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Prognostic implications of CD-57 expression in oral squamous cell carcinoma cases

1 Department of Oral Pathology and Microbiology, Vananchal Dental College and Hospital, Garhwa, Jharkhand, India
2 Department of Oral Pathology and Microbiology, Faculty of Dental Sciences, Institute of Medical Sciences, BHU, Varanasi, Uttar Pradesh, India
3 Department of Oral Pathology and Microbiology, King George's Medical University, Lucknow, Uttar Pradesh, India
4 Private Practitioner, Patna, Bihar, India

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Date of Submission09-Sep-2021
Date of Decision11-Jul-2022
Date of Acceptance13-Jul-2022
Date of Web Publication03-Feb-2023


Background: Natural killer (NK) cells are one of the most important effector T -lymphocytes having antitumor effect. Only a few studies are present in the literature, correlating the expression of cluster of differentiation 57 (CD-57) in NK cells in oral squamous cell carcinoma (OSCC). Aims: To analyze the effect of the presence of NK cells in the stroma of OSCC and their effect on the tumor progression and prognosis. Materials and Methods: A retrospective study was performed over 122 OSCC patients who had undergone surgical treatment between 2014 and 16. Wax blocks were obtained and subjected to immunohistochemical (IHC) analysis for CD-57 expression. Patients were followed up for 3 years and 100 cases were finally included in the study and divided into three groups. Statistical Analysis Used: The association between variable were studied through the Chi-square test, Kruskal–Wallis test, and Mann–Whitney U test. Result: A total of 91% of cases showed positive staining for CD-57. Maximum positive expression (96.6%) was observed in the patients who were alive and without recurrence (Group-II) as compared to dead patients (Group-I) and in well-differentiated squamous cell carcinoma (WDSCC) cases (96.8%) than in poorly differentiated cases. Also, the mean CD-57 positive cells count was found to be highest in Group-II and WDSCC cases. Conclusion: A significant correlation was observed between CD-57 expression and patients' health status. As the expression of CD-57 increased in the tumor stroma of OSCC, the chances for the patient to be alive were increased; therefore, it may serve as a good prognostic marker.

Keywords: CD-57, immunohistochemistry, natural killer cells, oral squamous cell carcinoma, OSCC, prognostic marker

How to cite this URL:
Kumar S, Chandra A, Agrawal R, Singh P, Pallavi M. Prognostic implications of CD-57 expression in oral squamous cell carcinoma cases. Indian J Pathol Microbiol [Epub ahead of print] [cited 2023 Mar 27]. Available from:

   Introduction Top

Oral squamous cell carcinoma (OSCC) is the commonest epithelial malignancy of the oral cavity. Carcinomas are heterogeneous cellular entities whose growth is dependent on reciprocal interactions between genetically altered cells and the microenvironment in which they reside. The innate immune cells present in this microenvironment include granulocytes, dendritic cells, macrophages, natural killer cells (NK cells), and mast cells.[1]

NK cells are one of the most important effectors of T-lymphocytes with an antitumor effect. They play a critical role both in innate and adaptive immunity. These cells defend against viral infections as well as help in tumor surveillance.[2],[3] Most important feature of NK cells is their capacity to distinguish stressed cells from healthy cells such as tumor cells, infected cells, and cells that have undergone physical or chemical injuries.[4]

NK cells and cytotoxic T-lymphocytes (CTLs) play complementary roles in immune responses against tumor cells.[5] CTLs are antigen-specific and recognize tumor antigens presented by major histocompatibility complex (MHC) class I molecules, but NK cells, as specialized innate immune lymphocytes, can detect the lack of MHC class I molecules by their surface receptors and kill tumor cells that have down-regulated MHC class I molecules.[6]

It has been found that NK cells activity decreases in patients suffering from breast cancer. Therefore these cells could be effective for evaluating the prognosis of treatment.[7] Various prognostic markers used in OSCC are CD-57, S-100, CD-56, CD-16+, CD-8+, CD-94, CD-54, and CD-1d.[2],[8],[9]

The cluster of differentiation 57 (CD-57) is a marker of terminally differentiated human CD-8+ T cells, that is, NK cells. Only a few studies are present correlating the expression of CD-57 in NK cells in OSCC. Therefore, this study was planned to analyze the presence of NK cells in the tumor stroma and their effect on tumor progression by assessing the expression of CD-57 in OSCCs of various histological grades and with a 3-year survival of these patients.

   Materials and Methods Top

A retrospective study was performed on OSCC patients after obtaining approval from the ethical committee of the institute. Tissue samples for the study were obtained from paraffin-embedded wax blocks of histologically proven OSCC cases of various grades from the archives of the department which had undergone surgery within the year 2014–16.

Out of a total of 122 cases analyzed, 100 OSCC cases were finally selected for the study. Demographic data and mobile numbers of all patients were obtained. They were followed up every 3 months after treatment through telephonic conversation regarding their health and any recurrence. If there was any indication of reappearance of the lesion, then the patient was immediately instructed to visit the hospital for proper checkups and investigations for validation of recurrence. Follow-up was done for 3 years or till death, whichever reached first. Based on the follow-up, the samples were divided into three groups, Group-I (Dead Patients), Group-II (Patients alive, without recurrence), and Group-III (Patients alive, with recurrence).

Inclusion criteria for the study were patients willing to participate in the study, the only primary lesions (OSCCs), cases with a proper record, biopsy samples properly fixed, processed, and stained, proper communication with patients, and proper follow-up of patients till the end of the study.

Paraffin blocks were sectioned on a rotary semiautomatic soft tissue microtome. Two slides were prepared for each case. One slide with a 5 μm thick section was stained with hematoxylin and eosin stain, while the other slide with a 3 μm thick section was taken on a poly-L-Lysine-coated glass slide for immunohistochemical (IHC) staining.[10]

IHC staining was done according to the manufacturer's guidelines including incubation with CD-57 mouse monoclonal primary antibody (BioGenex, USA) for 60 min, followed by secondary antibody, Super Sensitive Polymer-HRP reagent IHC Detection System (BioGenex, USA) for 30 min. When distinct brown staining was confined to the cytoplasm of the cell, CD-57 expression was considered positive.

The sections were first analyzed for the expression of CD-57 at low power (10×). Selected fields were then used for the assessment of the CD-57 positive cells. In each slide, 500 cells were counted in five different fields (high power field) using a magnification of 40× and if the cytoplasm of the cell was stained with CD-57 regardless of the intensity of the stain, then it was included in the counting.

   Statistics and Result Top

At the time of completion of this study, 33% of patients were dead (Group-I), 58% of patients were alive and showed no recurrence (Group-II), and 9% of patients were alive but had recurrence (Group-III) [Table 1].
Table 1: Distribution of OSCC patients according to histological grading within the study groups

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Among 31 well-differentiated squamous cell carcinoma (WDSCC) patients, 5 were in Group-I and 26 were in Group-II. Out of 41 moderately differentiated squamous cell carcinoma (MDSCC) patients, 9 were in Group-I, 26 were in Group-II, and 6 were in Group-III. Among the 28 poorly differentiated squamous cell carcinoma (PDSCC) patients, 19 were in Group-I, 6 were in Group-II, and 3 were in Group-III. A highly significant association was observed between survival and OSCC grades of patients (P < 0.001) [Table 1].

For CD-57 expression 91.0% of cases showed positive staining [Table 2]. Among the 33 Group-I patients, 27 (81.8%) showed positive staining. Out of 58 Group-II patients, 56 (96.6%) were positive. Among the 9 Group-III patients, 8 (88.9%) showed positive staining. However, no significant association (P = 0.06) was observed between CD-57 staining and the participants of the study groups.
Table 2: Distribution of OSCC patients according to CD-57 expression

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The expression of CD-57 among different OSCC grade patients' showed that, out of 31 WDSCC patients, 30 (96.8%) showed positive staining. Out of 41 MDSCC patients, 39 (95.1%) were positive, and among the 28 PDSCC patients, 22 (78.6%) showed positive staining [Figure 1]. A significant association was observed between CD-57 staining and OSCC grades (P = 0.03) [Table 2].
Figure 1: Photomicrograph showing positive CD-57 expression in PDSCC (×400)

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   Discussion Top

The key role of the human immune system is to confront cells undergoing carcinogenesis. It has been observed that the absence or reduced MHC class I molecule expression in malignant cells activates NK cells. So, CD-57 may be a more reliable marker to evaluate NK cells in tumor tissue.[11]

Out of a total of 100 patients, 33% patients in the present study were dead till the time of completion of the study. Among the remaining survived 67% patients, 58% showed no recurrence. Therefore, the disease-free survival rate after 3 years was 58% which is in accordance with the previously documented studies. Forastiere et al. found that 35–55% of patients remained disease-free after 3 years post-standard treatment, whereas Agarwal et al. documented that the average 3 years survival rate was 56%.[1],[12]

Also, disease-free survival was highest for WDSCC patients with 83.9% of cases found to be in study Group-II, while it was lowest for PDSCC (21.4%) cases. A highly significant association was observed between survival and different OSCC grades of the patients (P < 0.001). The results are similar to the study conducted by Coca et al.[13] who studied the influence of NK cell intratumoral infiltration on the survival of patients. The authors found that the survival difference between patients with little or moderate versus those with extensive NK cell infiltration was very significant in patients with tumor-node-metastasis stage-III disease.

In this study, 91% of patients were showing positive staining for CD-57. The results correspond with the previously documented study conducted by Agarwal et al.[1] in which 94% of samples were positive for CD-57 expression. The group-wise distribution shows that 96.6% of Group-II patients, 88.9% of Group-III, and 81.8% of Group-I patients were showing positive staining. However, no significant association (P = 0.06) was observed between the CD-57 staining and patients within different study groups.

According to histological grading, 96.8% of WDSCC cases, 95.1% of MDSCC cases, and 78.6% of PDSCC cases were showing positive staining for CD-57 expression. A statistically significant association (P = 0.03) was observed between CD-57 staining and different OSCC grades. However, de Sousa Lopes et al.[14] in their study found that OSCC's microenvironment is highly infiltrated by CD-57 immune cells and is a consisting finding, regardless of patient's age, tumor stage, or morphological grading.

The mean CD-57 positive cells count was found to be highest for Group-II patients (32.64 ± 10.78) and lowest for Group-I patients (7.98 ± 4.63) with a highly significant difference (P < 0.001) between the patients of different study groups. CD-57 positive cells count was less in dead patients, as compared to the patients who had survived more. Apart from that, the mean CD-57 positive cells count was found to be highest in WDSCC cases and least in PDSCC. Fang et al.[15] in their study also found that CD-57 expression in OSCC was associated with features of better prognosis including no lymph node metastasis and early clinical stages. CD-57 provided the highest predictive accuracy. Ishigami et al.[9] found the same results in gastric carcinoma. The positive correlation of CD-57 expression with survival may be because NK cells play a major role in eliminating neoplastic cells.

   Conclusion Top

Patients with a high rate of NK cell infiltration may have sufficient antitumor effect, effective surveillance of NK cells, and controlled progression of the disease, particularly in the early stages. Thus increase in the expression of CD-57 in the tumor stroma of OSCC increases the survival rate and may serve as a good prognostic marker.

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Conflicts of interest

There are no conflicts of interest.

   References Top

Agarwal R, Chaudhary M, Bohra S, Bajaj S. Evaluation of natural killer cell (CD-57) as a prognostic marker in oral squamous cell carcinoma: An immunohistochemistry study. J Oral Maxillofac Pathol 2016;20:173-77.  Back to cited text no. 1
[PUBMED]  [Full text]  
Lopez-Verges S, Milush JM, Pandey S, York VA, Arakawa-Hoyt J, Pircher-H, et al. CD-57 defines a functionally distinct population of mature NK cells in the human CD-56dim CD-16+ NK cell subset. Blood 2010;116:3865-74.  Back to cited text no. 2
Zamai L, Ponti C, Mirandola P, Gobbi G, Papa S, Galeotti L, et al. NK cells and cancer. J Immunol 2007;178:4011-16.  Back to cited text no. 3
Vivir E, Ugolini S, Blaise D, Chabannon C, Brossay L. Targeting natural killer cells and natural killer T cells in cancer. Nat Rev 2012;12:239-52.  Back to cited text no. 4
da Silveira EJ, Miguel MC, Lima KC, Freitas Rde A, de Morais Mde L, Queiroz LM. Analysis of local immunity in squamous cell carcinoma of the tongue and lower lip. Exp Mol Pathol 2010;88:171-5.  Back to cited text no. 5
Zancope E, Costa NL, Junqueira- Kipnis AP, Valadares MC, Silva TA, Leles CR, et al. Differential infiltration of CD8+ and NK cells in lip and oral cavity squamous cell carcinoma. J Oral Pathol Med 2010;39:162-7.  Back to cited text no. 6
Piroozmand A, Hassan ZM. Evaluation of natural killer cell activity in pre and post treated breast cancer patients. J Can Res Ther 2010;6:478-81.  Back to cited text no. 7
[PUBMED]  [Full text]  
Villegas FR, Coca S, Villarrubia VG, Jimenez R, Chillon MJ, Jareno J, et al. Prognostic significance of tumor infiltrating natural killer cells subset CD57 in patients with squamous cell lung cancer. Lung Cancer 2002;35:23-8.  Back to cited text no. 8
Ishigami S, Natsugoe S, Tokuda K, Nakajo A, Che X, Lwashige H, et al. Prognostic value of intratumoral natural killer cells in gastric carcinoma. Cancer 2000;88:577-83.  Back to cited text no. 9
Chandra A, Singh A, Sebastian BT, Agnihotri A, Bali R, Verma PK, et al. Oral squamous cell carcinomas in age distinct population: A comparison of p53 immunoexpression. J Can Res Ther 2013;9:587-91.  Back to cited text no. 10
[PUBMED]  [Full text]  
Taghavi N, Bagheri S, Akbarzadeh A. Prognostic implication of CD57, CD16, and TGF-β expression in oral squamous cell carcinoma. J Oral Pathol Med 2016;45:58-62.  Back to cited text no. 11
Forastiere AA, Goepfert H, Maor M, Pajak TF, Weber R, Morrison W, et al. Concurrent chemotherapy and radiotherapy for organ preservation in advanced laryngeal cancer. N Engl J Med 2003;349:2091-8.  Back to cited text no. 12
Coca S, Perez-Piqueras J, Martinez D, Colmenarajo A, Saez MA, Vallejo C, et al. The prognostic significance of intratumoral natural killer cells in patients with colorectal carcinoma. Cancer 1997;79:2320-8.  Back to cited text no. 13
de Sousa Lopes MLD, da Silva Barros CC, de Souto Medeiros MR, da Costa Miguel MC, de Souza LB, Alves PM, et al. Evaluation of CD57+ cells in oral squamous cells carcinoma and their relationship with clinicopathological parameters. J Oral Diag 2017;2:e20170010.  Back to cited text no. 14
Fang J, Li X, Ma D, Liu X, Chen Y, Wang Y, et al. Prognostic significance of tumor infiltrating immune cells in oral squamous cell carcinoma. BMC Cancer 2017;17:375.  Back to cited text no. 15

Correspondence Address:
Akhilesh Chandra,
Faculty of Dental Sciences, IMS, BHU, Varanasi - 221 005, Uttar Pradesh
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Source of Support: None, Conflict of Interest: None

DOI: 10.4103/ijpm.ijpm_906_21


  [Figure 1]

  [Table 1], [Table 2]


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