Immunocytochemical Profiling of CD4+, CD8+, and CD3+ T Cell Subsets: Clinical Implications in Rheumatic Diseases, Respiratory Disorders, and Oncology-Scilight

Trends in Immunotherapy

Review

Immunocytochemical Profiling of CD4+, CD8+, and CD3+ T Cell Subsets: Clinical Implications in Rheumatic Diseases, Respiratory Disorders, and Oncology

Downloads

Chen, J., Gong, L., Jiang, C., Xie, Y., Wang, Y., Lu, L., & Liu, Q. (2025). Immunocytochemical Profiling of CD4+, CD8+, and CD3+ T Cell Subsets: Clinical Implications in Rheumatic Diseases, Respiratory Disorders, and Oncology. Trends in Immunotherapy, 9(2), 129–142. https://doi.org/10.54963/ti.v9i2.1222

Authors

  • Junlin Chen

    Pathology of Department, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan 430061, China
  • Liangju Gong

    Pathology of Department, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan 430061, China
  • Can Jiang

    Pathology of Department, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan 430061, China
  • Yuxuan Xie

    Pathology of Department, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan 430061, China
  • Ying Wang

    Pathology of Department, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan 430061, China
  • Lu Lu

    Pathology of Department, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan 430061, China
  • Qing Liu

    Pathology of Department, Hubei Provincial Hospital of Traditional Chinese Medicine, Wuhan 430061, China

Received: 9 May 2025; Revised: 19 May 2025; Accepted: 3 June 2025; Published: 8 June 2025

Immunocytochemical staining has emerged as a cost-effective and minimally invasive method for profiling T cell subsets (CD4+, CD8+, and CD3+) in the clinical setting. The quantification of T lymphocyte subsets  has become a critical tool for assessing immune dysregulation in chronic inflammatory and autoimmune diseases. This study evaluates the utility of this method in monitoring immune dysregulation across rheumatic diseases (systemic lupus erythematosus—SLE, rheumatoid arthritis—RA, Sjögren’s syndrome—SS), respiratory disorders, and malignancies. Peripheral blood samples from 12 patients (10 of whom had rheumatic diseases, one with lung cancer tested seven times and one with liver cancer tested five times) were collected in EDTA-K2 tubes and analyzed using standardized immunocytochemical protocols. Key findings include: (1) CD4+/CD8+ ratio inversion (<1.0) correlates with disease activity in SLE (OR = 3.2 for lupus nephritis) and predicts lymphoma risk in SS (HR = 2.8). (2) In RA, reduced Th17 proportions post-methotrexate therapy (12.3%→6.8%, p < 0.01) reflect inflammation suppression, while TNF-α inhibitors improve CD4+/CD8+ ratios alongside DAS28 scores (p = 0.003). (3) In oncology, longitudinal CD3+ T cell counts (136–504 cells/μL) and CD8+ dynamics mirror tumor burden and immunotherapy efficacy. Compared to flow cytometry, immunocytochemical staining offers practical advantages for routine clinical use, though limitations in functional subpopulation resolution persist. Integrating these assays with multi-omics approaches may refine personalized treatment strategies. This study underscores the pivotal role of T cell subset analysis in bridging mechanistic immunology with precision medicine.

Keywords:

Immunocytochemical Staining; CD4+/CD8+ Ratio; T Cell Subsets

References

  1. Abbas, A.K.; Lichtman, A.H.; Pillai, S. Basic Immunology: Functions and Disorders of the Immune System; Elsevier Health Sciences: Philadelphia, PA, USA, 2015.
  2. Getz, G.S. Bridging the Innate and Adaptive Immune Systems. J. Lipid Res. 2005, 46, 619–622.
  3. Sun, L.; Su, Y.; Jiao, A.; et al. T Cells in Health and Disease. Signal Transduct. Target. Ther. 2023, 8, 235.
  4. Wang, M.; Windgassen, D.; Papoutsakis, E.T. Comparative Analysis of Transcriptional Profiling of CD3+, CD4+ and CD8+ T Cells Identifies Novel Immune Response Players in T-Cell Activation. BMC Genom. 2008, 9, 1–16.
  5. Li, T. K.; Bao, Y.; Chen, Z.; et al. Changes and Clinical Significance of Cellular Immune Function in Oral Cancer Patients Before and After Surgery. Chin. J. Gerontol. 2018, 38, 827–829.
  6. Tao, C.; Liu, C.; Ge, P.; et al. T Cells Immune Imbalance Presents in Patients with Multiple Intracranial Aneurysms. Clin. Exp. Immunol. 2025, 219, uxae058.
  7. Chen, C.; Liu, Y.M.; Xuan, S.X.; et al. Establishment and Clinical Application of a Method for Detecting T Lymphocyte Subsets by Cellular Immunochip Technology. J. Inflamm. Res. 2021, 14, 7529–7537.
  8. Rybtsova, N.; Berezina, T.N.; Rybtsov, S. Molecular Markers of Blood Cell Populations Can Help Estimate Aging of the Immune System. Int. J. Mol. Sci. 2023, 24, 5708.
  9. Liarski, V.M.; Sibley, A.; van Panhuys, N.; et al. Quantifying In Situ Adaptive Immune Cell Cognate Interactions in Humans. Nat. Immunol. 2019, 20, 503–513.
  10. Casalegno Garduño, R.; Spitschak, A.; Pannek, T.; et al. CD8+ T Cell Subsets as Biomarkers for Predicting Checkpoint Therapy Outcomes in Cancer Immunotherapy. Biomedicines 2025, 13, 930.
  11. Yan, H.L.; Ju, X.L.; Huang, A.L.; et al. Advancements in Technology for Characterizing the Tumor Immune Microenvironment. Int. J. Biol. Sci. 2024, 20, 21–51.
  12. Zhou, J. M.; Zhang, Y. Plant Immunity: Danger Perception and Signaling. Cell 2020, 181, 978–989.
  13. Datta, P.; Ukey, R.; Bruiners, N.; et al. Highly Versatile Antibody Binding Assay for the Detection of SARS-CoV-2 Infection and Vaccination. J. Immunol. Methods 2021, 499, 113–165.
  14. Knuutila, S.; Teerenhovi, L. Immunophenotyping of Aneuploid Cells. Cancer Genet. Cytogenet. 1989, 41, 1–17.
  15. Kazazi, F.; Mathijs, J.-M.; Foley, P.; et al. Variations in CD4 Expression by Human Monocytes and Macrophages and Their Relationship to Infection with the Human Immunodeficiency Virus. J. Gen. Virol. 1989, 70, 2661–2672.
  16. Sahir, F.; Mateo, J.M.; Steinhoff, M.; et al. Development of a 43 Color Panel for the Characterization of Conventional and Unconventional T‐Cell Subsets, B Cells, NK Cells, Monocytes, Dendritic Cells, and Innate Lymphoid Cells Using Spectral Flow Cytometry. Cytom. Part A 2024, 105, 404–410.
  17. Zhang, H.; Wu, T. CD4+ T, CD8+ T Counts and Severe COVID-19: A Meta-Analysis. J. Infect. 2020, 81, e82–e84.
  18. Zhu, X.; Gadgil, A.S.; Givelber, R.; et al. Peripheral T Cell Functions Correlate with the Severity of Chronic Obstructive Pulmonary Disease. J. Immunol. 2009, 182, 3270–3277.
  19. Mikko, M.; Fredriksson, K.; Wahlström, J.; et al. Human T Cells Stimulate Fibroblast-Mediated Degradation of Extracellular Matrix in Vitro. Clin. Exp. Immunol. 2008, 151, 317–325.
  20. Søyseth, V.; Šaltytė Benth, J.; Stavem, K. The Association Between Hospitalisation for Pneumonia and the Diagnosis of Lung Cancer. Lung Cancer 2007, 57, 152–158.
  21. Nair, G.B.; Niederman, M.S. Community-Acquired Pneumonia: An Unfinished Battle. Med. Clin. 2011, 95, 1143–1161.
  22. Zhang, S.; Asquith, B.; Szydlo, R.; et al. Peripheral T Cell Lymphopenia in COVID-19: Potential Mechanisms and Impact. Immunother. Adv. 2021, 1, ltab015.
  23. Folds, J.D.; Schmitz, J.L. 24. Clinical and Laboratory Assessment of Immunity. J. Allergy Clin. Immunol. 2003, 101, 702–711.
  24. Yang, L.; Zheng, R.; Wang, N.; et al. Incidence and Mortality of Stomach Cancer in China, 2014. Chin. J. Cancer Res. 2018, 30, 291.
  25. Grivennikov, S.I.; Greten, F.R.; Karin, M. Immunity, Inflammation, and Cancer. Cell 2010, 140, 883–899.
  26. Mellman, I.; Coukos, G.; Dranoff, G. Cancer Immunotherapy Comes of Age. Nature 2011, 480, 480–489.
  27. Sadeghi Rad, H.; Monkman, J.; Warkiani, M.E.; et al. Understanding the Tumor Microenvironment for Effective Immunotherapy. Med. Res. Rev. 2021, 41, 1474–1498.
  28. Lu, W.; Mehraj, V.; Vyboh, K.; et al. CD4: CD8 Ratio as a Frontier Marker for Clinical Outcome, Immune Dysfunction and Viral Reservoir Size in Virologically Suppressed HIV‐Positive Patients. J. Int. AIDS Soc. 2015, 18, 20–52.
  29. Panos, R.; Eschenbacher, W. A COPD Primer; Walter de Gruyter GmbH & Co KG: Berlin, Germany, 2015.
  30. Arida, A.; Protogerou, A.D.; Kitas, G.D.; et al. Systemic Inflammatory Response and Atherosclerosis: The Paradigm of Chronic Inflammatory Rheumatic Diseases. Int. J. Mol. Sci. 2018, 19, 18–90.
  31. Zamanpoor, M. The Genetic Pathogenesis, Diagnosis and Therapeutic Insight of Rheumatoid Arthritis. Clin. Genet. 2019, 95, 547–557.
  32. Sakkas, L.I.; Bogdanos, D.P. Infections as a Cause of Autoimmune Rheumatic Diseases. Autoimm. Highlights 2016, 7, 1–8.
  33. Keogan, M.; Wallace, E.M.; O'Leary, P. Concise Clinical Immunology for Healthcare Professionals; Routledge: London, UK, 2006.
  34. Sun, C.; Pan, Q.; Du, M.; et al. Decoding the Roles of Heat Shock Proteins in Liver Cancer. Cytokine Growth Factor Rev. 2024, 75, 81–92.
  35. Sivalingam, A.M. Emerging Mechanisms and Biomarkers Associated with T-Cells and B-Cells in Autoimmune Disorders. Clin. Rev. Allergy Immunol. 2025, 68, 14.