HIV-1 gp120-specific Antibody Dependent Cell-mediated ... · HIV-1+ Elite Controllers have higher...
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Mariana M Mata 1 , Fareeha Mahmood 1 , Elizabeth T Golub 2 , Ruth M Greenblatt 3 , Kathryn Anastos 4 , Mareck Nowicki 5 , Mary Young 6 , Helen Durkin 7 , Seema N Desai 1 , Linda L Baum 1 . 1 Department of Immunology and Microbiology, Rush University Medical Center, Chicago, IL; 2 Department of Epidemiology, Johns Hopkins University, Baltimore, MD; 3 Department of Clinical Pharmacy, Medicine, Epidemiology and Biostatistics, University of California, San Francisco, CA; 4 Deparment of Medicine, Epidemiology and Population Health, Montefiore Medical Center, Bronx, NY; 5 Department of Medicine, University of Southern California, Los Angeles, CA; 6 Department of Medicine, Division of Infectious Disease, Georgetown University Medical Center, Washington, DC; 7 Department of Pathology, SUNY Downstate Medical Center, Brooklyn, NY. Background: The results of the RV144 HIV vaccine trial in Thailand indicates that HIV specific antibody dependent-cell mediated cytotoxicity (ADCC) contributes to host defense against HIV infection. This suggests that protective HIV specific ADCC can be stimulated by vaccination. We hypothesize that natural production of HIV-specific ADCC antibodies may also be protective and that these antibodies may be at least partially responsible for HIV viral control observed in women who are Elite controllers (EC). i.e., those who naturally control the virus without antiretroviral drugs. Methodology: We compared the ADCC response of antibodies in the serum and cervicovaginal lavage (CVL) from 35 EC, 34 HIV-infected HAART initiators, and 35 HIV-uninfected women in a standard 51Cr- release assay. All were participants in the Women’s Interagency HIV Study (WIHS), the largest ongoing prospective cohort study of HIV among US women. The 35 EC in our study had stable or increasing CD4 T cell numbers, were not on antiretroviral therapy and had undetectable HIV RNA for ≥1.5 years. HAART initiators were evaluated at three visits: visit 1, an asymptomatic visit with comparable CD4 T cell count to the EC group; visit 2, the last visit before HAART; and visit 3, a progressing visit during treatment. Statistical comparisons were conducted using the Mann-Whitney or Wilcoxon Paired Test. Significant differences were considered when p<0.05. Results: EC had significantly higher serum (see Fig.1) and CVL ADCC activity than HAART initiators (p≤0.009 at 1:10 CVL dilution); this difference was most prominent at visit 1, the asymptomatic visit. Another measure of this activity, serum ADCC antibody titer, was significantly higher in EC than in HAART Initiators: visit 1, p≤0.003; visit 2, p≤0.04; visit 3, p≤0.006. While 97% of EC and 77% of HAART Initiators (Visit1) had a detectable serum ADCC titer, CVL titers were present in 40% of the EC and 27% of the HAART Initiators (Visit 1). CVL ADCC antibody titers were detected in 17% of the women immediately prior to HAART and increased to 32% during HAART treatment. The HIV-1 seronegative women had no detectable gp120- specific ADCC. Conclusions: HIV EC have higher serum and CVL ADCC antibodies against HIV-gp120 than HAART initiators. ADCC may contribute to the ability of EC to control viral load. Evidence that the proportion of women with CVL ADCC activity increases after the initiation of treatment suggests that HAART may facilitate a rebound in ADCC antibodies. We used a 3.5 hr 51 Cr-release assay to compare the HIV-1 gp120-specific ADCC response of antibodies in the serum and cervicovaginal lavage (CVL) among HIV-infected EC, HIV-infected Progressors, and HIV-uninfected women (Table 1). All subjects were participants in the Women’s Interagency HIV Study (WIHS). The EC in our study were not on antiretroviral therapy and had undetectable HIV RNA for ≥1.5 years. Table 1. Study Participants from WIHS cohort HIV-1 + Elite Controllers have higher serum and CVL ADCC antibodies against HIV-1 gp120 than HIV-1 + Progressors on HAART treatment. ADCC may contribute to the ability of Elite Controllers to control viral load. Evidence that the proportion of women with CVL ADCC activity increases after the initiation of HAART treatment suggests that HAART may facilitate a rebound in ADCC antibodies. This work was supported by NIH PO1AI082971 and by the Chicago Developmental Center for AIDS Research P30AI082151. Samples in this study were provided by the Women Interagency HIV Study (WIHS). HIV-1 gp120-specific Antibody Dependent Cell-mediated Cytotoxicity in HIV-1 + Elite Controllers Participants Sample Timing Sample Type N HIV-1 + Elite Controllers Controlling virus Serum and CVL 35 HIV-1 + progressors Asymptomatic Visit, “Visit 1” Serum and CVL 34 HIV-1 + progressors Last Visit before HAART treatment, “Visit 2” Serum and CVL 34 HIV-1 + progressors Progressing Visit on HAART, “Visit 3” Serum and CVL 34 HIV-1 seronegatives ---- Serum and CVL 35 Results Serum ADCC CVL ADCC Group Undetected 10 10 2 10 3 10 4 Total of Participants with an ADCC titer n, ( %) n, ( %) n, (%) n, (%) n, (%) HIV-1 Seronegative 33 (94.3%) 1 (3%) 0 (0%) 1 (3%) 1 (3%) 3 (8.6%) HIV-1 Elite Controllers 21 (60%) 7 (20%) 3(8.6%) 1 (3%) 3(8.6% 14 (40%) HIV-1 Progressors Asymptomatic Visit 25 (73.5%) 2 (6%) 5 (15%) 0 (0%) 1 (3%) 9 (26.5%) HIV-1 Progressors Last Visit before HAART 28 (82%) 1 (3%) 3 (9%) 2 (6%) 1 (3%) 6 (17.6%) HIV-1 Progressors Progressing Visit on HAART 23 (3%) 4 (12%) 4 (12%) 1 (3%) 2 (6% 11 (32.4 %) Group Undetecte d 10 10 2 10 3 10 4 Total of Participants with an ADCC titer p-value Relative to EC n, ( %) n, ( %) n, (%) n, (%) n, (%) HIV-1 Seronegative 32 (91%) 1 (3%) 1 (3%) 1 (3%) 0, (0%) 3 (8.6%) 0.0001 HIV-1 Elite Controllers 1 (3%) 0 (0%) 0 (0%) 1 (3%) 33 (94,3%) 34 (97%) --- HIV-1 Progressors Asymptomatic Visit 8 (23.5%) 1 (3%) 0, (0%) 0,( 0%) 25 (73.5%) 26 (76.5%) 0.003 HIV-1 Progressors Last Visit before HAART 5 (15%) 0 (0%) 0 (0%) 0 (0%) 29 (85%) 29 (85.3%) 0.044 HIV-1 Progressors Progressing Visit on HAART 7 (20.6%) 0 (0%) 1 (3%) 0 (0%) 26 (76.5%) 27 (79.4 %) 0.006 A. B. C. A. B. C. Results Table 2. High Serum ADCC Antibody Titers are more prevalent in HIV-1 + Elite controllers than HIV-1 + Progressors. Using a standard 3.5 hr 51 Cr-release assay, Antibody Dependent Cell-mediated Cytotoxicity (ADCC) serum antibody titers were determined from percent specific release (%SR) values and compared among HIV-1 + Elite Controllers (EC), HIV-1 + Progressors at visit 1, 2 and 3, and to HIV-1 seronegative samples. All women were participants of the Women’s Interagency HIV Study (WIHS). Statistical significant differences were considered when p-values were less than 0.05 using Mann-Whitney or Wilcoxon Paired Test. Table 3. CVL ADCC Antibody Titers are more prevalent in HIV-1 + Elite controllers than HIV-1 + Progressors. Using a standard 3.5 hr 51 Cr-release assay, Antibody Dependent Cell-mediated Cytotoxicity (ADCC) antibody titers in the cervicovaginal fluid (CVL) were determined from percent specific release (%SR) values and compared among HIV-1 + Elite Controllers (EC), HIV-1 + Progressors at visit 1, 2 and 3, and to HIV-1 seronegative samples. All women were participants of the Women’s Interagency HIV Study (WIHS). Figure 1. Serum Antibody Dependent Cell-mediated Cytotoxicity (ADCC) in HIV-1 Elite Controllers. ADCC effector cells from uninfected healthy donors were incubated for 3.5 hours with 51 Cr-labeled HIV-1 gp120 bearing CEM.NKR target cells in the presence of serum (10-fold serial dilutions) from HIV-1 + Elite Controllers (EC), HIV-1 + Progressors and seronegatives. All women were participants of the Women’s Interagency HIV Study (WIHS). Serum ADCC activity shown as percent specific release (%SR) values of HIV-1 + EC is compared to HIV-1 + Progressors at: A) Visit 1; B) Visit 2; C) Visit 3. Significant differences were considered when p-values were less than 0.05 using Mann-Whitney or Wilcoxon Paired Test. Figure 2. CVL Antibody Dependent Cell-mediated Cytotoxicity (ADCC) in HIV-1 Elite Controllers. ADCC effector cells from uninfected healthy donors were incubated for 3.5 hours with 51 Cr-labeled HIV-1 gp120 bearing CEM.NKR target cells in the presence of cervicovaginal fluid (CVL) (10-fold serial dilutions) from HIV-1 + Elite Controllers (EC), HIV-1 + Progressors and seronegatives. All women were participants of the Women’s Interagency HIV Study (WIHS). A) CVL ADCC activity shown as percent specific release (%SR) values of HIV-1 + EC is compared to HIV-1 + Progressors at: Visit 1; B) Visit 2; and C) Visit 3. Differences were considered statistically significant when p-values were less than 0.05 using Mann-Whitney or Wilcoxon Paired Test. ADCC Titer ADCC Titer
Transcript of HIV-1 gp120-specific Antibody Dependent Cell-mediated ... · HIV-1+ Elite Controllers have higher...
Mariana M Mata1, Fareeha Mahmood1, Elizabeth T Golub2, Ruth M Greenblatt3, Kathryn Anastos4, Mareck Nowicki5, Mary Young6, Helen Durkin7, Seema N Desai1, Linda L Baum1. 1Department of Immunology and Microbiology, Rush University Medical Center, Chicago, IL; 2Department of Epidemiology, Johns Hopkins University, Baltimore, MD; 3Department of Clinical Pharmacy, Medicine, Epidemiology and Biostatistics, University of California, San Francisco, CA; 4Deparment of Medicine, Epidemiology and Population Health, Montefiore Medical Center, Bronx, NY; 5Department of Medicine, University of Southern California,
Los Angeles, CA; 6Department of Medicine, Division of Infectious Disease, Georgetown University Medical Center, Washington, DC; 7Department of Pathology, SUNY Downstate Medical Center, Brooklyn, NY.
Background: The results of the RV144 HIV vaccine trial in Thailand indicates that HIV specific antibody dependent-cell mediated cytotoxicity (ADCC) contributes to host defense against HIV infection. This suggests that protective HIV specific ADCC can be stimulated by vaccination. We hypothesize that natural production of HIV-specific ADCC antibodies may also be protective and that these antibodies may be at least partially responsible for HIV viral control observed in women who are Elite controllers (EC). i.e., those who naturally control the virus without antiretroviral drugs. Methodology: We compared the ADCC response of antibodies in the serum and cervicovaginal lavage (CVL) from 35 EC, 34 HIV-infected HAART initiators, and 35 HIV-uninfected women in a standard 51Cr-release assay. All were participants in the Women’s Interagency HIV Study (WIHS), the largest ongoing prospective cohort study of HIV among US women. The 35 EC in our study had stable or increasing CD4 T cell numbers, were not on antiretroviral therapy and had undetectable HIV RNA for ≥1.5 years. HAART initiators were evaluated at three visits: visit 1, an asymptomatic visit with comparable CD4 T cell count to the EC group; visit 2, the last visit before HAART; and visit 3, a progressing visit during treatment. Statistical comparisons were conducted using the Mann-Whitney or Wilcoxon Paired Test. Significant differences were considered when p<0.05. Results: EC had significantly higher serum (see Fig.1) and CVL ADCC activity than HAART initiators (p≤0.009 at 1:10 CVL dilution); this difference was most prominent at visit 1, the asymptomatic visit. Another measure of this activity, serum ADCC antibody titer, was significantly higher in EC than in HAART Initiators: visit 1, p≤0.003; visit 2, p≤0.04; visit 3, p≤0.006. While 97% of EC and 77% of HAART Initiators (Visit1) had a detectable serum ADCC titer, CVL titers were present in 40% of the EC and 27% of the HAART Initiators (Visit 1). CVL ADCC antibody titers were detected in 17% of the women immediately prior to HAART and increased to 32% during HAART treatment. The HIV-1 seronegative women had no detectable gp120-specific ADCC. Conclusions: HIV EC have higher serum and CVL ADCC antibodies against HIV-gp120 than HAART initiators. ADCC may contribute to the ability of EC to control viral load. Evidence that the proportion of women with CVL ADCC activity increases after the initiation of treatment suggests that HAART may facilitate a rebound in ADCC antibodies.
We used a 3.5 hr 51Cr-release assay to compare the HIV-1 gp120-specific ADCC response of antibodies in the serum and cervicovaginal lavage (CVL) among HIV-infected EC, HIV-infected Progressors, and HIV-uninfected women (Table 1). All subjects were participants in the Women’s Interagency HIV Study (WIHS). The EC in our study were not on antiretroviral therapy and had undetectable HIV RNA for ≥1.5 years.
Table 1. Study Participants from WIHS cohort
HIV-1+ Elite Controllers have higher serum and CVL ADCC antibodies against HIV-1 gp120 than HIV-1+ Progressors on HAART treatment. ADCC may contribute to the ability of Elite Controllers to control viral load. Evidence that the proportion of women with CVL ADCC activity increases after the initiation of HAART treatment suggests that HAART may facilitate a rebound in ADCC antibodies.
This work was supported by NIH PO1AI082971 and by the Chicago Developmental Center for AIDS Research P30AI082151. Samples in this study were provided by the Women Interagency HIV Study (WIHS).
HIV-1 gp120-specific Antibody Dependent Cell-mediated
Cytotoxicity in HIV-1+ Elite Controllers
Participants Sample Timing Sample Type N
HIV-1+
Elite Controllers Controlling virus Serum and CVL
35
HIV-1+ progressors Asymptomatic Visit, “Visit 1” Serum and
CVL
34
HIV-1+ progressors Last Visit before HAART treatment, “Visit 2”
Serum and CVL
34
HIV-1+ progressors Progressing Visit on HAART, “Visit 3”
Serum and CVL
34
HIV-1 seronegatives ---- Serum and
CVL 35
Results Serum ADCC
CVL ADCC
Group Undetected 10 102 103 104
Total of Participants with an
ADCC titer
n, ( %) n, ( %) n, (%) n, (%) n, (%)
HIV-1
Seronegative 33 (94.3%) 1 (3%) 0 (0%) 1 (3%) 1 (3%) 3 (8.6%)
HIV-1 Elite
Controllers 21 (60%) 7 (20%) 3(8.6%) 1 (3%) 3(8.6% 14 (40%)
HIV-1
Progressors
Asymptomatic
Visit
25 (73.5%) 2 (6%) 5 (15%) 0 (0%) 1 (3%) 9 (26.5%)
HIV-1
Progressors Last
Visit before
HAART
28 (82%) 1 (3%) 3 (9%) 2 (6%) 1 (3%) 6 (17.6%)
HIV-1
Progressors
Progressing Visit
on HAART
23 (3%) 4 (12%) 4 (12%) 1 (3%) 2 (6% 11 (32.4 %)
Group Undetecte
d 10 102 103 104
Total of Participants with an ADCC titer
p-value Relative to
EC
n, ( %) n, ( %) n, (%) n, (%) n, (%)
HIV-1 Seronegative 32 (91%) 1 (3%) 1 (3%) 1 (3%) 0, (0%) 3 (8.6%) 0.0001
HIV-1 Elite
Controllers 1 (3%) 0 (0%) 0 (0%) 1 (3%) 33 (94,3%) 34 (97%) ---
HIV-1 Progressors
Asymptomatic Visit 8 (23.5%) 1 (3%) 0, (0%) 0,( 0%) 25 (73.5%) 26 (76.5%) 0.003
HIV-1 Progressors
Last Visit before
HAART
5 (15%) 0 (0%) 0 (0%) 0 (0%) 29 (85%) 29 (85.3%) 0.044
HIV-1 Progressors
Progressing Visit on
HAART
7 (20.6%) 0 (0%) 1 (3%) 0 (0%) 26 (76.5%) 27 (79.4 %) 0.006
A. B. C.
A. B. C.
Results
Table 2. High Serum ADCC Antibody Titers are more prevalent in HIV-1+ Elite controllers than HIV-1+ Progressors. Using a
standard 3.5 hr 51Cr-release assay, Antibody Dependent Cell-mediated Cytotoxicity (ADCC) serum antibody titers were determined from percent specific release (%SR) values and compared among HIV-1+ Elite Controllers (EC), HIV-1+ Progressors at visit 1, 2 and 3, and to HIV-1 seronegative samples. All women were participants of the Women’s Interagency HIV Study (WIHS). Statistical significant differences were considered when p-values were less than 0.05 using Mann-Whitney or Wilcoxon Paired Test.
Table 3. CVL ADCC Antibody Titers are more prevalent in HIV-1+ Elite controllers than HIV-1+ Progressors. Using a standard 3.5 hr 51Cr-release assay, Antibody Dependent Cell-mediated Cytotoxicity (ADCC) antibody titers in the cervicovaginal fluid (CVL) were determined from percent specific release (%SR) values and compared among HIV-1+ Elite Controllers (EC), HIV-1+ Progressors at visit 1, 2 and 3, and to HIV-1 seronegative samples. All women were participants of the Women’s Interagency HIV Study (WIHS).
Figure 1. Serum Antibody Dependent Cell-mediated Cytotoxicity (ADCC) in HIV-1 Elite Controllers. ADCC
effector cells from uninfected healthy donors were incubated for 3.5 hours with 51Cr-labeled HIV-1 gp120 bearing CEM.NKR target cells in the presence of serum (10-fold serial dilutions) from HIV-1+ Elite Controllers (EC), HIV-1+ Progressors and seronegatives. All women were participants of the Women’s Interagency HIV Study (WIHS). Serum ADCC activity shown as percent specific release (%SR) values of HIV-1+ EC is compared to HIV-1+ Progressors at: A) Visit 1; B) Visit 2; C) Visit 3. Significant differences were considered when p-values were less than 0.05 using Mann-Whitney or Wilcoxon Paired Test.
Figure 2. CVL Antibody Dependent Cell-mediated Cytotoxicity (ADCC) in HIV-1 Elite Controllers. ADCC effector
cells from uninfected healthy donors were incubated for 3.5 hours with 51Cr-labeled HIV-1 gp120 bearing CEM.NKR target cells in the presence of cervicovaginal fluid (CVL) (10-fold serial dilutions) from HIV-1+ Elite Controllers (EC), HIV-1+ Progressors and seronegatives. All women were participants of the Women’s Interagency HIV Study (WIHS). A) CVL ADCC activity shown as percent specific release (%SR) values of HIV-1+ EC is compared to HIV-1+ Progressors at: Visit 1; B) Visit 2; and C) Visit 3. Differences were considered statistically significant when p-values were less than 0.05 using Mann-Whitney or Wilcoxon Paired Test.
ADCC Titer
ADCC Titer
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