Distribution of M1 and M2 macrophages in tumor islets and stroma in relation to prognosis of non-small cell lung cancer

Jurgita Jackute, Marius Zemaitis, Darius Pranys, Brigita Sitkauskiene, Skaidrius Miliauskas, Simona Vaitkiene, Raimundas Sakalauskas, Jurgita Jackute, Marius Zemaitis, Darius Pranys, Brigita Sitkauskiene, Skaidrius Miliauskas, Simona Vaitkiene, Raimundas Sakalauskas

Abstract

Background: Non-small cell lung cancer (NSCLC) remains the most common cause of cancer related death worldwide. Tumor-infiltrating macrophages are believed to play an important role in growth, progression, and metastasis of tumors. In NSCLC, the role of macrophages remains controversial; therefore, we aimed to evaluate the distribution of macrophages (M1 and M2) in tumor islets and stroma and to analyze their relations to patients' survival.

Methods: Lung tissue specimens from 80 NSCLC patients who underwent surgical resection for NSCLC (pathological stage I-III) and 16 control group subjects who underwent surgery because of recurrent spontaneous pneumothorax were analyzed. Immunohistochemical double staining of CD68/iNOS (markers for M1 macrophages) and CD68/CD163 (markers for M2 macrophages) was performed and evaluated in a blinded manner. The numbers of M1 and M2 macrophages in tumor islets and stroma were counted manually.

Results: Predominant infiltration of M1 and M2 macrophages was observed in the tumor stroma compared with the tumor islets. M2 macrophages predominated over M1 macrophages in the tumor tissue. Tumor islets-infiltrating M1 macrophages and the number of total tumor-infiltrating M2 macrophages were independent predictors of patients survival: high infiltration of M1 macrophages in tumor islets was associated with increased overall survival in NSCLC (P < 0.05); high infiltration of total M2 macrophages in tumor (islets and stroma) was associated with reduced overall survival in NSCLC (P < 0.05).

Conclusions: This study demonstrated that high infiltration of M1 macrophages in the tumor islets and low infiltration of total tumor-infiltrating M2 macrophages were associated with improved NSCLC patients' survival.

Trial registration: ClinicalTrials.gov NCT01955343 , registered on September 27, 2013.

Conflict of interest statement

Ethics approval and consent to participate

All study procedures were approved by Kaunas Regional Ethics Committee for Biomedical Research (No. BE-2-20). Written informed consent was obtained from each study individual.

Consent for publication

Not applicable.

Competing interests

The authors declare that they do not have any competing interests.

Publisher’s Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Figures

Fig. 1
Fig. 1
Study flow chart
Fig. 2
Fig. 2
Immunohistochemical staining of macrophages in non-small cell lung cancer tissue. M1 macrophages double stained with the anti-CD68 and anti-iNOS (arrow). Original magnification: 400×
Fig. 3
Fig. 3
Immunohistochemical staining of macrophages in non-small cell lung cancer tissue. M2 macrophages double stained with the anti-CD68 and anti-CD163 (arrow). Original magnification: 400×
Fig. 4
Fig. 4
Distribution of total M1 and M2 macrophages in lung tissue of NSCLC and control group subjects
Fig. 5
Fig. 5
Distribution of M1 and M2 macrophages in tumor islets and stroma in NSCLC patients
Fig. 6
Fig. 6
Kaplan-Meier survival curves demonstrate M1 macrophages density in islets in correlation to overall survival
Fig. 7
Fig. 7
Kaplan-Meier survival curves demonstrate total M2 macrophages density in correlation to overall survival
Fig. 8
Fig. 8
Serum INF-γ (a), IL-10 (b) and TNF-α (c) concentration in NSCLC and control group patients

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