Cellular Immunology Research Articles

As its discovery nears a half century, the widespread use of F4/80 antigen as a differentiation marker of tissue macrophages of the mouse, continues to raise questions in and beyond experimental cellular immunology. Its structure as a 7 transmembrane G Protein-Coupled Receptor initiated the discovery of a diverse family of plasma membrane receptors. This review will trace milestones of research into the expression of F4/80, also known as Emr1, its value as a marker in formulation of the Mononuclear Phagocyte System and its function in a model of peripheral immune tolerance in the anterior chamber of the eye. Human EMR1 is closely related to a primate-restricted myeloid mechanoreceptor, EMR2, with a novel autocatalytic activation mechanism. We describe their relation to structurally related members of the group E adhesion GPCR subfamily and their contributions to homeostatic systems of the body through local plasma membrane cellular interactions.

Abstract Objective: Principal component analysis is a powerful dimensionality reduction tool that can uncover hidden patterns in complex biological data. In cellular immunology research, principal component analysis may help identify meaningful relationships between various biomarkers. This study aims to investigate the applicability of principal component analysis for exploring immunometabolic cellular pathways and behaviors in the context of human peripheral blood monocytes. Methods: This methodological case study analyzed data from 19 healthy young individuals, including body mass index, fasting lipid profiles, and multiparametric flow cytometry of monocyte subsets. Monocytes were classified as classical, intermediate, or nonclassical based on CD14/CD16 expression, and surface markers, cell size, granularity, and intracellular lipid content were assessed. Principal component analysis was performed to explore clusters of correlated parameters and their possible biological significance. Results: In classical and intermediate monocytes, principal component analysis revealed consistent patterns linking decreased CD14 expression with increased cell size, granularity, and lipid accumulation, reflecting known monocyte maturation processes from CD16– to CD16+. An inverse relationship between body mass index and LDL receptor expression was consistently observed, suggesting metabolic influences on monocyte phenotype. Strong positive loadings for CD11b and CD36 further indicated a link between immune activation and lipid uptake pathways. Conclusions: This methodological case study demonstrates that principal component analysis can reveal biologically plausible clusters in multiparametric flow cytometry data, offering new perspectives on immunometabolic interactions. While the small sample size limits general-izability, the findings highlight the potential of principal component analysis for hypothesis generation and pathway discovery in immune cell research.

Abstract Diffuse alveolar damage (DAD) is recognized as a deadly type of acute inflammatory lung injury caused by toxic inhalants, but its cellular and molecular pathogenesis remains largely unclear. In this study, by using a mouse model of ricin-induced DAD, we explored the heterogeneity of recruited monocyte (Mono) and Mono-derived interstitial macrophage (IM) in the DAD lung. There was the development of 2 distinct IM subsets, namely IMpi (pro-inflammatory) and IMai (anti-inflammatory), from recruited Monopi. A subset of recruited Monopi could get the proliferating phenotype (namely pMonopi), and meanwhile pMonopi served as the intermediate of Monopi-to-IMai transition. The presence of growth differentiation factor 15 (GDF15) facilitated Monopi-to-pMonopi-to-IMai transition, whereas GDF15 deficiency exerted the negative feedback effect of enhancing Monopi-to-IMpi shift. These findings provided a cell differentiation landscape for Mono and IM in the DAD lung, which would promote a deeper understanding of cellular immunology of DAD and offer a theoretical basis for developing novel therapeutic strategies against acute lung injury.

Initial success with B cell-targeted chimeric antigen receptor (CAR) T cells for the treatment of systemic lupus erythematosus and other rheumatic diseases has generated enthusiasm for the broad application of this technology outside of the field of oncology. Paediatric patients with severe rheumatic diseases require lifelong therapy with a substantial toxicity burden and a high cost of care. Paradigm-shifting treatments, including CAR T cells, are desperately needed. Although CAR T cell therapy shows promise for paediatric rheumatic diseases, there are unique aspects of care compared with adults, whichrequire careful consideration and expertise. In response, we established the Integrated Multidisciplinary Paediatric Autoimmunity and Cell Therapy (IMPACT) working group, comprising international experts in the fields of paediatric rheumatology, oncology and cellular therapy, immunology and nephrology, to address the challenges of introducing cell therapies to patients with paediatric-onset autoimmune diseases. Given the possible benefits, we advocate for the study of CAR T cells in paediatric patients with rheumatic diseases who carry a lifelong risk of morbidity and mortality from chronic illness and medication toxicity. As this patient population is relatively small, consensus around definitions of success, robust study of predictors of response and uniform assessment and reporting of toxicities are critical to advancing the field.

Tribute to Mike Cancro – Former editor-in-chief at Cellular Immunology

ABSTRACTAnticipating the evolution of septic patients with community‐acquired pneumonia (CAP) is challenging for front‐line physicians in the Emergency Department (ED). Prognosis depends mainly on early identification, antibiotics, organ support, but also immune status. The objective of this proof‐of‐concept study was to perform a cluster analysis to investigate whether specific phenotypes, including cellular immunology parameters, are associated with the prognosis in patients with CAP presenting to the ED. We performed an exploratory study in the ED of Limoges university hospital (France) on patients with a confirmed CAP. Deterioration was defined by a composite criterion monitored during 7 days following admission: (i) acute respiratory failure with a high flow oxygen requirement, (ii) subsequent ICU admission, (iii) shock, (iv) worsening of organ dysfunction, and (v) in‐hospital mortality. Multicolor Flow Cytometry (MFC) was performed within 12 h after ED admission. Monocyte HLA‐DR (mHLA‐DR) panels consisting of 11 colors for neutrophils and eight colors for lymphocytes were utilized. Phenotypes were defined using non‐supervised hierarchical clustering, including 65 clinical, biological, and immunological variables. During 5 months, 63 patients were prospectively studied (age = 66 ± 19 years; 38 men [60%]; SOFA score = 2.6 ± 1.5; Sepsis = 71%; in‐hospital mortality = 5%) of whom 11 patients (17%) were assigned to the deterioration group. Upon admission, we observed no differences in any markers or in the demographic or clinical characteristics of the patients. In contrast, by performing hierarchical clustering, we identified three groups: Cluster #1 corresponded to a population with a low deterioration (5%) compared with Clusters #2 (23%) and #3 (31%). Markers from the myeloid lineage, including mHLA‐DR, immature neutrophils, and CD64+ neutrophils, were among the parameters discriminating for cluster construction. Cluster #3 displayed the most severe profile, characterized by elevated markers such as CRP, PCT, and immature granulocytes, along with reduced mHLA‐DR levels. A clustering strategy, based on myeloid markers obtained through flow cytometry, provided prognostic insights by identifying three phenotypes with distinct outcomes, while none of the individual markers studied (n = 65, both clinical and biological) showed similar prognostic value. A panel of myeloid markers, alongside clinical management, could optimize patient triage and resource allocation upon ED admission.

Development of molecular genetics and cellular immunology has fundamentally changed the understanding of the relationship between mother and child during pregnancy. For a successful pregnancy, it is necessary for the mother's immune system to recognize and accept the semi-allogeneic fetus, ensuring effective penetration of fetal trophoblasts into the uterine body and protecting the fetus from external pathogens and the mother's microbiota. This is achieved through the exchange of immune and hematopoietic stem cells of the mother and the child with the formation of a cellular chimera called feto-maternal microchimerism as well as vice versa, the formation of the innate immunity of the fetus, ensuring its tolerance to the microbiota and chronic infections of the mother. Innate immunity and hematopoiesis are formed from the cells of the yolk sac mesoderm and extraembryonic mesenchymal tissue due to granule-macrophage-like precursors and pluripotent erythroid cells. During the first trimester of pregnancy and prior to the formation of the thymus and bone marrow hematopoiesis and, therefore, the implementation of the adaptive immune response, the fetus has a formed innate immune defense and cellular control over the biology of its own development represented by neutrophils, eosinophils, basophils, monocytes, macrophages, dendritic cells and the five types of innate lymphoid cells. Innate immune cells are activated by molecular pattern molecules mediated by pattern recognition receptors which provides rapid protection against microbiota pathogens and chronic infections and helps get rid of dying and/or damaged innate cells. Analysis of the interaction between the microbiome, acute and chronic maternal infections, development and functioning of innate lymphoid cells in the fetal and neonatal period is of a great clinical importance aiming at detailing the pathogenesis and developing pathogenetically based therapy, prevention and treatment of eclampsia, pregnancy complications, such as premature birth and other problems associated with placental abruption, bronchopulmonary dysplasia, necrotizing enterocolitis, pneumonia and sepsis in premature and newborn infants. Author presents an analysis of the recent data on the role of innate immunity in terms of evidence-based links with perinatal disorders in newborns and proposals for planning of further scientific research in this important area.

Myocarditis is an inflammatory disease of the myocardium with heterogeneous etiology, clinical presentation, and prognosis; when it is associated with myocardial dysfunction, this identifies the entity of inflammatory cardiomyopathy. In the last few decades, the relevance of the immune system in myocarditis onset and progression has become evident, thus having crucial clinical relevance in terms of treatment and prognostic stratification. In fact, the advances in cardiac immunology have led to a better characterization of the cellular subtypes involved in the pathogenesis of inflammatory cardiomyopathy, whether the etiology is infectious or autoimmune/immune-mediated. The difference in the clinical course between spontaneous recovery to acute, subacute, or chronic progression to end-stage heart failure may be explained not only by classical prognostic markers but also through immune-pathological mechanisms at a cellular level. Nevertheless, much still needs to be clarified in terms of immune characterization and molecular mechanisms especially in biopsy-proven myocarditis. The aims of this review are to (1) describe inflammatory cardiomyopathy etiology, especially immune-mediated/autoimmune forms, (2) analyze recent findings on the role of different immune cells subtypes in myocarditis, (3) illustrate the potential clinical relevance of such findings, and (4) highlight the need of further studies in pivotal areas of myocarditis cellular immunology.

The recent study from the Pogge von Strandmann group published in Cellular and Molecular Immunology, by Alashkar Alhamwe et al., combined for the first time the Cre-LoxP recombination system with single-cell sequencing. The group monitored the tumor-derived extracellular vesicle (EV) uptake and the EV functions in the recipient non-malignant cells in a pancreatic ductal adenocarcinoma mouse model. Recombination events and EV uptake, together with resulting gene expression changes in macrophages, neutrophils, and mast cells, were detected by single-cell sequencing technology of the tumor tissue. This new approach is highly specific, as it can identify single EV recipient cells without interfering with the EV biogenesis or the phenotype.

Genetic link between gut microbiota, immune cells, and rheumatoid arthritis: Mechanism of action of CD28 proteins

Through multidisciplinary research, translational medicine allows a comprehensive approach to diseases in order to provide patients with adequate treatment, whose aim is personalized and precision medicine. In this context, the Cellular Immunology Laboratory of the Immunochemistry Research Unit, and the Dermatology Service of the 21st Century National Medical Center's Specialties Hospital (Hospital de Especialidades del Centro Médico Nacional Siglo XXI) have achieved a partnership that aspires to provide better quality patient care. This objective is achieved by evaluating the inflammatory skin infiltrates in different pathologies to observe changes in local immunity related to the response to treatment, and with the disease progression distinguish between dermatological diseases, highlight the importance of comprehensive skin care, and make decisions about treatment choice. This article will describe the results obtained from different research protocols (derived from this collaboration) in patients with different dermatoses such as warts, squamous cell carcinoma, mycosis fungoides and psoriasis. The evidence presented highlights the importance of collaborative work between clinical services and research laboratories so that scientific knowledge can be translated towards the comprehensive management of patients and towards personalized and precision medicine.

Born in 1940 in Rasht, Iran, Dr. Farrokh Modabber earned his B.A. in Bacteriology and Ph.D. in Microbiology from University of California, Los Angeles (UCLA). He joined the Harvard Medical School as a fellow before transitioning to a faculty role at Harvard School of Public Health (HSPH) and played a pivotal role in advancing Cellular Immunology. In the early 1970s, he returned to Iran as an Associate Professor at Pahlavi University in Shiraz. Subsequently, he rejoined HSPH before embarking on a tenure at Tehran University. As the head of the Pathobiology Department at Tehran University School of Public Health, he initiated the Tehran/Harvard joint M.Sc. program in Immunology, which played a crucial role in shaping the careers of numerous Iranian immunologists over the following decades. Dr. Modabber went on to hold esteemed positions such as Director General of the Pasteur Institute of Iran, visiting immunology lecturer at various universities, Coordinator of Research Capability Strengthening of WHO's Special Program for Research and Training in Tropical Diseases (TDR), Director of the Infectious Disease Research Institute (IDRI), and Senior Advisor of Drugs for Neglected Diseases initiative (DNDi), to name a few. This article highlights Dr. Modabber's impactful career, focusing on his efforts to combat global leishmaniasis.

At one time considered a possible form of neuromyelitis optica (NMO) spectrum disorder (NMOSD), it is now accepted that myelin oligodendrocyte glycoprotein (MOG) antibody (Ab)-associated disorder (MOGAD) is a distinct entity from either NMO or multiple sclerosis (MS) and represents a broad spectrum of clinical phenotypes. Whereas Abs targeting aquaporin-4 (AQP4) in NMO are pathogenic, the extent that anti-MOG Abs contribute to CNS damage in MOGAD is unclear. Both AQP4-specific Abs in NMO and MOG-specific Abs in MOGAD are predominantly IgG1, a T cell-dependent immunoglobulin (Ig) subclass. Key insights in neuroimmunology and MOGAD pathogenesis have been learned from MOG experimental autoimmune encephalomyelitis (EAE), described 2 decades before the term MOGAD was introduced. MOG-specific T cells are required in MOG EAE, and while anti-MOG Abs can exacerbate EAE and CNS demyelination, those Abs are neither necessary nor sufficient to cause EAE. Knowledge regarding the spectrum of MOGAD clinical and radiologic presentations is advancing rapidly, yet our grasp of MOGAD pathogenesis is incomplete. Understanding both the humoral and cellular immunology of MOGAD has implications for diagnosis, treatment, and prognosis.

Macrolide antibiotic azithromycin is widely used in clinical practice to treat respiratory tract infections and inflammatory diseases. However, its mechanism of action is not fully understood. Given the involvement of the CD27 pathway in the pathophysiology of various T-lymphocyte-mediated inflammatory, autoimmune, and lymphoproliferative diseases, we examined the impact of AZM on CD27 regulation and potential consequences on CD4+ and CD8+ T-cell phenotypes. Using cellular immunology approaches on healthy donors' peripheral blood mononuclear cells, we demonstrate AZM-mediated downregulation of surface CD27 expression as well as its extracellular release as soluble CD27. Notably, AZM-exposed CD27high (hi) cells were defective in their ability to expand compared to CD27intermediate (Int) and CD27low (lo) subsets. The defective CD27hi subset expansion was found to be associated with impaired cell proliferation and cell division. At the molecular level, the CD27hi subset exhibited lower mTOR activity than other subsets. Functionally, AZM treatment resulted in marked depletion of helper CD4+ (Th1) and cytotoxic CD8+ T-lymphocyte (Tc1)-associated CXCR3+CD27hi effector cells and inhibition of inflammatory cytokine IFN-γ production. These findings provide mechanistic insights on immunomodulatory features of AZM on T-lymphocyte by altering the CD27 pathway. From a clinical perspective, this study also sheds light on potential clinical benefits observed in patients on prophylactic AZM regimens against various respiratory diseases and opens avenues for future adjunct therapy against Th1- and Tc1-dominated inflammatory and autoimmune diseases.

Avrion Mitchison, universally known as Av, made enormous contributions to the field of cellular immunity. Working under Peter Medawar FRS during his PhD in Oxford, Av made the crucial discovery that viable lymphocytes were responsible for rejection of grafts of foreign tissue, initiating the field of cellular immunology, which was to preoccupy him throughout his career. In 1962 he was appointed head of the Division of Experimental Biology at the National Institute for Medical Research and began using proteins decorated with small synthetic molecules termed haptens, enabling him to make his second major discovery. He showed that optimal antibody responses to a hapten required the cooperation of two cell populations: bone-marrow-derived ‘B’ lymphocytes, which synthesized the antibody, and thymus-derived ‘T’ lymphocytes, which recognized moieties in the carrier protein molecule and provided help to the B cells. In 1971 Av became the Jodrell Professor of Zoology at University College London and head of the Imperial Cancer Research Fund Tumour Immunology Unit (TIU), where he continued to investigate how antibody and cytotoxic T cell responses are regulated and the role of major histocompatibility genes in this process. The TIU was a mecca for immunologists from around the world, and Av inspired those who passed through to make major contributions to cellular immunity, neuroimmunology, leukaemogenesis, tumour immunology and autoimmune disease. In 1991 he became the first director of the Rheumatism Research Centre in Berlin, and in a few short years after the fall of the Berlin wall established a flourishing institute. The annual award of the Mitchison Prize for Young Researchers honours his legacy.

Glioblastoma multiforme (GBM) is the most aggressive malignant brain tumour. The average survival time for a patient diagnosed with GBM, using standard treatment methods, is several months. Authors of the article pose a direct question: Is it possible to treat GBM solely with radioactive iodine (¹³¹I) therapy without employing the sodium iodide symporter (NIS) gene? After all, NIS has been detected not only in the thyroid but also in various tumours. The main author of this article (A.C.), with the assistance of her colleagues (physicians and pharmacologists), underwent ¹³¹I therapy after prior iodine inhibition, resulting in approximately 30% reduction in tumour size as revealed by magnetic resonance imaging (MRI). Classical therapy for GBM encompasses neurosurgery, conventional radiotherapy, and chemotherapy (e.g. temozolomide). Currently, tyrosine kinase inhibitors (imatinib, sunitinib, and sorafenib) are being used. Additionally, novel drugs such as crizotinib, entrectinib, or larotrectinib are being applied. Recently, personalised multimodal immunotherapy (IMI) based on anti-tumour vaccines derived from oncolytic viruses has been developed, concomitant with the advancement of cellular and molecular immunology. Thus, ¹³¹I therapy has been successfully employed for the first time in the case of GBM recurrence.

The investigation of pathological diseases largely relies on laboratory examinations. The ability to identify and characterise cells is an essential process for clinicians to reach an accurate diagnosis and inform appropriate treatments. There is currently a gap between the advancement of scientific knowledge on cellular and molecular pathways and the development of novel techniques capable of detecting subtle cellular changes associated with disease. Biospectroscopy is the use of spectroscopy techniques to investigate biological materials. Within a biological sample, important molecules such as lipids, carbohydrates, nucleic acids, and proteins are held together by chemical bonds; these bonds will vibrate following excitation with infrared light. By measuring the vibrational energy of each molecule present in a biological sample, a unique spectrum, known as the “molecular fingerprint” is generated. As disease-related changes in biological samples will be reflected in the molecular fingerprint, biospectroscopy is a well-placed candidate for the investigation of disease. Biospectroscopy has been gaining wider acceptance and application in the clinical setting over the past decade; however, it has yet to reach diagnostic laboratories and healthcare clinics as a routine platform for clinical assessment. Immunological disorders are complex, often demonstrating interaction across multiple molecular pathways which results in delayed diagnosis. Vibrational spectroscopy is being applied in many fields, and here we present a review of its use in cellular immunology. Potential benefits, including an enhanced definition of molecular processes and the use of spectroscopy in disease diagnosis, monitoring, and treatment response, are discussed. The translation of vibrational spectroscopic techniques into clinical practice offers rapid, noninvasive, and inexpensive methods to obtain information on the molecular composition of biological samples. The potential clinical benefits of biospectroscopy include providing a more prompt and accurate disease diagnosis, thus improving patient care and resulting in better health outcomes.

Molecular Attributes of CAR T Cell Fitness in Patients with Chronic Lymphocytic Leukemia

Rapid and noninvasive detection of macrophage polarization can facilitate the acquisition of crucial information for pre- and post-evaluation in cancer immunotherapy, unraveling tumor immune escape mechanisms and discovering drugs. Herein, we demonstrate an approach to identify macrophage polarization using the metasurface-based terahertz biosensor of quasi-bound state in the continuum. Non-polarized macrophages (M0) are driven into a pro-inflammatory phenotype (M1) or an anti-inflammatory phenotype (M2) in response to lipopolysaccharide or interleukin-4 stimulation. It is found that the M1 macrophages result in a slight resonance frequency blueshift, while the M2 results in an obvious resonance frequency blueshift as well as a broader resonance linewidth. The permittivity change reveals the aforementioned frequency change. Flow cytometry and gene expression measurements reveal distinct differences of phenotypes between M1 and M2 polarizations, consistent with the results of terahertz biosensing. Our results manifest a cost-effective approach for detecting macrophage polarization status and their functional activations, which holds significant implications for clinical and cellular immunology research.

In solid and malignant tumors, innate and adaptive immunity are combined in antitumor responses. This study aimed to analyze the activation of plasma cells and the correlation between the infiltration of B and T lymphocytes with the degree of malignancy or Gleason grade in human prostate biopsies diagnosed with cancer. Prostate cancer biopsies were obtained from the Clinical Hospital of Universidad de Chile (n=70), according to the bioethical norms of the institution. Histological sections of 5µm thickness were processed for immunohistochemistry with primary antibodies against BL and total TL (HRP/DAB). Recognition and quantification were performed under a Leica DM750 optical microscope. Microsoft Excel and GraphPad software were used for the statistical study. Correlation coefficient (Pearson) and mean comparison tests (Kruskal-Wallis and Dunn) and p≤ 0.05 were developed. B and T lymphocyte populations were inversely interregulated in prostate cancer (Gleason) (r= -0.46). Their relationship with Gleason grade is variable according to lymphocyte type (LB vs. Gleason r= -0.0.47 and LT vs. Gleason r= -0.21). Histological diagnosis of prostate cancer correlates with a predominance of LT. The malignancy of the pathology correlates with a predominance of LTs, according to the Gleason grade. The increased knowledge of B and T lymphocyte infiltration and plasma cell activation could be used to better target clinical trials on treatments based on immune system responses. Immunotherapy could be a new paradigm to apply better antitumor therapy strategies.