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Authors are the souls of any journal, and deserve much respect. To publish a journal manuscripts are needed from authors. Authors have a great responsibility for producing facts of their work in terms of number and results truthfully and an individual honesty is expected from authors in this regards. Both ways its true "No authors-No manuscripts-No journals" and "No journals–No manuscripts–No authors". Reviewing a manuscript is also a very responsible and important task of any peer-reviewed journal and to be taken seriously. It needs knowledge on the subject, sincerity, honesty and determination. Although the process of reviewing a manuscript is a time consuming task butit is expected to give one's best remarks within the time frame of the journal.
Salient features of the JCDR: It is a biomedical, multidisciplinary (including all medical and dental specialities), e-journal, with wide scope and extensive author support. At the same time, a free text of manuscript is available in HTML and PDF format. There is fast growing authorship and readership with JCDR as this can be judged by the number of articles published in it i e; in Feb 2007 of its first issue, it contained 5 articles only, and now in its recent volume published in April 2011, it contained 67 manuscripts. This e-journal is fulfilling the commitments and objectives sincerely, (as stated by Editor-in-chief in his preface to first edition) i e; to encourage physicians through the internet, especially from the developing countries who witness a spectrum of disease and acquire a wealth of knowledge to publish their experiences to benefit the medical community in patients care. I also feel that many of us have work of substance, newer ideas, adequate clinical materials but poor in medical writing and hesitation to submit the work and need help. JCDR provides authors help in this regards.
Timely publication of journal: Publication of manuscripts and bringing out the issue in time is one of the positive aspects of JCDR and is possible with strong support team in terms of peer reviewers, proof reading, language check, computer operators, etc. This is one of the great reasons for authors to submit their work with JCDR. Another best part of JCDR is "Online first Publications" facilities available for the authors. This facility not only provides the prompt publications of the manuscripts but at the same time also early availability of the manuscripts for the readers.
Indexation and online availability: Indexation transforms the journal in some sense from its local ownership to the worldwide professional community and to the public.JCDR is indexed with Embase & EMbiology, Google Scholar, Index Copernicus, Chemical Abstracts Service, Journal seek Database, Indian Science Abstracts, to name few of them. Manuscriptspublished in JCDR are available on major search engines ie; google, yahoo, msn.
In the era of fast growing newer technologies, and in computer and internet friendly environment the manuscripts preparation, submission, review, revision, etc and all can be done and checked with a click from all corer of the world, at any time. Of course there is always a scope for improvement in every field and none is perfect. To progress, one needs to identify the areas of one's weakness and to strengthen them.
It is well said that "happy beginning is half done" and it fits perfectly with JCDR. It has grown considerably and I feel it has already grown up from its infancy to adolescence, achieving the status of standard online e-journal form Indian continent since its inception in Feb 2007. This had been made possible due to the efforts and the hard work put in it. The way the JCDR is improving with every new volume, with good quality original manuscripts, makes it a quality journal for readers. I must thank and congratulate Dr Hemant Jain, Editor-in-Chief JCDR and his team for their sincere efforts, dedication, and determination for making JCDR a fast growing journal.
Every one of us: authors, reviewers, editors, and publisher are responsible for enhancing the stature of the journal. I wish for a great success for JCDR."
Thanking you
With sincere regards
Dr. Rajendra Kumar Ghritlaharey, M.S., M. Ch., FAIS
Associate Professor,
Department of Paediatric Surgery, Gandhi Medical College & Associated
Kamla Nehru & Hamidia Hospitals Bhopal, Madhya Pradesh 462 001 (India)
E-mail: drrajendrak1@rediffmail.com
On May 11,2011
Dr. Shankar P.R.
"On looking back through my Gmail archives after being requested by the journal to write a short editorial about my experiences of publishing with the Journal of Clinical and Diagnostic Research (JCDR), I came across an e-mail from Dr. Hemant Jain, Editor, in March 2007, which introduced the new electronic journal. The main features of the journal which were outlined in the e-mail were extensive author support, cash rewards, the peer review process, and other salient features of the journal.
Over a span of over four years, we (I and my colleagues) have published around 25 articles in the journal. In this editorial, I plan to briefly discuss my experiences of publishing with JCDR and the strengths of the journal and to finally address the areas for improvement.
My experiences of publishing with JCDR: Overall, my experiences of publishing withJCDR have been positive. The best point about the journal is that it responds to queries from the author. This may seem to be simple and not too much to ask for, but unfortunately, many journals in the subcontinent and from many developing countries do not respond or they respond with a long delay to the queries from the authors 1. The reasons could be many, including lack of optimal secretarial and other support. Another problem with many journals is the slowness of the review process. Editorial processing and peer review can take anywhere between a year to two years with some journals. Also, some journals do not keep the contributors informed about the progress of the review process. Due to the long review process, the articles can lose their relevance and topicality. A major benefit with JCDR is the timeliness and promptness of its response. In Dr Jain's e-mail which was sent to me in 2007, before the introduction of the Pre-publishing system, he had stated that he had received my submission and that he would get back to me within seven days and he did!
Most of the manuscripts are published within 3 to 4 months of their submission if they are found to be suitable after the review process. JCDR is published bimonthly and the accepted articles were usually published in the next issue. Recently, due to the increased volume of the submissions, the review process has become slower and it ?? Section can take from 4 to 6 months for the articles to be reviewed. The journal has an extensive author support system and it has recently introduced a paid expedited review process. The journal also mentions the average time for processing the manuscript under different submission systems - regular submission and expedited review.
Strengths of the journal: The journal has an online first facility in which the accepted manuscripts may be published on the website before being included in a regular issue of the journal. This cuts down the time between their acceptance and the publication. The journal is indexed in many databases, though not in PubMed. The editorial board should now take steps to index the journal in PubMed. The journal has a system of notifying readers through e-mail when a new issue is released. Also, the articles are available in both the HTML and the PDF formats. I especially like the new and colorful page format of the journal. Also, the access statistics of the articles are available. The prepublication and the manuscript tracking system are also helpful for the authors.
Areas for improvement: In certain cases, I felt that the peer review process of the manuscripts was not up to international standards and that it should be strengthened. Also, the number of manuscripts in an issue is high and it may be difficult for readers to go through all of them. The journal can consider tightening of the peer review process and increasing the quality standards for the acceptance of the manuscripts. I faced occasional problems with the online manuscript submission (Pre-publishing) system, which have to be addressed.
Overall, the publishing process with JCDR has been smooth, quick and relatively hassle free and I can recommend other authors to consider the journal as an outlet for their work."
Dr. P. Ravi Shankar
KIST Medical College, P.O. Box 14142, Kathmandu, Nepal.
E-mail: ravi.dr.shankar@gmail.com
On April 2011 Anuradha
Dear team JCDR, I would like to thank you for the very professional and polite service provided by everyone at JCDR. While i have been in the field of writing and editing for sometime, this has been my first attempt in publishing a scientific paper.Thank you for hand-holding me through the process.
Dr. Anuradha
E-mail: anuradha2nittur@gmail.com
On Jan 2020
Original article / research
Full Version
Immunohistochemical Expression of FOXP3 and CD8 in Oral Squamous Cell Carcinoma: A Cross-sectional Study
Correspondence Address :
Dr. TN Suresh,
Professor and Head, Department of Pathology, Sri Devaraj Urs Medical College,
Tamaka-563103, Kolar, Karnataka, India.
E-mail: Sureshtn.path@gmail.com
Introduction: In India, Oral Squamous Cell Carcinoma (OSCC) is among the top three malignancies. Tumour-Infiltrating Immune Cells (TIIC), as key elements of tumour microenvironment have recently been demonstrated to be crucial to the development and spread of cancer. The immune infiltrates of tumours is predominantly lymphocytes with T cells being major cell type.
Aim: To determine FOXP3 and CD8 immunohistochemical expression in OSCC and its correlation with Tumour -NodeMetastasis (TNM) staging and lymph node metastases.
Materials and Methods: The present cross-sectional laboratory based study was conducted in the Department of Pathology, Sri Devaraj Urs Medical College, Tamaka, Kolar, Karnataka, India from April 2023 to April 2025. A total of 86 histologically confirmed cases of OSCC were screened for lymph node metastases, graded and TNM staged using the current American Joint Committee on Cancer (AJCC) staging criteria. The cases were then immunohistochemically stained using FOXP3 and CD8. Data analysis was done using Microsoft Excel and Statistical Package for Social Sciences (SPSS) version 22 (IBM SPSS Statistics, Somers, NY, USA). Chi-square test/Fischer’s exact test was employed as the test of significance. The Pearson’s correlation coefficient was used to perform correlations. A p-value <0.05 was deemed statistically significant.
Results: The study patients were predominantly females and the mean age of the patients was 52 years. Nodal metastases were present in 37 (43%) of patients. Among the cases studied, 3 (3.5%), 18 (20.9%), 23 (26.7%),30 (34.9%) and 12 (14%) belonged to stage I, II, III, IVA and IVB, respectively. High CD8+IM (invasive margin) was associated with node negative cases (p=0.0074). Low CD8+IM (invasive margin) was associated with higher TNM stages (p=0.0092). Increased CD8+IM/FOXP3+ ratio was associated with lower TNM stages (p=0.023).
Conclusion: The quantity, location and roles of Tumour Infiltrating Lymphocytes (TIL) are dynamic in nature. CD8+ IM (invasive margin)/FOXP3+ ratio is a promising biomarker in cancer research. Modulating this ratio could enhance antitumour immunity and improve treatment outcomes.
Cluster of differentiation 8, Forkhead box P3, Lymph node metastasis, Oral cancer, Tumour infiltrating lymphocytes, Tumour microenvironment
Cancer of oral cavity is the most frequent among men in India and second among women. It accounts for 29.66% of cancer cases in Kolar, Karnataka, India (1). Majority of the cases are Squamous cell Carcinoma (SCC) (2). Despite a number of treatment options which includes immunotherapy, chemotherapy, radiation and surgery, OSCC has 50% overall survival (3),(4). In order to improve oncological risk stratification, it is noteworthy that recent research indicates an increasing interest in combining the pathogenic and immunological characteristics of different malignancies (5). Also, the infiltration of immune cells in different compartments of tissue is ambiguous and not uniform in OSCC.
The tumour cells surround the stromal cells causing an inflammatory response that recruits various immune cells to aid in the cancer progression (6). The immune system has a complex relationship with cancer cells and can either inhibit or promote tumour growth (7). TIICs are myeloid and lymphoid cells that enter the tumour from the vasculature. Majority of TILs are T-lymphocytes with T-helper cells, cytotoxic T-cells and regulatory T-cells (Treg) subpopulations (8). These are primarily cytotoxic T lymphocytes that are CD8 positive. Nodal negativity and better outcomes are all favourably connected with high CD8+T cells in a number of human malignancies, including SCC of head and neck (9).
Diverse subsets of immunosuppressive T cells make up regulatory T cells (Treg). FOXP3, a regulatory protein belonging to the fork head/ winged-helix family contributes to the development and function of Treg. By inhibiting self-antigen reactive T cells, Tregs can mediate peripheral tolerance (10). T lymphocytes that are receptive to tumour antigens are inhibited by Tregs posing as a barrier to antitumour immunity because majority of tumour antigens are self-antigens (11). Some studies have linked increased intratumoural FOXP3+ expression to worse clinical outcomes (12), while other studies showed contradictory findings with increased infiltration of FOXP3+ regulatory T-cells in OSCC, particularly among never-smokers and never-drinkers, which was paradoxically associated with improved survival (13). Therefore, the present study was conducted to evaluate the immunohistochemical expression of FOXP3 and CD8 in OSCC and to analyse their association with clinicopathological parameters, particularly including tumour stage and nodal involvement.
The present cross-sectional laboratory based study was done in the Department of Pathology, Sri Devaraj Urs Medical College, Tamaka, Kolar, Karnataka, India from April 2023 to April 2025 (Institutional Ethical Clearance number: DMC/KLR/IEC/11/2023-24). The study included a total of 86 histopathologically diagnosed cases of OSCC.
Sample size calculation: Sample size was estimated based on CD8 expression of 66.6% in OSCC as reported in the study done by De Meulenaere A et al., and considering an absolute error of 10% with 95% confidence interval (14). These cases were screened for lymph node metastases, graded and TNM staged using the current AJCC staging criteria followed by immunohistochemical staining using FOXP3 and CD8 (15),(16).
Inclusion and Exclusion criteria: The study included histopathologically diagnosed cases of OSCC treated with composite resection and cervical lymph node dissection.Patients who underwent neoadjuvant radiation and/or chemotherapy before surgery and having second primary cancers were excluded.
Study Procedure
Tissue sections fixed in 10% formalin were subjected to Immunohistochemistry (IHC) staining. Incubation with primary rabbit monoclonal antibody against CD8 (Clone:EP 334, PathnSitu) and primary rabbit monoclonal antibody against FOXP3 (Clone:EP 340, PathnSitu) were done. Sections from tonsil served as positive control for both CD8 and FOXP3.
CD8+ and FOXP3+ IHC scoring: Grading of immunostaining was done according to the criteria used by Mukherjee G et al., where CD8 was scored based on the percentage-based method (16). However, for FOXP3, universally accepted or standardised scoring system was not identified in the literature. To maintain methodological uniformity and enable direct comparison between immune cell populations within the same tumour microenvironment, the same percentage-based scoring approach for both CD8 and FOXP3 expression was adopted. Membranous staining and nuclear staining were considered positive for CD8 and FOXP3, respectively. In each slide, two compartments of the tissue sections were considered i.e., invasive margin of tumour (tumour and normal tissue interface) and the tumour centre (any region within tumour other than the invasive margin). Each section was first inspected at low power magnification to select five hotspots, then viewed under 400x magnification and a mean percentage of immunopositivity score was calculated for each of the sites. It was scored as: low (0-25%), intermediate (26-50%) and high (51-100%). Association of CD8+ and FOXP3+ immunohistochemical score were done with tumour thickness (<40 mm, ≥40 mm) (17) grade of tumour, depth of invasion, lymph node metastases, tumour staging, perineural invasion, bone invasion and lymphovascular invasion.
STATISTICAL ANALYSIS
Microsoft Excel and SPSS version 22 (IBM SPSS Statistics, Somers, NY, USA) was used for analysis of data. Chi-square test/Fischer’s exact test was employed as the test of significance. The Pearson’s correlation coefficient was used to perform correlations. A p-value <0.05 was deemed statistically significant.
The age of the patients included in the present study ranged from 33 to 94 years (mean 52 years), with the majority 62 (72.1%) being female. The tumour subsites were classified as the buccal mucosa 53 (61.6%), gingivobuccal sulcus 21 (24.4%), tongue 9 (10.5%) and retromolar trigone 3 (3.5%). 74 (86%) cases were well-differentiated and 12 (14%) cases moderately-differentiated. Among the cases studied, 3 (3.5%) cases, 29 (33.7%) cases, 30 (34.9%) cases and 24 (27.9%) cases belonged to AJCC classification T1, T2, T3 and T4a stages respectively. 37 (43%) cases had presence of nodal metastasis. 3 (3.5%) cases, 18 (20.9%) cases, 23 (26.7%) cases and 42 (48.9%) cases belonged to stage I, II, III, IV, respectively. CD8+ (IM: invasive margin and TC: tumour centre) and FOXP3+ scoring were correlated with different clinicopathological parameters such as tumour thickness, depth of invasion, histological grade, lymph node status, tumour staging, pathological TNM staging, lymphovascular invasion, perineural invasion and bone invasion (Table/Fig 1), (Table/Fig 2). Low CD8+IM scoring 33 (80.4%) was noted in higher TNM stages (stage III & IV) with a p-value of 0.0092. High CD8+IM (invasive margin) was associated with node negative cases (p=0.0074). No significant correlation was noted among the other parameters. The IHC staining of CD8 and FOXP3 is shown in (Table/Fig 3), (Table/Fig 4) respectively.
Significant association (p=0.023) was noted between the CD8+IM/ FOXP3+ ratio and pathological TNM stage. As the CD8+ IM/ FOXP3+ ratio increases, the tumour stage tends to decrease (Table/Fig 5). This inverse relationship between the immune ratio (CD8+ IM/ FOXP3+) and tumour stage could mean that higher immune activity is associated with lower stage tumours, suggesting a better prognosis or stronger immune response in earlier stages.
No statistically significant correlation (p=0.055, r=0.209) was found between CD8+TC/FOXP3+ ratio and tumour stage. The correlation analysis suggests a weak negative correlation between the CD8+IM/ FOXP3+ ratio and lymph node positivity. However, it was found to be not significant (p=0.093, r=0.181).
Over the past few decades, research on a variety of prognostic histopathological indicators has been captivated by the aggressiveness of OSCC. It also influences the choice of adjuvant treatment. Tumour thickness, depth of invasion, nodal metastasis and pathological TNM stage are the most well-known evaluable important variables linked to survival in OSCC (18).
There is mounting evidence that distinct types of TILs are present in various locations, including peritumoural or intratumoural sites. Thus, the prognosis of individuals may also be influenced by the placement of CD8+T cells. A patient’s baseline tumour immune profile is a major factor for immunotherapy effectiveness (19). Three categories of TME are currently recognised: “immune hot, immune cold and immune altered” (20). CD3+ and CD8+T cells accumulation is seen in the invasive margin and tumour centre in hot immune tumours, only in invasive margin for immune altered tumours whereas cold immune tumours have no T cells (21).
It was observed in the present study that both CD8+IM and CD8+TC low scoring were noted in higher T stages. Even though it was not statistically significant, similar findings were noted in studies done by Lequerica-Fernandez P et al., (22) and Mukherjee G et al., with regards to CD8+ infiltration in invasive margin (16). Meanwhile, CD8+ scoring in tumour centre showed opposite findings to a study done by Lequerica-Fernandez P et al., (22). This could be attributed to difference in the scoring method used. Present study findings on CD8+ TC with regards to T stage correlates with a study done by Mukherjee G et al., (16). These findings suggest how cytotoxic CD8+ T cells contribute to host anti-tumour response.
On comparing FOXP3+ results of the present study, it was not concurrent with other studies done by Lequerica-Fernandez P et al., Song JJ et al., and Hayashi T et al., (22),(23),(24). Present study observed low FOXP3+ scoring found in higher T stages meanwhile other studies suggested otherwise. This could be due to the distinct scoring systems used and FOXP3+ expression varying within tumours affecting the results which can also explain tumour heterogeneity.
High CD8+ scoring at invasive margin was associated with node negative cases (p=0.0074). Similar findings were noted in study done by Mukherjee G et al., (16). In the present study, we observed that low FOXP3+ expression was associated with node negative cases (p=0.2052) which was a contradictory finding when compared to studies done by Lequerica-Fernandez P et al., Song JJ et al., Hayashi T et al., and Ni Y et al., (22),(23),(24),(25). Since there is no established standard method for scoring FOXP3, different studies have used different methods of scoring. It was observed that both CD8+IM (p=0.0092) and CD8+ TC (p=0.5271) low scoring was associated with higher TNM stages. This finding was consistent with Lequerica-Fernandez P et al., study findings when scoring was done in invasive margins (22).
Also, increased CD8+ IM/FOXP3+ ratio was found to be associated with lower TNM stages (p=0.023). This finding strongly concurred with a study done by Ni Y et al., (25). The CD8+IM/FOXP3+ ratio is a promising biomarker in cancer research. Modulating this ratio could enhance anti-tumour immunity and improve treatment outcomes. Understanding the relationship between FOXP3+ expression and tumour stage can help in better understanding of cancer biology.
Lequerica-Fernandez P et al., in their study of 125 OSCC cases evaluated both stromal and tumoural FOXP3+ and emphasised on superiority of FOXP3 with respect to clinical outcomes (22). A considerably better prognosis was linked to higher concentrations of stromal and tumoural FOXP3 (p=0.03 and p=0.03, respectively) among his study group. Meanwhile, low tumoural CD8+/FOXP3+ ratio was also found to significantly associated with a better prognosis (p=0.04) and a low stromal CD8+/FOXP3+ ratio was significantly associated with well-differentiated tumours (p=0.03). These findings were different from studies done by Kirchner J et al., and Kakkar A et al., where they observed high CD8+/FOXP3+ ratio correlating with good clinical outcomes (26),(27).
In the present study, CD8+IM correlated with nodal metastasis and TNM staging. Meanwhile, when considered as a ratio between the two, increased CD8+IM/FOXP3+ ratio associated with lower TNM stages (p=0.023). This shifts the focus on importance of estimating the ratio between the different subpopulations of TILs rather than considering them each independently.
These results underline the notion that the quantity, location and roles of TILs are dynamic in nature and support the existence of various TIL subtypes in OSCC tumour microenvironment interacting with one another to exercise their effects.
Limitation(s)
Being an Institutional based study, the patient population included may not fully represent the broader demographic or clinical spectrum of OSCC, as regional factors could influence disease characteristics. The small sample size may also limit the study’s generalisability and differences in methods of immunohistochemical scoring may affect the results. Multicenter studies involving diverse populations are needed to validate these findings and assess their broader applicability.
FOXP3 and CD8 expression in OSCC may serve as prognostic biomarkers and their interplay in different sites of tumour can shape the TME. With CD8+IM being significantly correlated with nodal metastasis and pathological TNM staging, it signifies its prognostic value in oral cancer. Positive correlation between high CD8+IM/ FOXP3+ ratio and lower TNM stages may also shed light on how tumour cells evade the immune system, with FOXP3 Treg cells potentially contributing to immune suppression and CD8+T cells playing a role in anti-tumour immunity.
DOI: 10.7860/JCDR/2026/86053.24237
Date of Submission: Dec 24, 2025
Date of Peer Review: Feb 27, 2026
Date of Acceptance: May 04, 2026
Date of Publishing: Sep 01, 2026
AUTHOR DECLARATION:
• Financial or Other Competing Interests: None
• Was Ethics Committee Approval obtained for this study? Yes
• Was informed consent obtained from the subjects involved in the study? Yes
• For any images presented appropriate consent has been obtained from the subjects. Yes
PLAGIARISM CHECKING METHODS:
• Plagiarism X-checker: Dec 28, 2025
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ETYMOLOGY: Author Origin
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