The implication of interleukin-2 on the expression of CD56<sup>bright</sup>, CD56<sup>dim</sup>, and interferon-γ in patients with systemic lupus erythematosus

Dwi Soelistyoningsih; Hani Susianti; Handono Kalim; Kusworini Handono; Jonny Karunia Fajar

doi:10.12688/f1000research.130553.1

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Research Article

The implication of interleukin-2 on the expression of CD56^bright, CD56^dim, and interferon-γ in patients with systemic lupus erythematosus

[version 1; peer review: 2 approved with reservations, 1 not approved]

Dwi Soelistyoningsih ^1,2, Hani Susianti³, Handono Kalim⁴, Kusworini Handono³, Jonny Karunia Fajar ⁵

Dwi Soelistyoningsih ^1,2, Hani Susianti³, [...] Handono Kalim⁴, Kusworini Handono³, Jonny Karunia Fajar ⁵

PUBLISHED 28 Nov 2023

Author details Author details

¹ Doctoral Program, Faculty of Medicine, Universitas Brawijaya, Malang, 65145, Indonesia
² Department of Nursing, STIKES Widyagama Husada, Malang, 65145, Indonesia
³ Department of Clinical Pathology, Faculty of Medicine, Universitas Brawijaya, Malang, 65145, Indonesia
⁴ Division of Reumatology & Immunology, Department of Internal Medicine, Faculty of Medicine, Universitas Brawijaya, Malang, 65145, Indonesia
⁵ Brawijaya Internal Medicine Research Center, Department of Internal Medicine, Faculty of Medicine, Universitas Brawijaya, Malang, 65145, Indonesia

Dwi Soelistyoningsih
Roles: Conceptualization, Data Curation, Formal Analysis, Investigation, Methodology, Project Administration, Resources, Writing – Original Draft Preparation

Hani Susianti
Roles: Supervision, Validation

Handono Kalim
Roles: Supervision, Validation

Kusworini Handono
Roles: Supervision, Validation

Jonny Karunia Fajar
Roles: Formal Analysis, Methodology, Software, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing

OPEN PEER REVIEW

REVIEWER STATUS

This article is included in the Cell & Molecular Biology gateway.

Abstract

Background: Interleukin-2 (IL-2) stimulation had been reported as having a beneficial impact to the expression of CD56^bright, CD56^dim, and interferon-γ (IFN-γ) in the case of immunological dysfunction diseases. However, in the case of systemic lupus erythematosus (SLE), the role of IL-2 had never been investigated. The objective of this study was to assess the impact of IL-2 on the expression of CD56^bright, CD56^dim, and IFN-γ in SLE patients.
Methods: An experimental study was conducted by involving peripheral blood mononuclear cells isolated from six SLE patients. The study consisted of four groups based on IL-2 stimulation: D0 (0U/ml), D1 (50U/ml), D2 (150U/ml), and D3 (250U/ml); and they were then cultured for 72 hours. The levels of CD56^bright and CD56^dim were measured by FACSMelody^TM, while the levels of IFN-γ were measured using ELISA.
Results: In group D0, D1, D2, and D3; the levels of CD56^bright were 57.27±37.27, 241.16±64.41, 256.94±50.95, and 259.37±36.44 x1000 cells/mm3 respectively. Moreover, the levels of CD56^dim were 812.85±167.37, 631.98±129.90, 616.42±157.97, and 615.90±155.57 x1000 cells/mm3 respectively. On the other hand, the levels of IFN-γ were 24.01±2.56, 26.09±4.79, 30.11±5.34, and 32.43±7.14 pg/ml respectively. Our analysis elucidated that the administration of IL-2 provided potential impact to the levels of CD56^bright, but not to the levels of CD56^dim and IFN-γ. Our findings indicated that the increased dosage of IL-2 resulted in a more significant impact on CD56^bright.
Conclusions: Our study clarifies that IL-2 provides a beneficial impact on CD56^bright expression in SLE patients.

Keywords

systemic lupus erythematosus, interleukin – 2, natural killer cells, CD56bright, CD56dim, interferon – γ

Corresponding authors: Dwi Soelistyoningsih, Jonny Karunia Fajar

Competing interests: No competing interests were disclosed.

Grant information: The author(s) declared that no grants were involved in supporting this work.

Copyright: © 2023 Soelistyoningsih D et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

How to cite: Soelistyoningsih D, Susianti H, Kalim H et al. The implication of interleukin-2 on the expression of CD56^bright, CD56^dim, and interferon-γ in patients with systemic lupus erythematosus [version 1; peer review: 2 approved with reservations, 1 not approved]. F1000Research 2023, 12:1525 (https://doi.org/10.12688/f1000research.130553.1) First published: 28 Nov 2023, 12:1525 (https://doi.org/10.12688/f1000research.130553.1) Latest published: 28 Nov 2023, 12:1525 (https://doi.org/10.12688/f1000research.130553.1)

Introduction

Systemic lupus erythematosus (SLE) remains a serious health issue. The global morbidity of SLE was predicted to be between 9.0 and 366.6 cases per 100,000 individuals, and of them, the mortality was reported ranging from 6.7 to 37.8%.¹ The management of SLE presents considerable difficulties, both in terms of diagnosing the condition and providing effective treatment. It has been widely known that SLE is a disease with 1000 faces, suggesting that SLE has a wide variety of clinical manifestation, and therefore, the diagnosis of SLE requires the carefully holistic investigation.² On the other hand, the treatment of SLE remains a challenging global issue. The current treatment for SLE patients only targets remission, no single effective treatment has been proven in curing SLE. This challenging treatment issue is the implication of the complicated pathogenesis of SLE.³ Theoretically, the pathogenesis of SLE is complex and may involve interferon regulatory factor, toll-like receptor, protein tyrosine phosphatase, natural killer (NK) cells, and interferon-γ (IFN-γ).⁴ Studies suggested that, in SLE patients, NK cells dysfunction was observed, and the activity of NK cells was governed by the balance expression between CD56^bright (NK cells regulator) and CD56^dim (NK cells cytotoxic).⁵^–⁷ Moreover, CD56^bright had been proven to have the ability to initiate the production of IFN-γ.⁸ Therefore, a therapeutic modality by targeting the activity of NK cells might provide a beneficial impact.

Interleukin-2 (IL-2) is one of the primary cytokines having pleiotropic impact on the immune system.⁹ IL-2 has been reported as the potential treatment for the management of several diseases with immunological dysfunction such as: chronic graft-versus-host disease (GVHD)¹⁰ and acute myeloid leukemia.¹¹ The possible mechanism of action of IL-2 in NK cells is by binding to NK cell activating receptors to affect the expression of NK cell subsets such as CD56^bright and CD56^dim, and as a result, the proliferation and the production IFN-γ may occur.⁵ In the case of GVHD and acute myeloid leukemia; IL-2 had been reported to affect the expression of CD56^bright and CD56^dim NK cells as well as the levels of IFN-γ.¹⁰^,¹¹ On the other hand, in the case of SLE, since a previous report revealed that the levels of IL-2 was decreased,¹² the potential treatment of IL-2 in SLE patients by assessing NK cells subsets has never been investigated. Therefore, our present study sought to assess the implication of IL-2 administration to the expression of CD56^bright and CD56^dim as well as the levels of IFN-γ.

Methods

Ethical considerations

The study was approved by the local Ethical Committee of Saiful Anwar Malang Hospital (No. 400/231/K.3/302/2019; approved on 22 October 2019)¹³ and conducted in concordance with the Declaration of Helsinki.¹⁴ Before being involved in the study, patients provided written informed consent. A total of 18 mL of peripheral venous blood was collected and placed in a vacutainer tube containing EDTA.

Study design and sample preparation

During January–July 2020, lymphocytes were collected from six female SLE patients in Saiful Anwar Hospital, Indonesia. SLE patients in our present study conformed with the classification criteria of the 2019 European League Against Rheumatism and the American College of Rheumatology (2019 EULAR/ACR).¹⁵ The baseline characteristics of SLE patients in our study were as follows: age 18–49 years old (mean = 36,17±6,494 years) and in active disease with Mexico-SLE Disease Activity Index (MEX-SLEDAI) score > 5 (mean = 9,5±2,345 points). The subjects were divided into four different groups of IL-2 stimulation: D0 (control/0 U/ml), D1 (50 U/ml), D2 (150 U/ml), and D3 (250 U/ml).

Peripheral blood mononuclear cell (PBMC) incubation

Peripheral blood mononuclear cells (PBMCs) were collected by Lymphoprep™ Density Gradient Medium 1.07 (Axis – Shield, Oslo, Norway) within 6 hours of blood drawing. PBMCs were then incubated in the medium of Roswell Park Memorial Institute (RPMI) 1640 (Sigma-Aldrich, Missouri, US, RRID:SCR_013728) containing L-Glutamine and 10% Gibco™ Fetal Bovine Serum (Sigma-Aldrich, Missouri, US, RRID:SCR_013728) and supplemented with penicillin (100 U/ml)/streptomycin (100 mg/ml; Sigma-Aldrich, Missouri, US, RRID:SCR_013728) and Hepes (Sigma-Aldrich, Missouri, US, RRID:SCR_013728) overnight. Subsequently, PBMCs were stimulated with rh-IL-2 (BioLegend, California, US, RRID:SCR_001134), with four different doses (0, 50, 150, 250 U/ml) and incubated for 72 hours at 37°C with 5% CO₂.

Flow cytometry analysis

Following incubation with four different doses of IL-2, cells were collected, washed, and rehydrated for coloration. PBMCs obtained from the incubation were collected and distributed into four separate Eppendorf tubes. Subsequently, they were subjected to centrifugation at 4000 rpm for a duration of 3 minutes at a temperature of 22°C. The pellets were then washed using cell staining buffer (CSB) and centrifuged once again at 4000 rpm for 3 minutes at 22°C. Cells were colored at 4°C for 30 minutes with anti-CD3 antibody (Biolegend, SanDiego, CA, RRID:SCR_001134) conjugated with Peridinin-Chlorophyll-Protein (PerCP) (Biolegend, SanDiego, CA, RRID:SCR_001134) and anti-CD56 antibody (Biolegend, SanDiego, CA, RRID:SCR_001134) conjugated with Fluorescein Isothiocyanate (FITC) (BioLegend, San Diego, CA, RRID:SCR_001134). Cells were washed twice and analyzed using Fluorescence-activated Cell Sorting (FACS) Melody (BD Bioscience, Haryana, India, RRID:SCR_018091). The cell number was analyzed by the BD Cell Quest software (BD Biosciences, San Jose, CA, US, RRID:SCR_014489). The analysis yielded a percentage (%) of the cells. The lymphocyte population was targeted to identify the CD3⁺ and CD3^- lymphocyte populations. The CD3-negative lymphocyte populations were then further analyzed to determine the CD56 expression patterns. By using the FACSMelody (BD Bioscience, Haryana, India, RRID:SCR_018091), the lymphocyte population was analyzed and separated into CD3-positive and CD3-negative subgroups. Following that, only the CD3-negative lymphocyte population was selected to evaluate the expression levels of CD56. The results were expressed as percentages of isolated NK cells (CD3^-CD56⁺). The isolated NK cells were further sorted as CD56^dim and CD56^bright (see the Underlying data).¹³

Enzyme-linked immunosorbent assay (ELISA) for cytokines

Supernatants from IL-2 stimulated by PBMC incubation were collected and stored at -80°C to determine the levels of cytokines. The secretion of IFN-γ (BioLegend, San Diego, CA, RRID:SCR_001134) was measured with the ELISA kits according to the instructions (Legend Max™ Human IFN-γ ELISA Kit, Catalog no: 430107, BioLegend, San Diego, CA, RRID:SCR_001134). Measurement of IFN-γ levels was conducted on all culture plates, taken from the culture supernatant. The cell culture results were centrifuged first, and the supernatant present at the top was isolated, followed by measurement using ELISA. A 500 μL volume of the 1,000 pg/mL top standard was prepared by diluting 25 μL of the standard stock solution in 475 μL of Assay Buffer A. Six two-fold serial dilutions of the 1,000 pg/mL top standard were conducted in separate tubes, using Assay Buffer A as the diluent. This resulted in the following concentrations of IFN-γ in the tubes: 1,000 pg/mL, 500 pg/mL, 250 pg/mL, 125 pg/mL, 62.5 pg/mL, 31.3 pg/mL, and 15.6 pg/mL. Assay Buffer A was utilized as the zero standard (0 pg/mL). To prepare the plate, it was washed four times with a minimum of 300 μL of 1X Wash Buffer per well. Any residual buffer was removed by tapping the plate firmly upside down on absorbent paper. Subsequent washes were carried out in the same manner. A 50 μL volume of Assay Buffer A was added to each well designated for standard dilutions and samples. Similarly, a 50 μL volume of the appropriate standard dilutions or samples was added to their respective wells. The plate was sealed using the provided Plate Sealer and incubated at room temperature for 2 hours while shaking at 200 rpm. After the incubation, the contents of the plate were discarded into a sink, and the plate was washed four times with 1× Wash Buffer. A 100 μL volume of Human IFN-γ Detection Antibody solution was added to each well, the plate was sealed, and incubated at room temperature for 1 hour with shaking. The contents were then discarded into a sink, and the plate was washed four times with 1× Wash Buffer. Subsequently, a 100 μL volume of Avidin - Horseradish Peroxidase (HRP) A solution was added to each well, the plate was sealed, and incubated at room temperature for 30 minutes with shaking. The contents were discarded into a sink, and the plate was washed five times with 1× Wash Buffer. During the final wash, the wells were soaked in 1× Wash Buffer for 30 seconds per wash to minimize background. Furthermore, a 100 μL volume of Substrate Solution F was added to each well and incubated for 15 minutes in the dark. Wells containing human IFN-γ exhibited a blue color, with intensity correlating to their concentration. To stop the reaction, a 100 μL volume of Stop Solution was added to each well. The color of the solution changed from blue to yellow. Within 30 minutes, the absorbance was measured at 450 nm.

Statistical analysis

The difference of CD56^bright NK cells, CD56^dim NK cells, and IFN-γ levels between groups was analyzed using ANOVA, and the post hoc Tukey was used to analyze the comparison between groups and to determine effect estimate. A p-value less than 0.05 was considered significant impact. The effect estimate was presented using mean difference (MD) and 95% confidence interval (95%CI). We used Statistical Product and Service Solution 18 (SPSS 18, SPSS inc. Chicago, IL, USA, RRID:SCR_002865) to analyze the data. A scatter plot was used to describe the comparison of CD56^bright NK cells, CD56^dim NK cells, and IFN-γ between groups.

Results

CD56^bright NK cells

The levels of CD56^bright NK cells after the administration of different dose of IL-2 such as D0, D1, D2, and D3 were as follows: 57.27 ± 37.27, 241.16 ± 64.41, 256.94 ± 50.95, and 259.37 ± 36.44 ×1000 cells/mm³ respectively (Table 1). We found that the levels of CD56^bright NK cells were higher in D1, D2, and D3 compared to D0 and the MDs were 183.89, 199.67, and 202.10 x1000 cells/mm³; respectively (Figure 1).

Table 1. The levels of CD3-CD56^bright NK cells, CD3-CD56^dim NK cells, and IFN-γ after the administration of IL-2.

Parameters	D0	D1	D2	D3	p
CD3-CD56^bright (×1000 cells/mm³)	57.27 ± 37.27	241.16 ± 64.41	256.94 ± 50.95	259.37 ± 36.44	<0.0001
CD3-CD56^dim (×1000 cells/mm³)	812.85 ± 167.37	631.98 ± 129.90	616.42 ± 157.97	615.90 ± 155.57	0.1030
IFN-γ (pg/ml)	24.01 ± 2.56	26.09 ± 4.79	30.11 ± 5.34	32.43 ± 7.14	0.0560

Figure 1. The summary of comparison between the different dose of IL-2 administration and the levels of NK CD3-CD56^bright.

(A) D1 vs. D0 [MD: 183.89; CI: 105.29, 262.48; p<0.0001]. (B) D2 vs. D0 [MD: 199.67; CI: 121.08, 278.26; p<0.0001]. (C) D3 vs. D0 [MD: 202.10; CI: 123.51, 280.70; p<0.0001].

CD56^dim NK cells

After the administration of IL-2, the levels of CD56^dim NK cells in D0, D1, D2, and D3 groups were 812.85 ± 167.37, 631.98 ± 129.90, 616.42 ± 157.97, and 615.90 ± 155.57 ×1000 cells/mm³ respectively. We were unable to determine the impact of IL-2 administrations on the levels of CD56^dim NK cells when comparing D1, D2, or D3 to D0 (Table 1).

IFN-γ levels

The levels of IFN-γ in groups D0, D1, D2, and D3 were 24.01 ± 2.56, 26.09 ± 4.79, 30.11 ± 5.34, and 32.43 ± 7.14 pg/ml respectively. In our analysis, we found that the administration of IL-2 had no significant impact on the levels of IFN-γ (Table 1).

Discussion

Our current study found that the administration of IL-2 had potential impact to the levels of CD56^bright NK cells, but not to the levels of CD56^dim NK cells and IFN-γ. Compared to the control group, we elucidated that the higher the dose of IL-2 administration the higher the impact to the levels of CD56^bright. Our present study was the first report on the role of IL-2 on the regulation of CD56^bright in the case of SLE. However, our current findings were consistent to what has been reported by previous studies in other disease settings and healthy individuals. In the case of chronic GVHD, Hirakawa et al. conducted a study to assess the administration of IL-2 and its impact on the expression of CD56^bright NK cells. They found that a low dose of IL-2 was sufficiently effective to affect the expression of CD56^bright NK cells.¹⁰ Additionally, similar findings on the impact of IL-2 administration to the regulation of CD56^bright NK cells in the case of chronic GVHD were also reported by Kubo et al.¹⁶ On the other hand, in the case of healthy individuals, the induction of CD56^bright NK cells regulation after the administration of IL-2 has also been reported by McQuaid et al.¹⁷ Similar findings in the case of healthy individuals has also been reported by Saito et al.¹⁸ Moreover, in the case of acute myeloid leukemia, the treatment by using the combination of histamine dihydrochloride and IL-2 provided the reconstitution of a deficient NK cell fraction through the specific amplification of CD56^bright NK cells.¹¹ Thus, the results of our study might add insight into the role of IL-2 in the regulation of CD56^bright in various diseases.

The theoretical basis for our findings regarding the impact of IL-2 on the regulation of CD56^bright in SLE cases remains inadequately explained. However, several possible mechanisms might help to elucidate the potential role of IL-2 on CD56^bright. IL-2 is a cytokine required for the activation and proliferation of immune cells, including NK cells, and NK cells have the receptors to interact with IL-2 subunits, i.e: IL-2Rα, IL-2Rβ, and IL-2Rγ.¹⁹ To establish the interaction, the high affinity of T cells and activated NK cells was needed, and the affinity of T cells is expressed by a small T cell subunit and CD56^bright NK cells. IL-2 received by the receptor complex in the NK cells may activate Janus tyrosine-kinase (JAK)-1 and JAK-3 which is required for the signal transduction. Furthermore, the activation of JAK-1 and JAK-3 may initiate the phosphorylation of signal transducer and activator of transcription (STAT)-5, the control center and regulation of specific gene transcription.²⁰ On the other hand, IL-2 signal may also induce phosphorylation from the adapter protein Shc which will activate ras-raf map kinase and PI3K signals,²¹ and subsequently, this circumstance may cause the transmission of IL-2 signals from the membrane to the core.²²^,²³ In addition, the complex interaction between IL-2 and IL-2Rα/β/γ may induce the phosphorylation of STAT-5 and increase CD25 and Foxp3 expression in Treg, and as a result, this condition may activate its suppression activity.²⁴ In SLE patients, it has been known that dysfunction of NK cells may develop,²⁵ and has been associated with an impaired release of a soluble cytotoxic factor.²⁶ Therefore, the administration of IL-2 may help to alleviate the number and function of Tregs cells, and as expected, this condition may control autoreactive cells by modulating the activation of NK cells.²⁷ IL-2 may increase NK cell reactivity, while Tregs cells can limit the intrinsic production of IL-2 from T cells, and thereafter, they may regulate the cytotoxicity of NK cells. This mechanism was likely to underlie our findings showing that the administration of IL-2 was associated with upregulation of CD56^bright.

To the best of our knowledge, our current investigation is the first study reporting the role of IL-2 on CD56^bright NK cells, CD56^dim NK cells, and IFN-γ in the case of SLE; and we revealed that only CD56^bright NK cells were affected by the administration of IL-2. Our current finding might add new insight on the impact of IL-2 administration to CD56^bright NK cells. Moreover, the findings of our present study might serve as the initial reference for further studies in the context of the implication of IL-2 administration to the expression of NK cells in the case of SLE. Based on several previous studies suggesting that a therapeutic option by targeting NK cells might provide benefits for SLE patients,²⁸^,²⁹ we expected that our present findings could serve as a foundation for future studies, potentially leading to the establishment of SLE treatment strategies that target NK cells.

Our study had several important limitations. First, in our present study, some potential factors involved in the pathogenesis of SLE that might affect the final findings were not analyzed, for example: human leukocyte antigen, STAT-5, JAK-1, and JAK-3.²⁰ Second, the design of our study was a post-test only control group design. A study by applying a pre-test post-test control group design might be required to obtain better evidence. Third, we only assessed IL-2 as the potential cytokine in our study. Further studies involving other cytokines might be required to obtain more comprehensive findings.

Conclusion

Our study reveals that the administration of IL-2 provides the beneficial impact in the expression of CD56^bright NK cells in the case of SLE. Our current study may establish the additional insight on the role of IL-2 in the treatment of SLE by targeting NK cells.

Authors contributions

Conceptualization: DS; Data Curation: DS; Formal Analysis: DS, JKF; Investigation: DS; Project Administration: DS; Resources: DS; Methodology: DS, JKF; Software: JKF; Visualization: JKF; Supervision: HS, HK, KH; Validation: HS, HK, KH; Writing – Original Draft Preparation: DS, JKF; Writing – Review & Editing: JKF. All authors have critically reviewed and approved the final draft and are responsible for the content and similarity index of the manuscript.

Data availability

Underlying data

Figshare: Supplementary files: The implication of interleukin – 2 on the expression of CD56bright, CD56dim, and interferon – γ in patients with systemic lupus erythematosus, https://doi.org/10.6084/m9.figshare.21821670 .¹³

This project contains the following underlying data:

• Ethical Clearance_NK_2019.jpeg (The ethical clearance of our study)
• Data set.xlsx (The levels of IFN-γ and the expression of CD56^bright and CD56^dim in each study group)
• FIGURE FLOWCITOMETRY.jpg (Flowcitometry of each study group)
• Supplementary files.docx (Summary of the supplementary files, including the levels of IFN-γ and the expression of CD56^bright and CD56^dim in each study group and the plots of their mean)

Data are available under the terms of the Creative Commons Attribution 4.0 International license (CC-BY 4.0).

Acknowledgement

We would like to thank to all patients who had participated in this study. We also would like to thank for the gratitude to dr. Rifa’i, dr. Putra, dr. Singgih, dr. Ramadi, Mr. Yudha, Ns. Elvira, & Mrs. Lilis for their support and technical assistance in hospital and laboratory.

References

1. Barber MRW, Drenkard C, Falasinnu T, et al.: Global epidemiology of systemic lupus erythematosus. Nat. Rev. Rheumatol. 2021; 17: 515–532. PubMed Abstract | Publisher Full Text | Free Full Text
2. Levy DM, Peschken CA, Tucker LB, et al.: Influence of ethnicity on childhood-onset systemic lupus erythematosus: results from a multiethnic multicenter Canadian cohort. Arthritis Care Res. 2013; 65: 152–160. PubMed Abstract | Publisher Full Text | Free Full Text
3. Yildirim-Toruner C, Diamond B: Current and novel therapeutics in the treatment of systemic lupus erythematosus. J. Allergy Clin. Immunol. 2011; 127: 303–312; quiz 13-4. PubMed Abstract | Publisher Full Text | Free Full Text
4. Moulton VR, Suarez-Fueyo A, Meidan E, et al.: Pathogenesis of Human Systemic Lupus Erythematosus: A Cellular Perspective. Trends Mol. Med. 2017; 23: 615–635. PubMed Abstract | Publisher Full Text | Free Full Text
5. Caligiuri MA: Human natural killer cells. Blood. 2008; 112: 461–469. PubMed Abstract | Publisher Full Text | Free Full Text
6. Schepis D, Gunnarsson I, Eloranta ML, et al.: Increased proportion of CD56bright natural killer cells in active and inactive systemic lupus erythematosus. Immunology. 2009; 126: 140–146. PubMed Abstract | Publisher Full Text | Free Full Text
7. Liu M, Liu J, Zhang X, et al.: Activation status of CD56(dim) natural killer cells is associated with disease activity of patients with systemic lupus erythematosus. Clin. Rheumatol. 2021; 40: 1103–1112. PubMed Abstract | Publisher Full Text
8. Fogel LA, Yokoyama WM, French AR: Natural killer cells in human autoimmune disorders. Arthritis Res. Ther. 2013; 15: 216. PubMed Abstract | Publisher Full Text | Free Full Text
9. Jiang T, Zhou C, Ren S: Role of IL-2 in cancer immunotherapy. Oncoimmunology. 2016; 5: e1163462. PubMed Abstract | Publisher Full Text | Free Full Text
10. Hirakawa M, Matos TR, Liu H, et al.: Low-dose IL-2 selectively activates subsets of CD4(+) Tregs and NK cells. JCI Insight. 2016; 1: e89278. Publisher Full Text
11. Cuapio A, Post M, Cerny-Reiterer S, et al.: Maintenance therapy with histamine plus IL-2 induces a striking expansion of two CD56bright NK cell subpopulations in patients with acute myeloid leukemia and supports their activation. Oncotarget. 2016; 7: 46466–46481. PubMed Abstract | Publisher Full Text | Free Full Text
12. Lieberman LA, Tsokos GC: The IL-2 defect in systemic lupus erythematosus disease has an expansive effect on host immunity. J. Biomed. Biotechnol. 2010; 2010: 740619.
13. Fajar JK: Supplementary files: The implication of interleukin – 2 on the expression of CD56bright, CD56dim, and interferon – γ in patients with systemic lupus erythematosus. Fighshare. 2023; 1: 1.
14. World Medical A: World Medical Association Declaration of Helsinki: ethical principles for medical research involving human subjects. JAMA. 2013; 310: 2191–2194. Publisher Full Text
15. Aringer M, Costenbader K, Daikh D, et al.: 2019 European League Against Rheumatism/American College of Rheumatology Classification Criteria for Systemic Lupus Erythematosus. Arthritis Rheumatol. 2019; 71: 1400–1412. PubMed Abstract | Publisher Full Text | Free Full Text
16. Kubo T, Romee R, Koreth J, et al.: Low-dose interleukin-2 therapy enhances cytotoxicity of CD56bright NK cells in patients with chronic GvHD. Blood. 2018; 132: 606. Publisher Full Text
17. McQuaid SL, Loughran ST, Power PA, et al.: Low-dose IL-2 induces CD56(bright) NK regulation of T cells via NKp44 and NKp46. Clin. Exp. Immunol. 2020; 200: 228–241. PubMed Abstract | Publisher Full Text | Free Full Text
18. Saito S, Morii T, Enomoto M, et al.: The effect of interleukin 2 and transforming growth factor-beta 2 (TGF-beta 2) on the proliferation and natural killer activity of decidual CD16- CD56bright natural killer cells. Cell. Immunol. 1993; 152: 605–613. PubMed Abstract | Publisher Full Text
19. Malek TR, Castro I: Interleukin-2 receptor signaling: at the interface between tolerance and immunity. Immunity. 2010; 33: 153–165. PubMed Abstract | Publisher Full Text | Free Full Text
20. Carson WE, Fehniger TA, Caligiuri MA: CD56bright natural killer cell subsets: characterization of distinct functional responses to interleukin-2 and the c-kit ligand. Eur. J. Immunol. 1997; 27: 354–360. PubMed Abstract | Publisher Full Text
21. Ravichandran KS, Burakoff SJ: The adapter protein Shc interacts with the interleukin-2 (IL-2) receptor upon IL-2 stimulation. J. Biol. Chem. 1994; 269: 1599–1602. PubMed Abstract | Publisher Full Text
22. Meazza R, Azzarone B, Orengo AM, et al.: Role of common-gamma chain cytokines in NK cell development and function: perspectives for immunotherapy. J. Biomed. Biotechnol. 2011; 2011: 861920.
23. Ye C, Brand D, Zheng SG: Targeting IL-2: an unexpected effect in treating immunological diseases. Signal Transduct. Target. Ther. 2018; 3: 2. PubMed Abstract | Publisher Full Text | Free Full Text
24. Zhou X, Xing J, Tang X, et al.: Interleukin-2 (IL-2) Interacts With IL-2 Receptor Beta (IL-2Rbeta): Its Potential to Enhance the Proliferation of CD4+ T Lymphocytes in Flounder (Paralichthys olivaceus). Front. Immunol. 2020; 11: 531785. PubMed Abstract | Publisher Full Text | Free Full Text
25. Henriques A, Teixeira L, Ines L, et al.: NK cells dysfunction in systemic lupus erythematosus: relation to disease activity. Clin. Rheumatol. 2013; 32: 805–813. PubMed Abstract | Publisher Full Text
26. Sibbitt WL Jr, Mathews PM, Bankhurst AD: Natural killer cell in systemic lupus erythematosus. Defects in effector lytic activity and response to interferon and interferon inducers. J. Clin. Invest. 1983; 71: 1230–1239. PubMed Abstract | Publisher Full Text | Free Full Text
27. Gasteiger G, Hemmers S, Firth MA, et al.: IL-2-dependent tuning of NK cell sensitivity for target cells is controlled by regulatory T cells. J. Exp. Med. 2013; 210: 1167–1178. PubMed Abstract | Publisher Full Text | Free Full Text
28. Green MR, Kennell AS, Larche MJ, et al.: Natural killer cell activity in families of patients with systemic lupus erythematosus: demonstration of a killing defect in patients. Clin. Exp. Immunol. 2005; 141: 165–173. PubMed Abstract | Publisher Full Text | Free Full Text
29. Green MR, Kennell AS, Larche MJ, et al.: Natural killer T cells in families of patients with systemic lupus erythematosus: their possible role in regulation of IGG production. Arthritis Rheum. 2007; 56: 303–310. PubMed Abstract | Publisher Full Text

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VERSION 1 PUBLISHED 28 Nov 2023

Author details Author details

¹ Doctoral Program, Faculty of Medicine, Universitas Brawijaya, Malang, 65145, Indonesia
² Department of Nursing, STIKES Widyagama Husada, Malang, 65145, Indonesia
³ Department of Clinical Pathology, Faculty of Medicine, Universitas Brawijaya, Malang, 65145, Indonesia
⁴ Division of Reumatology & Immunology, Department of Internal Medicine, Faculty of Medicine, Universitas Brawijaya, Malang, 65145, Indonesia
⁵ Brawijaya Internal Medicine Research Center, Department of Internal Medicine, Faculty of Medicine, Universitas Brawijaya, Malang, 65145, Indonesia

Dwi Soelistyoningsih
Roles: Conceptualization, Data Curation, Formal Analysis, Investigation, Methodology, Project Administration, Resources, Writing – Original Draft Preparation

Hani Susianti
Roles: Supervision, Validation

Handono Kalim
Roles: Supervision, Validation

Kusworini Handono
Roles: Supervision, Validation

Jonny Karunia Fajar
Roles: Formal Analysis, Methodology, Software, Visualization, Writing – Original Draft Preparation, Writing – Review & Editing

Competing interests

No competing interests were disclosed.

Grant information

The author(s) declared that no grants were involved in supporting this work.

Article Versions (1)

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Published: 28 Nov 2023, 12:1525

https://doi.org/10.12688/f1000research.130553.1

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© 2023 Soelistyoningsih D et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Soelistyoningsih D, Susianti H, Kalim H et al. The implication of interleukin-2 on the expression of CD56^bright, CD56^dim, and interferon-γ in patients with systemic lupus erythematosus [version 1; peer review: 2 approved with reservations, 1 not approved]. F1000Research 2023, 12:1525 (https://doi.org/10.12688/f1000research.130553.1)

NOTE: If applicable, it is important to ensure the information in square brackets after the title is included in all citations of this article.

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ApprovedThe paper is scientifically sound in its current form and only minor, if any, improvements are suggested

Approved with reservations A number of small changes, sometimes more significant revisions are required to address specific details and improve the papers academic merit.

Not approvedFundamental flaws in the paper seriously undermine the findings and conclusions

Version 1

VERSION 1

PUBLISHED 28 Nov 2023

Views

3

Reviewer Report 25 May 2024

Guo-Min Deng, Wuhan Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China

Not Approved

https://doi.org/10.5256/f1000research.143321.r248096

This manuscript displays the implication of IL-2 on the expression of CD56bright, CD56dim, and interferon-gamma in patients with SLE. There are some concerns in this manuscript.
1, In the abstract, author wrote that the role of IL-2 had never ... Continue reading

This manuscript displays the implication of IL-2 on the expression of CD56bright, CD56dim, and interferon-gamma in patients with SLE. There are some concerns in this manuscript.
1, In the abstract, author wrote that the role of IL-2 had never been investigated in the the case of SLE, this is not appropriate. Actually, there are some papers published that displayed effect of low dose of IL-2 treatment in SLE.
2. In study of design and sample preparation in methods, during January -July 2020, lymphocytes were collected from six female SLE, here lymphocytes should be PUMC but not lymphocytes.
3. Figure 1 shows the summary of comparison between the different doses of IL-2 administration. The results of flow cytometry should display dot picture and let reviewer to evaluate the results.
4. Table 1 shows that IL-2 significantly enhanced expression of CD56 bright, but did not significantly enhanced expression of IFN-gamma. These results are against each other because author in introduction of manuscript described that CD56bright had been proven to have the ability to initiate the production of IFN-γ(ref 8) .
5. Writing should be improved.

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Partly
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

Not applicable
Are all the source data underlying the results available to ensure full reproducibility?

Partly
Are the conclusions drawn adequately supported by the results?

Partly

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Pathogenesis and treatment of rheumatoid arthritis and systemic lupus erythematosus

I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above.

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4

Reviewer Report 17 May 2024

Stevent Sumantri, Department of Internal Medicine, Pelita Harapan University, Tangerang, Banten, Indonesia

Approved with Reservations

https://doi.org/10.5256/f1000research.143321.r266850

Dwi Soelistyoningsih et al. discuss the implications of interleukin-2 (IL-2) on the expression of CD56 bright, CD56 dim, and interferon-γ in patients with systemic lupus erythematosus (SLE), highlighting the potential beneficial impact of IL-2 administration on CD56 bright NK cells ... Continue reading

Dwi Soelistyoningsih et al. discuss the implications of interleukin-2 (IL-2) on the expression of CD56 bright, CD56 dim, and interferon-γ in patients with systemic lupus erythematosus (SLE), highlighting the potential beneficial impact of IL-2 administration on CD56 bright NK cells in SLE.

The research has been carried out using scientific methods in accordance with appropriate immunological investigations. The results show that CD56 bright NK cells in the peripheral blood of lupus patients can be successfully increased by interleukin-2 therapy, which is a new and very interesting finding. The results of this study are expected to provide input for potential therapy in lupus patients, especially those whose pathogenesis is related to NK cell imbalance and/or dysfunction.

The authors have described some of the study's shortcomings, including the fact that other immuno pathogenesis factors were not analyzed in addition to NK cell levels of CD56 bright, CD56 dim, and interferon-gamma. The authors have also pointed out another shortcoming: there was no pre-treatment evaluation before treatment with interleukin-2, which we believe is one of the major shortcomings of this study.

The researchers have briefly described the characteristics of the research subjects, but given that there are only six subjects, we suggest that a description of each subject's organ involvement could also be further explained.
In addition, we suggest updating the citations and discussing the relationship between lupus and NK cells with some recent literature. We attach the citations below.
We also suggest that the authors reread and improve the quality of the English writing.

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Yes
Are sufficient details of methods and analysis provided to allow replication by others?

Yes
If applicable, is the statistical analysis and its interpretation appropriate?

I cannot comment. A qualified statistician is required.
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

Yes

References

1. Thangjam N, Dey B, Khonglah Y, Tiewsoh I, et al.: Natural Killer Cell Count in Systemic Lupus Erythematosus Patients: A Flow Cytometry-Based Study.Cureus. 2023; 15 (10): e46885 PubMed Abstract | Publisher Full Text
2. Chen N, Dai Y, Li H, Long X, et al.: Increased ILT2+ natural killer T cells correlate with disease activity in systemic lupus erythematosus.Clin Rheumatol. 2023; 42 (11): 3113-3121 PubMed Abstract | Publisher Full Text
3. Soelistyoningsih D, Susianti H, Kalim H, Handono K: The phenotype of CD3-CD56bright and CD3-CD56dim natural killer cells in systemic lupus erythematosus patients and its relation to disease activity.Reumatologia. 2022; 60 (4): 258-265 PubMed Abstract | Publisher Full Text

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Autoimmune Disease, Systemic Lupus Erythematosus, Rheumatoid Arthritis, Clinical Immunology

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.

CITE

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6

Reviewer Report 24 Apr 2024

Yong-Fei Wang, The Chinese University of Hong Kong, Shenzhen, Guangdong, China

Jinglu Zhao, The Chinese University of Hong Kong, Shenzhen, Guangdong, China

Approved with Reservations

https://doi.org/10.5256/f1000research.143321.r258820

The paper presents a novel exploration of the effects of interleukin-2 (IL-2) administration on the expression of CD56 bright, CD56 dim, and interferon-γ in systemic lupus erythematosus (SLE) patients. It is a topic of interest to researchers in immunology, rheumatology, ... Continue reading

The paper presents a novel exploration of the effects of interleukin-2 (IL-2) administration on the expression of CD56 bright, CD56 dim, and interferon-γ in systemic lupus erythematosus (SLE) patients. It is a topic of interest to researchers in immunology, rheumatology, and related areas, but the paper needs very significant improvement. My major comments are as follows:

The abstract's claim that IL-2's role in case of SLE has not been investigated contradicts recent literature documenting the significance and function of IL-2 in SLE([1],[2],[3].). Thus, it is essential to integrate a comprehensive review of this related literature to emphasize the study's novelty and significance.
While some characteristics of the study participants are provided, presenting this information in a table would offer clearer insights. It is recommended to include a comprehensive table detailing the characteristics of all clinical samples, encompassing sex distribution, age range, MEX-SLEDAI scores, presence of SLE-related antibodies (e.g., anti-dsDNA), average disease duration, percentage of patients under regular corticosteroid treatment, and cell population data.
The small sample size of six patients and the lack of a healthy control group considerably limits the findings' generalizability. To improve the study's robustness and credibility, it is recommended to increase the sample size and incorporate a diverse patient cohort as well as healthy controls.
Although the study includes a control group, a more detailed explanation regarding the selection criteria for IL-2 dosages (D1, D2, D3) is essential for transparency and to substantiate the study's validity.
Expanding on methodological details, including IL-2 administration procedures and sample handling and analysis, will improve reproducibility.
The graphical results are poorly displayed, and it is difficult to correlate the figures with the text. Providing compensated plots with detailed gating strategies in supplementary materials, especially for flow cytometry figures, is advised.
The results indicate a significant impact of IL-2 on the levels of CD56bright but not on CD56dim and IFN-γ. Including a discussion on why IL-2 stimulation did not affect CD56dim and IFN-γ levels as expected would be insightful. Are there other pathways or regulatory mechanisms specific to SLE patients that could explain these findings?

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Partly
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

Yes
Are all the source data underlying the results available to ensure full reproducibility?

Partly
Are the conclusions drawn adequately supported by the results?

Partly

References

1. He J, Zhang R, Shao M, Zhao X, et al.: Efficacy and safety of low-dose IL-2 in the treatment of systemic lupus erythematosus: a randomised, double-blind, placebo-controlled trial.Ann Rheum Dis. 2020; 79 (1): 141-149 PubMed Abstract | Publisher Full Text
2. Lieberman LA, Tsokos GC: The IL-2 defect in systemic lupus erythematosus disease has an expansive effect on host immunity.J Biomed Biotechnol. 2010; 2010: 740619 PubMed Abstract | Publisher Full Text
3. Mak A: T Cells, Interleukin-2 and Systemic Lupus Erythematosus-From Pathophysiology to Therapy.Cells. 2022; 11 (6). PubMed Abstract | Publisher Full Text

Competing Interests: No competing interests were disclosed.

Reviewer Expertise: Immunogenetics

We confirm that we have read this submission and believe that we have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however we have significant reservations, as outlined above.

CITE

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VERSION 1 PUBLISHED 28 Nov 2023

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Version 1 28 Nov 23	read	read	read

Yong-Fei Wang, The Chinese University of Hong Kong, Shenzhen, Guangdong, China

Jinglu Zhao, The Chinese University of Hong Kong, Shenzhen, China
Stevent Sumantri, Pelita Harapan University, Tangerang, Indonesia
Guo-Min Deng, Huazhong University of Science and Technology, Wuhan, China

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Reviewer Report

3 Views

25 May 2024 | for Version 1

Guo-Min Deng, Wuhan Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China

3 Views Cite this report Responses(0)

Not Approved

This manuscript displays the implication of IL-2 on the expression of CD56bright, CD56dim, and interferon-gamma in patients with SLE. There are some concerns in this manuscript.
1, In the abstract, author wrote that the role of IL-2 had never been investigated in the the case of SLE, this is not appropriate. Actually, there are some papers published that displayed effect of low dose of IL-2 treatment in SLE.
2. In study of design and sample preparation in methods, during January -July 2020, lymphocytes were collected from six female SLE, here lymphocytes should be PUMC but not lymphocytes.
3. Figure 1 shows the summary of comparison between the different doses of IL-2 administration. The results of flow cytometry should display dot picture and let reviewer to evaluate the results.
4. Table 1 shows that IL-2 significantly enhanced expression of CD56 bright, but did not significantly enhanced expression of IFN-gamma. These results are against each other because author in introduction of manuscript described that CD56bright had been proven to have the ability to initiate the production of IFN-γ(ref 8) .
5. Writing should be improved.

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Partly
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

Not applicable
Are all the source data underlying the results available to ensure full reproducibility?

Partly
Are the conclusions drawn adequately supported by the results?

Partly

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Pathogenesis and treatment of rheumatoid arthritis and systemic lupus erythematosus

I confirm that I have read this submission and believe that I have an appropriate level of expertise to state that I do not consider it to be of an acceptable scientific standard, for reasons outlined above.

Respond to this report

Responses (0)

Back to all reports

Reviewer Report

4 Views

17 May 2024 | for Version 1

Stevent Sumantri, Department of Internal Medicine, Pelita Harapan University, Tangerang, Banten, Indonesia

4 Views Cite this report Responses(0)

Approved With Reservations

Dwi Soelistyoningsih et al. discuss the implications of interleukin-2 (IL-2) on the expression of CD56 bright, CD56 dim, and interferon-γ in patients with systemic lupus erythematosus (SLE), highlighting the potential beneficial impact of IL-2 administration on CD56 bright NK cells in SLE.

The research has been carried out using scientific methods in accordance with appropriate immunological investigations. The results show that CD56 bright NK cells in the peripheral blood of lupus patients can be successfully increased by interleukin-2 therapy, which is a new and very interesting finding. The results of this study are expected to provide input for potential therapy in lupus patients, especially those whose pathogenesis is related to NK cell imbalance and/or dysfunction.

The authors have described some of the study's shortcomings, including the fact that other immuno pathogenesis factors were not analyzed in addition to NK cell levels of CD56 bright, CD56 dim, and interferon-gamma. The authors have also pointed out another shortcoming: there was no pre-treatment evaluation before treatment with interleukin-2, which we believe is one of the major shortcomings of this study.

The researchers have briefly described the characteristics of the research subjects, but given that there are only six subjects, we suggest that a description of each subject's organ involvement could also be further explained.
In addition, we suggest updating the citations and discussing the relationship between lupus and NK cells with some recent literature. We attach the citations below.
We also suggest that the authors reread and improve the quality of the English writing.

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Yes
Are sufficient details of methods and analysis provided to allow replication by others?

Yes
If applicable, is the statistical analysis and its interpretation appropriate?

I cannot comment. A qualified statistician is required.
Are all the source data underlying the results available to ensure full reproducibility?

Yes
Are the conclusions drawn adequately supported by the results?

Yes

References

1. Thangjam N, Dey B, Khonglah Y, Tiewsoh I, et al.: Natural Killer Cell Count in Systemic Lupus Erythematosus Patients: A Flow Cytometry-Based Study.Cureus. 2023; 15 (10): e46885 PubMed Abstract | Publisher Full Text
2. Chen N, Dai Y, Li H, Long X, et al.: Increased ILT2+ natural killer T cells correlate with disease activity in systemic lupus erythematosus.Clin Rheumatol. 2023; 42 (11): 3113-3121 PubMed Abstract | Publisher Full Text
3. Soelistyoningsih D, Susianti H, Kalim H, Handono K: The phenotype of CD3-CD56bright and CD3-CD56dim natural killer cells in systemic lupus erythematosus patients and its relation to disease activity.Reumatologia. 2022; 60 (4): 258-265 PubMed Abstract | Publisher Full Text

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Autoimmune Disease, Systemic Lupus Erythematosus, Rheumatoid Arthritis, Clinical Immunology

I confirm that I have read this submission and believe that I have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however I have significant reservations, as outlined above.

Respond to this report

Responses (0)

Back to all reports

Reviewer Report

6 Views

24 Apr 2024 | for Version 1

Yong-Fei Wang, The Chinese University of Hong Kong, Shenzhen, Guangdong, China

Jinglu Zhao, The Chinese University of Hong Kong, Shenzhen, Guangdong, China

6 Views Cite this report Responses(0)

Approved With Reservations

The paper presents a novel exploration of the effects of interleukin-2 (IL-2) administration on the expression of CD56 bright, CD56 dim, and interferon-γ in systemic lupus erythematosus (SLE) patients. It is a topic of interest to researchers in immunology, rheumatology, and related areas, but the paper needs very significant improvement. My major comments are as follows:

The abstract's claim that IL-2's role in case of SLE has not been investigated contradicts recent literature documenting the significance and function of IL-2 in SLE([1],[2],[3].). Thus, it is essential to integrate a comprehensive review of this related literature to emphasize the study's novelty and significance.
While some characteristics of the study participants are provided, presenting this information in a table would offer clearer insights. It is recommended to include a comprehensive table detailing the characteristics of all clinical samples, encompassing sex distribution, age range, MEX-SLEDAI scores, presence of SLE-related antibodies (e.g., anti-dsDNA), average disease duration, percentage of patients under regular corticosteroid treatment, and cell population data.
The small sample size of six patients and the lack of a healthy control group considerably limits the findings' generalizability. To improve the study's robustness and credibility, it is recommended to increase the sample size and incorporate a diverse patient cohort as well as healthy controls.
Although the study includes a control group, a more detailed explanation regarding the selection criteria for IL-2 dosages (D1, D2, D3) is essential for transparency and to substantiate the study's validity.
Expanding on methodological details, including IL-2 administration procedures and sample handling and analysis, will improve reproducibility.
The graphical results are poorly displayed, and it is difficult to correlate the figures with the text. Providing compensated plots with detailed gating strategies in supplementary materials, especially for flow cytometry figures, is advised.
The results indicate a significant impact of IL-2 on the levels of CD56bright but not on CD56dim and IFN-γ. Including a discussion on why IL-2 stimulation did not affect CD56dim and IFN-γ levels as expected would be insightful. Are there other pathways or regulatory mechanisms specific to SLE patients that could explain these findings?

Is the work clearly and accurately presented and does it cite the current literature?

Partly
Is the study design appropriate and is the work technically sound?

Partly
Are sufficient details of methods and analysis provided to allow replication by others?

Partly
If applicable, is the statistical analysis and its interpretation appropriate?

Yes
Are all the source data underlying the results available to ensure full reproducibility?

Partly
Are the conclusions drawn adequately supported by the results?

Partly

References

1. He J, Zhang R, Shao M, Zhao X, et al.: Efficacy and safety of low-dose IL-2 in the treatment of systemic lupus erythematosus: a randomised, double-blind, placebo-controlled trial.Ann Rheum Dis. 2020; 79 (1): 141-149 PubMed Abstract | Publisher Full Text
2. Lieberman LA, Tsokos GC: The IL-2 defect in systemic lupus erythematosus disease has an expansive effect on host immunity.J Biomed Biotechnol. 2010; 2010: 740619 PubMed Abstract | Publisher Full Text
3. Mak A: T Cells, Interleukin-2 and Systemic Lupus Erythematosus-From Pathophysiology to Therapy.Cells. 2022; 11 (6). PubMed Abstract | Publisher Full Text

Competing Interests

No competing interests were disclosed.

Reviewer Expertise

Immunogenetics

We confirm that we have read this submission and believe that we have an appropriate level of expertise to confirm that it is of an acceptable scientific standard, however we have significant reservations, as outlined above.

Respond to this report

Responses (0)

[1] 1. Barber MRW, Drenkard C, Falasinnu T, et al.: Global epidemiology of systemic lupus erythematosus. Nat. Rev. Rheumatol. 2021; 17: 515–532. PubMed Abstract | Publisher Full Text | Free Full Text

[2] 2. Levy DM, Peschken CA, Tucker LB, et al.: Influence of ethnicity on childhood-onset systemic lupus erythematosus: results from a multiethnic multicenter Canadian cohort. Arthritis Care Res. 2013; 65: 152–160. PubMed Abstract | Publisher Full Text | Free Full Text

[3] 3. Yildirim-Toruner C, Diamond B: Current and novel therapeutics in the treatment of systemic lupus erythematosus. J. Allergy Clin. Immunol. 2011; 127: 303–312; quiz 13-4. PubMed Abstract | Publisher Full Text | Free Full Text

[4] 4. Moulton VR, Suarez-Fueyo A, Meidan E, et al.: Pathogenesis of Human Systemic Lupus Erythematosus: A Cellular Perspective. Trends Mol. Med. 2017; 23: 615–635. PubMed Abstract | Publisher Full Text | Free Full Text

[5] 5. Caligiuri MA: Human natural killer cells. Blood. 2008; 112: 461–469. PubMed Abstract | Publisher Full Text | Free Full Text

[6] 6. Schepis D, Gunnarsson I, Eloranta ML, et al.: Increased proportion of CD56bright natural killer cells in active and inactive systemic lupus erythematosus. Immunology. 2009; 126: 140–146. PubMed Abstract | Publisher Full Text | Free Full Text

[7] 7. Liu M, Liu J, Zhang X, et al.: Activation status of CD56(dim) natural killer cells is associated with disease activity of patients with systemic lupus erythematosus. Clin. Rheumatol. 2021; 40: 1103–1112. PubMed Abstract | Publisher Full Text

[8] 8. Fogel LA, Yokoyama WM, French AR: Natural killer cells in human autoimmune disorders. Arthritis Res. Ther. 2013; 15: 216. PubMed Abstract | Publisher Full Text | Free Full Text

[9] 9. Jiang T, Zhou C, Ren S: Role of IL-2 in cancer immunotherapy. Oncoimmunology. 2016; 5: e1163462. PubMed Abstract | Publisher Full Text | Free Full Text

[10] 10. Hirakawa M, Matos TR, Liu H, et al.: Low-dose IL-2 selectively activates subsets of CD4(+) Tregs and NK cells. JCI Insight. 2016; 1: e89278. Publisher Full Text

[11] 11. Cuapio A, Post M, Cerny-Reiterer S, et al.: Maintenance therapy with histamine plus IL-2 induces a striking expansion of two CD56bright NK cell subpopulations in patients with acute myeloid leukemia and supports their activation. Oncotarget. 2016; 7: 46466–46481. PubMed Abstract | Publisher Full Text | Free Full Text

[12] 12. Lieberman LA, Tsokos GC: The IL-2 defect in systemic lupus erythematosus disease has an expansive effect on host immunity. J. Biomed. Biotechnol. 2010; 2010: 740619.

[13] 13. Fajar JK: Supplementary files: The implication of interleukin – 2 on the expression of CD56bright, CD56dim, and interferon – γ in patients with systemic lupus erythematosus. Fighshare. 2023; 1: 1.

[14] 14. World Medical A: World Medical Association Declaration of Helsinki: ethical principles for medical research involving human subjects. JAMA. 2013; 310: 2191–2194. Publisher Full Text

[15] 15. Aringer M, Costenbader K, Daikh D, et al.: 2019 European League Against Rheumatism/American College of Rheumatology Classification Criteria for Systemic Lupus Erythematosus. Arthritis Rheumatol. 2019; 71: 1400–1412. PubMed Abstract | Publisher Full Text | Free Full Text

[16] 16. Kubo T, Romee R, Koreth J, et al.: Low-dose interleukin-2 therapy enhances cytotoxicity of CD56bright NK cells in patients with chronic GvHD. Blood. 2018; 132: 606. Publisher Full Text

[17] 17. McQuaid SL, Loughran ST, Power PA, et al.: Low-dose IL-2 induces CD56(bright) NK regulation of T cells via NKp44 and NKp46. Clin. Exp. Immunol. 2020; 200: 228–241. PubMed Abstract | Publisher Full Text | Free Full Text

[18] 18. Saito S, Morii T, Enomoto M, et al.: The effect of interleukin 2 and transforming growth factor-beta 2 (TGF-beta 2) on the proliferation and natural killer activity of decidual CD16- CD56bright natural killer cells. Cell. Immunol. 1993; 152: 605–613. PubMed Abstract | Publisher Full Text

[19] 19. Malek TR, Castro I: Interleukin-2 receptor signaling: at the interface between tolerance and immunity. Immunity. 2010; 33: 153–165. PubMed Abstract | Publisher Full Text | Free Full Text

[20] 20. Carson WE, Fehniger TA, Caligiuri MA: CD56bright natural killer cell subsets: characterization of distinct functional responses to interleukin-2 and the c-kit ligand. Eur. J. Immunol. 1997; 27: 354–360. PubMed Abstract | Publisher Full Text

[21] 21. Ravichandran KS, Burakoff SJ: The adapter protein Shc interacts with the interleukin-2 (IL-2) receptor upon IL-2 stimulation. J. Biol. Chem. 1994; 269: 1599–1602. PubMed Abstract | Publisher Full Text

[22] 22. Meazza R, Azzarone B, Orengo AM, et al.: Role of common-gamma chain cytokines in NK cell development and function: perspectives for immunotherapy. J. Biomed. Biotechnol. 2011; 2011: 861920.

[23] 23. Ye C, Brand D, Zheng SG: Targeting IL-2: an unexpected effect in treating immunological diseases. Signal Transduct. Target. Ther. 2018; 3: 2. PubMed Abstract | Publisher Full Text | Free Full Text

[24] 24. Zhou X, Xing J, Tang X, et al.: Interleukin-2 (IL-2) Interacts With IL-2 Receptor Beta (IL-2Rbeta): Its Potential to Enhance the Proliferation of CD4+ T Lymphocytes in Flounder (Paralichthys olivaceus). Front. Immunol. 2020; 11: 531785. PubMed Abstract | Publisher Full Text | Free Full Text

[25] 25. Henriques A, Teixeira L, Ines L, et al.: NK cells dysfunction in systemic lupus erythematosus: relation to disease activity. Clin. Rheumatol. 2013; 32: 805–813. PubMed Abstract | Publisher Full Text

[26] 26. Sibbitt WL Jr, Mathews PM, Bankhurst AD: Natural killer cell in systemic lupus erythematosus. Defects in effector lytic activity and response to interferon and interferon inducers. J. Clin. Invest. 1983; 71: 1230–1239. PubMed Abstract | Publisher Full Text | Free Full Text

[27] 27. Gasteiger G, Hemmers S, Firth MA, et al.: IL-2-dependent tuning of NK cell sensitivity for target cells is controlled by regulatory T cells. J. Exp. Med. 2013; 210: 1167–1178. PubMed Abstract | Publisher Full Text | Free Full Text

[28] 28. Green MR, Kennell AS, Larche MJ, et al.: Natural killer cell activity in families of patients with systemic lupus erythematosus: demonstration of a killing defect in patients. Clin. Exp. Immunol. 2005; 141: 165–173. PubMed Abstract | Publisher Full Text | Free Full Text

[29] 29. Green MR, Kennell AS, Larche MJ, et al.: Natural killer T cells in families of patients with systemic lupus erythematosus: their possible role in regulation of IGG production. Arthritis Rheum. 2007; 56: 303–310. PubMed Abstract | Publisher Full Text

The implication of interleukin-2 on the expression of CD56bright, CD56dim, and interferon-γ in patients with systemic lupus erythematosus

Abstract

Keywords

Introduction

Methods

Ethical considerations

Study design and sample preparation

Peripheral blood mononuclear cell (PBMC) incubation

Flow cytometry analysis

Enzyme-linked immunosorbent assay (ELISA) for cytokines

Statistical analysis

Results

CD56bright NK cells

Table 1. The levels of CD3-CD56bright NK cells, CD3-CD56dim NK cells, and IFN-γ after the administration of IL-2.

Figure 1. The summary of comparison between the different dose of IL-2 administration and the levels of NK CD3-CD56bright.

CD56dim NK cells

IFN-γ levels

Discussion

Conclusion

Authors contributions

Data availability

Underlying data

Acknowledgement

References

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The implication of interleukin-2 on the expression of CD56^bright, CD56^dim, and interferon-γ in patients with systemic lupus erythematosus

CD56^bright NK cells

Table 1. The levels of CD3-CD56^bright NK cells, CD3-CD56^dim NK cells, and IFN-γ after the administration of IL-2.

Figure 1. The summary of comparison between the different dose of IL-2 administration and the levels of NK CD3-CD56^bright.

CD56^dim NK cells