Role of Autoreactive Tc17 Cells in the Pathogenesis of Experimental Autoimmune Encephalomyelitis (EAE)

doi:10.21203/rs.3.rs-3382546/v1

Download PDF

Research Article

Role of Autoreactive Tc17 Cells in the Pathogenesis of Experimental Autoimmune Encephalomyelitis (EAE)

https://doi.org/10.21203/rs.3.rs-3382546/v1

This work is licensed under a CC BY 4.0 License

Journal Publication

published 18 Mar, 2024

Read the published version in Neuroprotection →

Version 1

posted

You are reading this latest preprint version

Background

The pathogenesis of multiple sclerosis (MS) and its animal model, experimental autoimmune encephalomyelitis (EAE), is primarily mediated by T cells. However, recent studies have only focused on CD4 + T-helper cells that secrete interleukin-17 (IL-17), also known as Th17 cells. This study aims to determine the similarities and differences between Th17 cells and CD8⁺ T-cytotoxic cells that secrete IL-17 (Tc17) in the context of MS/EAE.

Methods

Female C57BL/6 mice (n = 20) were immunized with myelin oligodendrocyte glycoprotein peptides 35–55 (MOG_{35 − 55}), pertussis toxin, and Complete Freund’s adjuvant to establish the EAE animal model. T-cells were isolated from the spleen (12–14 days post-immunization) and purified into CD4⁺ and CD8⁺ using flow cytometry. These cells were differentiated into Tc17 and Th17 using MOG₃₅-₅₅ and IL-23. Secretion levels of interferon-γ (IFN-γ) and IL-17 were measured via enzyme-linked immunosorbent assay (ELISA) using cultured CD4⁺ and CD8⁺ T-cells supernatant. Pathogenicity of Tc17 and Th17 cells was tested through adoptive transfer (tEAE), with the clinical course assessed using an EAE score (0–5). Hematoxylin and eosin and Luxol fast blue staining were used to examine the spinal cord.

Results

Purified CD8⁺CD3⁺ and CD4⁺CD3⁺ cells were differentiated into Tc17 and Th17 cells, and then stimulated with MOG_{35 − 55} peptide for proliferation assays. The results showed that Tc17 cells exhibited a weaker response to MOG_{35 − 55} compared to Th17 cells. However, this response was not dependent on Th17 cells. Tc17 cells secreted lower levels of IFN-γ and IL-17. In the tEAE mouse model, similar EAE scores and slight inflammation and demyelination were observed in Tc17 cell-induced tEAE mice compared to Th17 cell-induced tEAE mice.

Conclusion

Although Tc17 cells were pathogenic in EAE, their degree of pathogenicity was lower than that of Th17 cells. Tc17 cells secreted similar levels of IL-17 to Th17 cells after antigen stimulation, but their IFN-γ secretion was significantly lower.

Tc17 cells

Experimental autoimmune myelitis

Multiple sclerosis

CD8+ T cells

IL-17

IFN- γ

Multiple sclerosis (MS) is an inflammatory disease that primarily damages the white matter of the central nervous system (CNS), yet its pathogenesis remains unclear.[1, 2] In the past few decades, MS and experimental autoimmune encephalomyelitis (EAE) have been recognized as typical CD4⁺ T cell-mediated autoimmune diseases.[3] Multiple studies, including our previous research, have demonstrated the involvement of CD8⁺ T cells in different stages of MS/EAE,[4] and have shown that the number of CD8⁺ T cells in the brain lesion area, perivascular cuff, and white matter area is greater than that of CD4+ T cells.[3, 5] Given their critical role in the pathogenesis of MS, CD8⁺ T cells may be a significant target for MS/EAE treatment.

Interleukin (IL)-17⁺ CD4⁺ T helper (Th17) cells and their related antibodies (Abs) have the potential for treating autoimmune diseases, including MS/EAE.[6-13] Although secukinumab, ixekizumab, and brodalumab have been approved for the treatment of other diseases,[14-18] clinical trials of these antibodies for MS have failed.[19-22] Therefore, other T cell subsets, particularly CD8+ T cells and their subsets, should be considered.

Recent studies have identified several subsets of CD8⁺T cells, including T cytotoxic (Tc) cells such as Tc1 (dominant IFN-g-secretion), Tc2 (dominant IL-4 and IL-5-secretion), Tc9 (dominant IL-9-secretion), Tc17 (dominant IL-17-secretion), and regulatory T cells (Tregs, dominant IL-10 and/or Foxp3 expression).[23, 24] Tc17 cells, discovered in 2009 in humans and mice;[25-27] are now a focal point in global immunology research, with an emphasis on their role in pathogenesis and potential new treatment methods for tumor immunity, rejection in organ transplantation, AIDS, and autoimmune diseases.[28-34]

Few studies have also reported on the role of Tc17 cells in MS/EAE.[35-38] These cells have been detected in the peripheral blood (PB), cerebrospinal fluid (CSF), and active lesions in patients with MS.[39-41] Dimethyl fumarate (DMF) treatment for MS can reduce the frequency of Tc17 expression in PB mononuclear cells, decrease the ratio of the regulatory factor RORC to TBX21, and boost its expression.[42-47] Unlike Tc1 cells, Tc17 cells are not cytotoxic; however, they can reduce the expression of T-bet and Eomes (Tc1 cell-specific transcription factors) and increase the expression of IFN-g, cytotoxic granzyme B, and perforin.[48-50]

In our previous study, we demonstrated that CD8⁺ T cells purified from patients with EAE had antigenic specificity and induced encephalitis,[4] similar to our findings on CD8⁺ T cells in experimental autoimmune uveitis (EAU).[51-53] Interestingly, antigen-specific CD8⁺ T cells also showed expression of IL-17 in EAU. While IL-23 can induce antigen-specific Th17 cells in EAU, TGF-β, and IL-6 can only induce non-antigen-specific Th17 cells and are not pathogenic.[54] However, our previous study had several limitations, which required further research:(1) The study of EAE was limited to the cytological characteristics and pathogenicity of CD8+ T cells and did not involve subsets, particularly Tc17 cells, and their associated antibodies and antagonists. (2) Although our research on EAU provided a good idea and methodological basis for this study, the results on EAE may differ due to differences in target organs and antigens. Therefore, further research and validation using the EAE model are necessary.

In this study, we aimed to investigate the role of Tc17 cells in EAE and examine the effects of different antagonists on their cytological characteristics and pathogenicity. Furthermore, we compared the similarities, differences, and interactions between Tc17 and Th17 cells based on these characteristics. Our objective was to provide new research directions for understanding the pathogenesis of MS/EAE and developing targeted cell therapy. As a result, this study holds significant theoretical implications and potential applications.

2.1. Ethics approval

All experiments were conducted at the Science and Technology Innovation Center of the Hunan University of Chinese Medicine in Changsha, China. The experimental procedures were approved by the Ethics Committee of the Hunan University of Chinese Medicine (Approval No. LL2022050501). The animal treatments were conducted in accordance with the guidelines provided by the National Institutes of Health Guide for the Care and Use of Laboratory Animals, and all necessary steps were taken to minimize the pain and suffering of the animals.

2.2. Selection of animal participants

Specific pathogen-free female C57BL/6 mice (n = 20, aged 10–14 weeks) were purchased from Hunan Slake Jingda Laboratory Animal Co., Ltd. (Changsha, Hunan, China; License no. SCXK [Hunan] 2019-0004) and housed in the Center of Experimental Animals at the Hunan University of Chinese Medicine (License no. SYXK [Hunan] 2019-0009). The mice were housed individually in cages with independent ventilation systems at a temperature of 20–26°C and relative humidity of 40–60%. They were provided with free access to food and water, and steps were taken to ensure minimal pain and suffering. The sample size was determined based on the experimental design requirements.

2.3. Antigen-induced EAE (aEAE) models

The aEAE models were established in our previous study.[4] To actively induce disease in mice, we injected a 200 mL emulsion containing 200 µg of myelin oligodendrocyte glycoprotein peptides 35–55 (MOG_35–55; amino acids 35–55 of bovine MOG; Sigma-Aldrich, St. Louis, MO, USA) purified using high-performance liquid chromatography with a purity of > 95%, and Complete Freund’s adjuvant (CFA) (Sigma-Aldrich, USA) subcutaneously over six sites at the tail base and on the flank. At 0 and 24 h after immunization, mice were injected intraperitoneally with pertussis toxin (List Biological Laboratories, Campbell, CA). EAE scores were assigned on a scale of 0–5 as follows: 0, no obvious changes in motor functions of the mouse compared with non-immunized mice; 1, limp tail; 2, limp tail and weakness of hind limbs; 3, limp tail and complete paralysis of hind limbs (most common) or limp tail with paralysis of one front and one hind limb; 4, complete hind limb and partial front limb paralysis; and 5, death or euthanasia due to severe paralysis.[55]

2.4. Purification of MOG_35–55-specific CD4⁺ and CD8⁺ T cells

This procedure was performed according to our previously published protocol.[4, 54] The EAE mice were sacrificed 14 days after immunization using sodium pentobarbital (50 mg/kg; Sinopharm Chemical Reagent Co., Beijing, China), and T cells were isolated from the spleen by passing them through a nylon wool column (Kisker, Steinfurt, Germany). CD4⁺ and CD8⁺ T cells were purified from the spleen using CD4 and CD8 isolation kits (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany), respectively. The spleen cells were incubated in 10 mL of CD8 MicroBeads and 90 mL of buffer per 1 × 10⁷ cells for 15 min in a refrigerator (2–8°C). The buffer was added to make up a final volume of 500 mL, and 5 × 10⁷ cells were obtained. Then, the cells were separated into bound and unbound cells on a magnetic separator column (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) and washed with 15 mL of medium which is according to the manufacturer’s protocol. The flow-through fraction containing CD4- or CD8-enriched cells was collected, and the purity of the isolated cell fraction was determined by flow cytometry.

In accordance with our previously published protocol,[4] we double-stained aliquots of 2 x 10⁵ cells with combinations of monoclonal antibodies against CD3, CD4, or CD8, conjugated with allophycocyanin (APC)–fluorescein isothiocyanate (FITC) or phycoerythroprotein (PE), following the manufacturer's instructions (BioLegend, San Diego, CA, USA). Data collection and analysis were performed using a BD LSRFortessa 4 flow cytometer (BD Biosciences, San Jose, CA, USA), and data were analyzed using FlowJo software (FlowJo Co., San Diego, CA, USA).

2.5. Preparation of MOG_35–55-specific Th1, Tc1, Th17 and Tc17 cells

For the preparation of Th1 and Tc1 cells, 1 × 10⁷ purified CD4⁺ and CD8⁺ T cells were mixed with 2 mL of Roswell Park Memorial Institute (RPMI) 1640 medium per well in a six-well plate (Costar; Corning, Corning, NY, USA) were stimulated with 20 mg/mL MOG_35–55, in the presence of 1 × 10⁷ mitomycin C (MMC; Med Chem Express, NJ, USA)-treated syngeneic spleen cells as antigen-presenting cells (APCs).[4] Activated lymphoblasts were isolated after 2 days using density gradient centrifugation (Lymphocyte Separation, Tianjin, China) and cultured in RPMI 1640 medium containing 10 ng/mL IL-2 (USCN Co., Wuhan, Hubei, China).

Similarly, for preparation of Th17 and Tc17 cells, 1 × 10⁷ purified CD4⁺ and CD8⁺ T cells in 2 mL RPMI 1640 medium per well in a six-well plate were stimulated with 20 mg/mL MOG_35–55 in the presence of 1 × 10⁷ MMC-treated syngeneic APCs plus 10 ng/mL IL-23 (R&D Systems, Minneapolis, MN, USA). Activated lymphoblasts were isolated after 2 days using density gradient centrifugation and cultured in RPMI 1640 medium containing 10 ng/mL IL-23.[54]

2.6. Proliferation assay

As per our previously published protocol,[4, 54] MOG_35–55-specific Th1, Tc1, Th17, and Tc17 cells were prepared and seeded in 96-well plates, at 4 × 10⁵ cells/ well. The cells were cultured in 200 mL medium with or without MOG_35–55, in the presence of MMC-treated syngeneic APCs (1 × 10⁵), at 37°C for 48 h. [³H]-thymidine incorporation during the last 8 h was assessed using a microplate scintillation counter (Packard; PerkinElmer, Meriden, CT, USA). The proliferative response was expressed as the mean count per minute ± standard deviation (SD) of triplicate determinations.

2.7. Enzyme-linked immunosorbent assays (ELISA)

As previously described,[4, 54] for ELISA, 1 × 10⁶ MOG_35–55-specific Th1, Tc1, Th17, and Tc17 cells mixed in 1 mL of RPMI 1640 medium per well in a 24-well plate (Costar; Corning, Corning, NY, USA) were stimulated with 20 mg/mL MOG_35–55 in the presence of MMC-treated syngeneic spleen cells as APCs. After 48, 96, and 144 h, IFN-γ and IL-4 secretion in the supernatants of cultured MOG_35–55-specific T cells of four types were measured using commercially available ELISA kits (USCN Co., Wuhan, Hubei, China).

2.8. Adoptive-transferred EAE models (tEAE)

For adoptive transfer, C57BL/6 mice were injected via the caudal vein with 5 × 10⁶ MOG_35–55-specific Th1, Tc1, Th17, and Tc17 cells, prepared in 0.2 mL phosphate buffer saline (PBS), as described previously.[4, 54]

2.9. Histological study

Spinal cord sections were prepared from tissues collected from tEAE animals, that were adoptively transferred with MOG_35–55-specific Th1, Tc1, Th17, and Tc17 cells from aEAE animals.[4, 54] Spinal cord tissues were fixed in 4% ice-cold paraformaldehyde/methanol solution and embedded in paraffin, before preparing them into microtome sections (5 mm thick) for staining with hematoxylin and eosin (H&E) and Luxol Fast Blue (LFB) to stain myelin.

2.10. Statistical analysis

Statistical analyses were performed using SPSS 25.0 statistical software (IBM, Armonk, NY, USA). All data were statistically analyzed using an unpaired Student’s t-test. Data for the proliferation assay, ELISA, and EAE score were expressed as mean ± SD. Each experiment was repeated at least thrice. P-value < 0.05 was considered statistically significant.

3.1. Tc17 cells are MOG_35–55 specific

The four types of T cells (Th1, Tc1, Th17, and Tc17) were assessed for their antigen-specific functions using a proliferation assay. The results presented in Fig. 1 and Table 1 demonstrated that the MOG_35–55 peptides exhibited a potent stimulatory effect on all four types of T cells at the highest dose (20 µg/mL, P = 0.000) but not at lower doses (0.2 or 2 µg/mL, P = 0.741 or P = 0.502) or in the absence of peptides (0 µg/mL P = 0.262). This suggests that all four types of T cells were specific to MOG_35–55, which is in line with our earlier reports.[51, 53, 54, 56]

At the highest dose (20 µg/mL) of MOG_35–55 peptide, all four types of T cells displayed distinct responses, as depicted in Table 1. The counts per minute (cpm) of Th1 (55630 ± 4077) and Th17 cells (55709 ± 4196)—derived from CD4⁺ T cells—were much higher than those of Tc1 (15368 ± 3533; Th1 vs Tc1, P = 0.00) and Tc17 (15951 ± 1985; Th17 vs Tc17, P = 0.00)—derived from CD8⁺ T cells (Fig. 1C and 1D). These findings are consistent with those of our previous reports.[4, 51, 53, 54, 56] Interestingly, there were no significant differences between Th1 and Th17 cells (Th1 vs Th17: P = 0.979), or between Tc1 and Tc17 cells (Tc1 vs Tc17: P = 0.846) (Fig. 1A and 1B). This leads us to question the distinction between Th1 and Th17, as well as Tc1 and Tc17. To address this, we examined the cytokine profiles of all four types of T cells.

3.2. Cytokine profiles of MOG _35–55 -specific Tc17 and Th17 cells are similar, but Tc17 profiles are weaker than Th17 cells

To determine the cytokine profiles of activated Th1, Tc1, Th17, and Tc17 cells, we used ELISAs to measure the cytokine levels in the culture supernatants of these cells at three-time points (48, 96, and 144 h) post-stimulation.

3.2.1. IFN-γ secretion

At the three time points, the IFN-γ secretions from all four types of MOG_35–55-peptide-activated T cells were increased only at the highest dose (20 µg/mL) of MOG_35–55 peptide (P = 0.00), but not at the lower doses (0.2 or 2 µg/mL, P = 0.144 or P = 0.170) or at no peptide dose (0 µg/mL, P = 0.135) (P༞0.05, for example, at 48 h) (Fig. 2A–C). These results are similar to those of our previous studies.[51, 53, 54, 56]

Interestingly, after 48 h post-stimulation, IFN-γ secretions (pg/mL) of Th1 (1113.33 ± 64.30) and Th17 cells (556.67 ± 15.28) were much higher than Tc1 (563.35 ± 15.28; Th1 vs Tc1: P = 0.00) and Tc17 (280.00 ± 15.00; Th17 vs Tc17: P = 0.00) cells, respectively (Table 2A and Fig. 2A). In addition, similar patterns were seen in IFN-γ secretions at 96 and 144 h (Table 2B and 2C, and Fig. 2B and 2C, respectively). These results are similar to those of our previous studies.[4, 51, 53, 54, 56]

Moreover, at the highest dose (20 µg/mL) of MOG_35–55 peptide, IFN-γ secretion by Th1 cells was much higher than that by Th17 cells (Th1 vs Th17: P = 0.00, at 48, 96, and 144 h)), as well as by Tc1 and Tc17 (Tc1 vs Tc17: P = 0.00 at 48, 96, and 144 h) (Fig. 2G).

3.2.2 IL-17 secretion

At the three time points, IL-17 secretion from all four types of MOG_35–55-peptide-activated T cells was increased only at the highest dose (20 µg/mL) of MOG_35–55 peptide (P = 0.00), but not at lower doses (0.2 or 2 µg/mL,) or at no peptide (0 µg/mL, P = 0.382) (P༞0.05, for example, at 48 h). These results are similar to those of our previous study.[54]

After 48 h, IL-17 secretions (pg/mL) of Th1 (238.67 ± 25.72) and Th17 cells (288.33 ± 12.58) were much higher than Tc1 (96.00 ± 5.29; Th1 vs Tc1: P = 0.00) and Tc17 (102.67 ± 5.86; Th17 vs Tc17: P = 0.00) cells, respectively (Table 2D and Fig. 2D). Similar patterns were observed in IFN-γ secretion at 96 and 144 h, as shown in Table 2E and 2F, and Fig. 2E and 2F.

In addition, at the highest dose (20 µg/mL) of the MOG_35–55 peptide, IL-17 secretion from Th1 cells was much lower than that from Th17 cells (Th1 vs Th17: P = 0.00 at 48, 96, and 144 h), and from Tc1 and Tc17 cells (Tc1 vs Tc17: P = 0.00 at 48, 96, and 144 h) (Fig. 2H).

3.3. Adoptive transfer of MOG_35–55-specific Tc17 cells to naïve mice induces tEAE

To determine the role of MOG_35–55-specific Tc17 cells in the pathogenesis of EAE, we induced the disease in wild-type B6 naive mice by adoptive transfer of MOG_35–55-specific Th1, Tc1, Th17, and Tc17 cells from B6 aEAE mice, and then measured the clinical signs using the EAE score assessed by daily checks and histopathology. As shown in Fig. 3A, the EAE score after transferring Tc17 cells from immunized mice to naive mice (Tc17-tEAE) was similar to that after transferring Tc1 cells (Tc17-tEAE). However, these scores were slightly lower than the EAE score of Th17-tEAE and Th1-tEAE.

Histopathological examination revealed slight inflammation [H&E staining, Fig. 3B–3E] and obvious demyelination [LFB staining, Fig. 3F–3I] in naïve B6 mice transferred with activated Th1, Tc1, Th17, and Tc17 cells from immunized B6 mice, which is consistent with our previous observations.

Our previous reports have shown that in the EAE model, MOG_35–55 peptide-specific CD8⁺ T cells were antigen-specific and encephalitogenic, with IFN-γ production, but were weaker than their CD4⁺ counterparts. However, the subtype of CD8⁺ T cells that plays a critical role in the pathogenesis of EAE remained unclear. Currently, the major subsets of CD8⁺ T cells are Tc1, Tc2, Tc9, Tc17, and Treg cells.[23, 24] Among these subsets, Tc17 cells are strongly associated with autoimmune diseases in both human and animal models.[28–34]

In the present study, we used our already-established strategy for the preparation of the EAE model to study the role of MOG_35–55-specific Tc17 cells in the pathogenesis of EAE and MS.[51, 53, 54, 56] Here, we focused on Tc17 cells in EAE and used Th17 cells (CD4⁺ counterparts of Tc17) and Tc1 cells (different subsets of CD8⁺) as controls. We found that Tc17 cells responded to MOG_35–55 peptides and secreted IFN-γ and IL-17, as well as could induce tEAE in naïve mice models. However, we observed that the proliferation assay results, cytokine secretion, and encephalitogenic activities of Tc17 cells are weaker than those of their Th17 counterparts. Compared with MOG_35–55-specific Tc1 cells which we previously explored, MOG_35–55-specific Tc17 cells showed similar proliferation assay results, lower IFN-γ and higher IL-17 secretion, and weaker encephalitogenic. The results of this study are consistent with our previous findings on MOG_35–55-specific Th1 and Tc1 cells in the EAE model, and interphotoreceptor retinoid-binding protein (IRBP)_1–20-specific Th1 and Th17 cells in the EAU model, both of which were studied on female C57BL/6 mice.

As mentioned previously, similar functional profiles of CD4⁺ and CD8⁺ autoreactive T cells and their subsets were observed in closely related autoimmune disease models, such as MS, uveitis, and type I diabetes, all of which were also female-dominated animal models. Hence, future research into the common mechanisms of these autoimmune diseases, including thymus development,[57] local microenvironments,[57, 58] and estrogen relationships[59, 60] would be beneficial.

Wagner et al. suggested that classic EAE was dominated by CD4⁺ T cells, whereas atypical EAE was CD8⁺ T cells dominant, explaining why the classic EAE model did not mimic all features of MS very well.[5] Unfortunately, only a few studies have reported about CD8⁺ T cells dominant atypical EAE model, which uses myelin basic protein (MBP) on TCR-transgenic 8.8 mice.[5] This is a major drawback for research on CD8⁺ T cells, particularly Tc17 cells, in EAE. In addition, studies on different types of EAE models—such as myelin antigen-specific TCR-transgenic mouse EAE models,[5, 61–64] spontaneous EAE models,[61, 65, 66] humanized mouse EAE models,[63, 67–69] and non-human primate EAE models[70–72]—might provide new insights into the pathogenesis of the disease.

Although Tc17 cells have cytokine profiles similar to those of Th17 cells, there are still some differences between the two cell types.[24] In the skin, Tc17 and Th17 cells co-express retinoic acid receptor-related orphan nuclear receptor gamma (RORγt) and GATA-3 only under tissue challenge.[73] RORγt regulates IL-6 and IL-23 production via signal transducer and activator of transcription 3 (STAT3), and regulates IL-17A, IL-17F, IL-23R, and IL-21 production via epigenetic mechanisms.[74] In the thymus, T cell factor (TCF)-1 suppresses Tc17 cells only at the double-positive stage and suppresses Th17 cells before the stage of CD4 co-receptor expression.[75] Interferon regulator factor 3 (IRF3), via RORγt, inhibits Tc17 cell development much more than Th17 development.[76] Moreover, DMF affects IL-17 production in murine and human Tc17 cells, but to a much lesser extent than in Th17 cells. Tc17 and Th17 cells may have different AKT/mTOR signaling requirements, as well as divergent metabolic requirements.[77] Finally, the plasticity and related functions of Tc17 and Th17 cells are different.[24] Th17 cells can switch to a Th1-like phenotype, resulting in a more pathogenic profile, which has been confirmed by studies on pathological inflammation in the CNS[78] and intestine during bacterial infection or colitis.[79] In contrast, Tc17 cells shift to a Tc1-like phenotype only during CNS autoimmunity.[77] This suggests that the functional specificity of Tc17 and Th 17 cells differs in terms of plasticity towards the type 1 phenotype.[80]

Murine Tc17 cells exhibit phenotypic similarities to Th17 cells; however, the cytokine profile of human Tc17 cells remains unclear. A report has shown that some cytokines—such as IFN-γ, tumor necrosis factor-alpha (TNF-α), IL-21, IL-22, GM-CSF, RORγt, and its subfamily homolog RORα—are co-expressed with IL-17 in cultured Tc17 cells.[47] RORγt is a key factor in the development, maintenance, and function of IL-17-producing cells, and plays a role in regulating thymopoiesis. Clinical trials of several RORγ inhibitors in patients are ongoing. Other TFs, including STAT3, IRF3, and IRF4 also promote Tc17 cell differentiation.[47, 81]

Different subsets of CD8⁺ T cells are sometimes not stable, and differ between Tc1 and Tc17 phenotypes—a phenomenon called “plasticity of Tc17 cells”.[24] For example, the lack of CTLA-4 drives Tc17 cells to downregulate RORγt and IL-17 expression and shift towards Tc1.[82] In contrast, in a mouse model, Tc17 cells expressed a stable phenotype and mediated protection against fungi and bacteria.[83–85] Tc17 cells can also switch to Tc2 phenotype by increasing IL-5 and IL-13 cytokine production.[73] Thus, the plasticity of Tc17 cells can be an intriguing and promising target for future studies. TGF-β inhibits various functions of Tc1 cells, however, in presence of IL-6, TGF-β induces Tc17 cells.[86] In HKx31 (H3N2) influenza A virus-infected Il2^−/−, Il2ra^−/−, Il12^−/−, Blimp1^GFP, and Blimp1^fl/fl mice, Tc17 cells were non-cytotoxic and downregulated the expression of T-bet and Eomes, compared with Tc1 cells.[48]

In summary, our results suggest that autoreactive Tc17 cells have a lower encephalitogenic function, but are unique and independent of the pathogenicity of EAE compared to their Th17 counterparts and the Tc1 subset. The pathogenesis of MS is still unclear, and many factors—such as different cells and their subsets, and different kinds of EAE models—need to be investigated to determine the underlying mechanisms of MS.

Ab, antibody

APC, allophycocyanin

APCs, antigen-presenting cells

CFA, complete Freund’s adjuvant

CNS, central nervous system

CSF, cerebrospinal fluid;

DMF, dimethyl fumarate

EAE, experimental autoimmune encephalomyelitis ()

EAU, experimental autoimmune uveitis

ELISA, enzyme-linked immunosorbent assay

FITC, fluorescein isothiocyanate

HE, hematoxylin and eosin

IFN- g, Interferon g

LFB, Luxol fast blue staining

MS, multiple sclerosis

PA, psoriatic arthritis

PB, peripheral blood

PE, phycoerythroprotein

PPP, primary platelet purpura

PTX, pertussis toxin

RA, rheumatoid arthritis

RPMI, Roswell Park Memorial Institute

RRMS, remission relapsing MS

SLE, systemic lupus erythematous

STZ, streptozotocin

TID, type I diabetes

MG, myasthenia gravis

MOG_35-55, myelin oligodendrocyte glycoprotein peptides 35-55

Tc1, dominated IFN-g-secretionCD8⁺

Tc2, dominated IL-4 and IL-5-secretion CD8⁺

Tc9, dominated IL-9-secretion CD8⁺

Tc17, dominated IL-17-secretionCD8⁺

Th1, dominated IL-17-secretionCD4⁺ T helper cells

Th17, dominated IL-17-secretionCD4⁺ T helper cells

Tregs CD8, CD8⁺ Tregs, dominated IL10 and/or Foxp3 expression CD8⁺

RORgt, Retinoic acid receptor-related orphan nuclear receptor gamma ()

STAT3, signal transducer and activator of transcription 3 ()

TCF1, T cell factor 1

IRF3, Interferon regulator factor 3

-Ethics approval and consent to participate

-Consent for publication

We agreed to publish in BMC Neurology, if this manuscript will be accepted.

-Availability of data and materials

All are submitted to Journal

-Competing interests

The authors declare no conflict of interest.

-Funding

This work was supported by the Scientific Research Project of Hunan Provincial Health Commission, China (No. C2023030765 to YP), Key Plans of Hunan Administration Traditional Chinese Medicine, China (No. A2023039 to YP), University-Hospital Joint-Fund of Hunan University of Chinese Medicine, China(No. 2022XYLH198 to YP), Fund for Creative Research Group of Affiliated First Hospital of Hunan Traditional Chinese Medical College, China (No. 2021B-003 to YP), and Technology Plan Project of Zhuzhou City, Hunan Province, China (No. 2021-009 to YP).

-Authors' contributions

Y.P. received funding support and developed the research hypothesis. Y.P. X.L.Z., Y.D.T., S.Q.H., G.L.R., Q. C., Y.H.X., H.J., S.L., Z.Y.Z., Y.X. finished the experiments, analysis the data. Y.P. X.L.Z., wrote the main manuscript. The final manuscript is the product of the joint writing efforts of all authors.

-Acknowledgements (optional)

Not applicable.

-Authors' information (optional)

Not applicable.

Data sharing: no additional data available

Thompson AJ, Banwell BL, Barkhof F, Carroll WM, Coetzee T, Comi G, Correale J, Fazekas F, Filippi M, Freedman MS et al: Diagnosis of multiple sclerosis: 2017 revisions of the McDonald criteria. Lancet Neurol 2017, 17(2):162-173.
Nana A, Ruth AM, Christina B, Rochelle G, Douglas GM, Ron W, Philippe F, Julie B, Karen T, Kim R: Multiple sclerosis in Canada 2011 to 2031: results of a microsimulation modelling study of epidemiological and economic impacts. Health Promotion and Chronic Disease Prevention in Canada : Research, Policy and Practice 2017, 37(2):37-48.
Kutzelnigg A, Lassmann, H: Pathology of multiple sclerosis and related inﬂammatory demyelinating diseases. In: D.S., G. (Ed.), Handbook of Clinical Neurology. Elsevier,. DS, G (Ed), Handbook of Clinical Neurology 2014:15-58.
Peng Y, Zhu FZ, Chen ZX, Zhou JX, Gan L, Yang SS, Gao S, Liu QQ: Characterization of myelin oligodendrocyte glycoprotein (MOG)35-55-specific CD8+ T cells in experimental autoimmune encephalomyelitis. Chinese medical journal 2019, 132(24):2934-2940.
Wagner CA, Roqué PJ, Mileur TR, Liggitt D, Goverman JM: Myelin-specific CD8 T cells exacerbate brain inflammation in CNS autoimmunity. J Clin Invest 2020, 130(1):203-213.
Yamada H: Current perspectives on the role of IL-17 in autoimmune disease. J Inflamm Res 2010, 3:33-44.
Mondal S, Kundu M, Jana M, Roy A, Rangasamy SB, Modi KK, Wallace J, Albalawi YA, Balabanov R, Pahan K: IL-12 p40 monomer is different from other IL-12 family members to selectively inhibit IL-12Rβ1 internalization and suppress EAE. Proc Natl Acad Sci U S A 2020, 117(35):21557-21567.
Othy S, Jairaman A, Dynes JL, Dong TX, Tune C, Yeromin AV, Zavala A, Akunwafo C, Chen F, Parker I et al: Regulatory T cells suppress Th17 cell Ca(2+) signaling in the spinal cord during murine autoimmune neuroinflammation. Proc Natl Acad Sci U S A 2020, 117(33):20088-20099.
Rodica Balasa LB, Adrian Balasa,Anca Motataianu,Corina Roman-Filip,Doina Manu. : The action of TH17 cells on blood brain barrier in multiple sclerosis and experimental autoimmune encephalomyelitis[J]. Human Immunology 2020.
Miyauchi E, Kim SW, Suda W, Kawasumi M, Onawa S, Taguchi-Atarashi N, Morita H, Taylor TD, Hattori M, Ohno H: Gut microorganisms act together to exacerbate inflammation in spinal cords. Nature 2020, 585(7823):102-106.
Zhong S-S, Xiang Y-J, Liu P-J, He Y, Yang T-T, Wang Y-Y, Rong A, Zhang J, Liu G-Z: Effect of Cordyceps sinensis on the Treatment of Experimental Autoimmune Encephalomyelitis: A Pilot Study on Mice Model. Chinese medical journal 2017, 130(19):2296-2301.
Prinz I, Sandrock I, Mrowietz U: Interleukin-17 cytokines: Effectors and targets in psoriasis-A breakthrough in understanding and treatment. J Exp Med 2020, 217(1).
Benschop RJ, Chow CK, Tian Y, Nelson J, Barmettler B, Atwell S, Clawson D, Chai Q, Jones B, Fitchett J et al: Development of tibulizumab, a tetravalent bispecific antibody targeting BAFF and IL-17A for the treatment of autoimmune disease. MAbs 2019, 11(6):1175-1190.
Campa M, Mansouri B, Warren R, Menter A: A Review of Biologic Therapies Targeting IL-23 and IL-17 for Use in Moderate-to-Severe Plaque Psoriasis. Dermatol Ther (Heidelb) 2016, 6(1):1-12.
Wasilewska A, Winiarska M, Olszewska M, Rudnicka L: Interleukin-17 inhibitors. A new era in treatment of psoriasis and other skin diseases. Postepy Dermatol Alergol 2016, 33(4):247-252.
Baeten D, Sieper J, Braun J, Baraliakos X, Dougados M, Emery P, Deodhar A, Porter B, Martin R, Andersson M et al: Secukinumab, an Interleukin-17A Inhibitor, in Ankylosing Spondylitis. N Engl J Med 2015, 373(26):2534-2548.
McInnes IB, Mease PJ, Kirkham B, Kavanaugh A, Ritchlin CT, Rahman P, van der Heijde D, Landewé R, Conaghan PG, Gottlieb AB et al: Secukinumab, a human anti-interleukin-17A monoclonal antibody, in patients with psoriatic arthritis (FUTURE 2): a randomised, double-blind, placebo-controlled, phase 3 trial. Lancet 2015, 386(9999):1137-1146.
Mease PJ, McInnes IB, Kirkham B, Kavanaugh A, Rahman P, van der Heijde D, Landewé R, Nash P, Pricop L, Yuan J et al: Secukinumab Inhibition of Interleukin-17A in Patients with Psoriatic Arthritis. N Engl J Med 2015, 373(14):1329-1339.
Bittner S, Wiendl, H., : Neuroimmunotherapies targeting T cells: from pathophysiology to therapeutic applications. Neurotherapeutics 2016, 13:4-19.
Lubrano E, Perrotta FM: Secukinumab for ankylosing spondylitis and psoriatic arthritis. Ther Clin Risk Manag 2016, 12:1587-1592.
van Langelaar J, van der Vuurst de Vries RM, Janssen M, Wierenga-Wolf AF, Spilt IM, Siepman TA, Dankers W, Verjans G, de Vries HE, Lubberts E et al: T helper 17.1 cells associate with multiple sclerosis disease activity: perspectives for early intervention. Brain 2018, 141(5):1334-1349.
Comi G, Kappos L, Selmaj KW, Bar-Or A, Arnold DL, Steinman L, Hartung HP, Montalban X, Kubala Havrdová E, Cree BAC et al: Safety and efficacy of ozanimod versus interferon beta-1a in relapsing multiple sclerosis (SUNBEAM): a multicentre, randomised, minimum 12-month, phase 3 trial. Lancet Neurol 2019, 18(11):1009-1020.
Mousset CM, Hobo W, Woestenenk R, Preijers F, Dolstra H, van der Waart AB: Comprehensive Phenotyping of T Cells Using Flow Cytometry. Cytometry A 2019, 95(6):647-654.
Peng Y, Deng X, Zeng Q, Tang Y: Tc17 cells in autoimmune diseases. Chinese medical journal 2022, 135(18):2167-2177.
Kondo T, Takata H, Matsuki F, Takiguchi M: Cutting edge: Phenotypic characterization and differentiation of human CD8+ T cells producing IL-17. Journal of immunology (Baltimore, Md : 1950) 2009, 182(4):1794-1798.
Intlekofer AM, Banerjee A, Takemoto N, Gordon SM, Dejong CS, Shin H, Hunter CA, Wherry EJ, Lindsten T, Reiner SL: Anomalous type 17 response to viral infection by CD8+ T cells lacking T-bet and eomesodermin. Science 2008, 321(5887):408-411.
Yen HR, Harris TJ, Wada S, Grosso JF, Getnet D, Goldberg MV, Liang KL, Bruno TC, Pyle KJ, Chan SL et al: Tc17 CD8 T cells: functional plasticity and subset diversity. Journal of immunology (Baltimore, Md : 1950) 2009, 183(11):7161-7168.
Liang Y, Pan HF, Ye DQ: Tc17 Cells in Immunity and Systemic Autoimmunity. Int Rev Immunol 2015, 34(4):318-331.
Volarić I, Vičić M, Prpić-Massari L: The Role of CD8+ T-Cells and their Cytokines in the Pathogenesis of Psoriasis. Acta Dermatovenerol Croat 2019, 27(3):159-162.
Casciano F, Pigatto PD, Secchiero P, Gambari R, Reali E: T Cell Hierarchy in the Pathogenesis of Psoriasis and Associated Cardiovascular Comorbidities. Frontiers in immunology 2018, 9:1390.
Gonzalez SM, Taborda NA, Rugeles MT: Role of Different Subpopulations of CD8(+) T Cells during HIV Exposure and Infection. Frontiers in immunology 2017, 8:936.
Majchrzak K, Nelson MH, Bailey SR, Bowers JS, Yu XZ, Rubinstein MP, Himes RA, Paulos CM: Exploiting IL-17-producing CD4+ and CD8+ T cells to improve cancer immunotherapy in the clinic. Cancer Immunol Immunother 2016, 65(3):247-259.
Dagur PK, McCoy JP, Jr.: Endothelial-binding, proinflammatory T cells identified by MCAM (CD146) expression: Characterization and role in human autoimmune diseases. Autoimmun Rev 2015, 14(5):415-422.
Konduri V, Oyewole-Said D, Vazquez-Perez J, Weldon SA, Halpert MM, Levitt JM, Decker WK: CD8(+)CD161(+) T-Cells: Cytotoxic Memory Cells With High Therapeutic Potential. Frontiers in immunology 2020, 11:613204.
Salou M, Nicol B, Garcia A, Baron D, Michel L, Elong-Ngono A, Hulin P, Nedellec S, Jacq-Foucher M, Le Frère F et al: Neuropathologic, phenotypic and functional analyses of Mucosal Associated Invariant T cells in Multiple Sclerosis. Clinical Immunology 2016, 166-167:1-11.
Nicol B, Salou M, Vogel I, Garcia A, Dugast E, Morille J, Kilens S, Charpentier E, Donnart A, Nedellec S et al: An intermediate level of CD161 expression defines a novel activated, inflammatory, and pathogenic subset of CD8+ T cells involved in multiple sclerosis. Journal of Autoimmunity 2018, 88:61-74.
Magdalena Huber SH, Axel Pagenstecher, Katharina Reinhard, Josephine Ritter, Alexander Visekruna, Anna Guralnik, Nadine Bollig, Katharina Jeltsch, Christina Heinemann, Eva Wittmann, Thorsten Buch, Olivia Prazeres da Costa,Anne Brüstle,6 Dirk Brenner, Tak W. Mak, Hans-Willi Mittrücker, Björn Tackenberg,10 Thomas Kamradt,2 and Michael Lohoff: IL-17A secretion by CD8+ T cells supports Th17-mediated autoimmune encephalomyelitis. The Journal of clinical investigation 2013, 123:247-260.
Ifergan I, Kebir H, Alvarez JI, Marceau G, Bernard M, Bourbonnière L, Poirier J, Duquette P, Talbot PJ, Arbour N et al: Central nervous system recruitment of effector memory CD8+ T lymphocytes during neuroinflammation is dependent on α4 integrin. Brain 2011, 134(12):3560-3577.
Peelen E, Thewissen M, Knippenberg S, Smolders J, Muris AH, Menheere P, Tervaert JW, Hupperts R, Damoiseaux J: Fraction of IL-10+ and IL-17+ CD8 T cells is increased in MS patients in remission and during a relapse, but is not influenced by immune modulators. Journal of neuroimmunology 2013, 258(1-2):77-84.
Huber M HS, Pagenstecher A, Reinhard K, Ritter J, Visekruna A, et al. : IL-17A secretion by CD8+ T cells supports Th17-mediated autoimmune encephalomyelitis. J Clin Invest 2013, 123:247-260.
Tzartos JS, Friese MA, Craner MJ, Palace J, Newcombe J, Esiri MM, Fugger L: Interleukin-17 production in central nervous system-infiltrating T cells and glial cells is associated with active disease in multiple sclerosis. The American journal of pathology 2008, 172(1):146-155.
Snyder-Cappione JE, Tincati C, Eccles-James IG, Cappione AJ, Ndhlovu LC, Koth LL, Nixon DF: A comprehensive ex vivo functional analysis of human NKT cells reveals production of MIP1-α and MIP1-β, a lack of IL-17, and a Th1-bias in males. PLoS One 2010, 5(11):e15412.
Garcillán B, Marin AV, Jiménez-Reinoso A, Briones AC, Muñoz-Ruiz M, García-León MJ, Gil J, Allende LM, Martínez-Naves E, Toribio ML et al: γδ T Lymphocytes in the Diagnosis of Human T Cell Receptor Immunodeficiencies. Frontiers in immunology 2015, 6:20.
Norde WJ, Maas F, Hobo W, Korman A, Quigley M, Kester MG, Hebeda K, Falkenburg JH, Schaap N, de Witte TM et al: PD-1/PD-L1 interactions contribute to functional T-cell impairment in patients who relapse with cancer after allogeneic stem cell transplantation. Cancer Res 2011, 71(15):5111-5122.
Wong MT, Ye JJ, Alonso MN, Landrigan A, Cheung RK, Engleman E, Utz PJ: Regulation of human Th9 differentiation by type I interferons and IL-21. Immunol Cell Biol 2010, 88(6):624-631.
Annunziato F, Romagnani S: Heterogeneity of human effector CD4+ T cells. Arthritis Res Ther 2009, 11(6):257.
Chemin K, Ramsköld D, Diaz-Gallo LM, Herrath J, Houtman M, Tandre K, Rönnblom L, Catrina A, Malmström V: EOMES-positive CD4(+) T cells are increased in PTPN22 (1858T) risk allele carriers. Eur J Immunol 2018, 48(4):655-669.
Xin A, Masson F, Liao Y, Preston S, Guan T, Gloury R, Olshansky M, Lin JX, Li P, Speed TP et al: A molecular threshold for effector CD8(+) T cell differentiation controlled by transcription factors Blimp-1 and T-bet. Nat Immunol 2016, 17(4):422-432.
Intlekofer AM, Takemoto N, Kao C, Banerjee A, Schambach F, Northrop JK, Shen H, Wherry EJ, Reiner SL: Requirement for T-bet in the aberrant differentiation of unhelped memory CD8+ T cells. J Exp Med 2007, 204(9):2015-2021.
Sullivan BM, Juedes A, Szabo SJ, von Herrath M, Glimcher LH: Antigen-driven effector CD8 T cell function regulated by T-bet. Proc Natl Acad Sci U S A 2003, 100(26):15818-15823.
Peng Y, Shao H, Ke Y, Zhang P, Han G, Kaplan HJ, Sun D: Minimally Activated CD8 Autoreactive T Cells Specific for IRBP Express a High Level of Foxp3 and Are Functionally Suppressive. Investigative ophthalmology & visual science 2007, 48(5):2178-2184.
Han G, Shao H, Peng Y, Zhang P, Ke Y, Kaplan HJ, Sun D: Suppressor role of Rat CD8(+)CD45RC(low) T cells in Experimental Autoimmune Uveitis (EAU). Journal of neuroimmunology 2007, 183(1-2):81-88.
Peng Y, Shao H, Ke Y, Zhang P, Xiang J, Kaplan HJ, Sun D: In Vitro Activation of CD8 Interphotoreceptor Retinoid-Binding Protein-Specific T Cells Requires not only Antigenic Stimulation but also Exogenous Growth Factors. The Journal of Immunology 2006, 176(8):5006-5014.
Peng Y, Han G, Shao H, Wang Y, Kaplan HJ, Sun D: Characterization of IL-17(+) Interphotoreceptor Retinoid-Binding Protein-Specific T Cells in Experimental Autoimmune Uveitis. Investigative ophthalmology & visual science 2007, 48(9):4153-4161.
LaMothe RA, Kolte PN, Vo T, Ferrari JD, Gelsinger TC, Wong J: Tolerogenic Nanoparticles Induce Antigen-Specific Regulatory T Cells and Provide Therapeutic Efficacy and Transferrable Tolerance against Experimental Autoimmune Encephalomyelitis. Frontiers in immunology 2018, 9(281).
Shao H, Peng Y, Liao T, Wang M, Song M, Kaplan HJ, Sun D: A Shared Epitope of the Interphotoreceptor Retinoid-Binding Protein Recognized by the CD4+ and CD8+Autoreactive T Cells. The Journal of Immunology 2005, 175(3):1851-1857.
Waisman A, Johann L: Antigen-presenting cell diversity for T cell reactivation in central nervous system autoimmunity. J Mol Med (Berl) 2018, 96(12):1279-1292.
Macri C, Pang ES, Patton T, O’Keeffe M: Dendritic cell subsets. Seminars in Cell & Developmental Biology 2018, 84:11-21.
Sciarra F, Campolo F, Franceschini E, Carlomagno F, Venneri MA: Gender-Specific Impact of Sex Hormones on the Immune System. International journal of molecular sciences 2023, 24(7).
Peckham H, Webb K, Rosser EC, Butler G, Ciurtin C: Gender-Diverse Inclusion in Immunological Research: Benefits to Science and Health. Frontiers in medicine 2022, 9:909789.
Salvador F, Deramoudt L, Leprêtre F, Figeac M, Guerrier T, Boucher J, Bas M, Journiac N, Peters A, Mars LT et al: A Spontaneous Model of Experimental Autoimmune Encephalomyelitis Provides Evidence of MOG-Specific B Cell Recruitment and Clonal Expansion. Frontiers in immunology 2022, 13:755900.
Luu T, Cheung JF, Baccon J, Waldner H: Priming of myelin-specific T cells in the absence of dendritic cells results in accelerated development of Experimental Autoimmune Encephalomyelitis. PLoS One 2021, 16(4):e0250340.
Leno-Durán E, Ng SL, Strominger JL: Regulation of EAE by spontaneously generated IL-10-secreting regulatory T cells in HLA-DR15/TCR.Ob1A12 double transgenic mice. Immunology 2021, 163(3):338-343.
Yeola AP, Ignatius Arokia Doss PM, Baillargeon J, Akbar I, Mailhot B, Balood M, Talbot S, Anderson AC, Lacroix S, Rangachari M: Endogenous T Cell Receptor Rearrangement Represses Aggressive Central Nervous System Autoimmunity in a TcR-Transgenic Model on the Non-Obese Diabetic Background. Frontiers in immunology 2019, 10:3115.
Cossu D, Yokoyama K, Sakanishi T, Sechi LA, Hattori N: Bacillus Calmette-Guérin Tokyo-172 vaccine provides age-related neuroprotection in actively induced and spontaneous experimental autoimmune encephalomyelitis models. Clinical and experimental immunology 2023, 212(1):70-80.
Nitsch L, Petzinna S, Zimmermann J, Getts DR, Becker A, Müller M: MOG-Specific T Cells Lead to Spontaneous EAE with Multilocular B Cell Infiltration in the GF-IL23 Model. Neuromolecular medicine 2022, 24(4):415-423.
Tacke S, Chunder R, Schropp V, Urich E, Kuerten S: Effects of a Fully Humanized Type II Anti-CD20 Monoclonal Antibody on Peripheral and CNS B Cells in a Transgenic Mouse Model of Multiple Sclerosis. International journal of molecular sciences 2022, 23(6).
Wemlinger SM, Parker Harp CR, Yu B, Hardy IR, Seefeldt M, Matsuda J, Mingueneau M, Spilker KA, Cameron TO, Larrick JW et al: Preclinical Analysis of Candidate Anti-Human CD79 Therapeutic Antibodies Using a Humanized CD79 Mouse Model. Journal of immunology (Baltimore, Md : 1950) 2022, 208(7):1566-1584.
Breakell T, Tacke S, Schropp V, Zetterberg H, Blennow K, Urich E, Kuerten S: Obinutuzumab-Induced B Cell Depletion Reduces Spinal Cord Pathology in a CD20 Double Transgenic Mouse Model of Multiple Sclerosis. International journal of molecular sciences 2020, 21(18).
t Hart BA: A Tolerogenic Role of Cathepsin G in a Primate Model of Multiple Sclerosis: Abrogation by Epstein-Barr Virus Infection. Archivum immunologiae et therapiae experimentalis 2020, 68(4):21.
Dunham J, van de Vis R, Bauer J, Wubben J, van Driel N, Laman JD, t Hart BA, Kap YS: Severe oxidative stress in an acute inflammatory demyelinating model in the rhesus monkey. PloS one 2017, 12(11):e0188013-e0188013.
Carvalho RHF, Real CC, Cinini S, Garcez AT, Duran FLS, Marques FLN, Mello LE, Busatto Filho G, de Vries EFJ, de Britto LRG et al: [(11)C]PIB PET imaging can detect white and grey matter demyelination in a non-human primate model of progressive multiple sclerosis. Multiple sclerosis and related disorders 2019, 35:108-115.
Harrison OJ, Linehan JL, Shih HY, Bouladoux N, Han SJ, Smelkinson M, Sen SK, Byrd AL, Enamorado M, Yao C et al: Commensal-specific T cell plasticity promotes rapid tissue adaptation to injury. Science 2019, 363(6422).
Stadhouders R, Lubberts E, Hendriks RW: A cellular and molecular view of T helper 17 cell plasticity in autoimmunity. J Autoimmun 2018, 87:1-15.
Mielke LA, Liao Y, Clemens EB, Firth MA, Duckworth B, Huang Q, Almeida FF, Chopin M, Koay HF, Bell CA et al: TCF-1 limits the formation of Tc17 cells via repression of the MAF-RORγt axis. J Exp Med 2019, 216(7):1682-1699.
Ysebrant de Lendonck L, Tonon S, Nguyen M, Vandevenne P, Welsby I, Martinet V, Molle C, Charbonnier LM, Leo O, Goriely S: Interferon regulatory factor 3 controls interleukin-17 expression in CD8 T lymphocytes. Proc Natl Acad Sci U S A 2013, 110(34):E3189-3197.
Lückel C, Picard F, Raifer H, Campos Carrascosa L, Guralnik A, Zhang Y, Klein M, Bittner S, Steffen F, Moos S et al: IL-17(+) CD8(+) T cell suppression by dimethyl fumarate associates with clinical response in multiple sclerosis. Nat Commun 2019, 10(1):5722.
Duhen R, Glatigny S, Arbelaez CA, Blair TC, Oukka M, Bettelli E: Cutting edge: the no additional data available (Baltimore, Md : 1950) 2013, 190(9):4478-4482.
Ahlfors H, Morrison PJ, Duarte JH, Li Y, Biro J, Tolaini M, Di Meglio P, Potocnik AJ, Stockinger B: IL-22 fate reporter reveals origin and control of IL-22 production in homeostasis and infection. Journal of immunology (Baltimore, Md : 1950) 2014, 193(9):4602-4613.
Lückel C, Picard FSR, Huber M: Tc17 biology and function: Novel concepts. Eur J Immunol 2020, 50(9):1257-1267.
Mittrücker HW, Visekruna A, Huber M: Heterogeneity in the differentiation and function of CD8⁺ T cells. Archivum immunologiae et therapiae experimentalis 2014, 62(6):449-458.
Arra A, Lingel H, Kuropka B, Pick J, Schnoeder T, Fischer T, Freund C, Pierau M, Brunner-Weinzierl MC: The differentiation and plasticity of Tc17 cells are regulated by CTLA-4-mediated effects on STATs. Oncoimmunology 2017, 6(2):e1273300.
Nanjappa SG, McDermott AJ, Fites JS, Galles K, Wüthrich M, Deepe GS, Jr., Klein BS: Antifungal Tc17 cells are durable and stable, persisting as long-lasting vaccine memory without plasticity towards IFNγ cells. PLoS Pathog 2017, 13(5):e1006356.
Nanjappa SG, Mudalagiriyappa S, Fites JS, Suresh M, Klein BS: CBLB Constrains Inactivated Vaccine-Induced CD8(+) T Cell Responses and Immunity against Lethal Fungal Pneumonia. Journal of immunology (Baltimore, Md : 1950) 2018, 201(6):1717-1726.
Loures FV, Araújo EF, Feriotti C, Bazan SB, Costa TA, Brown GD, Calich VL: Dectin-1 induces M1 macrophages and prominent expansion of CD8+IL-17+ cells in pulmonary Paracoccidioidomycosis. J Infect Dis 2014, 210(5):762-773.
Liu SJ, Tsai JP, Shen CR, Sher YP, Hsieh CL, Yeh YC, Chou AH, Chang SR, Hsiao KN, Yu FW et al: Induction of a distinct CD8 Tnc17 subset by transforming growth factor-beta and interleukin-6. J Leukoc Biol 2007, 82(2):354-360.

No competing interests reported.

Download PDF

Journal Publication

published 18 Mar, 2024

Read the published version in Neuroprotection →

Version 1

posted

You are reading this latest preprint version

Role of Autoreactive Tc17 Cells in the Pathogenesis of Experimental Autoimmune Encephalomyelitis (EAE)

Status:

Journal Publication

Version 1

Abstract

Figures

1 Introduction

2. Methods

2.1. Ethics approval

2.2. Selection of animal participants

2.3. Antigen-induced EAE (aEAE) models

2.4. Purification of MOG_35–55-specific CD4⁺ and CD8⁺ T cells

2.5. Preparation of MOG_35–55-specific Th1, Tc1, Th17 and Tc17 cells

2.6. Proliferation assay

2.7. Enzyme-linked immunosorbent assays (ELISA)

2.8. Adoptive-transferred EAE models (tEAE)

2.9. Histological study

2.10. Statistical analysis

3. Results

3.1. Tc17 cells are MOG_35–55 specific

3.2.1. IFN-γ secretion

3.2.2 IL-17 secretion

3.3. Adoptive transfer of MOG_35–55-specific Tc17 cells to naïve mice induces tEAE

4. Discussion

Abbreviations

Declarations

References

Tables

Additional Declarations

Status:

Journal Publication

Version 1

Role of Autoreactive Tc17 Cells in the Pathogenesis of Experimental Autoimmune Encephalomyelitis (EAE)

Status:

Journal Publication

Version 1

Abstract

Figures

1 Introduction

2. Methods

2.1. Ethics approval

2.2. Selection of animal participants

2.3. Antigen-induced EAE (aEAE) models

2.4. Purification of MOG35–55-specific CD4+ and CD8+ T cells

2.5. Preparation of MOG35–55-specific Th1, Tc1, Th17 and Tc17 cells

2.6. Proliferation assay

2.7. Enzyme-linked immunosorbent assays (ELISA)

2.8. Adoptive-transferred EAE models (tEAE)

2.9. Histological study

2.10. Statistical analysis

3. Results

3.1. Tc17 cells are MOG35–55 specific

3.2.1. IFN-γ secretion

3.2.2 IL-17 secretion

3.3. Adoptive transfer of MOG35–55-specific Tc17 cells to naïve mice induces tEAE

4. Discussion

Abbreviations

Declarations

References

Tables

Additional Declarations

Status:

Journal Publication

Version 1

2.4. Purification of MOG_35–55-specific CD4⁺ and CD8⁺ T cells

2.5. Preparation of MOG_35–55-specific Th1, Tc1, Th17 and Tc17 cells

3.1. Tc17 cells are MOG_35–55 specific

3.3. Adoptive transfer of MOG_35–55-specific Tc17 cells to naïve mice induces tEAE