An Immune Atlas of Nephrolithiasis: Single-Cell Mass Cytometry on SIRT3 Knockout and Calcium Oxalate-Induced Renal Injury

Background. As a common urological disease with a high recurrence rate, nephrolithiasis caused by CaOx may elicit a strong immunologic response. We present a CyTOF-based atlas of the immune landscape in nephrolithiasis models to understand how the immune system contributes to, and is affected by, the underlying response caused by SIRT3 knockout and CaOx inducement. Materials and Methods. We performed a large-scale CyTOF analysis of immune cell abundance profiles in nephrolithiasis. The immunophenotyping data were collected from four different mouse models, including the SIRT3 wild-type or knockout, including and excluding CaOx inducement. Unsupervised analysis strategies, such as SPADE and viSNE, revealed the intrarenal resident immune components and the immune alterations caused by SIRT3 knockout and CaOx-induced renal injury. Results. An overview analysis of the immune landscape identified T cells and macrophages as the main immune cell population in nephrolithiasis models. Highly similar phenotypes were observed among CD4+ and CD8+ T cell subsets, including cells expressing Ki67, Ly6C, Siglec-F, and TCRβ. Macrophages expressed a characteristic panel of markers with varied expression levels including MHC II, SIRPα, CD11c, Siglec-F, F4/80, CD64, and CD11b, indicating more subtle differences in marker expression than T cells. The SIRT3KO/CaOx and SIRT3WT/CaOx groups exhibited global differences in the intrarenal immune landscape, whereas only small differences existed between the SIRT3KO/CaOx and SIRT3KO/Ctrl groups. Among the major immune lineages, the response of CD4+ T cells, NK cells, monocytes, and M1 to CaOx inducement was regulated by SIRT3 expression in contrast to the expression changes of B cells, DCs, and granulocytes caused by CaOx inducement. The panel of immune markers influenced by CaOx inducement significantly varied with and without SIRT3 knockout. Conclusion. In a CaOx-induced nephrolithiasis model, SIRT3 has a critical role in regulating the immune system, especially in reducing inflammatory injury. The characteristic panel of altered immune clusters and markers provides novel insights leading to improved prediction and management of nephrolithiasis.

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Human Tissue-Resident Memory T Cells in the Maternal–Fetal Interface. Lost Soldiers or Special Forces?

Cells ◽

10.3390/cells9122699 ◽

2020 ◽

Vol 9 (12) ◽

pp. 2699

Author(s):

Caitlin S. DeJong ◽

Nicholas J. Maurice ◽

Stephen A. McCartney ◽

Martin Prlic

Keyword(s):

T Cells ◽

Immune System ◽

Cell Function ◽

Immune Cell ◽

Memory T Cells ◽

Critical Role ◽

Immune Cell Function ◽

Histocompatibility Complex ◽

Resident Memory T Cells ◽

Maternal Fetal Interface

The immune system plays a critical role during pregnancy, but the specific mechanisms and immune cell function needed to support pregnancy remain incompletely understood. Despite decades of research efforts, it is still unclear how the immune system maintains tolerance of fetal-derived tissues, which include most cells of the placenta and of course the fetus itself, without forfeiting the ability to protect against harmful infections. T cells recognize antigen in the context of major histocompatibility complex (MHC) encoded proteins, but classical MHC class I and II expression are diminished in fetal-derived cells. Can T cells present at the maternal–fetal interface (MFI) protect these cells from infection? Here we review what is known in regard to tissue-resident memory T (Trm) cells at the MFI. We mainly focus on how Trm cells can contribute to protection in the context of the unique features of the MFI, such as limited MHC expression as well as the temporary nature of the MFI, that are not found in other tissues.

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Molecular Bases for Pharmacotherapy of COVID -19

Medical & Clinical Research ◽

10.33140/mcr.06.024 ◽

2021 ◽

Vol 6 (5) ◽

Keyword(s):

T Cells ◽

Immune System ◽

Mortality Rate ◽

Large Scale ◽

Tissue Injury ◽

Diffuse Alveolar Damage ◽

New Drugs ◽

Patient Death ◽

Double Positive ◽

Host Cell Membrane

The most large-scale challenge aroused at the beginning of Y2020 was the global spread of the coronavirus disease 2019 (COVID-19), caused by a zoonotic beta-coronavirus. One year after we have nearly 270 thousand confirmed cases with mortality rate 1.3% in Georgia, and almost 120 billion confirmed cases with mortality rate 2.2% worldwide. As it is known, COVID-19 is triggered by coronavirus species 2 or SARS-CoV-2, which inters in the human body by binding to the angiotensin-converting enzyme 2 (ACE2) molecule on the host cell membrane via the viral spike protein and expresses complex pathological changes in many organs linked with vascular injuries. The most severe expression of this disease exposed by microscopic examination is bilateral diffuse alveolar damage with fibroblasts exudates, indicating Acute Respiratory Distress Syndrome (ARDS). Immune system plays crucial role in tissue damage. As clinical researches showed, the number of peripheral CD4+ and CD8 + T cells were significantly reduced, while their activity was hyper-expressed as evidenced by the high proportions of HLADR (CD4 3•47%) and CD38 (CD8 39•4%) double-positive fractions. Moreover, there was identified an amplified concentration of highly pro inflammatory CCR6+ Th17 in CD4 T cells. This date explains that severe tissue injury in later stages of COVID-19 is depend on the immune system abnormalities, but not on SARS-CoV-2 direct cell destruction. In the same time the scientists and doctors found out abnormalities in coagulation function in most of the severe COVID-19 patients, which were expressed in elevation of D-Dimer level and prolongation of prothrombin time, some of whom terminated in disseminated intravascular coagulation (DIC), deep venous thrombosis (DVT) or fatal pulmonary thromboembolism (PTE). At the later stage in some severe patients it was identified thrombocytopenia as a result of excessive platelets consuming, which significantly affected on treatment and prognosis. More than 300 drugs are used for the treatment of COVID-19 worldwide. Now, the most popular treatments include Remdesivir, Hydroxychloroquine, Betamethasone, Tocilizumab, anti HIV drugs, and convalescent plasma. In the same time, WHO supports vaccines distribution for immunization. Currently, almost 8 vaccines are approved by different countries and more than 180 vaccines are under the clinical trails. Conclusion & Significance: Up till now it is challenging problem to combat SARS-CoV-2 with not well-defined origin and inexplicable biological characteristics as well as to control a pandemic of COVID-19 with such a high R0, a long incubation period and different disease outcomes. Unfortunately, we have limited understandings of particular mechanisms running to abnormal expression of immune system and coagulation processes. In the same time, we don’t have complete picture of vasculopathy leading to the tissue injury and patient death. Therefore, it is problematic to manage SARS-CoV-2 induced processes successfully using available drugs with no significant restoring effect on the organ damages in severe COVID-19 patients. So, we need new targets and new drugs for the prophylaxes and treatment of COVID-19 even we have vaccines available.

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The impact of body mass index on adaptive immune cells in the human bone marrow

10.21203/rs.2.22852/v2 ◽

2020 ◽

Author(s):

Luca Pangrazzi ◽

Erin Naismith ◽

Carina Miggitsch ◽

Jose’ Antonio Carmona Arana ◽

Michael Keller ◽

...

Keyword(s):

Body Mass Index ◽

Bone Marrow ◽

T Cells ◽

Immune System ◽

T Cell ◽

Immune Cells ◽

Memory T Cells ◽

T Cell Subsets ◽

Adaptive Immune ◽

Adaptive Immune Cells

Abstract Background. Obesity has been associated with chronic inflammation and oxidative stress. Both conditions play a determinant role in the pathogenesis of age-related diseases, such as immunosenescence. Adipose tissue can modulate the function of the immune system with the secretion of molecules influencing the phenotype of immune cells. The importance of the bone marrow (BM) in the maintenance of antigen-experienced adaptive immune cells has been documented in mice. Recently, some groups have investigated the survival of effector/memory T cells in the human BM. Despite this, whether high body mass index (BMI) may affect immune cells in the BM and the production of molecules supporting the maintenance of these cells it is unknown.Methods. Using flow cytometry, the frequency and the phenotype of immune cell populations were measured in paired BM and PB samples obtained from persons with different BMI. Furthermore, the expression of BM cytokines was assessed. The influence of cytomegalovirus (CMV) on T cell subsets was additionally considered, dividing the donors into the CMV- and CMV+ groups.Results. Our study suggests that increased BMI may affect both the maintenance and the phenotype of adaptive immune cells in the BM. While the BM levels of IL-15 and IL-6, supporting the survival of highly differentiated T cells, and oxygen radicals increased in overweight persons, the production of IFNγ and TNF by CD8+ T cells was reduced. In addition, the frequency of B cells and CD4+ T cells positively correlated with BMI in the BM of CMV- persons. Finally, the frequency of several T cell subsets, and the expression of senescence/exhaustion markers within these subpopulations, were affected by BMI. In particular, the levels of bona fide memory T cells may be reduced in overweight persons.Conclusion. Our work suggests that, in addition to aging and CMV, obesity may represent an additional risk factor for immunosenescence in adaptive immune cells. Metabolic interventions may help in improving the fitness of the immune system in the elderly.

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Prolonged activation of nasal immune cell populations and development of tissue-resident SARS-CoV-2 specific CD8 T cell responses following COVID-19

10.1101/2021.04.19.21255727 ◽

2021 ◽

Author(s):

Anna H.E. Roukens ◽

Marion König ◽

Tim Dalebout ◽

Tamar Tak ◽

Shohreh Azimi ◽

...

Keyword(s):

T Cells ◽

Immune System ◽

T Cell ◽

Nasal Mucosa ◽

Immune Cell ◽

Inflammatory Responses ◽

Upper Airway ◽

Effector Memory ◽

Long Term Effects ◽

Activated T Cells

AbstractThe immune system plays a major role in Coronavirus Disease 2019 (COVID-19) pathogenesis, viral clearance and protection against re-infection. Immune cell dynamics during COVID-19 have been extensively documented in peripheral blood, but remain elusive in the respiratory tract. We performed minimally-invasive nasal curettage and mass cytometry to characterize nasal immune cells of COVID-19 patients during and 5-6 weeks after hospitalization. Contrary to observations in blood, no general T cell depletion at the nasal mucosa could be detected. Instead, we observed increased numbers of nasal granulocytes, monocytes, CD11c+ NK cells and exhausted CD4+ T effector memory cells during acute COVID-19 compared to age-matched healthy controls. These pro-inflammatory responses were found associated with viral load, while neutrophils also negatively correlated with oxygen saturation levels. Cell numbers mostly normalized following convalescence, except for persisting CD127+ granulocytes and activated T cells, including CD38+ CD8+ tissue-resident memory T cells. Moreover, we identified SARS-CoV-2 specific CD8+ T cells in the nasal mucosa in convalescent patients. Thus, COVID-19 has both transient and long-term effects on the immune system in the upper airway.

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Specific expression of the human CD4 gene in mature CD4+ CD8- and immature CD4+ CD8+ T cells and in macrophages of transgenic mice

Molecular and Cellular Biology ◽

10.1128/mcb.14.2.1084-1094.1994 ◽

1994 ◽

Vol 14 (2) ◽

pp. 1084-1094

Author(s):

Z Hanna ◽

C Simard ◽

A Laperrière ◽

P Jolicoeur

Keyword(s):

T Cells ◽

T Cell ◽

Transgenic Mice ◽

Cd8 T Cells ◽

Transcription Initiation ◽

Critical Role ◽

Regulatory Elements ◽

T Cell Subsets ◽

Double Positive ◽

Specific Expression

The CD4 protein plays a critical role in the development and function of the immune system. To gain more insight into the mechanism of expression of the human CD4 gene, we cloned 42.2 kbp of genomic sequences comprising the CD4 gene and its surrounding sequences. Studies with transgenic mice revealed that a 12.6-kbp fragment of the human CD4 gene (comprising 2.6 kbp of 5' sequences upstream of the transcription initiation site, the first two exons and introns, and part of exon 3) contains the sequences required to support the appropriate expression in murine mature CD4+ CD8- T cells and macrophages but not in immature double-positive CD4+ CD8+ T cells. Expression in CD4+ CD8+ T cells was found to require additional regulatory elements present in a T-cell enhancer fragment recently identified for the murine CD4 gene (S. Sawada and D. R. Littman, Mol. Cell. Biol. 11:5506-5515, 1991). These results suggest that expression of CD4 in mature and immature T-cell subsets may be controlled by distinct and independent regulatory elements. Alternatively, specific regulatory elements may control the expression of CD4 at different levels in mature and immature T-cell subsets. Our data also indicate that mouse macrophages contain the regulatory factors necessary to transcribe the human CD4 gene.

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Single-Cell Analysis of the Neonatal Immune System Across the Gestational Age Continuum

Frontiers in Immunology ◽

10.3389/fimmu.2021.714090 ◽

2021 ◽

Vol 12 ◽

Author(s):

Laura S. Peterson ◽

Julien Hedou ◽

Edward A. Ganio ◽

Ina A. Stelzer ◽

Dorien Feyaerts ◽

...

Keyword(s):

T Cells ◽

Immune System ◽

Single Cell ◽

Gestational Age ◽

Immune Cell ◽

Single Cell Analysis ◽

Inflammatory Diseases ◽

Interferon Α ◽

Cytotoxic Lymphocytes ◽

Neonatal Immune System

Although most causes of death and morbidity in premature infants are related to immune maladaptation, the premature immune system remains poorly understood. We provide a comprehensive single-cell depiction of the neonatal immune system at birth across the spectrum of viable gestational age (GA), ranging from 25 weeks to term. A mass cytometry immunoassay interrogated all major immune cell subsets, including signaling activity and responsiveness to stimulation. An elastic net model described the relationship between GA and immunome (R=0.85, p=8.75e-14), and unsupervised clustering highlighted previously unrecognized GA-dependent immune dynamics, including decreasing basal MAP-kinase/NFκB signaling in antigen presenting cells; increasing responsiveness of cytotoxic lymphocytes to interferon-α; and decreasing frequency of regulatory and invariant T cells, including NKT-like cells and CD8+CD161+ T cells. Knowledge gained from the analysis of the neonatal immune landscape across GA provides a mechanistic framework to understand the unique susceptibility of preterm infants to both hyper-inflammatory diseases and infections.

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Evolution during primary HIV infection does not require adaptive immune selection

10.1101/2020.12.07.20245480 ◽

2020 ◽

Author(s):

David A Swan ◽

Morgane Rolland ◽

Joshua Herbeck ◽

Joshua T Schiffer ◽

Daniel B Reeves

Keyword(s):

Viral Load ◽

Immune System ◽

Hiv Infection ◽

Evolutionary Dynamics ◽

Immune Cell ◽

Adaptive Immune System ◽

Viral Fitness ◽

Immune Cell Population ◽

Adaptive Immune

AbstractModern HIV research depends crucially on both viral sequencing and population measurements. To directly link mechanistic biological processes and evolutionary dynamics during HIV infection, we developed multiple within-host phylodynamic (wi-phy) models of HIV primary infection for comparative validation against viral load and evolutionary dynamics data. The most parsimonious and accurate model required no positive selection, suggesting that the host adaptive immune system reduces viral load, but does not drive observed viral evolution. Rather, random genetic drift primarily dictates fitness changes. These results hold during early infection, and even during chronic infection when selection has been observed, viral fitness distributions are not largely different from in vitro distributions that emerge without adaptive immunity. These results highlight how phylogenetic inference must consider complex viral and immune-cell population dynamics to gain accurate mechanistic insights.One sentence summaryThrough the lens of a unified population and phylodynamic model, current data show the first wave of HIV mutations are not driven by selection by the adaptive immune system.

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CD4+ T cells are essential for the development of destructive thyroiditis induced by anti–PD-1 antibody in thyroglobulin-immunized mice

Science Translational Medicine ◽

10.1126/scitranslmed.abb7495 ◽

2021 ◽

Vol 13 (593) ◽

pp. eabb7495

Author(s):

Yoshinori Yasuda ◽

Shintaro Iwama ◽

Daisuke Sugiyama ◽

Takayuki Okuji ◽

Tomoko Kobayashi ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Lymph Nodes ◽

Mononuclear Cells ◽

Critical Role ◽

T Cell Subsets ◽

Effector Memory ◽

Histopathological Analysis ◽

Activated T Cells ◽

Cervical Lymph Nodes

Immune-related adverse events induced by anti–programmed cell death–1 antibodies (PD-1-Ab), including destructive thyroiditis (thyroid-irAE), are thought to be caused by activated T cells. However, the T cell subsets that are directly responsible for damaging self-organs remain unclear. To clarify which T cell subsets are involved in the development of thyroid-irAE, a mouse model of thyroid-irAE was analyzed. PD-1-Ab administration 2.5 months after immunization with thyroglobulin caused destructive thyroiditis. Thyroiditis was completely prevented by previous depletion of CD4+ T cells and partially prevented by depleting CD8+ T cells. The frequencies of central and effector memory CD4+ T cell subsets and the secretion of interferon-γ after stimulation with thyroglobulin were increased in the cervical lymph nodes of mice with thyroid-irAE compared with controls. Histopathological analysis revealed infiltration of CD4+ T cells expressing granzyme B in thyroid glands and major histocompatibility complex class II expression on thyrocytes in mice with thyroid-irAE. Adoptive transfer of CD4+ T cells from cervical lymph nodes in mice with thyroid-irAE caused destruction of thyroid follicular architecture in the irradiated recipient mice. Flow cytometric analyses showed that the frequencies of central and effector memory CD4+ T cells expressing the cytotoxic marker CD27 were higher in peripheral blood mononuclear cells collected from patients with thyroid-irAE induced by PD-1-Ab versus those without. These data suggest a critical role for cytotoxic memory CD4+ T cells activated by PD-1-Ab in the pathogenesis of thyroid-irAE.

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Body mass index affects adaptive immune cells in the human bone marrow

10.21203/rs.2.22852/v1 ◽

2020 ◽

Author(s):

Luca Pangrazzi ◽

Erin Naismith ◽

Carina Miggitsch ◽

Jose’ Antonio Carmona Arana ◽

Michael Keller ◽

...

Keyword(s):

Body Mass Index ◽

Bone Marrow ◽

T Cells ◽

Immune System ◽

T Cell ◽

Body Mass ◽

Immune Cells ◽

T Cell Subsets ◽

Adaptive Immune ◽

Adaptive Immune Cells

Abstract Background. Obesity has been associated with chronic inflammation and oxidative stress. Both conditions play a determinant role in the pathogenesis of age-related diseases, such as immunosenescence. Adipose tissue can modulate the function of the immune system with the secretion of molecules influencing the phenotype of immune cells. Recently, the importance of the bone marrow (BM) in the maintenance of antigen-experienced adaptive immune cells has been documented. Despite this, whether high body mass index (BMI) may affect immune cells in the BM and the production of molecules supporting the maintenance of these cells it is unknown. Methods. Using flow cytometry, the frequency and the phenotype of immune cell populations were measured in paired BM and PB samples obtained from persons with different BMI. Furthermore, the expression of BM cytokines was assessed. The influence of cytomegalovirus (CMV) on T cell subsets was additionally considered, dividing the donors into the CMV - and CMV + groups. Results. Our study suggests that increased BMI may affect both the maintenance and the phenotype of adaptive immune cells in the BM. While the BM levels of IL-15 and IL-6, supporting the survival of highly differentiated T cells, and oxygen radicals increased in overweight persons, the production of IFNγ and TNF by CD8 + T cells was reduced. In addition, the frequency of B cells and CD4 + T cells positively correlated with BMI in the BM of CMV - persons. Finally, the frequency of several T cell subsets, and the expression of senescence/exhaustion markers within these subpopulations, were affected by BMI. In particular, the levels of bona fide memory T cells may be reduced in overweight persons. Conclusion. Our work suggests that obesity may represent an independent risk factor supporting immunosenescence, in addition to aging and CMV. Metabolic interventions may help in improving the fitness of the immune system in the elderly.

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Adaptive and innate immune cell responses in tendons and lymph nodes after tendon injury and repair

Journal of Applied Physiology ◽

10.1152/japplphysiol.00682.2019 ◽

2020 ◽

Vol 128 (3) ◽

pp. 473-482 ◽

Cited By ~ 6

Author(s):

Andrew C. Noah ◽

Thomas M. Li ◽

Leandro M. Martinez ◽

Susumu Wada ◽

Jacob B. Swanson ◽

...

Keyword(s):

T Cells ◽

Lymph Node ◽

Immune System ◽

Innate Immune System ◽

Immune Cell ◽

Tendon Healing ◽

Tendon Injury ◽

Innate Immune ◽

Adaptive Immune ◽

Immune Cell Response

Tendon injuries are a common clinical condition with limited treatment options. The cellular components of the innate immune system, such as neutrophils and macrophages, have been studied in tendon injuries. However, the adaptive immune system, comprising specialized lymphocytes, plays an important role in orchestrating the healing of numerous tissues, but less is known about these cells in tendon healing. To gain a greater understanding of the biological processes that regulate tendon healing, we determined how the cellular components of the adaptive and innate immune system respond to a tendon injury using two-month-old male mice. We observed that lymphatic vasculature is present in the epitenon and superficial regions of Achilles tendons, and that the lymphatics drain into the popliteal lymph node. We then created an acute Achilles tenotomy followed by repair, and collected tendons and popliteal lymph nodes 1, 2, and 4 wk after injury. Tendon injury resulted in a robust adaptive immune cell response that followed an initial innate immune cell response in tendons and lymph nodes. Monocytes, neutrophils, and macrophages initially accumulated at 1 wk after injury in tendons, while dendritic cells and CD4+ T cells peaked at 2 wk after injury. B cells and CD8+ T cells progressively increased over time. In parallel, immune cells of the popliteal lymph node demonstrated a similarly coordinated response to the injury. These results suggest that there is an adaptive immune response to tendon injury, and adaptive immune cells may play a role in regulating tendon healing. NEW & NOTEWORTHY While the innate immune system, consisting of macrophages and related hematopoietic cells, has been studied in tendon injury, less is known about the adaptive immune system. Using a mouse model of Achilles tendon tenotomy and repair, we observed an adaptive immune cell response, consisting of CD4+ and CD8+ T cells, and B cells, which occur through 4 wk after tendon injury. This response appeared to be coordinated by the draining popliteal lymph node.

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