IL-27 Concentration in Systemic Circulation and Tumor Micro-Circulation Samples of Sclc and Nsclc Patients; Association with Tumor Size, Histological Type and Presence of Metastases

Research Article

Austin J Clin Immunol. 2016; 3(1):1030.

IL-27 Concentration in Systemic Circulation and Tumor Micro-Circulation Samples of Sclc and Nsclc Patients; Association with Tumor Size, Histological Type and Presence of Metastases

Karlicic V¹, Vukovic J¹, Stanojevic I2,9, Sotirovic J3, Peric A3, Jovic M4, Cvijanovic V5, Djukic M6, Banovic T7, Pantic Bisevac J8 and Vojvodic Danilo2,9*

¹Clinic for Lung Disease, Military Medical Academy, Serbia1Clinic for Lung Disease, Military Medical Academy, Serbia

²Institute for Medical Research Military Medical Academy, Serbia

³Clinic for Ear, Nose and Throat, Military Medical Academy, Serbia

4Institute of Pathology and Forensic Medicine Military Medical Academy, Serbia

5Clinic for Thoracic Surgery, Military Medical Academy, Serbia

6Department of Toxicology, Belgrade University, Serbia

7Department of Immunology, Australia

8Institute for Medical Biochemistry, Military Medical Academy, Serbia

9Medical Faculty, University of Defense, Serbia

*Corresponding author: Vojvodic D, Institute for Medical Research, Military Medical Academy, Medical Faculty, University of Defense, Crnotravska 17, 11000 Belgrade, Serbia

Received: July 08, 2016; Accepted: August 05, 2016; Published: August 09, 2016


IL-27 Concentration in systemic circulation and tumor micro-circulation samples of sclc and nsclc patients; association with tumor size, histological type and presence of metastases

Introduction: Advanced lung carcinoma is charasterized with fast disease progression. IL-27 is considered as a cytokine with anti-tumor activities and its role in lung carcinoma pathogenesis and in the anti-tumor response has not yet been elucidated.

Aim: To correlate IL-27 concentration in serum and lung tumor microcirculation with clinical stage, disease outspread, pathohistological features and TNM stadium.

Patients and Methods: The study included 41 lung tumor patients in III and IV clinical stage. Histological type was determined immunohistochemically, while tumor size, localization and dissemination were determined radiologically (MSCT). IL-27 concentration was quantified with commercial ELISA test in serum and lung tumor microcirculation samples.

Results: IL-27 concentration was significantly decreased in samples of all patients comparing to controls. In the patient’s samples, IL-27 was significantly increased in tumor microcirculation comparing to sera samples. Patients with NSCLC large cell histology type had the lowest IL-27 concentration, both in tumors and sera samples. Degree of disease spread was significantly associated with IL-27 levels, because concentration was significantly decreased in patient’s clinical stage IV or those with metastases, in both types of samples. Both in tumor microcirculation and sera samples, there was significant association of tumor size and IL-27 concentration, with the highest value of IL-27 in patients with the smallest lung tumors.

Conclusion: IL-27 concentration is significantly elevated in tumor microenvironment of investigated lung cancer patients. IL-27 values significantly differ between lung cancer patients according to tumor histology, degree of disease extent and size of tumor. Significant association of higher IL-27 concentration with smaller tumors, earlier clinical stage and absence of metastases indicates that IL-27 cytokine have anti-tumor functions in these patients.

Keywords: NSCLC; SCLC; IL-27; Serum; Tumor microcirculation; Tumor size


SCLC: Small Cell Lung Cancer; NSCLC: Non Small Cell Lung Cancer; NSCLC Ad: NSCLC Adenocarcinoma; NSCLC Sq: NSCLC Squamous Carcinoma; NSCLC LC: NSCLC Large Cell Carcinoma; EBI3: Epstein Barr Induced 3 gene


Interleukin 27 is a relatively recently characterized cytokine mainly recognized by its anti tumor effects [1]. By structure it belongs to IL-12 cytokine family that includes heterodimeric cytokines IL- 12 (composed of IL-12p35 + IL-12p40), IL-23 (composed of IL- 23p19 + IL-12p40), IL-27 (composed of IL-27p28 + EBI3) and IL-35 (composed of IL-12p35 + EBI3) [2]. IL-27 exerts its effect by binding specific IL-27 receptor constructed of two subunits, WSX and gp130 [3]. Unique characteristic of this receptor that every subunit is bound to a distinct transcription factor (WSX1 subunit to STAT1 and gp130 subunit to STAT3) so IL-27 have capacity to efficiently activate both STAT1 and STAT3 cell transcription factors. First demonstrated function of IL-27 was its inflammatory role, based on study where mice deficient for alpha subunit of IL27R showed significant susceptibility for L. Monocytogenes and L. Mayor Infections [4]. Pflanz et al. also showed that IL-27 supported proliferation and stimulation of IFN-γ secretion from CD4+T lymphocytes. IL-27 induces the TH1 T cells expansion [5] and inhibits differentiation of inducible regulatory T lymphocytes [6]. Beyond CD4+T lymphocytes IL-27 stimulates proliferation and the expression of IFN-γ and beta subunit of IL12 on CD8+T lymphocytes [7]. IL-27 potently affect Cytotoxic T Lymphocyte response (CTL) by inducing the expression of granzyme B [8,9].

Biological functions of IL-27 are mainly characterized from studies in different tumor models, where IL-27 has demonstrated antitumor activities. IL-27 suppressed tumor growth in metastatic murine neuroblastoma tumors [10], murine colon carcinoma cells [11,12], melanoma cell lines [13,14], head and neck squamous cell carcinoma [15], lung carcinoma cell lines [16] and multiple myeloma [17]. Some of this anti tumor effects were mediated by CD8+ T lymphocytes, as shown in models of metastatic murine neuroblastoma tumors [10] and murine colon carcinoma cells [11]. In other tumor models IL-27 acted through different mechanisms, enhanced NK cell anti tumor activity [13], inhibited angiogenesis [12], directly suppressed tumor cell proliferation [14,17] and induced antibody dependent tumor cell cytotoxicity [15]. Data from all these studies demonstrated anti tumor effects of IL-27, executed through various effector mechanisms.

But not all biological activities of IL-27 are stimulatory for immune cells. IL-27 inhibits the development of Th17 T lymphocytes and induces IL-10 secretion [18]. In some conditions IL-27 is an important factor for differentiation of induced regulatory T lymphocytes (Treg) with potent IL-10 producing capacity [19-22], and transforms dendritic cells into immunosuppressive by induction of B7-H1 expression [23]. In vitro study in a model of a human epithelial ovarian cells additionally demonstrated that IL-27 induced expression of IL18 inhibitor, together with Programmed Death Ligand -1 (PD-1L) and Indoleamine 2,3 Dioxigenase (IDO), potent immunosuppressive check points in activation of adaptive immune response [1].

Studies that analyze IL-27 significance in lung cancer patients are still rare [24-27]. We have shown that high IL-27 sera levels are associated with early clinical stage and limited disease in patients with melanoma [28]. However, because still little is known about IL-27 importance in lung cancer patients, in this study we have investigated association of IL-27 levels in systemic circulation and tumor microcirculation with clinical parameters and tumor characteristics in our SCLC and NSCLC patients.

Material and Methods


The study included 41 patients, diagnosed for lung cancer and 30 healthy controls (Table 1). Patients were diagnosed and treated at Clinic for lung diseases, Military Medical Academy, Belgrade, Serbia, in period from March till December 2015. All necessary diagnostic procedures (histological, laboratory and radiological) were performed at Military Medical Academy, Belgrade, Serbia. All patients and healthy controls were consented and this study was approved by the local Research Ethics Committee, Military Medical Academy (11-03/2014).


Blood samples were taken from cubital vein. Tumor microcirculation samples were taken from accessible pathological blood vessels with needle aspiration during diagnostic bronchoscopy. Serum was separated from the samples after centrifugation (3000g, 10 minutes, RT) and frozen at -70 °C until testing. IL-27 was quantified with commercial ELISA test (R&D Systems).

Statistical analysis

All statistical tests were performed with software GraphPad Prism 5, using ANOVA test (with Bonferroni post testing) for multiple group comparisons, Mann Whitney test for to compare differences between two independent groups, Wilcoxon for comparison of paired (serum/tumor) samples and Pearson correlation test.


IL-27 in control and lung cancer patient samples

Healthy control persons had significantly higher average IL- 27 concentration comparing to lung cancer patients (Table 1). This finding was constant whether we are analyzed all subjects or after we divided patients according to sex. Healthy men demonstrated higher average IL-27 concentration comparing to healthy women. This difference was not as pronounced in sera samples from lung cancer patients of different sex. Analysis of tumor microcirculation samples showed higher average IL-27 value in samples of women with lung tumors.

Lung cancer histology and IL-27 concentration

The lowest average serum IL-27 concentration was detected in samples of patients that had NSCLC Large Cell histology tumors, significantly less than all other investigated types (Table 1& Figure 1). The same relationship was found in the tumor microcirculation samples, again with the lowest average IL-27 concentration in same patient group. All tumor samples had increased IL-27 level comparing to corresponding sera samples, with the significant increase in tumor compartment in patients with SCLC and NSCLC adenocarcinoma tumors.