How to deplete CD8+ T Cells in vivo

1. Understanding CD8+ T Cell Depletion

The fall or removal of CD8+ T cells, a form of T cell depletion, leads to the recognition and eradicating of various abnormal cells, virally infected cells, and tumor cells because CD8+ T cells are essential components of the immune defense system [1]. There are several ways to eliminate T cells, but antibody-based depletion has become a breakthrough in the respective sector. The antibodies attached to CD8+ particles stimulate their exclusion or functionally inactivity on the T cell’s surface [2]. 

2. Importance of CD8+ T Cell Depletion in Vivo Research

The CD8+ T cell depletion studies support exploring their contributions to several pathological and physiological mechanisms and their effects on disease development, immune response, and complete health conditions by removing these T cells [3]. Furthermore, by modifying immune system components, preclinical research with CD8+ T cell depletion applications helps to investigate various immunotherapies’ effectiveness, including adoptive T cell transfer and cancer vaccines [4]. 

3. Antibody Clones for CD8+ T Cell Depletion

Among the antibodies used for CD8+ T cell depletion, clone 2.43 is one of the most widely used and well-characterized. This monoclonal antibody targets the CD8α chain, an essential protein on CD8+ T cells. Administering clone 2.43 in vivo successfully depletes CD8+ T cell populations, facilitating detailed investigations into the consequences of their absence [5, 6]. 

Advantages of Clone 2.43:

High specificity: 2.43 has a solid binding affinity for CD8α, ensuring selective depletion of CD8+ T cells without affecting other immune cells.

Established efficacy: Numerous preclinical studies have demonstrated the effectiveness of clone 2.43 in depleting CD8+ T cells across various models and species.

Versatility: 2.43 is compatible with multiple animal models, including mice and rats, making it suitable for diverse research applications.

Pitfalls of Clone 2.43:

Potential off-target effects: 2.43 may cause off-target binding or unintended immunomodulatory effects despite its specificity. 

Immunogenicity: Repeated administration of monoclonal antibodies, including clone 2.43, can elicit immune responses in the host, affecting long-term study reliability.

4. Dosing Range for Clone 2.43 in CD8+ T Cell Depletion

The dosing of clone 2.43 for effective CD8+ T cell depletion can vary based on factors such as the animal model, the route of administration, and the specific experimental conditions.

Generally, the following dosing ranges are recommended:

Mice: The typical dose range for depleting CD8+ T cells in mice using clone 2.43 is between 100 µg to 500 µg per mouse. This dosage is usually administered intraperitoneally or intravenously. Researchers often use 250 µg per mouse as a standard dose for effective depletion.

Rats: The dosing for rat models may need adjustment, and researchers typically use 0.5 mg to 2 mg per rat, administered intraperitoneally.

Frequency: The dosing schedule can also vary; standard protocols involve administering the antibody every 3 to 7 days to maintain effective depletion. Some studies use a single high dose followed by lower maintenance doses.

5. Alternative Antibody Clones

Researchers may consider other clones for CD8+ T cell depletion, particularly for different animal species or specific research needs.

Clone YTS 169.4: Commonly used in mouse models, it targets the CD8α chain and provides effective depletion.

Clone H35-17.2: Suitable for rat models, this antibody also targets the CD8α chain.

Clone OKT8: Frequently used in humanized mouse models, targeting the human CD8α molecule.

Clone 53-6.7: Another well-established clone for mouse models, targeting CD8α with high specificity.

6. Recent Studies on CD8+ T Cell Depletion Using Clone 2.43

Elong et al. (2017) demonstrated the impacts of antibody-oriented depletion of CD8+ T cells in a mouse model. Clone 2.43 was used to investigate the elimination of T cells in the virally infected and its effects on illness and immunological response due to the lack of CD8+ T cells [7].  

Another study by Zhu et al. (2024) and Ma et al. (2015) reported that CD8+ T cells can be targeted for monoclonal antibody 2.43 in treating autoimmune diseases. This investigation explains targeted immunomodulation efficiency and T cell-mediated pathophysiology regarding possible CD8+ T cell depletion [8, 9]. 

 According to Cross et al. (2019) and Liu et al. (2016), A comparative study evaluated CD8+ T cell depletion strategies using clones 2.43 and 3.56 in the animal model, offering insights into optimal depletion protocols for translational research applications [10, 11]. 

Therefore, using antibody clone 2.43 for CD8+ T cell depletion in vivo and considering its advantages, pitfalls, dosing strategies, and alternative options, researchers can design robust experiments to elucidate the complex roles of CD8+ T cells in health and disease.

7. Depleting Cells in vivo

For our other guides on how to deplete immune cells in vivo click here. 

8. References 

1.  Xie Q, Ding J, Chen Y. Role of CD8+ T lymphocyte cells: Interplay with stromal cells in tumor microenvironment. Acta Pharm Sin B. 2021 Jun;11(6):1365-1378. 

2.  Raskov H, Orhan A, Christensen JP, Gögenur I. Cytotoxic CD8+ T cells in cancer and cancer immunotherapy. Br J Cancer. 2021 Jan;124(2):359-367. 

3. Nath PR, Pal-Nath D, Kaur S, Gangaplara A, Meyer TJ, Cam MC, Roberts DD. Loss of CD47 alters CD8+ T cell activation in vitro and immunodynamics in mice. Oncoimmunology. 2022 Sep 6;11(1):2111909. 

4.  Restifo NP, Dudley ME, Rosenberg SA. Adoptive immunotherapy for cancer: harnessing the T cell response. Nat Rev Immunol. 2012 Mar 22;12(4):269-81. 

5.  Tavaré R, McCracken MN, Zettlitz KA, Knowles SM, Salazar FB, Olafsen T, Witte ON, Wu AM. Engineered antibody fragments for immuno-PET imaging of endogenous CD8+ T cells in vivo. Proc Natl Acad Sci U S A. 2014 Jan 21;111(3):1108-13. 

6.  White JM, Keinänen OM, Cook BE, Zeglis BM, Gibson HM, Viola NT. Removal of Fc Glycans from [89Zr]Zr-DFO-Anti-CD8 Prevents Peripheral Depletion of CD8+ T Cells. Mol Pharm. 2020 Jun 1;17(6):2099-2108.

7. Elong NA, Vizcarra EA, Tang WW, Sheets N, Joo Y, Kim K, Gorman MJ, Diamond MS, Shresta S. Mapping and Role of the CD8+ T Cell Response During Primary Zika Virus Infection in Mice. Cell Host Microbe. 2017 Jan 11;21(1):35-46. 

8.  Ma L, Simpson E, Li J, Xuan M, Xu M, Baker L, Shi Y, Yougbaré I, Wang X, Zhu G, Chen P, Prud'homme GJ, Lazarus AH, Freedman J, Ni H. CD8+ T cells are predominantly protective and required for effective steroid therapy in murine models of immune thrombocytopenia. Blood. 2015 Jul 9;126(2):247-56. 

9.  Zhu HX, Yang SH, Gao CY, Bian ZH, Chen XM, Huang RR, Meng QL, Li X, Jin H, Tsuneyama K, Han Y, Li L, Zhao ZB, Gershwin ME, Lian ZX. Targeting pathogenic CD8+ tissue-resident T cells with chimeric antigen receptor therapy in murine autoimmune cholangitis. Nat Commun. 2024 Apr 5;15(1):2936. 

10.  Cross EW, Blain TJ, Mathew D, Kedl RM. Anti-CD8 monoclonal antibody-mediated depletion alters the phenotype and behavior of surviving CD8+ T cells. PLoS One. 2019 Feb 8;14(2): e0211446. 

11.  Liu J, Xiao Q, Zhou R, Wang Y, Xian Q, Ma T, Zhuang K, Zhou L, Guo D, Wang X, Ho WZ, Li J. Comparative Analysis of Immune Activation Markers of CD8+ T Cells in Lymph Nodes of Different Origins in SIV-Infected Chinese Rhesus Macaques. Front Immunol. 2016 Sep 21; 7:371.