Single-Cell RNA Sequencing Reveals Expanded Clones of Islet Antigen-Reactive CD4+ T Cells in Peripheral Blood of Subjects with Type 1 Diabetes

ABSTRACTChicken peripheral blood lymphocytes (PBLs) exhibit wide-ranging cell types, but current understanding of their subclasses, immune cell classification, and function is limited and incomplete. Previously, we found that viremia caused by avian leukosis virus subgroup J (ALV‐J) was eliminated by 21 days post infection (DPI), accompanied by increased CD8+ T cell ratio in PBLs and low antibody levels. Here we performed single-cell RNA sequencing (scRNA-seq) of PBLs in ALV-J infected and control chickens at 21 DPI to determine chicken PBL subsets and their specific molecular and cellular characteristics, before and after viral infection. Eight cell clusters and their potential marker genes were identified in chicken PBLs. T cell populations (clusters 6 and 7) had the strongest response to ALV-J infection at 21 DPI, based on detection of the largest number of differentially expressed genes (DEGs). T cell populations of clusters 6 and 7 could be further divided into four subsets: activated CD4+ T cells (cluster A0), Th1-like cells (cluster A2), Th2-like cells (cluster A1), and cytotoxic CD8+ T cells. Hallmark genes for each T cell subset response to viral infection were initially identified. Furthermore, pseudotime analysis results suggested that chicken CD4+ T cells could potentially differentiate into Th1-like and Th2-like cells. Moreover, ALV-J infection probably induced CD4+ T cell differentiation into Th1-like cells in which the most immune related DEGs were detected. With respect to the control group, ALV-J infection also had an obvious impact on PBL cell composition. B cells showed inconspicuous response and their numbers decreased in PBLs of the ALV-J infected chickens at 21 DPI. Percentages of cytotoxic Th1-like cells and CD8+ T cells were increased in the T cell population of PBLs from ALV-J infected chicken, which were potentially key mitigating factors against ALV-J infection. More importantly, our results provided a rich resource of gene expression profiles of chicken PBL subsets for a systems-level understanding of their function in homeostatic condition as well as in response to viral infection.

Download Full-text

GAD65-Specific CD4+ T-Cells with High Antigen Avidity Are Prevalent in Peripheral Blood of Patients With Type 1 Diabetes

Diabetes ◽

10.2337/diabetes.53.8.1987 ◽

2004 ◽

Vol 53 (8) ◽

pp. 1987-1994 ◽

Cited By ~ 83

Author(s):

H. Reijonen ◽

R. Mallone ◽

A.-K. Heninger ◽

E. M. Laughlin ◽

S. A. Kochik ◽

...

Keyword(s):

T Cells ◽

Type 1 Diabetes ◽

Peripheral Blood ◽

Cd4 T Cells

Download Full-text

Immunological characterisations of peripheral blood memory CD4+ T cells in children with type 1 diabetes, their non-diabetic siblings and age- and gender-matched unrelated healthy control children

10.14264/uql.2015.809 ◽

2015 ◽

Author(s):

Michael Lei Bian

Keyword(s):

T Cells ◽

Type 1 Diabetes ◽

Peripheral Blood ◽

Cd4 T Cells ◽

Age And Gender ◽

Healthy Control ◽

And Gender ◽

Memory Cd4 T Cells

Download Full-text

GAD65- and proinsulin-specific CD4+ T-cells detected by MHC class II tetramers in peripheral blood of type 1 diabetes patients and at-risk subjects

Journal of Autoimmunity ◽

10.1016/j.jaut.2005.09.018 ◽

2005 ◽

Vol 25 (3) ◽

pp. 235-243 ◽

Cited By ~ 63

Author(s):

Viveka Öling ◽

Jane Marttila ◽

Jorma Ilonen ◽

William W. Kwok ◽

Gerald Nepom ◽

...

Keyword(s):

At Risk ◽

T Cells ◽

Type 1 Diabetes ◽

Mhc Class Ii ◽

Peripheral Blood ◽

Cd4 T Cells ◽

Class Ii ◽

Diabetes Patients

Download Full-text

A new hypothesis for type 1 diabetes risk: The at-risk allele at rs3842753 associates with increased beta cell INS mRNA in a meta-analysis of single cell RNA sequencing data

Canadian Journal of Diabetes ◽

10.1016/j.jcjd.2021.03.007 ◽

2021 ◽

Author(s):

Su Wang ◽

Stephane Flibotte ◽

Joan Camunas-Soler ◽

Patrick E. MacDonald ◽

James D. Johnson

Keyword(s):

Type 1 Diabetes ◽

Beta Cell ◽

Single Cell ◽

Rna Sequencing ◽

Risk Allele ◽

Meta Analysis ◽

Sequencing Data ◽

Single Cell Rna Sequencing ◽

New Hypothesis

Download Full-text

Phenotypic and Epigenetic Adaptations of Cord Blood CD4+ T Cells to Maternal Obesity

Frontiers in Immunology ◽

10.3389/fimmu.2021.617592 ◽

2021 ◽

Vol 12 ◽

Author(s):

Suhas Sureshchandra ◽

Norma Mendoza ◽

Allen Jankeel ◽

Randall M. Wilson ◽

Nicole E. Marshall ◽

...

Keyword(s):

T Cells ◽

Cord Blood ◽

Single Cell ◽

Rna Sequencing ◽

Cd4 T Cells ◽

Maternal Obesity ◽

Ex Vivo ◽

Microbial Infection ◽

Single Cell Rna Sequencing ◽

The Impact

Pregravid obesity has been shown to disrupt the development of the offspring’s immune system and increase susceptibility to infection. While the mechanisms underlying the impact of maternal obesity on fetal myeloid cells are emerging, the consequences for T cells remain poorly defined. In this study, we collected umbilical cord blood samples from infants born to lean mothers and mothers with obesity and profiled CD4 T cells using flow cytometry and single cell RNA sequencing at resting and following ex vivo polyclonal stimulation. We report that maternal obesity is associated with higher frequencies of memory CD4 T cells suggestive of in vivo activation. Moreover, single cell RNA sequencing revealed expansion of an activated subset of memory T cells with maternal obesity. However, ex vivo stimulation of purified CD4 T cells resulted in poor cytokine responses, suggesting functional defects. These phenotypic and functional aberrations correlated with methylation and chromatin accessibility changes in loci associated with lymphocyte activation and T cell receptor signaling, suggesting a possible link between maternal obesogenic environment and fetal immune reprogramming. These observations offer a potential explanation for the increased susceptibility to microbial infection in babies born to mothers with obesity.

Download Full-text

Single-Cell RNA Sequencing Revealed CD14+ Monocytes Increased in Patients With Takayasu’s Arteritis Requiring Surgical Management

Frontiers in Cell and Developmental Biology ◽

10.3389/fcell.2021.761300 ◽

2021 ◽

Vol 9 ◽

Author(s):

Gao Qing ◽

Wu Zhiyuan ◽

Yu Jinge ◽

Miao Yuqing ◽

Chen Zuoguan ◽

...

Keyword(s):

T Cells ◽

Dendritic Cells ◽

Single Cell ◽

Rna Sequencing ◽

Surgical Management ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Clinical Markers ◽

Gene Expressions ◽

Single Cell Rna Sequencing

Objectives: Takayasu Arteritis (TA) is a highly specific vascular inflammation and poses threat to patients’ health. Although some patients have accepted medical treatment, their culprit lesions require surgical management (TARSM). This study aimed at dissecting the transcriptomes of peripheral blood mononuclear cells (PBMCs) in these patients and to explore potential clinical markers for TA development and progression.Methods: Peripheral blood were collected from four TA patients requiring surgical management and four age-sex matched healthy donors. Single cell RNA sequencing (scRNA-seq) was adopted to explore the transcriptomic diversity and function of their PBMCs. ELISA, qPCR, and FACS were conducted to validate the results of the analysis.Results: A total of 29918 qualified cells were included for downstream analysis. Nine major cell types were confirmed, including CD14+ monocytes, CD8+ T cells, NK cells, CD4+ T cells, B cells, CD16+ monocytes, megakaryocytes, dendritic cells and plasmacytoid dendritic cells. CD14+ monocytes (50.0 vs. 39.3%, p < 0.05) increased in TA patients, as validated by FACS results. TXNIP, AREG, THBS1, and CD163 increased in TA patients. ILs like IL-6, IL-6STP1, IL-6ST, IL-15, and IL-15RA increased in TA group.Conclusion: Transcriptome heterogeneities of PBMCs in TA patients requiring surgical management were revealed in the present study. In the patients with TA, CD14+ monocytes and gene expressions involved in oxidative stress were increased, indicating a new treatment and research direction in this field.

Download Full-text