Serum levels of interleukin-10 and its receptor in the multiple myeloma type I patients

Rajabzadeh, Mehdi; Khadem ansari, Mohamad hassan; Khardmand, Fatemeh; Rasmi, Yousef; Gholinejad, Zafar

doi:10.61186/rabms.9.4.193

Volume 9, Issue 4 (10-2023) Journal of Research in Applied and Basic Medical Sciences 2023, 9(4): 193-199 | Back to browse issues page

‎ 10.61186/rabms.9.4.193

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Rajabzadeh M, Khadem ansari M H, Khardmand F, Rasmi Y, Gholinejad Z. Serum levels of interleukin-10 and its receptor in the multiple myeloma type I patients. Journal of Research in Applied and Basic Medical Sciences 2023; 9 (4) :193-199
URL: http://ijrabms.umsu.ac.ir/article-1-266-en.html

Serum levels of interleukin-10 and its receptor in the multiple myeloma type I patients

Mehdi Rajabzadeh ^*

, Mohamad hassan Khadem ansari

, Fatemeh Khardmand

, Yousef Rasmi

, Zafar Gholinejad

Department of Biochemistry, Faculty of Medicine, Urmia University of Medical Sciences, Urmia, Iran , mahdi.r7392@yahoo.com

Keywords: Biomarker, Interleukin 10, Interleukin 10 Receptor, Multiple Myeloma

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Introduction
Multiple myeloma is a malignancy of plasma cells that characterized by the presence of neoplastic plasma cells in the bone marrow and elevated monoclonal paraprotein levels (1). Multiple myeloma is the second most common hematologic malignancy and associated with high morbidity rate due to its end-organ failure (2). Evaluation of serum and urine proteins by capillary electrophoresis and bone marrow biopsy analysis are the main diagnostic tools. However, the pathogenesis and molecular mechanisms of multiple myeloma are not well understood (3). Inflammation is a common feature in malignancy and plays a pivotal role in the cancer pathogenesis (4). The role of inflammation and pro-inflammatory cytokines in the multiple myeloma were reported frequently (5). Bone marrow stromal cells produce pro-inflammatory cytokines by autocrine and paracrine mechanism which predict the duration of disease-free survival. Furthermore, cytokines proposed as biomarkers for diagnosis of multiple myeloma, patient's prognosis and proposed as tools for immunotherapy (6). The serum levels of TNF-α, IL-6, IL-6R and IL-1β are increased in the multiple myeloma patients (7-11). Interleukin-10 (IL-10) is a cytokine with immunomodulatory and potent anti-inflammatory properties that is produced by activated T cells , macrophages , dendritic cells and B-lymphocytes (12, 13). This cytokine belongs to class II cytokine family and plays a critical role in blood malignancy and other diseases (14-16). The biological function of IL-10 is mediated by IL-10 receptor (IL-10R), which has two isoform including IL-10R1 and IL-10R2. The IL-10R1 is a cell membrane associated receptor with transmembrane and extracellular domain that has higher affinity to IL-10 than IL-10R2 (17, 18). These receptors are expressed in several cell lines. Activation of IL-10R activates JAK/STAT signaling pathway as downstream mediator. Interaction of IL-10 and receptors lead to overexpression of anti-inflammatory cytokines as well as induce cell proliferation in the CD8+T and natural killer cells (19).
To the best of our knowledge, there is no comprehensive study on the relationship between IL-10 and its receptor with multiple myeloma currently. The aim of this case-control study was to determine the serum levels of IL-10 and IL-10R in the multiple myeloma patients and healthy subjects in order to describe the role of this cytokine and its receptor in the pathogenesis and the predictive value as biomarker.

Materials & Methods
Sampling:
In this case-control study, Inclusion criteria was patents with a confirmed multiple myeloma and 30 patients were enrolled. For control group, 40 age and sex matched subjects were selected. Exclusion criteria were subjects who had inflammatory disorder, infection and immunodeficiency and or who had received NSAIDs or anti-inflammatory drugs. Blood samples were obtained and sera were separated by centrifugation at room temperature and were stored at -70°C refrigerator.
ELISA Assay:
Serum IL-10 and IL-10R1 levels were determined using ELISA kits (bioassay technology laboratory Shanghai). The ELISA assays were performed according to the manufacturers' instructions.
Statistical analysis:
Data analysis was performed using SPSS software version 23. Descriptive analysis was performed and represented :as char:ts, tables and figures. Parametric and non-parametric statistical tests were used to compare of the mean of variables between case and control groups. The findings were expressed as mean ± SD, and p < 0.05 was considered as statistically significant.

Results
Serum IL-10 Levels:
The data analysis results of IL-10 are shown in table 1 and the figure 1A depicted IL-10 levels. The mean IL-10 levels did not show a significant difference between patients and control groups.

Table 1. Serum IL-10 levels in patients and control groups

IL-10(pg/ml)	Minimum	Maximum	Mean	SD	Range	p value
Control	37.30	86.60	50.93	8.76	49.30	0.481
Multiple myeloma	23.30	71.90	52.56	10.47	48.60	0.481

Fig. 1. A: Comparison of serum IL-10 levels in multiple myeloma and normal groups. There was no significant difference, B: Serum IL-10R levels in multiple myeloma was lower than normal groups significantly (p=0.041).

IL-10R Levels:
The descriptive analysis results of IL-10R are shown in able 2 and the figure 1B depicted IL-10R levels. The mean IL-10R levels showed significant difference between patients and control groups (0.041). The mean IL-10R of levels was lower in the multiple myeloma patients.

Table 2. Serum IL-10R levels in patients and control groups

IL-10R(pg/ml)	Minimum	Maximum	Mean	SD	Range	p value
Control	1.2	5.6	2.51	0.86	4.40	0.041
Multiple myeloma	1	4.40	2.1	0.76	3.40	0.041

Correlation analysis:
There was a significant positive correlation between IL-10 and IL-10R in patients, healthy subject and in total population. Figure 2 showed this correlation analysis between IL-10 and IL-10R levels.

Fig. 2. Correlation between serum IL-10 and IL-10R levels in patients(A) and healthy control(B)

Discussion
About two centuries ago, multiple myeloma w:as char:acterized but the molecular mechanisms of pathogenesis remains unclear (20, 21). Current knowledge supports the pathological role of inflammation in the multiple myeloma. Pro-inflammatory cytokines have double edge effects on multiple myeloma which increase cancer progression by induction of cell growth and apoptosis inhibition and anticancer effect by CD4+ T cells and natural killer cells (22).
In our study, IL-10 levels showed no significant difference between patients and healthy control. However, the previous study by Pappa et al. showed increased levels of IL-10 level in multiple myeloma (23). This inconsistency probably may be due to the patient's disease stage. Wang et al. showed that upregulation of IL-10 associated with poor prognosis (24). So, in the patients population who IL-10 levels remain unchanged, itwould have good prognosis that may be due to ethnicity and genetic properties (25).
In the multiple myeloma patients, IL-6 induces cell proliferation and cancer progression via IL-6R in bone marrow stromal cells(26).
The promoter sequences regulate the gene expression by modulation of transcription factors interaction. Mazur et al. showed that IL-10 promoter gene polymorphism does not associate with the susceptibility for multiple myeloma, which is in agreement with our finding (27). However, evaluation of serum IL-10 levels in the multiple myeloma patients with poor and good prognosis showed this cytokine is associated with better survival and outcome (24). In Wang et al. study, IL-10 mean was 201.96 pg/ml that was four times higher than our report (24). Because the race and ethnicity affect the cytokine levels and studied populations are different, we interpret this difference as racial variation (28).
The biological effects of ligands dependent on ligands concentration and receptors levels(29). Therefore, we conclude that the biological effects of IL-10 in the multiple myeloma patients are reduced due to downregulation of receptors. (30-32). We observed that IL-10R down regulated in multiple myeloma patients and probably be a good biomarker for diagnosis. The reductions of biological effects of an anti-inflammatory cytokine lead to inflammation, that would trigger the cancer progression. There are reports showing that IL-2R and IL-6R, the pro-inflammatory cytokines receptors were unregulated in multiple myeloma(6). These founding is in agreement with our finding because IL-10R is an anti-inflammatory signaling(33). Besides, Kasamatsu et al. found an association between IL-10R gene polymorphism and .multiple myeloma (34). However, another study found no significant changes in IL-10R gene expression(35).
Here, we report a positive correlation between IL-10 and IL-10R. Urbańska et al. showed there is a strong correlation between IL-10 and IL-6 receptor (36). We have no insight into the inducing effects of IL-10 on its receptors and other cytokines levels and their receptors regulation and need more studies (36). This study is the first report that measured the IL-10R levels in multiple myeloma patients type I. Shekarriz et al found a significant increase in the IL-10 of multiple myeloma patients where it was correlated with disease stage and markers of disease activity(37).
At the molecular level, toll like receptors is the main upstream receptors which induce the IL-10 production in myeloma and other cell lines(38, 39). Because toll like receptors is active in the multiple myeloma, absence of changes in the IL-10 levels in the type I may be due to the stage of disease or suppression of genes by other mechanisms such as microRNAs(40). Several studies showed that microRNAs have obvious effects on both expression of IL-10 and multiple myeloma progression(41).
In the downstream of IL-10 receptors, STAT3 transcription factor is located that express the anti-inflammatory gene(such as Bcl-xl). On the other hand, STAT3 is considered as a target for treatment of multiple myeloma(42). Therefore, the manipulation and regulation of IL-10 and its receptors could be considered as targets for treatment of disease and need more scrutiny.
Acknowledgments
We gratefully acknowledge the cooperation of all those who played a role in this research.
Conflict of interest
No conflict of interest declaration between the authors.

Funding/support
The Urmia Branch of the Islamic Azad University provided financial support for this study.

Type of Study: orginal article | Subject: Other

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