A Clinical Trial to Evaluate the Safety and Immunogenicity of Synthetic DNAs Encoding a Native-like HIV Env Trimer and Interleukin-12 (INO-6160), Alone or in a Prime-boost Regimen With 3M-052-AF + Alum Adjuvanted VRC HIV Env Trimer 4571 in Adult Participants Without HIV: Complete Statistical Analysis

1. Trial at a Glance

This page covers the registry results for A Clinical Trial to Evaluate the Safety and Immunogenicity of Synthetic DNAs Encoding a Native-like HIV Env Trimer and Interleukin-12 (INO-6160), Alone or in a Prime-boost Regimen With 3M-052-AF + Alum Adjuvanted VRC HIV Env Trimer 4571 in Adult Participants Without HIV (NCT05828095), a randomized phase 1 study. The primary outcome is Number of Participants Reporting Local Solicited Adverse Events Signs and Symptoms: Pain and/or Tenderness.

2. Clinical Question

The central comparison is between the two vaccine regimens in a randomized, open-label, parallel-group phase 1 trial. The registry lists 20 participants overall. The intervention descriptions distinguish INO-6160 alone from INO-6160 combined with the adjuvanted Env trimer. The trial is described as examining safety and immunogenicity; secondary outcomes include neutralizing antibody magnitude and response rates, HIV-1-specific IgG binding responses, and CD4+ and CD8+ T-cell responses.

The registry record does not report results for these immune-response outcomes. It also does not provide a single arm-level comparison for the primary local symptom outcome, which is represented across multiple categories or time points. Consequently, the trial question can be described from its design and registered outcomes, but the posted record does not establish a comparative estimate for either the primary safety measure or the stated immune-response hypothesis.

3. Trial Design

This was a phase 1, randomized, open-label clinical trial with a parallel-group model. It enrolled 20 adult participants without HIV and is marked completed in the registry record. The recorded start date is 2023-04-05, and the primary completion date is 2025-01-23. The sponsor is the National Institute of Allergy and Infectious Diseases.

One group received INO-6160 at 2 mg as 2 separate intradermal injections, one in each arm. The other group received that INO-6160 regimen together with Trimer 4571, described as 100 mcg, plus 3M-052-AF at 5 mcg and Alum at 500 mcg. The trimer regimen was administered as 2 intramuscular injections, one in each arm. The registry does not give a full administration schedule or the interval between doses in the information recorded for these interventions.

Randomization means that group assignment was determined through a chance-based process, but the method used to generate the sequence and conceal assignments is not reported in the registry record. Open-label means participants and trial personnel were not masked to group assignment. The registry does not describe masking of outcome assessors. The parallel design means the groups received their assigned regimens as separate groups rather than crossing over between regimens.

The listed secondary outcomes include neutralizing antibody magnitude and response rates against autologous and Tier 1a HIV-1 isolates following the third and fourth vaccinations; differential binding response rates and epitope specificity for HIV-1-specific IgG; and response rates and magnitudes for CD4+ and CD8+ T-cell responses. The record lists 27 outcome measures overall. It does not report a statistical analysis plan, a sample-size justification, or details of the randomization procedure.

4. Participant Flow and Accounting

The registry reports how participants moved through each stage of the study. All counts below are exactly as posted; missing counts are not inferred from other stages.

Overall Study

Reasons for not completing this period:

5. Baseline Characteristics

The table reproduces the baseline characteristics posted in the registry, which describe the participants analysed at baseline and help show whether the randomized groups were comparable.

6. Primary Endpoint

The registry lists the primary outcome as Number of Participants Reporting Local Solicited Adverse Events Signs and Symptoms: Pain and/or Tenderness, measured over Measured for 14 days after each injection.

The registered primary outcome is the number of participants reporting local solicited adverse-event signs and symptoms of pain and/or tenderness. The registry says these symptoms were measured for 14 days after each injection. Severity was graded using the Division of AIDS Table for Grading the Severity of Adult and Pediatric Adverse Events, Version 2.1, dated July 2017. For each symptom, the maximum grade observed during the relevant time frame is presented.

This endpoint captures participants’ reported local symptoms following vaccination, while the grading framework records symptom severity as well as whether a symptom was reported. In general, solicited symptom measures use a defined observation period and specified symptoms to make collection more consistent across participants and groups. They address a different aspect of a trial than laboratory measures of immune response.

The registered endpoint is expressed as a participant count, not as a continuous measurement. However, the registry record indicates that the primary outcome has multiple categories or time points and has not been reduced to one comparison. The available record therefore does not support a single numerical comparison of the primary endpoint between the randomized groups. The brief summary’s stated hypothesis concerns antibody and T-cell responses; the record does not explain why the listed primary outcome is the local pain-and-tenderness measure or how the two descriptions are linked.

7. Why No Re-analysis Is Shown

8. Statistical Interpretation

9. Statistical Methods Explained

What does a randomized parallel-group design contribute?

General: Random assignment is intended to make treatment groups comparable at the point of allocation, on average, by using chance rather than participant or investigator choice to determine assignment. It does not guarantee that every characteristic will be balanced in a small study. A parallel-group design keeps participants in their assigned group rather than having the same participants receive both regimens. In this trial, the registry identifies randomization and a parallel model, but the sequence-generation and allocation-concealment procedures are not reported in the registry record.

The open-label designation means assignment was not masked for participants and trial personnel. General: knowledge of assignment can affect subjective reporting, symptom assessment, or other trial procedures. That is a consideration when interpreting reported symptoms, not evidence that any particular assessment in this trial was biased. The registry does not say whether outcome assessors were masked.

How should a participant count be interpreted?

The primary outcome is a count of participants with local pain and/or tenderness, with the maximum grade during the specified observation window described. General: a count answers how many people met an outcome definition; it differs from a measurement of symptom intensity averaged across participants. For a binary outcome, an arm-level comparison commonly uses the number meeting the definition and the number assessed in each group. This record does not report one arm-level primary-outcome comparison, so such a comparison cannot be interpreted from the posted results.

A confidence interval, in general, describes a range of values compatible with the data under a specified statistical model and gives information about estimate precision. It is not a guarantee that the true effect lies within that range. No confidence interval for the primary endpoint or the listed immune-response analyses is reported in the registry record, so no trial-specific interval interpretation is possible.

What do the planning illustrations mean?

The stated sample-size assumptions describe planning illustrations based on the observed effect, rather than the trial’s own design justification. They use a two-sided significance level of 0.05, equal allocation between groups, and a normal approximation; the effect to detect is defined as a stated fraction of the observed effect. For outcomes other than time-to-event outcomes, the illustrations are expressed as participants per arm for 80% power or another stated power level; time-to-event illustrations use total events.

General: a planning calculation asks how much information a future study might need under explicit assumptions. It is not an observed result, does not change the number enrolled in this trial, and is not a measure of the probability that a hypothesis is true. Such calculations are sensitive to the assumed effect size and statistical model. Because the registry record does not report a sample-size justification for this trial, a planning illustration should not be presented as the rationale used to choose its enrollment. The posted record does not provide a trial-specific interpretation of any illustrated sample size.

Why do analysis details matter?

General: statistical conclusions depend on more than the outcome label. The analysis must specify which participants are included, how repeated observations or multiple outcome categories are handled, and what comparison is estimated. A prespecified approach can reduce ambiguity about which results are emphasized. The registry record lists 27 outcome measures but zero analyses, and it does not report the statistical plan, handling of missing data, or analysis populations. It is therefore not possible to determine from the registry which model or test was intended for each outcome.

Multiplicity arises when several hypotheses or outcomes are assessed. General: examining many outcomes can increase the chance of at least one apparently unusual result if each is tested separately, so an analysis plan may specify how results will be interpreted or adjusted. This trial lists several immunogenicity outcomes and a total of 27 outcome measures, but multiplicity procedures are not reported in the registry record. No adjustment should be assumed.

Over-dispersion is a general issue for count data when variation exceeds what a simple count model expects; if present, it can make standard errors too small unless the model accounts for it. The primary endpoint is a participant count, but no fitted model or dispersion estimate is reported, and the registry does not establish that over-dispersion was present. Likewise, it does not report a model for repeated measurements across injection periods. These are considerations for interpreting or planning analyses, not descriptions of analyses documented for this trial.

10. Multiplicity, Missing Data and Analysis Populations

The registry lists 27 outcome measures and zero analyses. Several secondary outcomes address different aspects of immunogenicity: neutralizing antibody magnitude and response rates, IgG binding response rates and epitope specificity, and CD4+ and CD8+ T-cell response rates and magnitudes. The record also describes neutralizing antibody measures following the third and fourth vaccinations. Assessing multiple outcomes and time points creates choices about which comparisons to emphasize and how to interpret chance variation across results.

General: multiplicity procedures can include prespecifying a primary comparison, adjusting statistical thresholds, or treating secondary outcomes as exploratory. The registry record does not report any multiplicity procedure or analysis hierarchy. It is not possible to tell whether these outcomes were planned as confirmatory or exploratory, or whether any adjustment was used. The reported hypothesis in the brief summary concerns envelope-specific antibody and T-cell responses, while the listed primary outcome concerns local pain and/or tenderness; the record does not explain how these were prioritized relative to one another.

Missing-data methods are not reported in the registry record. The record does not state how missing symptom observations, incomplete vaccination, or unavailable immune-response measurements were handled. Analysis populations, such as whether all randomized participants or only participants with particular measurements were included, are also not reported in the registry record. These omissions prevent assessment of how the analyzed groups may differ from the enrolled groups.

These are reporting gaps, not evidence that missing data or multiplicity problems occurred. They limit what can be concluded from the posted results because the analysis choices and their consequences cannot be evaluated. No adjustment, imputation method, or analysis population should be inferred from the outcome descriptions alone.

11. Safety Results

The registry reports serious adverse events as affected participants divided by participants at risk.

Deaths

Other (non-serious) adverse events

Participants with at least one non-serious event at or above the registry's 0% reporting threshold.

Most frequent non-serious adverse event terms

The registry’s adverse-event summary lists serious events and deaths separately from other adverse events. Group T1 has 10 participants at risk for serious events, with 0 affected, and 10 at risk for other adverse events, with 9 affected. Group T2 has 10 participants at risk for serious events, with 0 affected, and 10 at risk for other adverse events, with 10 affected. The death count is 0 in each group. These are participant counts in the posted summary.

General: “at risk” is the denominator used for the corresponding adverse-event category, and “affected” is the number of participants recorded with an event in that category. Serious events and other adverse events are distinct summary categories; the other-event counts should not be read as serious-event counts. The registry record does not list serious adverse-event terms, and no serious terms are included in the summary.

These figures describe the participants represented in the registry summary. They do not, by themselves, establish whether one regimen has a different safety profile, especially without definitions, event-level details, follow-up context, or a statistical comparison. The registry record also does not connect Group T1 and Group T2 explicitly to the intervention descriptions in the information. Zero recorded serious events and deaths in these groups is a description of the posted counts, not proof that an event is impossible or that risk is absent.

12. What the Result Does, and Does Not, Mean

The registry record establishes that this was a completed, randomized, open-label phase 1 trial enrolling 20 adults without HIV to examine INO-6160 alone or in a regimen with an adjuvanted HIV Env trimer. It defines local pain and/or tenderness as the primary outcome and names a range of antibody and T-cell measures as secondary outcomes. Its adverse-event summary reports the participant counts by group, including 0 participants affected by serious adverse events in each listed group and 0 deaths in each group.

Those details do not answer whether the regimens differed on the primary symptom outcome or on immunogenicity. The registry does not report analyses for its 27 outcome measures, and the primary outcome is not presented as one comparison across the groups. Immune-response results, effect estimates, confidence intervals, and p-values are not reported in the registry record. The intended hypothesis in the brief summary is not evidence that the hypothesis was supported or rejected.

General: a trial’s estimates apply most directly to the participants and conditions studied. A phase 1 study is commonly used to characterize early safety and biological responses, but that general role does not supply missing results or determine the outcome of a particular trial. The sample enrolled is 20, and the record offers no basis for making precise claims about outcomes beyond the trial participants.

The posted counts are descriptive and should not be turned into a claim that either regimen is safer, more immunogenic, or preferable. A comparative conclusion would require clearly reported outcome definitions, group-level results, analysis methods, and uncertainty. Those are not available for the key outcome comparisons in the registry record. The appropriate interpretation is limited to what the record states: the trial design and outcome intentions, the displayed adverse-event counts, and the absence of reported analyses for the outcome measures.

13. Why This Trial Matters Statistically

Statistically, this trial illustrates why a registry entry can identify important questions without supplying enough analyzed results to answer them. The study is randomized and parallel-group, so its design sets up a comparison between INO-6160 alone and a regimen combining INO-6160 with an adjuvanted Env trimer. Yet the record does not provide a single comparative analysis for its primary local symptom measure or analyses for the immune-response outcomes.

The distinction between a hypothesis and an endpoint is important. The brief summary describes a hypothesis about HIV-1 envelope-specific antibody and T-cell responses, whereas the registered primary outcome is a participant count for local pain and/or tenderness. The record does not explain the relationship between these descriptions. For statistical interpretation, readers need to know which outcomes are primary, how multiple outcomes are prioritized, and what estimand or comparison is intended. Those details are not reported in the registry record.

The entry also lists 27 outcome measures. This makes transparent analysis reporting especially important: immune-response measures differ in type, and some are recorded after specified vaccination numbers. Without the statistical plan, analysis populations, missing-data methods, and multiplicity approach, readers cannot evaluate the analysis choices or the strength of evidence behind any result. The registry lists zero analyses, so those questions remain open.

Finally, the adverse-event counts show how descriptive numbers can be informative while remaining limited. They record the numbers affected in each listed category and group, but do not establish comparative safety or precision. The record is useful for identifying what was studied and what was intended to be measured; its lack of reported outcome analyses limits conclusions about what the trial found.

14. Limitations

• The posted summary statistics are the only data these results rest on; patient-level data, full protocols and statistical analysis plans are not available.
• Recalculated results are simple, unadjusted calculations from posted summaries. They do not replace the sponsor's prespecified analysis.

The most important limitation for interpretation is that the registry lists 27 outcome measures but zero analyses. The primary endpoint’s definition is available, but the record says it has several categories or time points and is not reduced to one comparison. The registry does not report the corresponding symptom counts by grade and time point as a single comparative estimate, nor does it provide results for the listed immune-response outcomes.

The record does not report a statistical analysis plan, sample-size justification, randomization method, allocation concealment, analysis populations, or missing-data procedures. It also does not describe multiplicity handling across the many outcome measures. These omissions do not prove that such methods were absent from trial conduct; they mean they are not reported in the registry record, limiting evaluation of the planned and completed analyses.

The study is described as open-label, and masking of outcome assessors is not reported. Because the primary symptom endpoint is based on solicited local symptoms, awareness of assignment is a general consideration when interpreting subjective reports. The registry information does not establish that reporting or assessment was affected.

The adverse-event summary gives participant counts for serious and other adverse events, but does not supply serious-event terms or a comparative analysis. The registry record also does not explicitly map Group T1 and Group T2 to the detailed intervention descriptions. The total enrollment is 20, so the posted counts describe a limited group of participants; the record does not provide estimates of precision for broader populations. These limitations constrain interpretation without changing what the registry does report about the design and outcome definitions.

15. Related Tutorials

16. Related Statistical Calculators

17. Sources

• ClinicalTrials.gov: A Clinical Trial to Evaluate the Safety and Immunogenicity of Synthetic DNAs Encoding a Native-like HIV Env Trimer and Interleukin-12 (INO-6160), Alone or in a Prime-boost Regimen With 3M-052-AF + Alum Adjuvanted VRC HIV Env Trimer 4571 in Adult Participants Without HIV — NCT05828095.
• Registry record last updated 2025-09-02; status last verified 2025-07.
• Results first posted 2025-07-04.
• Data retrieved from ClinicalTrials.gov on 2026-11-09.
• Group sizes, means, event counts and sponsor-reported p-values are reproduced as posted. Effect estimates, confidence intervals and p-values not attributed to the sponsor are independently calculated from the posted summary statistics.

18. Record Summary

This completed phase 1 trial was randomized, open-label, and parallel-group, with 20 adult participants without HIV. It evaluated INO-6160 alone or with Trimer 4571 adjuvanted with 3M-052-AF and Alum.

The registered primary outcome was the number of participants reporting local pain and/or tenderness, measured for 14 days after each injection and graded using the Division of AIDS severity table. The brief summary states a hypothesis concerning HIV-1 envelope-specific antibody and T-cell responses; the secondary outcomes include antibody and T-cell measures.

The registry lists 27 outcome measures and zero analyses. The primary symptom outcome is not reduced to one comparison, and immune-response analyses are not reported in the registry record. The adverse-event summary reports 0 serious events and 0 deaths in both Group T1 and Group T2. The record does not report a statistical analysis plan, missing-data methods, or multiplicity procedures.