P095 PD-1 expressing T cells in patients with different types of colitis

Abstract Background Immunotherapy-related colitis is a frequent adverse event in patients with malignancies, treated with inhibitors targeting programmed death-1 (PD-1). This treatment leads to enhancement of lymphocyte activity, thereby generating antitumor T-cell activity. The role of PD-1+ T cells in the pathophysiology of different types of colitis is unclear. Therefore, we aimed to study the presence of PD-1+ T cells in inflammatory bowel disease (IBD), infectious colitis and healthy controls compared with anti-PD-1-related colitis. Methods We performed a prospective cohort study. Newly diagnosed patients with ulcerative colitis (UC, n = 51), Crohn’s disease (CD, n = 29), infectious colitis (n = 4), anti-PD-1-related colitis (n = 5) and healthy controls (n = 6) were included. Baseline colonic biopsy specimens were collected for immunohistochemistry identifying PD-1+ and PD-L1+ lymphocytes in the epithelium and lamina propria separately and for immunophenotyping by flow cytometry identifying PD-1+ T-cell subsets. Results Using immunohistochemistry, PD-1 was not present on lymphocytes in the epithelium of patients with any type of colitis, nor in healthy controls. Of all lymphocytes in the lamina propria, % PD-1 expression was 40% in UC, 5% in infectious colitis, 3% in anti-PD-1-related colitis and 0% in healthy controls. PD-L1 was expressed on lymphocytes in the epithelium and lamina propria of UC patients (12.5% and 40%) and in infectious colitis (1% and 30%), whereas in anti-PD-1-related colitis (0% and 15%) and healthy controls (0% and 15%) no PD-L1+ lymphocytes were demonstrated in the epithelium. Flowcytometry showed higher percentages of PD-1+ T cells in biopsy specimens of UC patients (25.2% (IQR 17.9–35.6)) compared with all other groups; CD patients (13.5% (5.0–25.3), p = 0.001), infectious colitis (9.8% (4.7–17.4), p = 0.005), anti-PD-1-related colitis (1.5% (1.1–2.1), p = 0.001) and healthy controls (14.3% (4.9–28.2), p = 0.08). In IBD and infectious colitis, the majority of PD-1+ T cells were CD4+ (84.8% (76.5–90.5), while in anti-PD-1-related colitis the majority of PD-1+ T cells were CD8+ (76.2% (68.5–82.1)). PD-1+ T cells in all patient groups were mainly effector T cells (CD45Ro+). Conclusion In patients with different types of colitis and in healthy controls, PD-1 was only expressed on T cells in the lamina propria and not in the epithelium. The percentage of PD-1+ T cells was significantly higher in patients with UC compared with patients with CD or infectious colitis and healthy controls. As expected PD-1+ T cells were nearly absent in patients with anti-PD-1-related colitis.

Download Full-text

CD3+CD4+gp130+ T Cells Are Associated With Worse Disease Activity in Systemic Lupus Erythematosus Patients

Frontiers in Immunology ◽

10.3389/fimmu.2021.675250 ◽

2021 ◽

Vol 12 ◽

Author(s):

Nur Diyana Mohd Shukri ◽

Aziz Farah Izati ◽

Wan Syamimee Wan Ghazali ◽

Che Maraina Che Hussin ◽

Kah Keng Wong

Keyword(s):

Systemic Lupus Erythematosus ◽

T Cells ◽

T Cell ◽

Disease Activity ◽

Lupus Erythematosus ◽

Lymphocyte Subpopulations ◽

Gene Set Enrichment Analysis ◽

T Cell Subsets ◽

Healthy Controls ◽

Systemic Lupus

The receptors for IL-35, IL-12Rβ2 and gp130, have been implicated in the inflammatory pathophysiology of autoimmune diseases. In this study, we set out to investigate the serum IL-35 levels and the surface levels of IL-12Rβ2 and gp130 in CD3+CD4+, CD3+CD4─ and CD3─CD4─ lymphocyte subpopulations in systemic lupus erythematosus (SLE) patients (n=50) versus healthy controls (n=50). The potential T cell subsets associated with gp130 transcript (i.e. IL6ST) expression in CD4+ T cells of SLE patients was also examined in publicly-available gene expression profiling (GEP) datasets. Here, we report that serum IL-35 levels were significantly higher in SLE patients than healthy controls (p=0.038) but it was not associated with SLEDAI-2K scores. The proportions of IL-12Rβ2+ and gp130+ cells in SLE patients did not differ significantly with those of healthy controls in all lymphocyte subpopulations investigated. Essentially, higher SLEDAI-2K scores were positively correlated with increased proportion of gp130+ cells, but not IL-12Rβ2+ cells, on CD3+CD4+ T cells (r=0.425, p=0.002, q=0.016). Gene Set Enrichment Analysis (GSEA) of a GEP dataset of CD4+ T cells isolated from SLE patients (n=8; GSE4588) showed that IL6ST expression was positively associated with genes upregulated in CD4+ T cells vs myeloid or B cells (q<0.001). In an independent GEP dataset of CD4+ T cells isolated from SLE patients (n=9; GSE1057), IL6ST expression was induced upon anti-CD3 stimulation, and that Treg, TCM and CCR7+ T cells gene sets were significantly enriched (q<0.05) by genes highly correlated with IL6ST expression (n=92 genes; r>0.75 with IL6ST expression) upon anti-CD3 stimulation in these SLE patients. In conclusion, gp130 signaling in CD3+CD4+ T cell subsets may contribute to increased disease activity in SLE patients, and it represents a promising therapeutic target for inhibition in the disease.

Download Full-text

High-Fat Diet Alters Immunogenic Properties of Circulating and Adipose Tissue-Associated Myeloid-Derived CD45+DDR2+ Cells

Mediators of Inflammation ◽

10.1155/2019/1648614 ◽

2019 ◽

Vol 2019 ◽

pp. 1-15

Author(s):

Sara J. Sidles ◽

Ying Xiong ◽

M. Rita I. Young ◽

Amanda C. LaRue

Keyword(s):

T Cells ◽

Adipose Tissue ◽

T Cell ◽

Peripheral Blood ◽

Cell Activity ◽

Cell Subset ◽

Total Body Weight ◽

T Cell Subsets ◽

Activation Markers

Chronic inflammation is evident in the adipose tissue and periphery of patients with obesity, as well as mouse models of obesity. T cell subsets in obese adipose tissue are skewed towards Th1- and Th17-associated phenotypes and their secreted cytokines contribute to obesity-associated inflammation. Our lab recently identified a novel, myeloid-derived CD45+DDR2+ cell subset that modulates T cell activity. The current study sought to determine how these myeloid-derived CD45+DDR2+ cells are altered in the adipose tissue and peripheral blood of preobese mice and how this population modulates T cell activity. C57BL/6 mice were fed with a diet high in milkfat (60%·kcal, HFD) ad libitum until a 20% increase in total body weight was reached, and myeloid-derived CD45+DDR2+ cells and CD4+ T cells in visceral adipose tissue (VAT), mammary gland-associated adipose tissue (MGAT), and peripheral blood (PB) were phenotypically analyzed. Also analyzed was whether mediators from MGAT-primed myeloid-derived CD45+DDR2+ cells stimulate normal CD4+ T cell cytokine production. A higher percentage of myeloid-derived CD45+DDR2+ cells expressed the activation markers MHC II and CD80 in both VAT and MGAT of preobese mice. CD4+ T cells were preferentially skewed towards Th1- and Th17-associated phenotypes in the adipose tissue and periphery of preobese mice. In vitro, MGAT from HFD-fed mice triggered myeloid-derived CD45+DDR2+ cells to induce CD4+ T cell IFN-γ and TNF-α production. Taken together, this study shows that myeloid-derived CD45+DDR2+ cells express markers of immune activation and suggests that they play an immune modulatory role in the adipose tissue of preobese mice.

Download Full-text

Intestinal CD103+CD4+ and CD103+CD8+ T-Cell Subsets in the Gut of Inflammatory Bowel Disease Patients at Diagnosis and During Follow-up

Inflammatory Bowel Diseases ◽

10.1093/ibd/izz049 ◽

2019 ◽

Vol 25 (9) ◽

pp. 1497-1509 ◽

Cited By ~ 8

Author(s):

Britt Roosenboom ◽

Peter J Wahab ◽

Carolijn Smids ◽

Marcel J M Groenen ◽

Elly van Koolwijk ◽

...

Keyword(s):

Inflammatory Bowel Disease ◽

T Cells ◽

T Cell ◽

Bowel Disease ◽

Cd8 T Cell ◽

T Cell Subsets ◽

Healthy Controls ◽

Inflammatory Bowel ◽

Active Inflammation

Abstract Background The integrin CD103 is proposed to be a potential therapeutical target in inflammatory bowel disease (IBD), as it can form a heterodimeric integrin with β7 (Etrolizumab, anti-β7 integrin) on epithelial T cells. Therefore, we aimed to study the frequencies of different intestinal CD103+T-cell subsets, both CD4+ and CD8+, in newly diagnosed, untreated IBD patients at baseline and during follow-up, compared with healthy controls. Methods Intestinal biopsies from inflamed segments during colonoscopy and peripheral blood samples were prospectively taken from IBD patients at diagnosis and during follow-up. Blood and single cell suspensions from biopsies were analyzed for CD103+ T-cell subpopulations by flow cytometry and expressed as median percentages of the total T-cell population. Results In total, 75 Crohn’s disease (CD) patients, 49 ulcerative colitis (UC) patients, and 16 healthy controls were included. At presentation, IBD patients displayed lower percentages of CD103+T-cell subsets in inflamed biopsies: 3% (1 to 5) CD103+CD4+ in IBD vs 5% (5 to 7) in healthy controls (P = 0.007) and 9% (4 to 15) CD103+CD8+ compared with 42% (23 to 57) in healthy controls (P = 0.001). The majority of intestinal T cells was composed of CD103-CD4+ T cells (65% [52 to 74]) in IBD compared with 30% (21 to 50) in healthy controls (P = 0.001). In patients with endoscopic remission during follow-up (n = 27), frequencies of CD103+ and CD103-T-cell subsets were comparable with healthy controls. Conclusion At diagnosis, active inflammation in IBD was associated with decreased percentages of both CD103+CD4+ and CD103+CD8+T-cell subsets in colon and ileum biopsies. In active disease during follow-up, these T-cell populations remained low but increased in remission to values comparable with healthy controls. A shift toward more CD103-T cells was observed during active inflammation.

Download Full-text

Regulatory functions of hapten-reactive helper and suppressor T lymphocytes. III. Amplification of a generation of tumor-specific killer T-lymphocyte activities by suppressor T-cell-depleted hapten-reactive T lymphocytes.

Journal of Experimental Medicine ◽

10.1084/jem.149.1.185 ◽

1979 ◽

Vol 149 (1) ◽

pp. 185-199 ◽

Cited By ~ 26

Author(s):

T Hamaoka ◽

H Fujiwara ◽

K Teshima ◽

H Aoki ◽

H Yamamoto ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

T Lymphocytes ◽

T Lymphocyte ◽

Lethal Dose ◽

Cell Activity ◽

Helper T Cell ◽

Relative Susceptibility ◽

Helper T Lymphocytes ◽

Lymphocyte Activity

2,4.6-trinitrophenyl (TNP)-reactive T-cell activities were raised in mice by immunization with TNP-isologous mouse gamma globulin. After establishing that TNP-reactive T lymphocytes can serve as amplifier cells for induction of killer T lymphocytes in allogeneic system, we explored the possibility of this hapten-reactive T-cell system to amplify tumor-specific killer T-lymphocyte activity in the syngeneic system. We utlized relatively weak immunogenic syngeneic plasmacytoma X5563 in C3H/He mice. Analysis of the TNP-reactive T-cell activities revealed that such T lymphocytes express the biological functions of both major subtypes of regulatory T cells, namely suppressors and helpers, and that TNP-reactive suppressor and helper T lymphocytes, respectively, differ in their relative susceptibility to specific inactivation by TNP conjugates of the nonimmunogenic D-amino acid copolymer, D-glutamic acid, and D-lysine (D-GL). By taking advantage of the relative susceptibility-difference to TNP-D-GL, selective inactivation of TNP-reactive suppressor T cells was induced by appropriate treatment with TNP-D-GL, and the generation of TNP-reactive helper T-cell activity was amplified. The supplement of augmented TNP-reactive helper T-cell activity to the system at the immunization with syngeneic X5563 with TNP-haptenation, resulted in a striking augmentation of induction of tumor-specific killer T-lymphocyte activity, and a considerable number of hosts survived after the challenge with lethal dose of viable tumor cells. Thus, appropriate manipulations designed to induce potent hapten-reactive helper T-lymphocytes provided the potential for a very effective mode of immunoprophylaxis against tumor.

Download Full-text

Rapid Acquisition of Tissue-specific Homing Phenotypes by CD4+ T Cells Activated in Cutaneous or Mucosal Lymphoid Tissues

Journal of Experimental Medicine ◽

10.1084/jem.20011502 ◽

2002 ◽

Vol 195 (1) ◽

pp. 135-141 ◽

Cited By ~ 374

Author(s):

Daniel J. Campbell ◽

Eugene C. Butcher

Keyword(s):

T Cells ◽

T Cell ◽

Lymph Nodes ◽

Lamina Propria ◽

Memory T Cells ◽

Lymphoid Organs ◽

T Cell Subsets ◽

Effector Memory ◽

Intestinal Lamina Propria ◽

Tissue Specific

Effector and memory T cells can be subdivided based on their ability to traffic through peripheral tissues such as inflamed skin and intestinal lamina propria, a property controlled by expression of ‘tissue-specific’ adhesion and chemoattractant receptors. However, little is known about the development of these selectively homing T cell subsets, and it is unclear whether activation in cutaneous versus intestinal lymphoid organs directly results in effector/memory T cells that differentially express adhesion and chemoattracant receptors targeting them to the corresponding nonlymphoid site. We define two murine CD4+ effector/memory T cell subsets that preferentially localize in cutaneous or intestinal lymphoid organs by their reciprocal expression of the adhesion molecules P-selectin ligand (P-lig) and α4β7, respectively. We show that within 2 d of systemic immunization CD4+ T cells activated in cutaneous lymph nodes upregulate P-lig, and downregulate α4β7, while those responding to antigen in intestinal lymph nodes selectively express high levels of α4β7 and acquire responsiveness to the intestinal chemokine thymus-expressed chemokine (TECK). Thus, during an immune response, local microenvironments within cutaneous and intestinal secondary lymphoid organs differentially direct T cell expression of these adhesion and chemoattractant receptors, targeting the resulting effector T cells to the inflamed skin or intestinal lamina propria.

Download Full-text

THU0039 DECIPHERING DISEASE-RELEVANT T CELL SUBSETS IN RHEUMATOID ARTHRITIS IDENTIFIES A NOVEL CELLULAR SUBSET OF PATHOGENETIC IMPORTANCE IN THERAPEUTIC RESISTANCE.

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2020-eular.5525 ◽

2020 ◽

Vol 79 (Suppl 1) ◽

pp. 232.1-233

Author(s):

M. Nyirenda ◽

I. Mcinnes ◽

C. Goodyear

Keyword(s):

Rheumatoid Arthritis ◽

T Cells ◽

T Cell ◽

Inflammatory Cytokines ◽

Regulatory T Cell ◽

Cell Subset ◽

T Cell Subsets ◽

Healthy Controls ◽

T Cell Subset ◽

Pro Inflammatory Cytokines

Background:Aberrant T cell responses are key in driving autoimmunity and are commonly associated with rheumatoid arthritis (RA). Unravelling pathways of importance in therapeutic partial response and failure is of critical importance, as this will potentially provide new insights into key drivers of immune-mediated pathogenesis.Objectives:To delineate disease-relevant T cell subsets in RA and assess their potential to act as cellular markers amenable to precision medicine approaches, particularly in the context of therapeutic partial or non-response.Methods:FACS-based immunophenotyping and ex-vivo functional response profiles of CD4+CD161+CCR2+CCR5+T cells were performed in peripheral blood mononuclear cells (PBMC) obtained from patients with RA and healthy controls, using previously characterised methodologies. RA patients fulfilled the 2010 ACR/EULAR criteria for RA. All samples were obtained after written consent, with the appropriate ethical approvals in place.Results:RA patients harboured a higher frequency of CCR2+CCR5+cells within the CD4+CD161+T cell compartment compared with healthy controls. In RA patients this T cell subset had a higher proportion of cells that secrete pro-inflammatory cytokines such as IL-17A, GM-CSF, IFN-γ, and TNF. Importantly, the CD4+CD161+CCR2+CCR5+T cell subset was significantly increased in DMARD non-responders compared to both responders and healthy controls. Moreover, in DMARD non-responders, these cells had a propensity to express increased proportions of pro-inflammatory cytokines. Notably, there was also a significant increase in the ratio of effector: regulatory T cell (Teff: Treg) compared to both responders and healthy controls. In addition, the CD4+CD161+CCR2+CCR5+T cell subset was less responsive to suppression by Tregs. In further support of a role for this T cell population in disease pathogenesis, the frequency of CD4+CD161+CCR2+CCR5+T cells significantly correlated with disease activity, as measured by the DAS28 (R2= 0.65; p = 0.003; n=11).Conclusion:Combined, our findings suggest that the CD4+CD161+CCR2+CCR5+T cell subset represents a substantially abnormal T cell subset in RA, exhibiting exaggerated pro-inflammatory responses, numerical abundance relative to Tregs, and resistant to regulation by Tregs. The CD4+CD161+CCR2+CCR5+T cell subset appears to be a marker of therapeutic response status in RA, via its contribution to disease pathology and highlights this subset as a potential therapeutic target in RA.References:[1]McInnes IB, Schett G. The pathogenesis of rheumatoid arthritis.N Engl J Med. 2011;365(23):2205-19.[2]Mexhitaj I, Nyirenda MH, Li R, O’Mahony J, Rezk A, Rozenberg A,et al. Abnormal effector and regulatory T cell subsets in paediatric-onset multiple sclerosis.Brain. 2019;142(3):617-32.[3]Cosmi L, Cimaz R, Maggi L, Santarlasci V, Capone M, Borriello F,et al. Evidence of the transient nature of the Th17 phenotype of CD4+CD161+T cells in the synovial fluid of patients with juvenile idiopathic arthritis.Arthritis Rheum. 2011;63(8):2504-15.Disclosure of Interests:Mukanthu Nyirenda: None declared, Iain McInnes Grant/research support from: Bristol-Myers Squibb, Celgene, Eli Lilly and Company, Janssen, and UCB, Consultant of: AbbVie, Bristol-Myers Squibb, Celgene, Eli Lilly and Company, Gilead, Janssen, Novartis, Pfizer, and UCB, Carl Goodyear: None declared

Download Full-text

Vaccine serologic responses among transplant patients associate with COVID-19 infection and T peripheral helper cells

10.1101/2021.07.11.21260338 ◽

2021 ◽

Author(s):

Jacob E. Lemieux ◽

Amy Li ◽

Matteo Gentili ◽

Cory A. Perugino ◽

Zoe F. Weiss ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

B Cell ◽

Cd4 T Cells ◽

Lung Transplant ◽

T Cell Subsets ◽

Healthy Controls ◽

Transplant Recipients ◽

Vaccine Response ◽

Lung Transplant Recipients

Background: Therapeutically immunosuppressed transplant recipients exhibit attenuated responses to COVID-19 vaccines. To better understand the immune alterations that determined poor vaccine response, we correlated quantities of circulating T and B cell subsets at baseline with longitudinal serologic responses to SARS-CoV-2 mRNA vaccination in heart and lung transplant recipients. Methods: Samples at baseline and at approximately 8 and 30 days after each vaccine dose for 22 heart and lung transplant recipients with no history of COVID-19, four heart and lung transplant recipients with prior COVID-19 infection, and 12 healthy controls undergoing vaccination were analyzed. Anti-spike protein receptor binding domain (RBD) IgG and pseudovirus neutralization activity were measured. Proportions of B and T cell subsets at baseline were comprehensively quantitated. Results: At 8-30 days post vaccination, healthy controls displayed robust anti-RBD IgG responses, whereas heart and lung transplant recipients showed minimally increased responses. A parallel absence of activity was observed in pseudovirus neutralization. In contrast, three of four (75%) transplant recipients with prior COVID-19 infection displayed robust responses at levels comparable to controls. Baseline levels of activated PD-1+ HLA-DR+ CXCR5- CD4+ T cells (also known as T peripheral helper [TPH] cells) and CD4+ T cells strongly predicted the ability to mount a response. Conclusions: Immunosuppressed patients have defective vaccine responses but can be induced to generate neutralizing antibodies after SARS-CoV-2 infection. Strong correlations of vaccine responsiveness with baseline TPH and CD4+ T cell numbers highlights a role for T helper activity in B cell differentiation into antibody secreting cells during vaccine response.

Download Full-text

Failure to demonstrate suppressor T cell activity in the lamina propria of Hymenolepis diminuta-infected rats

Journal of Helminthology ◽

10.1017/s0022149x00012803 ◽

1993 ◽

Vol 67 (1) ◽

pp. 17-23 ◽

Cited By ~ 1

Author(s):

C. Palmas ◽

D. Wakelin ◽

G. Bortoletti ◽

A. R. Ecca ◽

F. Gabriele

Keyword(s):

T Cells ◽

T Cell ◽

Lamina Propria ◽

Sandwich Elisa ◽

Cell Activity ◽

Hymenolepis Diminuta ◽

Suppressor T Cells ◽

Normal Peripheral Blood ◽

Suppressor T Cell ◽

Enzymatic Procedure

AbstractThe aim of this study was to examine whether there was an increase of suppressor T cells, relative to helper T cells, in the intestinal lamina propria of Hymenolepis diminuta-infected rats, a condition which might allow the parasite to survive for the life-span of the host. Lamina propria cells were isolated by enzymatic procedure. All lymphocytes were passed over nylon wool columns in order to remove B cells; then the T cells were purified by a panning technique using monoclonal anti-T cell antibodies. Changes in the mitogen-stimulated synthesis of 1gM, IgG and IgA by normal peripheral blood indicator lymphocytes, as measured by sandwich ELISA, was used as an index of help and/or suppression. No significant suppressor T cell activity was observed in the cultures, either from control or infected intestine.

Download Full-text

Dysfunctional Tregs with Absence of Cytokine-Secreting CD4+ T-Cell Subsets and Marked Expansion of Th1 Cells Is a Hallmark of Idiopathic Aplastic Anaemia (AA)

Blood ◽

10.1182/blood.v116.21.2233.2233 ◽

2010 ◽

Vol 116 (21) ◽

pp. 2233-2233

Author(s):

Shahram Kordasti ◽

Judith C. W. Marsh ◽

Pilar Perez Abellan ◽

Sufyan Alkhan ◽

Janet Hayden ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

B Cells ◽

Cd4 T Cells ◽

Cytokine Secretion ◽

T Cell Subsets ◽

Cd4 T Cell ◽

Th1 Cells ◽

Healthy Controls ◽

Ifn Γ

Abstract Abstract 2233 Introduction: Autoimmunity is an important contributor in the aetiology of AA. Although the expansion of oligoclonal CD8+ T-cells and their correlation with response to immunosuppressive therapy (IST) has been reported previously, the role of CD4+ in the pathogenesis is not elucidated. The focus of this study was to investigate the role of different CD4+ T-cell subsets, including regulatory T-cells (Tregs) and T helpers (Th1, Th2 and Th17) in the pathobiology of idiopathic AA. Patients and Methods: The percentage and absolute numbers of CD4+ and CD8+ T-cell subsets, NK & B cells and dendritic cells (DCs) in peripheral blood were assessed in 42 patients with idiopathic AA prior to any IST and 8 healthy age matched controls. T-cells were stimulated first and stained intracellularly for IFN-γ and TNF-a (Th1), IL-4 (Th2) and IL-17 (Th17). Serum levels of 30 cytokines were measured by 30 Plex bead analysis (Luminex). NK cells were defined as CD3– CD56+. B cells were defined as CD3-CD19+. CD3+ CD4+.T-cell subsets were defined as CD45RO–CD27+ naïve, CD45RO+ CD27+ CD62L+ central memory, CD45RO+ CD27+ CD62L– effector memory, CD45RO+CD27– effectors and CD45RO– CD27– terminal effectors. DCs were defined based on their BDCA 1,2, 3 & CD16 expression. CD4 Tregs were defined as CD3+CD4+ CD25high CD27+Foxp3+. Treg subsets were defined as (1) CD45RA+CD25lo resting Tregs, (2) CD45RA-CD25hi activated Tregs, and (3) cytokine-secreting CD45RA-CD25lo non-Tregs1. Treg function was evaluated by cytokine secretion of T effector cells (Te) with and without Tregs. IFN-γ secreting CD4+ T-cells (Th1) were enriched by magnetic beads followed by FACS sorting. The clonality of Th1 cells was evaluated based on the diversity of T-cell receptors by spectratyping as well as sequencing. Transcription factor expression was measured by qPCR. Results: There were no significant differences in the number or percentage of different CD8 T-cells compared to healthy controls. Surprisingly, despite a borderline decrease in the absolute number of naïve (p=0.19) and central memory (p=0.20) CD4+T-cells the number and percentage of Tregs were no different from healthy controls (1.36×107/L v 1.34×107/L, p=0.57). Although the ratio of Tregs to CD4+ T-effectors (Te) was higher than in healthy controls, the difference was not significant (0.49 v 0.12, p=0.86). The absolute numbers and percentages of Th1 cells and TNF-α + CD4+ T-cells were significantly higher in AA patients compared to healthy controls (4.2 × 107/L v 0.9 × 107/L & 2.44 × 108 v 1.26 × 108(p=0.001, p=0.004)). The diversity of T-cell receptor on Th1 cells was significantly lower compared to healthy age matched controls (on average 21 & 52 peaks). Amongst AA patients, the numbers of Th2, Th17, NK and B cells were not significantly different from healthy controls, whereas the absolute numbers of all DCs were reduced(p<0.01). The serum levels of proliferative cytokines, EGF (p=0.01), HGF (p=0.01), VEGF (p=0.01) and pro-inflammatory cytokines IL-13 (p=0.02), IL-8 (p<0.001) were significantly higher in AA patients. The percentage of cytokine secreting CD4+ CD25+ T-cells was markedly decreased in AA patients and the activated Treg subsets were predominantly of CD45RA+ phenotype, which was significantly different from healthy controls. Sorted Tregs from AA patients were unable to suppress cytokine secretion by Te cells in a 1:1 co-culture. However, IL-2, IFN-γ and TNF-α secretion of Te from AA patients was suppressible by allogeneic Tregs from healthy controls (on average 11 time suppression), whereas Tregs from AA patients were unable to suppress healthy Te cells. However, dysfunctional Tregs were not associated with abnormality of transcription factors, as judged by the levels of STAT1, 3, 4, 5 & 6, FoxP3 & T-bet of Tregs that were not significantly different from healthy age matched controls. Conclusion: Our data show that although FoxP3+ Tregs are normal in AA, a subset of these cells is markedly reduced and the activated Tregs aberrantly express CD45RA. Furthermore, unlike normal Tregs, the Tregs from AA patients do not suppress the inflammatory cytokine secretion by Te cells. The absence of DCs in the peripheral blood suggests their immigration to the inflammation site (e.g. bone marrow), which may play a role in the polarisation of T helpers toward a Th1 phenotype. Clonal expansion of Th1 cells may suggest potential antigen specificity that may lead to AA phenotype. 1. Miyara M, et al. Immunity. 2009. Disclosures: No relevant conflicts of interest to declare.

Download Full-text

T-Cell Subsets in Rheumatoid Arthritis Patients on Long-Term Anti-TNF or IL-6 Receptor Blocker Therapy

Mediators of Inflammation ◽

10.1155/2017/6894374 ◽

2017 ◽

Vol 2017 ◽

pp. 1-19 ◽

Cited By ~ 17

Author(s):

Sonja Dulic ◽

Zsófia Vásárhelyi ◽

Florentina Sava ◽

László Berta ◽

Balázs Szalay ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

T Cells ◽

T Cell ◽

T Cell Subsets ◽

Cell Phenotype ◽

Healthy Controls ◽

Biological Therapies ◽

Cd8 Cells ◽

The Impact

Data on the impact of biological therapies on the T-cell phenotype in rheumatoid arthritis are limited. Here, we prospectively measured the percentages of 15 circulating T-cell subtypes using flow cytometry. We obtained transversal and longitudinal data in 30 anti-TNF responders, 19 secondary anti-TNF nonresponders, and 43 IL-6R antagonist responders, before, 8 weeks and at least 6 months after biological therapy. Untreated RA patients and healthy controls were also included. The important findings are the following: (1) the proportion of regulatory T-cells (Tregs) which are decreased in untreated RA patients becomes normal in all long-term-treated groups; (2) in anti-TNF responders as well as in nonresponders, the frequencies of naïve CD4+ and CD8+ cells are lower, whereas those of proinflammatory Th1, Th2, and Th17 cells and HLA-DR+-activated cells are higher than those in untreated RA or healthy controls; (3) in IL-6R responders, Th1 proportion is decreased, while that of Th2 and Th17 is increased as compared to that in anti-TNF-treated patients and controls; (4) pending confirmation, a CD4CD69 ratio < 2.43 at baseline, could be useful to predict a good therapeutic response to anti-TNF therapy. This study provides comprehensive information regarding the long-term impacts of those biological therapies on the ecotaxis of T-cells in RA. The ClinicalTrials.gov registration number of our study is NCT03266822.

Download Full-text