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Status: OngoingResearch Proposal
Project Title: Baseline Ad26 Immunity and gp140 Responses in Mosaico: a Post-hoc Comparative Analysis Across Four Latin American Countries
Scientific Abstract:
Background: The Mosaico HIV vaccine trial showed no overall efficacy. Exploratory analyses suggested that pre-existing Ad26 vector immunity may attenuate vaccine-induced antibody responses, with an inverse association between baseline Ad26 neutralizing titers and Month-7 gp140 levels. Whether this association varies across countries remains unclear.
Objective: To evaluate whether baseline Ad26 neutralizing antibody titers differentially influence gp140 responses across the four Latin American countries in the Mosaico immunogenicity analysis set (Argentina, Brazil, Mexico, and Peru), and to descriptively explore HIV acquisition patterns among vaccine recipients stratified by baseline Ad26 titers.
Study Design: Post-hoc exploratory analysis using deidentified individual-level Mosaico data.
Participants: Vaccine recipients in the immunogenicity analysis set (IAS) with available baseline Ad26 titers and Month-7 gp140 concentrations. Acquisition analyses are restricted to vaccine recipients within the IAS.
Primary Outcome: Association between baseline Ad26 titers (log10 IC90) and Month-7 gp140 binding antibody concentrations, compared across four countries.
Secondary Outcomes: Country-level exposure-response curves; descriptive HIV acquisition patterns among vaccine recipients stratified by baseline Ad26 titers.
Statistical Analysis: Linear regression and spline-based models with country interaction terms. Kaplan-Meier estimates for acquisition analyses.
Brief Project Background and Statement of Project Significance:
Adenovirus serotype 26 (Ad26) is a widely used vaccine vector. Pre-existing vector-directed immunity is a known concern because baseline neutralizing antibodies can attenuate vaccine-induced responses. In the multicountry Mosaico HIV vaccine trial, there was no overall reduction in HIV acquisition, yet many participants mounted robust binding-antibody responses. Mosaico analyses (Figure S9 of the primary publication) suggested an inverse association between baseline Ad26 neutralizing titers and Month 7 gp140 responses, but they did not assess whether this association varied across regions.
Because Mosaico enrolled participants across multiple countries and the immunogenicity analysis set captures four Latin American countries (Argentina, Brazil, Mexico, and Peru) with notably different baseline characteristics, it is important to evaluate whether pre-existing Ad26 immunity varied geographically and whether the magnitude or shape of the association between baseline Ad26 titers and gp140 responses differs by country.
This project will address that gap with a post-hoc, individual-level, regional comparative analysis. We will model the continuous relationship between baseline Ad26 neutralizing antibody titers and Month 7 gp140 antibody concentrations, comparing exposure--response curves across countries, with a planned focus on Peru given its large enrollment. In exploratory, descriptive analyses--recognizing limited power--we will also summarize HIV acquisition by study arm within strata of baseline Ad26 titers.
By quantifying the relationship between baseline Ad26 neutralizing antibody titers and immune responses, this work will directly assess whether pre-existing vector immunity serves as a predictor of vaccine-induced antibody magnitude. Furthermore, given the regional differences observed in acquisition outcomes and immunogenicity, the analysis will help determine whether between-country differences in baseline Ad26 titers were sufficient to contribute to geographic variability in observed immune responses; descriptive summaries of acquisition by arm within Ad26 strata will provide additional context.
The study is descriptive and hypothesis-generating. By leveraging existing high-quality, de-identified Mosaico data, it will clarify whether anti-vector immunity contributes to geographic variability in immunogenicity. The information gained can materially enhance generalizable knowledge about how pre-existing vector immunity shapes antibody responses, informing future research directions such as evaluation of alternative vectors, heterologous boosting, or pre-screening strategies in subsequent studies. More broadly, insights on vector immunity derived here may extend to other adenoviral-vectored vaccines beyond HIV.
Specific Aims of the Project:
Study design: Post-hoc, individual-level, regional comparative analysis of Mosaico.
Exposure: Baseline Ad26 neutralizing antibody titers (IC90, log10).
Primary endpoint: Month-7 IgG anti-gp140.
Overall objective: Quantify the continuous association between baseline Ad26 immunity and vaccine-induced gp140 responses, and assess whether this association differs across countries (with planned focus on Peru). Descriptively summarize HIV acquisition by arm within Ad26 strata.
Aim 1 (primary): Estimate gp140 as a function of baseline Ad26 using flexible models. To compare regions, fit models that allow the Ad26--gp140 relationship to vary by country. Report country-level exposure--response curves and marginal effects.
- Hypothesis: Higher baseline Ad26 titers are associated with lower gp140 levels, and the magnitude and/or shape of this association differs by country, with stronger attenuation where baseline Ad26 titers are higher.
Aim 2 (Exploratory, descriptive): Within strata of baseline Ad26 titers (quartiles or tertiles), descriptively summarize HIV acquisition among vaccine recipients in the IAS using Kaplan--Meier estimates, recognizing that this analysis is limited to cases and controls within the four Latin American countries.
- Hypothesis: HIV acquisition patterns by arm, within strata of basal Ad26, will be directionally consistent with the immunogenicity gradient observed in Aim 1.
Study Design: Individual trial analysis
What is the purpose of the analysis being proposed? Please select all that apply.: New research question to examine treatment effectiveness on secondary endpoints and/or within subgroup populations Research on clinical prediction or risk prediction
Software Used: R, STATA
Data Source and Inclusion/Exclusion Criteria to be used to define the patient sample for your study:
Data Source: This study will use individual participant-level data from the Mosaico trial (HVTN 706/HPX3002), accessed through the YODA Project secure platform. Specifically, the Immunogenicity Analysis Set (IAS) will be analyzed, which includes participants with available baseline Ad26 neutralizing antibody (NAb) titers and Month 7 immune endpoints. HIV testing dates and results data from the trial will also be incorporated for exploratory analyses.
Analysis populations & criteria:
Aim 1 (Immunogenicity set)
Inclusion criteria: Randomized vaccine recipients with:
(i) available baseline Ad26 NAb titer
(ii) Month-7 gp140 measurement within the protocol window
(iii) HIV-negative up to the Month-7 sample
(iv) no protocol deviations that invalidate immunogenicity per CSR
*Placebo recipients are excluded from primary modeling.
Exclusion criteria: missing baseline Ad26 or Month-7 gp140; failed assay QC; confirmed HIV infection before Month-7 sampling; withdrawal of consent for data sharing.
Aim 2 (Acquisition set, descriptive)
Inclusion criteria: All randomized participants (both arms) with: (i) baseline Ad26 titer; (ii) at least one post-baseline HIV test result. Analyses will be as-randomized (ITT) and stratified by baseline Ad26.
Exclusion criteria: missing baseline Ad26; no post-baseline HIV testing; withdrawal of consent.
Additional Data Sources:
No external studies will be pooled. Analyses will be restricted to the Mosaico trial dataset provided through the YODA Project.
Analysis Platform:
All analyses will be conducted within the secure YODA platform environment, using R and/or Stata for statistical modeling.
Primary and Secondary Outcome Measure(s) and how they will be categorized/defined for your study:
Primary outcome (continuous):
Month-7 IgG anti-gp140 levels as a function of baseline Ad26 neutralizing antibody titers (IC90, log10), allowing comparison of exposure--response curves across countries.
Secondary outcomes:
- Country-level variation in the Ad26--gp140 association, reported as differences in curve shape and marginal effects across regions.
- HIV acquisition within strata of baseline Ad26 titers (quartiles or tertiles). This will be exploratory and descriptive, given the overall null efficacy of the Mosaico trial.
Main Predictor/Independent Variable and how it will be categorized/defined for your study:
Main Independent Variable: The main independent variable is baseline Ad26 neutralizing antibody (NAb) titers, measured as 90% inhibitory concentration (IC90) from virus neutralization assays.
Definition and Categorization:
- Continuous: Titers will be log10-transformed and analyzed as a continuous predictor of Month-7 IgG anti-gp140 and HIV acquisition.
- Categorical: For subgroup analyses, titers will be grouped into tertiles or quartiles of the observed distribution.
- Binary (serostatus): Defined by assay detection limit (seronegative [<=LOD] vs. seropositive [>LOD]).
Baseline Ad26 titers are hypothesized to directly modulate vaccine-induced gp140 responses and, indirectly, HIV acquisition patterns. Using both continuous and categorical approaches allows evaluation of dose--response relationships while maximizing statistical power.
Other Variables of Interest that will be used in your analysis and how they will be categorized/defined for your study:
In addition to baseline Ad26 neutralizing antibody titers (main predictor), several other variables will be used to characterize the study population, adjust for potential confounding, and conduct subgroup analyses.
Demographic variables:
- Age: Continuous (years) and categorical (<25, 25--34, >=35)
- Sex/gender identity: Male, female, transgender
- Region/Country: Country of enrollment.
Behavioral/clinical risk factors:
- PrEP use: Yes/No at baseline and during follow-up (time-varying where available)
- Sexually transmitted infections (STIs): Presence of gonorrhea, chlamydia, or syphilis at baseline (Yes/No)
- Sexual behavior variables (from the Mosaico questionnaire): number of sexual partners in the past month, receptive anal sex without condom (yes/no), insertive anal sex without condom (yes/no). These variables will be used individually as covariates, as no combined behavioral risk score exists in the Mosaico clinical database.
Trial-related variables:
- Reactogenicity measures: Local/systemic adverse events, graded as mild, moderate, or severe
- Dosing interval: Time between Ad26 prime and gp140 boost (continuous, weeks)
Statistical Analysis Plan:
Descriptive analyses: We will summarize baseline Ad26 neutralizing antibody titers, gp140 antibody levels, and demographic/clinical variables using appropriate measures of central tendency and dispersion. Regional distributions will be tabulated and visualized to characterize heterogeneity.
Primary analysis (Aim 1): The association between baseline Ad26 titers (log10 IC90) and Month-7 gp140 responses will be estimated using linear regression and flexible spline-based models. To evaluate heterogeneity by region, models will include interaction terms between baseline Ad26 titers and country. Exposure--response curves and marginal effects will be presented by region.
Secondary analyses (Aim 2):
Within strata of baseline Ad26 titers (e.g., quartiles), HIV acquisition will be summarized descriptively using Kaplan--Meier curves and Cox proportional hazards models, restricted to vaccine recipients within the IAS, given that baseline Ad26 titers are only available for this case-control subset from the four Latin American countries. These analyses will be exploratory given the lack of overall vaccine efficacy in Mosaico.
Covariate adjustment and subgroups:
Multivariable models will adjust for key demographic and clinical variables (e.g., age, sex/gender, PrEP use, baseline STIs, dosing interval). Subgroup analyses will explore consistency of findings across demographic and risk categories.
All analyses will be conducted within the secure YODA platform environment using R or Stata. Results will be reported with 95% confidence intervals and p-values, with emphasis on effect sizes and patterns rather than strict significance testing.
Narrative Summary: This post-hoc study will build on findings from the Mosaico trial showing that pre-existing Ad26 immunity can dampen antibody responses (gp140). Using individual-level data, we will assess whether this association differs across the four Latin American countries with available immunogenicity data (Argentina, Brazil, Mexico, and Peru) by modeling baseline Ad26 titers against Month-7 gp140 levels and comparing exposure--response curves, with particular focus on Peru.
Project Timeline:
- Project start date / Data access granted: January 2026
- Data preparation and descriptive analyses: February--March 2026
- Primary analysis (Aim 1, immunogenicity models): April--May 2026
- Secondary/exploratory analyses (Aim 2, HIV acquisition by strata): June 2026
- Drafting manuscript: July--August 2026
- Internal and collaborator review, revisions: September 2026
- First manuscript submission: October 2026
- Report results to YODA Project: November 2026
Dissemination Plan:
The primary product of this project will be a peer-reviewed manuscript presenting the association between baseline Ad26 neutralizing antibody titers and gp140 immune responses across regions in the Mosaico trial. Findings will be relevant to the HIV vaccine field and to broader adenovirus-vectored vaccine research.
We anticipate submission to high-impact infectious diseases or vaccine journals. In addition, results will be shared with the HIV Vaccine Trials Network (HVTN) and collaborators through internal presentations and working group discussions.
Key findings will be disseminated to the scientific community via presentations at major conferences (e.g., IAS HIV Science Conference, CROI, HIVR4P). A plain-language summary will be prepared for community audiences and shared with participating sites, including Peru, to ensure accessibility of results beyond the scientific community.
All results will be reported back to the YODA Project per data-sharing requirements.
Bibliography:
1. Baden LR, Stieh DJ, Sarnecki M, Walsh SR, Tomaras GD, Kublin JG, et al. Safety and immunogenicity of two heterologous HIV vaccine regimens in healthy, HIV-uninfected adults (TRAVERSE): a randomised, parallel-group, placebo-controlled, double-blind, phase 1/2a study. Lancet HIV. 2020;7(10):e688–e698. doi:10.1016/S2352-3018(20)30229-0.
2. Stephenson KE, Wegmann F, Tomaka F, Walsh SR, Tan CS, Lavreys L, et al. Comparison of shortened mosaic HIV-1 vaccine schedules: a randomised, double-blind, placebo-controlled phase 1 trial (IPCAVD010/HPX1002) and a preclinical study in rhesus monkeys (NHP 17–22). Lancet HIV. 2020;7(6):e410–e421. doi:10.1016/S2352-3018(20)30001-1.
3. Fausther-Bovendo H, Kobinger GP. Pre-existing immunity against Ad vectors: humoral, cellular, and innate response, what’s important? Hum Vaccin Immunother. 2014;10(10):2875–2884. doi:10.4161/hv.29594.
4. Gray G, Buchbinder S, Duerr A. Overview of STEP and Phambili trial results. Curr Opin HIV AIDS. 2010;5(5):357–61.
5. Sumida SM, Truitt DM, Lemckert AAC, Vogels R, Custers JHHV, Addo MM, et al. Neutralizing antibodies to adenovirus serotype 5 vaccine vectors are directed primarily against the adenovirus hexon protein. J Immunol. 2005;174(11):7179–7185. doi:10.4049/jimmunol.174.11.7179.
