Skip to main content
Download PDF

Prognostic value of CD66b positive tumor-infiltrating neutrophils in testicular germ cell tumor

BMC Cancer volume 16, Article number: 898 (2016) Cite this article

Abstract

Background

Prognostic value of immune cells is not clear in testicular germ cell tumors (TGCTs). We aimed to investigate the prognostic value of tumor-infiltrating neutrophils in TGCTs.

Methods

A total of 102 patients who underwent orchiectomy for TGCT were investigated for CD66b positive tumor-infiltrating neutrophils (CD66b + TINs). Immmunostaining for CD66b was performed in 102 sections as described. Clinicopathological parameters as well as cancer specific survival and overall survival were assessed for correlation with CD66b + TIN density.

Results

High density group was significantly correlated with tumor diameter ≥ 10 cm, presence of nodal/distant metastasis, S stage, diagnosis of nonseminomatous germ cell tumor (NGCT), and presence of venous invasion (p = 0.0198, p < 0.0001, p = 0.0275, p = 0.0004, and p = 0.0287, respectively). It was also significantly associated with cancer-specific and overall survival (logrank p = 0.0036, and p = 0.0002, respectively). Multivariate analysis showed that increased CD66b + TIN was an independent prognostic factor for overall survival (p = 0.0095).

Conclusions

Increased CD66b + TIN was significantly associated with presence of metastasis, S stage, and nonseminomatous germ cell tumor diagnosis. It was also an independent prognostic factor of overall survival in patients with TGCT.

Peer Review reports

Background

Inflammation is considered to play a significant role in tumor progression in many malignancies [1]. Tumors that produce various inflammatory cytokines recruit immune cells such as neutrophils, and activate them to favor tumor growth and progression [2]. Increased levels of neutrophils are observed both in peripheral bloods and tumor environment in various cancers [3–11]. However, to our knowledge, there has been no literature regarding association between neutrophils and testicular germ cell tumors (TGCTs).

The aim of this study was to investigate the prognostic value of tumor-infiltrating neutrophils (TINs) in TGCT. By assessing the intra-tumoral environment using immunohistochemistry, we considered that it would provide more direct information than parameters based on peripheral blood samples. Relationships between TIN and clinico-pathological parameters including prognosis in patients with TGCT are described in the present study.

Methods

Patient characteristics and tissue preparation

The study included 102 patients who underwent orchiectomy for TGCT at The Tokyo University Hospital between 1985 and 2008. Clinicopathological parameters were retrospectively investigated from clinical records. Peripheral blood white blood cell and neutrophil counts were not available in 22 and 36 patients, respectively. Tumors were restaged using the TNM 2009 staging system [12]. Patients with nodal and/or distant metastasis (30 cases) were classified according to the International Germ Cell Consensus Classification (IGCCC) [13]. No patients received chemotherapy or radiation before orchiectomy.

CD66b immunostaining was performed to evaluate intra-tumoral neutrophils, since CD66b is uniquely expressed by neutrophils and not expressed in monocytes or myeloid cells [14]. In addition, CD66b is a preferable marker of aggressiveness in cancer when compared with other markers such as myeloperoxidase [6]. Sections were available in all 102 patients for CD66b immunohistochemistry. Sections were obtained from the same tumor blocks used for routine pathological evaluation. Therefore, haematoxylin and eosin (H&E) stained sections were also available for reference regarding areas of tumors and vessels.

Informed consent was obtained from all individual participants included in the study. This study was approved by the Ethics Committee of the University of Tokyo Hospital (approval number #2283), and is in accordance with the Helsinki declaration.

Immunohistochemistry

Immunohistochemistry for CD66b staining was performed by the streptavidin-biotin method as previously described [15]. Six-micrometer-thick sections were deparaffinized with 2 changes of xylene for 3 min each, then dehydrated using decreasing concentrations of ethanol, and rinsed in Tris-buffered saline (TBS). Antigen retrieval was carried out immersing the sections in citric acid buffer (2 mM citric acid and 9 mM trisodium citrate dehydrate, pH 6.0) and autoclaved at 121 °C for 10 min. After cooling period of 3 min, the sections were washed with TBS and blocked with endogenous peroxidase with 0.3% H2O2. The sections were then incubated in 10% bovine serum albumin (BSA) for 30 min. The slides were incubated overnight at 4 °C with a primary mouse antibody for CD66b which was diluted at 1:200 (#305102, Biolegend®, San Diego, USA). After the sections were washed in TBS, they were incubated with CSA-2 kit (DAKO, Carpinteria, CA, USA). The antigen-antibody complex was visualized with 3,3′-diaminobenzidine tetrachloride (DAB) solution (1 mM DAB, 50 mM Tris-HCL buffer, pH 7.6, and 0.006% H2O2). All sections were counter-stained by Carazzi’s hematoxylin for 60 s. For negative controls, normal mouse IgG was used instead of primary antibodies.

Immunohistochemical assessment

The density of CD66b + TINs was assessed in immunostained sections as in previous literature [7, 16]. Stained cells with clear boundary and sufficient intensity were recognized as CD66b + neutrophils. Cells with blurry stains and unclear boundary were neglected. TINs were evaluated within the tumor, but not in the area showing necrosis or artifacts. TIN was counted from ten random microscopic fields (×200). TIN Counts of entire 102 sections were 14.8 ± 38.8 counts/microscopic field (mean ± SD), or median value of 1 (range 0 - 262). By using the receiver operating characteristic (ROC) curve analysis, cutoff value was selected from overall survival status from which the largest AUC was obtained (AUC = 0.80217, Additional file 1: Figure S1). Cutoff value was selected from the point which was closest to both maximum sensitivity and specificity (21.6 counts/microscopic field). Therefore, high and low TIN density was defined as counts ≤ 21 and counts > 21, respectively.

Two independent observers (YY and TN) evaluated the stained sections, blinded to the patients’ clinic-pathological details. The third observer (TF) determined the density (high or low) in case of disagreement between the 2 observers.

Statistical analyses

We used the statistical software JMP ®Pro version 10.0.2 (©2012 SAS Institute Inc., Cary, CA, USA) for data analysis. All continuous variables did not show normal distribution, and therefore Wilcoxon rank-sum test was used to compare differences between continuous variables between low and high TIN density groups. Pearson’s chi-square test and Fisher’s test (used when frequency was under 5) was used in analyzing differences of categorical variables between low and high TIN density groups in Table 2 and Table 3. Log-rank test was performed to analyze the statistical difference of cancer-specific and overall survival in low and high density groups. Multiple regression model was used to identify associated factors of cancer specific and overall survival. Variables that were significantly associated in univariate survival analysis were included in the multivariate analysis. Since the hypothesis ‘high TIN density increases the risk of cancer specific and overall mortality’ was considered unilateral, P value of < 0.025 was considered to be statistically significant in analysis regarding survival and multiple regression models. P value < 0.05 was considered to be statistically significant in statistical analysis evaluating association between other variables.

Results

Clinical characteristics of 102 patients with TGCT are presented in Table 1. Median value (interquartile range (IQR)) of patient age was 34 (26 - 40) years. Fifty-eight patients had pathological stage T1, and 44 had T2–T4. Median values (IQR) of LDH, βhCG, AFP levels were 240 (170 - 463) IU/ml, 1.7 (0 – 20.2) mIU/ml, and 5 (2 - 141) ng/ml, respectively. Eighty and sixty-eight patients had preoperative clinical records of white blood cell and neutrophil counts from peripheral blood samples, respectively. Median values (IQR) of white blood cell and neutrophil counts were 6800 (5900-8375) counts/μl and 4300 (3542-5937) counts/μl, respectively.

Table 1 Clinical characteristics of 102 patients with TGCT
Full size table

A total of 102 sections were examined for CD66b immunostaining. The number of intravascular neutrophils (Fig. 1) was considered negligible (mean count: 0.095 cells per microscopic field), and was not included in the TIN count. There were 81 cases for low, and 21 for high density group. There were no significant differences between high and low TIN density groups in terms of individual tumor markers and IGCCC risk (Table 2). TIN density did not show correlation with peripheral neutrophil counts (Wilcoxon rank-sum test p = 0.7947).

Fig. 1
figure 1

Representative examples of CD66b positive neutrophils in TGCT patients. Representative examples of CD66b + tumor-infiltrating neutrophils. Yellow arrow shows CD66b + neutrophil. Scale bars, 100 μm. a A patient with seminoma. b A patient with pure embryonal carcinoma. Abundant number of tumor-infiltrating neutrophil can be observed within the tumor

Full size image
Table 2 Relationships between TIN density and clinical characteristics in TGCT patients (N = 102)
Full size table

High density group was significantly correlated with diagnosis of NGCT (p = 0.0004), tumor diameter > 10 cm (p = 0.0198), and presence of venous invasion (p = 0.0287) (Table 3).

Table 3 Relationships between TIN density and pathological findings in TGCT patients (N = 102)
Full size table

Relationship of CD66b + TIN density to survival is shown in Fig. 2. High density group was significantly associated with poor survival for cancer-specific and overall survival in TGCT patients (logrank p = 0.0036, p = 0.0002, respectively). In addition, high TIN density group had lower cancer-specific and overall survival rates in SGCT patients (P = 0.0376 and P < 0.0001, respectively), whereas it showed tendency towards lower overall survival in NGCT patients (P = 0.0657).

Fig. 2
figure 2

Survival analysis of CD66b + TIN density in testicular germ cell tumor patients. a, b Cancer specific and overall survival in patients with non-seminomatous germ cell tumor. c, d Cancer specific and overall survival in patients with seminomatous germ cell tumor. e, f Cancer specific and overall survival in patients with testicular germ cell tumor. Log rank test was used to analyze the differences in survival

Full size image

In univariate analysis, clinical factors significantly associated with poor cancer specific and overall survival in TGCT patients were N stage (N1-3 vs N0), M stage (M1 vs M0), and TIN density status (High vs Low) (Table 4). In multivariate analysis, M stage was an independent factor of cancer-specific survival (P = 0.0126). High TIN density remained an independent prognostic factor of overall survival (p = 0.0095).

Table 4 Univariate and Multivariate analyses of risk factors predicting cancer specific and overall survival in patients with TGCT
Full size table

Discussion

To our knowledge, this study is the first report to show that increased CD66b + TIN is an independent prognostic factor for overall survival in patients with TGCT. In addition to this finding, our results also revealed that increased CD66b + TIN was significantly associated with diagnosis of non-seminomatous germ cell tumor, S stage of S2 and over, tumor size > 10 cm, presence of nodal and/or distant metastasis, and presence of venous invasion.

In general, neutrophils are viewed as one of the first recruited effectors involved in acute inflammatory response [1, 14]. Neutrophils that are recruited to the tumor environment are discriminated from naϊve neutrophils, since they are characterized with low amounts of granules, and reactive oxygen species (ROS) [17]. They also express elevated levels of CXCL1, CXCL2 [17], that are potent chemoattractant promoting neutrophil recruitment [18]. Neutrophil recruitment is also supported by cancer cells that produce granulocyte colony stimulating factor (GCSF), which leads to an increment of neutophils via stimulation of bone marrow granulocytic cells [19]. In addition, tumor microenvironment stabilizes and prolongs the survival of neutrophils [18].

The function of neutrophils in tumor environment is complex, since they have conflicting function in cancer environment according to their activation state [17]. Tumor cells produce immunosuppressive transforming growth factor β (TGFβ) that promotes the polarization of tumor-associated neutrophils to a pro-tumoral “N2 phenotype” [20]. This type of neutrophil may contribute to cancer progression, since it produces growth factors such as vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), MMP9, Bv8, and also have the capability to suppress cytotoxic lymphocytes [17, 18, 21, 22].

In this study, increased TINs correlated with presence of nodal and/or distant metastasis. This finding may be explained from the results of several studies that have identified the function of neutrophils to promote tumor migration and invasion. Head and neck squamous cell carcinoma (HNSCC) cells stimulated neutrophils to release proinflammatory cytokines which accelerated the migration of tumor cells [23]. Shamamian P et al. have shown that neutrophil serine proteases activated MMP2 via MT1-MMP, which lead to an invasion of fibrosarcoma cells [24].

In a clinical level, recent studies have shown that relationship between immune cells and tumor microenvironment is important in oncologic outcomes. In a study comprised of 121 patients undergoing nephrectomy for localized renal cell carcinoma, the presence of intra-tumoral neutrophils was an independent prognostic factor for cancer specific and overall survival [3]. In hepatocellular carcinoma patients, mean counts of intratumoral neutrophil were 27.3 ± 56.1 counts/microscopic field (×200), and presence of intratumoral neutrophil was a poor prognostic factor for hepatocellular carcinoma after resection [4]. Patients with low intratumoral neutrophils had a significantly longer 5-year recurrence free rate and overall survival (53% vs < 37% and 57% vs 18%, respectively). In colorectal cancer patients, increased intra-tumoral CD66b + neutrophil was not only positively correlated with pT status, M status, and clinical stage, but was also an independent prognostic factor in multivariate analysis [5]. Limitations include a possible bias in groups divided by TIN density because of the potential differences in comorbidity and treatment history.

Conclusions

Increased CD66b + TIN was an independent prognostic factor for overall survival in TGCT patients. Thus, evaluating density of TINs may be beneficial as an additional prognostic tool. However, larger and prospective studies are necessary to further elucidate the present findings.

Abbreviations

AFP:

Alpha-feto-protein

IGCCC:

International germ cell classification consensus

LDH:

Lactate dehydrogenase

NGCT:

Nonseminomatous germ cell tumor

TGCT:

Testicular germ cell tumor

TIN:

Tumor-infiltrating tumor

βhCG:

Beta human chorionic gonadotropin

References

  1. Coussens LM, Werb Z. Inflammation and cancer. Nature. 2002;420(6917):860–7.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  2. Galdiero MR, Bonavita E, Barajon I, Garlanda C, Mantovani A, Jaillon S. Tumor associated macrophages and neutrophils in cancer. Immunobiology. 2013;218(11):1402–10.

    CAS  Article  PubMed  Google Scholar 

  3. Jensen HK, Donskov F, Marcussen N, Nordsmark M, Lundbeck F, von der Maase H. Presence of intratumoral neutrophils is an independent prognostic factor in localized renal cell carcinoma. J Clin Oncol. 2009;27(28):4709–17.

    Article  PubMed  Google Scholar 

  4. Li YW, Qiu SJ, Fan J, Zhou J, Gao Q, Xiao YS, Xu YF. Intratumoral neutrophils: A poor prognostic factor for hepatocellular carcinoma following resection. J Hepatol. 2011;54(3):497–505.

    Article  PubMed  Google Scholar 

  5. Rao HL, Chen JW, Li M, Xiao YB, Fu J, Zeng YX, Cai MY, Xie D. Increased intratumoral neutrophil in colorectal carcinomas correlates closely with malignant phenotype and predicts patients’ adverse prognosis. PLOS One. 2012;7(1):e30806.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. Ilie M, Hofman V, Ortholan C, Bonnetaud C, Coëlle C, Mouroux J, Hofman P. Predictive clinical outcome of the intratumoral CD66b-positive neutrophil-to-CD8-positive T-cell ratio in patients with resectable nonsmall cell lung cancer. Cancer. 2012;118(6):1726–37.

    CAS  Article  PubMed  Google Scholar 

  7. Carus A, Ladekarl M, Hager H, Nedergaard BS, Donskov F. Tumour-associated CD66b + neutrophil count is an independent prognostic factor for recurrence in localized cervical cancer. Br J Cancer. 2013;108:2116–22.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  8. Viers BR, Boorjian SA, Franc I, Tarrell RF, Thapa P, Karnes RJ, Thompson RH, Tollefson MK. Pretreatment neutrophi-to- lymphocyte ratio is associated with advanced pathologic tumor stage and increased cancer-specific mortality among patients with urothelial carcinoma of the bladder undergoing radical cystectomy. Eur Urol. 2014;66:1157–64.

    Article  PubMed  Google Scholar 

  9. Hermanns T, Bhindi B, Wei Y, Yu J, Noon AP, Richard PO, Bhatt JR, Almatar A, Jewett MAS, Fleshner NE, Zlotta AR, Templeton AJ, Kulkarni GS. Pre-treatment neutrophil-to-lymphocyte ratio as predictor of adverse outcomes in patients undergoing radical cystectomy for urothelial carcinoma of the bladder. Br J Cancer. 2014;111:444–51.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  10. Gondo T, Nakashima J, Ohno Y, Ozu C, Horiguchi Y, Namiki K, Yoshioka K, Ohori M, Hatano T, Tachibana M. Prognostic value of neutrophil-to-lymphocyte ratio and establishment of novel preoperative risk stratification model in bladder cancer patients treated with radical cystectomy. Urology. 2012;79:1085–91.

    Article  PubMed  Google Scholar 

  11. Dutta S, Crumley ABC, Fullarton GM, Horgan PG, McMillan DC. Comparison of the prognostic value of tumour and patient related factors in patients undergoing potentially curative resection of gastric cancer. Am J Surg. 2012;204:294–9.

    Article  PubMed  Google Scholar 

  12. Albers P, Albrecht W, Algaba F, Bokemeyer C, Cohn-Cedermark G, Fizazi K, Horwich A, Laguna MP, Nicolai N, Oldenburg J; European Association of Urology. Guidelines on Testicular Cancer. 2015 http://uroweb.org/guideline/testicular-cancer. Accessed 10 Nov 2016.

  13. International Germ Cell Cancer Collaborative Group. International germ cell consensus classification: a prognostic factor-based staging system for metastatic germ cell cancers. J Clin Oncol. 1997;15(2):594–603. no authors listed.

    Google Scholar 

  14. Murdoch C, Muthana M, Coffelt SB, Lewis CE. The role of myeloid cells in the promotion of tumour angiogenesis. Nature Rev Cancer. 2008;8(8):618–31.

    CAS  Article  Google Scholar 

  15. Yamada Y, Fujimura T, Takahashi S, Takayama K, Urano T, Murata T, Obinata D, Ouchi Y, Homma Y, Inoue S. Clinical significance of amyloid precursor protein in patients with testicular germ cell tumor. Adv Urol. 2013; doi: 10.1155/2013/348438

  16. Ropponen KM, Eskelinen MJ, Lipponen PK, Alhava E, Kosma VM. Prognostic value of tumour-infiltrating lymphocytes (TILs) in colorectal cancer. J Pathol. 1997;182(3):318–24.

    CAS  Article  PubMed  Google Scholar 

  17. Sionov RV, Fridlender ZG, Granot Z. The multifaceted roles neutrophils play in the tumor microenvironment. Cancer Microenviron. 2014; doi: 10.1007/s12307-014-0147-5

  18. Dumitru CA, Lang S, Brandau S. Modulation of neutrophil granulocytes in the tumor microenvironment: Mechanisms and consequences for tumor progression. Semin Cancer Biol. 2013;23(3):141–8.

    CAS  Article  PubMed  Google Scholar 

  19. Lord BI, Bronchud MH. The kinetics of human granulopoiesis following treatment with granulocyte colony-stimulating factor in vivo. Proc Natl Acad Sci USA. 1989;86:9499–503.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  20. Fridlender ZG, Sun J, Kim S, Kapoor V, Cheng G, Ling L, Worthen GS, Albelda SM. Polarization of tumor-associated neutrophil phenotype by TGF-beta: “N1” versus “N2” TAN. Cancer Cell. 2009;16(3):183–94.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  21. Nozawa H, Chiu C, Hanahan D. Infiltrating neutrophils mediate the initial angiogenic switch in a mouse model of multistage carcinogenesis. PNAS. 2006;103(33):12493–8.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  22. Shojaei F, Wu X, Zhong C, Yu L, Liang XH, Yao J, Blanchard D, Bais C, Peale FV, van Bruggen N, Ho C, Ross J, Tan M, Carano RAD, Meng YG, Ferrara N. Bv8 regulates myeloid-cell-dependent tumour angiogenesis. Nature. 2007;450(7171):825–31.

    CAS  Article  PubMed  Google Scholar 

  23. Dumitru CA, Gholaman H, Trellakis S, Bruderek K, Dominas N, Gu X, Bankfalvi A, Whiteside TL, Lang S, Brandau S. Tumor-derived macrophage migration inhibitory factor modulates the biology of head and neck cancer cells via neutrophil activation. Int J Cancer. 2011;129:859–69.

    CAS  Article  PubMed  Google Scholar 

  24. Shamamian P, Schwartz JD, Pocock BJ, Monea S, Whiting D, Marcus SG, Mignatti P. Activation of progelatinase A (MMP-2) by neutrophil elastase, cathepsin G, and proteinase-3: a role for inflammatory cells in tumor invasion and angiogenesis. J Cell Phys. 2001;189:197–206.

    CAS  Article  Google Scholar 

Download references

Acknowledgements

The authors thank Ms. A Saito for her technical assistance in immunostaining.

Funding

None declared.

Availability of data and material

Data analyzed during the present study is available from the corresponding author on reasonable request.

Authors’ contributions

YY performed immunostaining procedure, analyzed the data, performed statistical analysis, and drafted the manuscript. TN assisted on data analysis, participated in the design of the study and total coordination. TS assisted on statistical analysis. TH, UY participated in investigation of the clinical records. TF participated in the design and analysis of immunostaining results. HF participated in the design of the study and coordination. KT and TU participated in study design and assisted on data analysis. HK participated in the design of the study, and assisted on statistical analysis. SI and YH participated in design and total coordination. All authors read and approved the final manuscript.

Competing interests

The authors declared that they have no competing interests.

Consent for publication

Not applicable.

Ethics approval and consent to participate

Informed consent was obtained from all individual participants included in the study. This study was approved by the institutional review board (#2283), and is in accordance with the Helsinki declaration.

Author information

Authors and Affiliations

  1. Department of Urology, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

    Yuta Yamada, Tohru Nakagawa, Toru Sugihara, Takamasa Horiuchi, Uran Yoshizaki, Tetsuya Fujimura, Hiroshi Fukuhara, Haruki Kume & Yukio Homma

  2. Department of Geriatric Medicine and Anti-Aging Medicine, Graduate School of Medicine, The University of Tokyo, Bunkyo-ku, Tokyo, Japan

    Tomohiko Urano

  3. Department of Functional Biogerontology, Tokyo Metropolitan Institute of Gerontology, Itabashi-ku, Tokyo, Japan

    Kenichi Takayama & Satoshi Inoue

Authors
  1. Yuta Yamada
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tohru Nakagawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Toru Sugihara
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Takamasa Horiuchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Uran Yoshizaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tetsuya Fujimura
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hiroshi Fukuhara
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Tomohiko Urano
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Kenichi Takayama
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Satoshi Inoue
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Haruki Kume
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Yukio Homma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Tohru Nakagawa.

Additional file

Additional file 1: Figure S1.

Receiver operating characteristic (ROC) curve analysis was performed to determine the cutoff value of TIN counts by using the 0, 1 criterion. In the present study, overall survival status had the largest AUC (0.80217), and we selected cutoff value determined by overall survival status. Cutoff value was selected from the point which was closest to both maximum sensitivity and specificity (21.6 counts/microscopic field). Therefore, high and low TIN density was defined as counts ≤ 21 and counts > 21, respectively. (TIF 217 kb)

Rights and permissions

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Yamada, Y., Nakagawa, T., Sugihara, T. et al. Prognostic value of CD66b positive tumor-infiltrating neutrophils in testicular germ cell tumor. BMC Cancer 16, 898 (2016). https://doi.org/10.1186/s12885-016-2926-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s12885-016-2926-5

Keywords

  • Tumor-inflitrating neutrophil
  • Testicular cancer
  • CD66b
  • Neutrophil

BMC Cancer

ISSN: 1471-2407

Contact us