Article Figures & Data
Figures
- FIG. 1.
Adsorption of anti-MPER antibodies from plasmas BB34, BB81, and BB105. MAb 4E10 and plasma samples were adsorbed with MPER-peptide-coated beads or blank beads or left untreated. (A) All samples were assayed by ELISA for binding to the MPER or V3 peptide and tested for neutralization of the HIV-2-HIV-1 MPER chimera C1C. OD, optical density; conc, concentration. (B) Adsorbed plasmas were tested for neutralization of the HIV-1 envelope-pseudotyped viruses COT6.15, CAP206.8, and Du156.12.
- FIG. 2.
Antibodies eluted from MPER-coated beads contain cross-neutralizing activity. (A) Neutralization of C1C by eluates from MPER-coated beads of plasmas BB34, CAP206, and SAC21 and MAbs 4E10, Z13e1, and 2F5. conc, concentration. (B) Neutralization of HIV-1 subtype C envelope-pseudotyped viruses COT6.15, ZM197M.PB7, Du156.12, and CAP206.8 and subtype B TRO.11 and JR-FL.
- FIG. 3.
Comparison of the IgG subclass profiles between original plasmas and eluates from MPER-coated beads. The pie charts represent the IgG subclasses found in the BB34, CAP206, and SAC21 plasmas and eluates. The table shows the IgG subclass concentrations in plasmas and in eluates. b.d, below detection level.
- FIG. 4.
Neutralizing anti-MPER antibodies are IgG3 in BB34 but not in CAP206. (A and B) IgG subclass profiles of total IgG, FTpA, and EpA of BB34 (A) and CAP206 (B). (C) BB34 fractions were tested for neutralization of C1C and HIV-1 envelope-pseudotyped viruses, as well as binding to the MPER peptide in ELISA. OD, optical density; conc, concentration. (D) CAP206 fractions were tested for neutralization of C1C and HIV-1 envelope-pseudotyped viruses.
Tables
- TABLE 1.
Screening for broadly cross-neutralizing plasma samples containing anti-MPER antibodies
Parametera Value in: BB cohort CAPRISA SAC cohort Total no. of plasmas 70 18 68 No. (%) BCN 16 (23)b 4 (22)c 17 (25)d No. of BCN anti-MPER antibodies positive 11 2 6 No. of BCN anti-MPER titers > 1:1,000 1 1 4 No. MPER peptide binding 1 1 1 Sample analyzed BB34 CAP206 SAC21 ↵a BCN, broadly cross-neutralizing. Anti-MPER activity was defined as neutralization of the HIV-2-HIV-1 MPER chimeric virus C1C.
↵b BCN plasmas were defined as able to neutralize at least 8 of 10 viruses tested (12).
↵c BCN plasmas were defined as able to neutralize at least 8 of 12 viruses from the tier 2 subtype C virus panel.
↵d BCN plasmas were defined as able to neutralize at least four of six viruses tested.
- TABLE 2.
Effect of anti-MPER antibody adsorbtions on neutralization breadth
Subtype Virus ID50 % Reductionc Blanka MPERb Adsorbed BB34 plasma HIV-2/HIV-1 MPER C1C 4,802 41 99 Subtype C COT6.15 1,350 65 95 CAP85 9 7,134 1,140 84 CAP88 B5 258 <40 84 CAP206 8 1,350 86 94 CAP210 B8 148 102 31 CAP228 51 245 73 70 CAP255 16 164 <40 76 Du151.2 484 636 0 Du422.1 155 <40 74 Du156.12 3,869 151 96 ZM197M.PB7 1,068 <40 96 ZM233M.PB6 219 66 70 ZM135M.PL10a 1,651 250 85 Subtype B 6535.3 549 102 81 QHO692.42 179 42 77 CAAN5342.A2 139 129 7 TRO.11 646 <40 94 SC422661.8 758 175 77 REJO4541.67 331 80 76 JR-FL 129 <40 69 Subtype A 92RW009 1,296 827 32 Subtype D 92UG024 1,480 1,006 32 Adsorbed CAP206 plasma HIV-2/HIV-1 MPER C1C 4,527 222 95 Subtype C COT6.15 1,236 109 91 CAP45 G3 4,720 193 96 CAP63 A9 180 132 27 CAP85 9 2,856 352 88 CAP88 B5 223 <40 82 CAP206 8 1,870 1,555 17 Du151.2 105 <40 62 Du422.1 165 47 72 Du156.12 692 57 92 Du172.17 234 <40 83 ZM197M.PB7 309 82 73 ZM135M.PL10a 248 91 63 Subtype B QHO692.42 383 66 83 AC10.0.29 111 47 58 WITO4160.33 144 99 31 TRO.11 491 <40 92 Subtype A 92RW009 915 793 13 Q23.17 320 340 0 Subtype D 92UG024 1,556 1,268 19 Adsorbed SAC21 plasma HIV-2/HIV-1 MPER C1C 3,157 246 92 Subtype C COT6.15 183 <40 78 CAP85 9 447 276 38 CAP88 B5 88 42 52 CAP206 8 361 140 61 CAP255 16 109 115 0 Du151.2 117 69 41 ZM197M.PB7 117 85 27 ZM233M.PB6 100 79 21 ZM135M.PL10a 1,114 301 73 Subtype B TRO.11 147 47 68 SC422661.8 88 <40 55 Subtype A 92RW009 1,665 1,045 37 Subtype D 92UG024 1,889 1,491 21 ↵a ID50 of plasmas adsorbed on blank beads. These titers were similar to the ID50 obtained with the untreated sera.
↵b ID50 of plasmas adsorbed on beads coated with the MPER peptide.
↵c Percentage reduction in ID50 due to adsorption on MPER-coated beads (1 − MPER/blank). Cases where the percent reduction was >50% are in boldface.
- TABLE 3.
Mapping of anti-MPER neutralizing antibodies
Chimera MPER sequencea Neutralizationb Plasma ID50 2F5 4E10 Z13e1 BB34 CAP206 SAC21 7312A NMYEL660QKLNSWDVFG670NWFDLASWVK680YIQYGVYIV − − − <20 21 <20 C1 NMYEL660LALDKWASLW670NWFDITKWLW680YIKYGVYIV ++ ++ ++ 5,560 3,903 3,871 C1C NMYEL660LALDSWKNLW670NWFDITKWLW680YIKYGVYIV − ++ ++ 3,945 2,867 2,733 C1C F/L NMYEL660LALDSWKNLW670NWLDITKWLW680YIKYGVYIV − − + 1,779 2,449 1,802 C3 NMYEL660LALDKWASLW670NWFDLASWVK680YIQYGVYIV ++ − − <20 <20 <20 C7(2F5) NMYEL660QALDKWAVFG670NWFDLASWVK680YIQYGVYIV ++ − − <20 <20 <20 C6(4E10) NMYEL660QKLNSWDVFG670NWFDITSWIK680YIQYGVYIV − ++ − <20 <20 <20 C4 NMYEL660QKLNSWDVFG670NWFDITKWLW680YIKYGVYIV − ++ +/− <20 723 189 C4GW NMYEL660QKLNSWDVFW670NWFDITKWLW680YIKYGVYIV − ++ ++ 7,482 3,067 2,987 C8 NMYEL660QKLNSWDSLW670NWFDITKWLW680YIKYGVYIV − ++ + 3,351 2,538 1,199 ↵a Grafted amino acids are indicated in italics, with the 7312A residues in lightface. Further mutations on the chimeras are in boldface.
↵b Neutralization by MAbs 2F5, 4E10, and Z13e1 are qualitatively indicated relative to the titers obtained with the C1 chimera. −, no neutralization; ++, neutralization similar to that of C1; +, neutralization within 3-fold of that of C1; +/−, neutralization within 10-fold of that of C1.
- TABLE 4.
Relative neutralization of pseudotyped COT6.15 envelope MPER mutantsa
COT6.15 4E10 BB34 CAP206 SAC21 IC50 Ratiob ID50 Ratioc ID50 Ratioc ID50 Ratioc Wild type 0.9 1.0 1,392 1.0 1,256 1.0 317 1.0 A662G 0.12 0.1 4,899 0.3 2,443 0.5 978 0.3 L663A 0.02 0.0 8,714 0.2 7,971 0.2 5,660 0.1 D664A 0.77 0.9 1,149 1.2 844 1.5 238 1.3 S665A 0.14 0.2 5,495 0.3 1,562 0.8 1,787 0.2 W666A 0.51 0.6 5,554 0.3 4,294 0.3 446 0.7 K667A 0.05 0.1 3,261 0.4 1,694 0.7 1,734 0.2 N668A 1.3 1.4 831 1.7 425 3.0 208 1.5 L669A 0.05 0.1 3,847 0.4 3,138 0.4 1,195 0.3 W670A 0.11 0.1 132 10.5 1,054 1.2 105 3.0 S671A 0.04 0.0 3,102 0.4 1,614 0.8 928 0.3 W672A >25 >25 2,959 0.5 2,244 0.6 468 0.7 F673A >25 >25 779 1.8 498 2.5 <50 >6.3 D674A 1.4 1.6 <50 >25 <50 >25 <50 >6.3 D674S 2.49 2.8 <50 >25 90 14.0 <50 >6.3 D674N 0.33 0.4 663 2.1 643 2.0 149 2.1 I675A 0.04 0.0 4,069 0.3 2,065 0.6 718 0.4 T676A 21.77 24.2 2,380 0.6 895 1.4 524 0.6 K677A 0.05 0.1 4,671 0.3 2,151 0.6 1154 0.3 W678A 0.05 0.1 3,842 0.4 1,885 0.7 1,007 0.3 L679A 0.09 0.1 2,085 0.7 1,448 0.9 225 1.4 W680A 10.89 12.1 731 1.9 904 1.4 142 2.2
Supplemental material
Files in this Data Supplement:
- Supplemental file 1
-
Fig. S1 (MPER sequences of the viruses used in this study.)
Table S1 (Neutralizing activity of the three plasmas used in this study.)
MS PowerPoint file, 72K.
- Supplemental file 1
-
Fig. S1 (MPER sequences of the viruses used in this study.)
