n n n n n.6 AG All B) Percent viruses neutrlized with n IC 5 < 5 μg/ml Clde n PGT PGT PGT PGT5 PGT6 PGT7 PGT8 PGT PGT PGT5 PGT6 PGT7 PGT4 PGT4 PGT4 PGT44 PGT45 VRC PGV4 b G 4E A B C D F G AE AG All C) Medin IC 5 (μg/ml) ginst ll viruses Clde n PGT PGT PGT PGT5 PGT6 PGT7 PGT8 PGT PGT PGT5 PGT6 PGT7 PGT4 PGT4 PGT4 PGT44 PGT45 VRC PGV4 b G 4E A B C D F G AE AG All D) Medin IC 9 (μg/ml) ginst viruses neutrlized with n IC 9 < 5 μg/ml Clde n PGT PGT PGT PGT5 PGT6 PGT7 PGT8 PGT PGT PGT5 PGT6 PGT7 PGT4 PGT4 PGT4 PGT44 PGT45 VRC PGV4 b G 4E A n B n C D F n n G n n 6.57 AE n n n n n n n n.96 AG n n n n All E) Percent viruses neutrlized with n IC 9 < 5 μg/ml Clde n PGT PGT PGT PGT5 PGT6 PGT7 PGT8 PGT PGT PGT5 PGT6 PGT7 PGT4 PGT4 PGT4 PGT44 PGT45 VRC PGV4 b G 4E A B C D F G AE AG All F) Medin IC 9 (μg/ml) ginst ll viruses Clde n PGT PGT PGT PGT5 PGT6 PGT7 PGT8 PGT PGT PGT5 PGT6 PGT7 PGT4 PGT4 PGT4 PGT44 PGT45 VRC PGV4 b G 4E A B C D F G AE AG All
3 SUPPLEMENTARY INFORMATION RESEARCH Supplementry Tble 4. Neutrlizing ctivity of PGT mabs ginst cross-clde 6-pseudovirus pnel. IC5 (µg/ml) IC5 (/dil'n) Isolte Subtype PGT- PGT- PGT- PGT-5 PGT-6 PGT-7 PGT-8 PGT- PGT- PGT-5 PGT-6 PGT-7 PGT-4 PGT-4 PGT-4 PGT-44 PGT-45 VRC PGV4 #7 #84 #6 #9 9RW > > RW >5 > >5 >5 > < 9RW >5.5 >5 > RW >5 >5.56 >5 >5 >5 > RW4 >5 >5 > >5 >5 >5 >5 >5 >5 > < RW > N/A UG >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > < UG >5 > RW9 >5 >5 >5 >5 >5 >5 > >5 >5 >5 > > < 589. < 7. 9UG >5 >5 >5 >5 >5 >5 >5 >5 > > UG >5 >5 >5 >5 >5 >5 > MGRM-A- >5 >5 >5.9 >5 >5 >5 >5 >5 >5 >5 > > >5 <.7 < 49. MGRM-A >5 >5 >5 > N/A MGRM-A- A >5 >5 >5 >5 >5 >5 >5 >5 > > < 6. < 75. MGRM-A-4 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > < < 9. MGRM-A-5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > < 5. < 8. MGRM-A-6.96 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > > MGRM-A-7 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > < < < 556. MGRM-A-8 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > >5 < < < 96. MGRM-A >5 >5 > MGRM-A >5 >5 > MGRM-A- >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > < 4. < 69. MGRM-A >5 >5 >5 >5 >5 >5 >5 >5 >5. > < MGRM-A >5 >5 >5 > > MGRM-A > > VLGCA >5 >5 >5.78 >5 >5.88 >5 >5 >5 >5 > > <.. < 94KE5 AC > TH >5 >5 > > >5 >5 > < CMU >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > >5 <. <. MGRM-AE- >5 >5 > > >5 >5 >5 >5 >5 >5 > < <.6. MGRM-AE- >5 >5 >5 >5 >5 > >5 >5 >5 >5 >5 >5 >5 > < MGRM-AE- >5 >5 >5.9 >5 > >5 >5 >5 >5 >5 >5 > < 5. < 8. MGRM-AE-4 AE >5 >5 >5 >5 >5 >5 >5.49 >5 >5 >5 > < MGRM-AE-5 >5 >5 >5..4 > >5 >5 >5 >5 >5 >5 > < MGRM-AE-6 >5 >5 > > >5 >5 >5 >5 >5 >5 >5 > >5.6 < MGRM-AE-7 >5 >5 >5.876 >5 >5 >5.8.6 >5 >5 > > < MGRM-AE-8 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > < 6.7 < 4. MGRM-AG >5 >5 >5 4.7 >5 >5 >5 >5 > > MGRM-AG- >5 >5 >5 >5 >5 > >5 >5 >5 >5 > MGRM-AG >5 >5 >5 > >5 >5 >5 >5 >5 >5 >5 > >5 < 55. <. MGRM-AG > >5 >5 > > > MGRM-AG >5 >5 >5 > > MGRM-AG-8 AG.494 >5 9.6 >5 >5 >5.668 >5 >5 >5 >5 > > < 69. MGRM-AG >5 >5 >5 >5 >5 >5 >5 >5 >5 >5.48 > MGRM-AG- >5 >5 >5 >5.45 >5.9 >5 >5 >5 >5 > < MGRM-AG- >5 >5 >5 >5 >5 >5 >5 >5 > >5 >5 >5 >5 >5 >5 > < MGRM-AG >5 >5 >5 >5.79 >5 > IC5 (µg/ml) IC5 (/dil'n) b Isolte Subtype PGT- PGT- PGT- PGT-5 PGT-6 PGT-7 PGT-8 PGT- PGT- PGT-5 PGT-6 PGT-7 PGT-4 PGT-4 PGT-4 PGT-44 PGT-45 VRC PGV4 #7 #84 #6 # >5 >5 >5 >5 >5 >5 >5 > BR > TH >5.4 >5 > > APV_ >5 >5 >5 >5 >5 > > APV_ >5 >5 >5 > APV_ >5 >5 >5 >5 > > CAAN.A >5 > >5 >5 >5 >5 >5 > JRFL >5 >5 >5 >5 >5 >5 > > MGRM-Chronic-B >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > > MGRM-Chronic-B >5 >5 >5 >5 >5 >5 >5 >5 > < MGRM-Chronic-B >5 >5 >5 >5 > > MGRM-Chronic-B >5 >5 > MGRM-Chronic-B > >5 >5 >5 >5 >5 >5 >5 > MGRM-Chronic-B >5 >5 >5 >5 >5 >5 >5 > MGRM-Chronic-B- >5 >5 > >5 >5 > > MGRM-Chronic-B >5 >5 >5 >5 > > B MGRM-Chronic-B-7 > >5 >5 >5 >5 >5 >5 >5 >5 >5 > < 8. MGRM-Chronic-B-8 >5 >5 >5 > >5 >5 >5 > < MGRM-Chronic-B >5.85 >5 >5 >5 >5 > > MGRM-Chronic-B >5 >5 >5 >5 >5 >5.9 >5 > > MGRM-Chronic-B >5 >5 >5 >5 > >5 >5 >5 > PVO >5 >5 >5 > QH >5 >5 >5 >5 >5 >5 >5 >5 >5 > >5 75. < SC >5 >5.75 >5 > > <. SF >5 >5 >5 >5 >5.4.8 > THR.8 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > > < TRJ > >5 >5 >5 >5 >5 >5 >5 > TRO VLGCB >5.4 >5.6 >5 >5 >5 >5 > NL4 >5 >5 >5 >5 >5 >5 >5 >5 > >5 >5 >5 > > JRCSF >5 > IN MW >5 >5 >5 > > ZA > >5 >5 >5 >5 >5 >5 > IN >5 > MGRM-C >5 >5 >5 >5 >5 > >5 >5 >5 >5 >5.69 >5 >5 < 7. < 74. MGRM-C >5 >5 >5 > > >5 > MGRM-C >5 >5 >5 >5 > MGRM-C >5 >5 > >5 > MGRM-C >5 >5 > > MGRM-C > >5. >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > < 44. MGRM-C >5.8 > >5 >5 >5 >5 >5 >5 >5 >5 > > < 9. MGRM-C-9 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > >5 < 46.9 < 4. MGRM-C- >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > >5 < 86.6 < 6. MGRM-C > >5 >5.7 >5. >5 >5 >5 > C MGRM-C >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 < < < 45. MGRM-C >5. >5 >5 >5 >5 >5 >5 >5 >5 > > > < 7.6 < < MGRM-C >5.4 >5.4 > MGRM-C >5 >5 >5 >5 > > MGRM-C-9 >5 >5 > > < MGRM-C- >5.955 >5 >5 >5 > >5 >5 >5 >5 > > < MGRM-C >5 >5 > MGRM-C MGRM-C >5 >5 >5 >5 >5 >5 >5 >5 > > < 7. MGRM-C >5.489 >5 >5 >5 >5 > > MGRM-C >5 >5 >5 >5 > MGRM-C > > >5 >5 > > MGRM-C >5 > > > < 5.
4 RESEARCH SUPPLEMENTARY INFORMATION IC5 (µg/ml) IC5 (/dil'n) b Isolte Subtype PGT- PGT- PGT- PGT-5 PGT-6 PGT-7 PGT-8 PGT- PGT- PGT-5 PGT-6 PGT-7 PGT-4 PGT-4 PGT-4 PGT-44 PGT-45 VRC PGV4 #7 #84 #6 #9 98CN9 CRF7_BC > CN6 CRF8_BC >5 >5 >5 >5 > > UG >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > >5 < < < 6. 9UG5 8.9 > > >5 >5 >5 >5 >5 >5 >5 > > < UG4 >5 >5 >5 >5 >5 >5 >5 >5 > >5 >5 >5 > < UG46 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > <.5 < 7. 94UG >5 >5.89 >5 >5 >5 >5 >5 > < MGRM-D >5 >5 >5 >5 >5 >5 >5 >5 >5.56 >5 >5 >5 < <.8 < MGRM-D >5 >5 >5 >5 >5 >5 >5 > < MGRM-D-.685 > >5 >5 >5 >5 >5 >5 > > MGRM-D-4 >5 >5 >5.7 >5 > >5 >5 >5 >5 >5 >5 > < MGRM-D-5 >5 >5 >5 > >5 >5 >5 >5 > >5 >5.8 < MGRM-D-8 >5 >5 >5 >5 4.9 >5. >5 >5 >5 >5 > < 48.8 < 56. MGRM-D > >5 >5 >5 > MGRM-D- >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > < D MGRM-D- >5 >5 >5 >5 >5 > >5 >5 > > < MGRM-D >5 > MGRM-D-6 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > < MGRM-D >5 5. >5 >5 >5 >5.65 > MGRM-D-9 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > > < MGRM-D >5 >5 >5 >5 >5 >5 >5 > < MGRM-D >5 >5 >5 >5 >5 >5 >5 >5.445 >5 >5 < < MGRM-D- >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 4.5 >5 >5 >5 < < < < MGRM-D > >5 >5 >5 >5 >5 >5 >5 >5 > > MGRM-D-6 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > > < 4.4. < MGRM-D >5 >5 >5 >5.85 >5 >5 >5 >5 >5 >5 >5 > < 56. MGRM-D-9 >5 >5 >5 >5 >5 >5 >5 >5 >5.849 >5 > >5 < 9.4 < 87. MGRM-F > MGRM-F >5 >5 >5 >5 > > < MGRM-F-8 >5 >5 >5 >5 >5 >5 > >5 >5 > >5 < 4. < 8. MGRM-F >5 >5 > >5 >5 >5 >5 >5 >5 >5 >5 > < 564. MGRM-F-.97 >5. >5. >5.7 >5 >5. >5 >5 >5 >5 >5 > MGRM-F >5 >5 >5.7 >5 >5.8 >5 > MGRM-F >5 >5 > >5 >5 >5 > < 58. MGRM-F >5 >5 >5 >5 >5 >5 >5.4 >5 >5 >5 >5 >5 > >5 < 45.7 < 4. F MGRM-F >5 >5 >5.9 >5 >5 >5 >5 >5 >5 >5 >5 > < 47. MGRM-F >5 >5 >5 >5 >5.659 >5 > >5 > > >5 < 79.4 < 8. MGRM-F > >5 >5 >5 >5 > > MGRM-F >5 >5 > > < 6. MGRM-F >5 >5 >5 >5 >5 >5.65 >5 >5 >5 >5 >5 > > MGRM-F >5 >5 > >5 > MGRM-F >5.88 >5 > > IC5 (µg/ml) IC5 (/dil'n) b Isolte Subtype PGT- PGT- PGT- PGT-5 PGT-6 PGT-7 PGT-8 PGT- PGT- PGT-5 PGT-6 PGT-7 PGT-4 PGT-4 PGT-4 PGT-44 PGT-45 VRC PGV4 #7 #84 #6 #9 MGRM-G >5.8.7 > MGRM-G >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > MGRM-G-6 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > < 548. < < MGRM-G >5 >5 >5 >5 >5 >5 >5 >5 > > < 45. MGRM-G >5 7.8 >5 8. >5 >5 >5 >5 > < 64. < 89. MGRM-G > >5 >5 >5 >5 >5 >5 >5 >5 >5 > > MGRM-G >5 >5 > MGRM-G-5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > < < < 66. G MGRM-G-6 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 > > < MGRM-G >5 >5 >5.9 >5 >5.7 > MGRM-G > > > MGRM-G >5 >5 > MGRM-G > >5 >5 >5 >5 >5 >5 > > MGRM-G > >5 >5 >5 > MGRM-G >5 >5 > > MLV negtive >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 >5 < < < < 4
5 SUPPLEMENTARY INFORMATION RESEARCH Supplementry Tble 5. Binding ctivity of PGT mabs. Donor mab # #6 8 5 #9 6 7 EC5 (µg/ml) WT JR-FL gp JR-FL gp V/ V JR-FL gp V Endo H treted JR-FL gp.. > >.. > >.. > >..6 > >.. > >..7 > > >. > > >.4 > > >... >... >... > Binding ws evluted by ELISA. EC5 vlues were derived by nonliner regression nlysis. Experiments were performed in duplicte, nd dt represent n verge of t lest two independent experiments. Supplementry Tble 6. Neutrlizing ctivity of PGT mabs ginst pnel of JR-CSF lnine mutnts. Fold IC5 increse reltive to wild-type c Muttion gp domin b PGT- PGT- PGT- PGT-5 PGT-6 PGT-7 PGT-8 PGT- PGT- PGT-5 DA ND.8 VA C L5A V7A N4A V N56A N6K T6A I65A R66A D67A K68A V E7A ND. Y77A L79A V8A D85A N88A N97A S99A C (V/V stem) TA FA N4A C N6A N76A
6 RESEARCH SUPPLEMENTARY INFORMATION Fold IC5 increse reltive to wild-type c Muttion gp domin b PGT- PGT- PGT- PGT-5 PGT-6 PGT-7 PGT-8 PGT- PGT- PGT-5 V9A N95A >8 T97A > P99A NA >5.7 > >7.5 NA TA.7.5. >5 >5 >5 >5 >5 >7. R4A K5A ND. S6A I7A >7. I9A > PA V R5A F7A T9A TA EA IA G4A ND ND >5 ND.4 D5A > >7.5 I6A R7A HA >5 NA > > >. 8 > >5 S4A >5 >5 >5.8 > > > Q7A N9A T4A K4A R5A C N55A N86A S87A T88A Fold IC5 increse reltive to wild-type c Muttion gp domin b PGT- PGT- PGT- PGT-5 PGT-6 PGT-7 PGT-8 PGT- PGT- PGT-5 N9Q >5 S9A T94A >75 W95A N96A N4A V T4A I44A Amino I45Acid numbering is bsed on the.6 sequence of HIV-HxB b C L46A refers to constnt domins nd V refers.6to vrible. loops c D47A Neutrliztion ctivity is reported s fold increse in IC5 vlue reltive to WT JR-CSF nd ws clculted using the eqution ( IC5 mutnt / IC5 WT). R49A Gry: substitutions which hd negligible effect on neutrliztion ctivity, yellow: -4 fold IC5 increse, red: >4 fold IC5 I4A increse. Experiments 9.8 were 5.4 performed K4Ain duplicte nd vlues represent. n verge. of t lest two.9independent. experiments C4 Q4A I4A > >8. I44A E466A F468A P47A G47A D474A V R476A D477A N478A R48A Amino cid numbering is bsed on the sequence of HIV-HxB. b C refers to constnt domins nd V refers to vrible loops. c Neutrliztion ctivity is reported s fold increse in IC5 vlue reltive to WT JR-CSF nd ws clculted using the eqution ( IC5 mutnt / IC5 WT). Experiments were performed in duplicte nd vlues represent n verge of t lest two independent experiments. 6
7 SUPPLEMENTARY INFORMATION RESEARCH OD OD 45 nm OD 45 OD dsdna ssdna gp4 histone ORI disilognglioside GD BL gp trnsferrin OVA potrnsferrin b 4E OD Supplementry Figure. Polyrectivity ELISA ssy. PGT mabs were tested for ELISA rectivity ginst pnel of ntigens. The bnabs b nd 4E were lso included for comprison. d.s, double-strnded; s.s, single-strnded. 7
8 RESEARCH SUPPLEMENTARY INFORMATION A bredth (%) IC 5 clde A... 5 IC 5 cut-off (μg/ml) b G 4E PGT PGT8 PGT5 PGT45 PGV4 VRC F bredth (%) IC 5 clde F... 5 IC 5 cut-off (μg/ml) b G 4E PGT PGT8 PGT5 PGT45 PGV4 VRC B bredth (%) IC 5 clde B... IC 5 cut-off (μg/ml) b G 4E PGT PGT8 PGT5 PGT45 PGV4 VRC G bredth (%) IC 5 clde G... 5 IC 5 cut-off (μg/ml) b G 4E PGT PGT8 PGT5 PGT45 PGV4 VRC C bredth (%) IC 5 clde C... IC 5 cut-off (μg/ml) b G 4E PGT PGT8 PGT5 PGT45 PGV4 VRC AE bredth (%) IC 5 clde AE... 5 IC 5 cut-off (μg/ml) b G 4E PGT PGT8 PGT5 PGT45 PGV4 VRC D bredth (%) IC 5 clde D... 5 IC 5 cut-off (μg/ml) b G 4E PGT PGT8 PGT5 PGT45 PGV4 VRC AG bredth (%) IC 5 clde AG... 5 IC 5 cut-off (μg/ml) b G 4E PGT PGT8 PGT5 PGT45 PGV4 VRC Supplementry Figure. Anlysis of neutrliztion ctivity by virus cldes. Cumultive frequency distribution of IC 5 vlues of brodly neutrlizing MAbs tested ginst 6 virus pnel seprted by cldes A, B, C, D, F, G, AE nd AG. 8
9 SUPPLEMENTARY INFORMATION RESEARCH donor 7 IC5 (μg/ml) PGT PGT PGT r = -.9 r = -.89 r = -.89 P <. n = 9 P <. n = 7 P <. n = 6 PGT5 PGT6 PGT7 PGT8 PGT PGT r = -.7 r = -.7 r = -.66 r = -.7 r = -.67 r = -.6 P <. P <. P <. P <. P <. P <. n = 5 n = 7 n = n = 5 n = 9 n = 4 PGT5 r = -.46 PGT6 r = -.7 PGT7 r = -.48 P <. P <. P <. n = 54 n = 54 n = 54 PGT4 PGT4 PGT4 PGT44 PGT45 P <. P <. P <. P <. P <. n = 45 n = 45 n = 45 n = 45 n = 5... NT5 (dilution) b donor 6 IC5 (μg/ml)... NT5 (dilution) c donor 9 IC5 (ug/ml)... NT5 (dilution) d donor 84 IC5 (μg/ml). r = -.59 r = -.58 r = -.59 r = -.4 r = NT5 (dilution) Supplementry Figure. MAb neutrliztion correltes strongly with serum neutrliztion. Correltion of IC5s of the MAbs nd serum NT5s of the corresponding donors 7 (), 6 (b), 9 (c) nd 84 (d) is shown. Spermn correltion ws used for sttisticl nlyses. Only viruses neutrlized by either the MAb (IC5 < 5 μg/ml) or the serum (NT5 > ) were included. W W W. N A T U R E. C O M / N A T U R E 9
10 RESEARCH SUPPLEMENTARY INFORMATION A) Recombinnt gp OD 45 JR-CSF gp... Hxb gp 4... SF6 gp... ADA gp 4... YU gp b G YU gp4 (Foldon) KNH44 gp4 (SOSIP) Trimerized gp4 OD JR-CSF KNH44 JR-FL E68K B) Cell surfce Env MFI Supplementry Figure 4. PGT 4-45 bind preferentilly to cell-surfce expressed trimers. A) Binding of PGTs 4-45 to monomeric gp nd rtificilly trimerized gp4 constructs s determined by ELISA. The bnabs b nd re included for comprison. OD, opticl density (bsorbnce t 45 nm). B) Binding of PGTs 4-45 to Env expressed on the surfce of 9T cells s determined by flow cytometry. The bnabs G nd re included for comprison.
11 SUPPLEMENTARY INFORMATION RESEARCH A) percent neutrliztion JR-CSF JR-CSF N6K JR-CSF + kifunensine G ntibody concentrtion (ug/ml) B).5 JR-FL E68K Normlized MFI concentrtion (μg/ml) Supplementry Figure 5. PGT mabs 4-45 bind to epitopes overlpping those of nd PG6. A) PGTs 4-45 re sensitive to the N6K muttion nd PGTs 4-44 fil to neutrlize pseudoviruses produced in the presence of kifunensine. The bnab G ws lso included for comprison. B) competes with PGTs 4-45 for binding to cell-surfce trimers. The bnab G ws included s negtive control.
12 RESEARCH SUPPLEMENTARY INFORMATION A) B) percent inhibition G -4 Mn 4 Dendron concentrtion (μm) -4 Mn 9 Dendron concentrtion (μm) Supplementry Figure 6. PGTs, nd competition with oligodendrons. Unlike PGTs 5, 6, 7, 8 nd, the binding of PGTs, nd to gp could not be competed by A) Mn4 or B) Mn9 dendrons.
13 SUPPLEMENTARY INFORMATION RESEARCH percent neutrliztion IgG Fb percent neutrliztion Ab Concentrtion (nm).. Ab Concentrtion (nm).. Ab Concentrtion (nm) Supplementry Figure 7. Neutrliztion ctivity of Fb frgments. Fb frgments of PGTs-5, 6, 7, 8, nd were generted by Lys-C digestion nd the neutrlizing ctivity tested ginst HIV-JR-CSF using single round of repliction pseudovirus ssy.
14 RESEARCH SUPPLEMENTARY INFORMATION 8 PGT or PGT8 PGT PGT8 8 PGT or PGT45 PGT PGT45 8 PGT or PGT PGT or PGV4 PGT PGV4 8 PGT 8 or PGV4 PGT8 PGV4 8 PGT8 or PGT PGT45 or PGV4 PGT45 PGV4 8 PGT45 or PGT45 8 PGV4 or PGV PGT or VRC PGT VRC 8 PGT8 or VRC PGT8 VRC 8 PGT45 or VRC PGT45 VRC or VRC VRC... 5 Supplementry Figure 8. Percent viruses covered by single MAbs (solid lines) or by t lest one of the MAbs in dul combintions (dshed blck lines) dependent on individul concentrtions. The grey re in ll pnels is the coverge of 6 MAbs tested on the 6-virus pnel (PGT-, PGT5-8, PGT-, PGT5-7, PGT4-45,, PG6, PGC4, VRC, PGV4, b, G, 4E, F5) nd depicts the theoreticl mximl chievble coverge known to dte. 4
15 SUPPLEMENTARY INFORMATION RESEARCH Supporting Notes Protocol G Principl Investigtors nd Institutions: G. Miiro nd J. Serwng (MRC/UVRI Ugnd Reserch Unit on AIDS, Ugnd Virus Reserch Institute, Entebbe, Ugnd), A. Poznik (St Stephens AIDS Trust, Chelse nd Westminster NHS Foundtion Trust, London UK), D. McPhee (NRL, St Vincent's Institute, Melbourne, Victori, Austrli), O. Mnigrt nd L. Mwnnynd (Zmbi Emory HIV Reserch Project, Lusk, Zmbi nd the Rwnd-Zmbi HIV Reserch Group, Emory University, Atlnt, GA, USA), E. Krit (Projet Sn Frncisco, Kigli, Rwnd nd the Rwnd-Zmbi HIV Reserch Group, Emory University, Atlnt, GA, USA), A. Inwoley (CeDReS/CHU Treichville, Abidjn, Côte d'ivoire), W. Joko (Keny AIDS Vccine Inititive, College of Helth Sciences, University of Nirobi, Nirobi, Keny), J. DeHovitz (SUNY Downstte Medicl Center, Brooklyn, NY, USA), L.G. Bekker (Desmond Tutu HIV Centre, University of Cpe Town, Cpe Town, South Afric), P. Pitisuttithum (Fculty of Tropicl Medicine, Mhidol University, Bngkok, Thilnd), R. Pris (Dept. of Retrovirology, Armed Forces Reserch Institute of Medicl Sciences, Bngkok, Thilnd), nd S. Allen (Rwnd- Zmbi HIV Reserch Group, Emory University, Atlnt, GA, USA). 5
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