Clinical Research and Public HealthAIDS/HIVInfectious disease Open Access | 10.1172/jci.insight.190831

Joshua Rhein,1 Jeffrey G. Chipman,2 Gregory J. Beilman,2 Ross Cromarty,3 Kevin Escandón,1 Jodi Anderson,1 Garritt Wieking,1 Jarrett Reichel,1 Rodolfo Batres,1 Alexander Khoruts,4 Christopher M. Basting,2 Peter Hinderlie,5 Zachary B. Davis,5 Anne Eaton,6 Byron P. Vaughn,4 Elnaz Eilkhani,7 Jeffrey T. Safrit,8 Patrick Soon-Shiong,8 Jason V. Baker,1,9 Nichole R. Klatt,2 Steven G. Deeks,7 Jeffrey S. Miller,3,5 and Timothy W. Schacker1

1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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1Division of Infectious Diseases and International Medicine, Department of Medicine;

2Department of Surgery;

3Masonic Cancer Center;

4Division of Gastroenterology, Department of Medicine;

5Division of Hematology, Oncology, and Transplantation, Department of Medicine; and

6Division of Biostatistics and Health Data Science, School of Public Health, University of Minnesota, Minneapolis, Minnesota, USA.

7Division of HIV, Infectious Diseases and Global Medicine, Department of Medicine, University of California, San Francisco, San Francisco, California, USA.

8ImmunityBio, Culver City, California, USA.

9Division of Infectious Diseases, Hennepin Healthcare Research Institute, Minneapolis, Minnesota, USA.

Address correspondence to: Timothy W. Schacker, University of Minnesota, MMC 250, 420 Delaware Street SE, Minneapolis, Minnesota 55455, USA. Phone: 612.624.9955; Email: schacker@umn.edu.

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Published June 5, 2025 - More info

Published in Volume 10, Issue 13 on July 8, 2025
JCI Insight. 2025;10(13):e190831. https://doi.org/10.1172/jci.insight.190831.
© 2025 Rhein et al. This work is licensed under the Creative Commons Attribution 4.0 International License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Published June 5, 2025 - Version history
Received: January 2, 2025; Accepted: May 23, 2025 View PDF Abstract

BACKGROUND. NK cell function is impaired in people with HIV (PWH), hindering their potential to reduce the lymphoid tissue (LT) reservoir. The IL-15 superagonist N-803 has been shown to enhance NK and T cell function and thus may reduce viral reservoirs.

METHODS. To determine the impact of N-803 on LTs, we conducted a clinical trial where 10 PWH on effective antiretroviral therapy (ART) were given 3 subcutaneous 6 mcg/kg doses of N-803. We obtained PBMCs and lymph node (LN) and gut biopsies at baseline and after the last N-803 dose.

RESULTS. We found a nonstatistically significant approximately 0.50 median log reduction in the frequency of viral RNA+ (vRNA+) and vDNA+ cells/g in the 6 participants with baseline and posttreatment LN biopsies. In the ileum, we observed reductions of vRNA+ cells in 8/10 participants and vDNA+ cells in all participants. We also found significant inverse correlations between NK cell proliferation and the frequency of vRNA+ cells and between NKG2A expression on NK cells and the frequency of vRNA+ cells.

CONCLUSION. Our findings suggest N-803 may reduce the HIV reservoir in LTs of PWH on ART, an effect likely mediated by enhanced NK cell function. Controlled studies assessing the impact of NK cell therapy on HIV LTs are needed.

TRIAL REGISTRATION. ClinicalTrials.gov NCT04808908.

FUNDING. NIH grants 5UM1AI126611, UL1TR002494, R01 AI147912, R35 CA283892, and UM1AI164561.

Introduction

HIV infection has become a chronic manageable condition due to the widespread use of antiretroviral therapy (ART) in people with HIV (PWH), but it is not currently curable because of the persistence of viral reservoirs. During primary infection, HIV establishes tissue reservoirs that include a large population of HIV-infected CD4+ T cells with a transcriptionally silent HIV provirus (the “latent” reservoir) (1). These cells are largely maintained via homeostatic proliferation (2, 3), and only a small fraction have an intact provirus that can reactivate from latency (4). Although most studies have focused on circulating CD4+ T cells harboring integrated genomes in PWH, the vast majority of these cells reside in tissue sanctuaries such as secondary lymph nodes (LNs) and gut-associated lymphoid tissues (GALTs), even in PWH receiving ART (5–8). Ideally, any strategy to cure HIV would seek to target both the latent reservoir and the reservoir of viral RNA+ (vRNA+) productively infected cells.

While NK cells are an important part of the innate immune system that target and kill infected cells, their function is frequently impaired in HIV infection (9–12). There are, however, immune-based therapies that are now available with the potential to improve or restore this function (13–15). One such possible intervention is the IL-15 superagonist N-803 (nogapendekin alfa inbakicept, ImmunityBio), which, in animal models of HIV, has been found to activate T cells and NK cells while reversing latency (14, 16–18). In a phase I study of ART-suppressed PWH, we found that N-803 induced virus production; activated CD4+ T cells, CD8+ T cells, and NK cells; and reduced the size of the reservoir in circulating PBMCs (19).

To assess the ability of N-803 to reduce viral reservoirs, we conducted a phase Ib clinical trial wherein PWH on ART received N-803 and provided both baseline and posttreatment tissue samples, including PBMCs and both LN and gut biopsies. In addition, we investigated NK cell and CD8+ T cell function in LNs and GALTs.

Results

Protocol. A phase Ib, single-arm, open-label study (ClinicalTrials.gov NCT04808908) was conducted. Participants were required to be on continuous ART for at least 2 years, to have undetectable plasma HIV RNA measures in the previous 12 months, and to have a screening CD4+ T cell count ≥350 cells/μL. Leukaphereses to obtain PBMCs, excisional inguinal LN biopsies, and colonoscopies with ileal biopsies were conducted at baseline (prior to the first dose of N-803) and again 1 week after the third dose of N-803. All 10 participants received 3 s.c. 6 μg/kg doses of N-803 at 21-day intervals. The dose and interval used were informed by our previous dose-escalation trial (19) and studies in people with cancer (20).

Participants. We screened and consented a total of 23 individuals (Figure 1), of whom 10 were eligible per our selection criteria and completed dosing. The remaining 13 participants included 11 participants who failed screening criteria and 2 participants who withdrew their consent after successful screening. Primary reasons for screen failure were detectable viral load at screening, evidence of currently active infections such as chronic hepatitis B and tuberculosis, and abnormalities in screening tests conducted per study protocol.

Figure 1

Study schematic. Flow diagram and design of the clinical trial.

All of the 10 enrolled participants received the planned 3 doses of 6 mcg/kg N-803 by s.c. injection. We performed leukapheresis and colonoscopy with biopsies at both baseline and 1 week after last N-803 dose in all 10 participants. Despite ultrasound guidance, we were unable to locate an LN at baseline in 4 of the individuals; hence, only 6 participants provided pre– and post–N-803 LN samples. Study participants were followed up for 3 months after completing N-803 dosing.

All participants were male, and their median age was 38 years (interquartile range [IQR], 32.3–46.3). Of the 10 participants, 8 identified as White and 2 identified as African American. Ethnicity was non-Hispanic for all participants. The median CD4+ T cell count at baseline was 593 cells/μL (IQR 450.3–949.3), and the median CD4/CD8 ratio was 0.87 (IQR 0.56–1.45) (Table 1).

Table 1

Demographic and clinical characteristics of study participants at baseline

Impact of N-803 on lymphoid tissue reservoirs of HIV RNA+ and DNA+ cells. We first measured the frequency of vRNA+ cells in LNs using RNAscope. The median log frequency of vRNA+ cells was 4.1 vRNA+ cells/g (IQR 3.9–4.6) in the 6 LNs obtained at baseline and 4.3 vRNA+ cells/g (IQR 3.8–4.8) in the 10 LNs from the post–N-803 time point (P = 0.8570, unpaired 2-tailed t test, Figure 2A). For the 6 individuals with LNs obtained at both baseline and post–N-803 time points, there was a median log decrease of 0.46 vRNA+ cells/g that was not statistically significant (IQR –0.33–1.20, P = 0.21, paired 2-tailed t test).

Figure 2

Analysis of changes in frequency of vRNA+ and vDNA+ cells in LTs. (A) Analysis of the frequency of vRNA+ cells in the 6 LNs obtained at baseline and the 10 LNs obtained at the post–N-803 time point using HIV RNA in situ hybridization by RNAscope. (B) Analysis of the frequency of vDNA+ cells by DNAscope. (C) Analysis of the frequency of vRNA+ cells in the ileum. (D) Analysis of the frequency of vDNA+ cells in the ileum. (E) Representative images from a participant’s LN showing vRNA+ and vDNA+ cells at the pre– and post–N-803 time points. Scale bar: 50 μm; inset, original magnification, 20×. Statistical analyses performed using an unpaired 2-tailed t test for A and B and paired 2-tailed t test for C and D; P values are shown above the pairings.

We next measured the frequency of vDNA+ cells in LNs using DNAscope (Figure 2B). The median log frequency of vDNA+ cells was 5.3 vDNA+ cells/g (IQR 5.0–5.5 vDNA+) in the 6 LNs obtained at baseline and 4.9 vDNA+ cells/g (IQR 4.7–5.3) in the 10 LNs from the post–N-803 time point (P = 0.1165, unpaired 2-tailed t test). Similar to what we observed for changes in vRNA+ cells in paired LNs, there was a median log decrease in the frequency of vDNA+ cells of 0.53 cells/g (IQR –0.12–0.70, P = 0.085, paired 2-tailed t test) that did not reach significance. The change for each of the 6 paired samples was –1.1, –0.56, –0.55, –0.52, +0.10, and +0.16 log vDNA+ cells/g LN.

We also measured the frequency of vRNA+ and vDNA+ cells/g in ileum at both time points for all 10 participants and found a modest decline in the frequency of vRNA+ cells in 8/10 participants (Figure 2C). The median change was –0.15 log (IQR –0.50–0.15 log). While this decline was small and of unknown biological significance, the differences were statistically significant (P = 0.0186, paired 2-tailed t test). There was also a decrease in the frequency of vDNA+ cells/g in all 10 participants in the ileum (Figure 2D); the median change was –0.21 log (IQR –0.003 to –0.5 log, P = 0.0007, paired 2-tailed t test). Figure 2E shows representative images of a participant’s LN analyzed for vRNA+ and vDNA+ cells at the pre– and post–N-803 time points.

To ensure that changes to vRNA+ and vDNA+ cell populations were not the result of a “dilutional effect” secondary to cells trafficking in and out of the LNs, we used quantitative image analysis (QIA) to calculate the frequency of CD4+ T cells in the T cell zone (TZ) of the paired LNs from 5 of the participants. Because the vRNA+ and vDNA+ measurements were on CD4+ T cells, we reasoned that a dilutional effect would be reflected by changes in the size of the CD4+ T cell population in the TZ. We found the median percentage change between time points was 0.6% (range 0.22%–0.87%); thus, any change in the size of the reservoir was not due to a dilutional effect from cells moving in and out of the LNs.

Measures and function of CD8+ T cells and NK cells. We next examined the impact of N-803 on CD8+ T cell and NK cell populations in LNs and GALTs, as N-803 is known to cause activation and proliferation of these effector cells. We stained tissues with antibodies directed against CD8+ T cells and used QIA to measure the frequency of CD8+ T cells in the parafollicular TZ and B cell follicles (identified by double-label staining with CD20 to mark B cells). We were particularly interested in the changes in these populations by anatomic location in the LNs because of a report of an increase in SIV-specific CD8+ T cells in follicles after N-803 treatment in a nonhuman primate (NHP) model of SIV infection (21). In contrast with that study, we found CD8+ T cells significantly decreased in follicles after N-803 therapy (P = 0.0169, paired 2-tailed t test, Figure 3A). However, when analyzing the whole tissue section, we detected no significant change in the frequency in LNs overall (Figure 3B, P = 0.5112, unpaired 2-tailed t test), with some individuals having an increase and some a decrease.

Figure 3

Measures and function of CD8+ T cells and NK cells in LNs. (A) Frequency of CD8+ T cells in B cell follicles in the 6 individuals with both a pre– and post–time point. We counted CD8+ T cells inside of B cell follicles and in the parafollicular T cell zone to provide an estimate of the percentage of those cells in the B cell. (B) Grouped analysis of the log frequency of CD8+ T cells in the group with 6 baseline LN samples and the group with 10 post–N-803 LN samples. (C) Grouped analysis of the log frequency of NKG2A+ cells in 6 pre– and 10 post–N-803 LNs. (D) Representative images from a participant’s LN with an increase in both CD8+ and NKG2A+ cells at the pre– and post–N-803 time points. Scale bar: 100 μm. Statistical analyses performed using a paired 2-tailed t test in A and an unpaired 2-tailed t test in B and C; P values shown above pairings.

The NK cell population was examined by using antibodies directed against NKG2A to detect NK cells in the LTs. We recognize that a very small population of T cells can express the NKG2A receptor, but for purposes of this histologic analysis (where double-label image analysis was not feasible), we restricted our analyses to using only the NKG2A marker. Further, NK cells in LNs are mostly of the CD56bright phenotype, which highly express NKG2A (22). There was no significant change in the overall frequency of NK cells in LNs (Figure 3C, P = 0.1724, unpaired 2-tailed t test). Among the paired samples (6 participants), there were 3 with a log increase of 1.06, 0.53, and 0.19 NK cells and 3 with a log decrease of 0.49, 0.22, an