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Topic:Immunology

The equine immune system is a complex network of cells, tissues, and organs that work collaboratively to defend against pathogens and maintain homeostasis. It consists of innate and adaptive components, each with distinct functions and mechanisms. The innate immune system provides the first line of defense through physical barriers, phagocytic cells, and the complement system. The adaptive immune system involves lymphocytes, such as B cells and T cells, which generate specific responses to antigens and provide immunological memory. Research in equine immunology explores the interactions between these components, the impact of genetic and environmental factors on immune function, and the development of vaccines and therapeutics. This page gathers peer-reviewed studies and scholarly articles focusing on the mechanisms, regulation, and clinical applications of the equine immune system in health and disease.
Comparison of in vitro degradation of immunoreactive adrenocorticotropic hormone and insulin in equine blood measured by two immunoassay methods.
Domestic animal endocrinology    May 2, 2026   Volume 96 107020 doi: 10.1016/j.domaniend.2026.107020
Durham AE, Gautam SS, Huber L.This study compared the in vitro stability of equine plasma adrenocorticotrophic hormone (ACTH) and serum insulin measured by chemiluminescent assay (CLA; Siemens Immulite 2000XPi) and immunofluorescent assay (IFA; Tosoh AIA-900) after storage at 4°C or room temperature (approximately 21°C) for clinically relevant periods. Fresh equine EDTA blood for ACTH and coagulated blood for insulin were aliquoted and analysed immediately (T0) and after storage. ACTH was assessed at 24, 48 and 72 hours; insulin was assessed at 24 hours, 72 hours and 7 days. Percentage recovery relative to T0 was analyse...
Unusual, fatal neck cancer in a foal: immunohistological investigations and differential diagnosis.
Journal of equine veterinary science    May 1, 2026   105921 doi: 10.1016/j.jevs.2026.105921
Romanucci M, Baghini S, Canonici F, Canonici P, Di Luzio Paparatti F, Marruchella G.Neck masses may represent a diagnostic challenge for equine veterinarians, since they may involve different structures with a similar clinical appearance. Whereas neck masses in foals mostly result from non-neoplastic conditions, including various congenital abnormalities, neoplastic cervical masses have been rarely documented. This report describes the unusual case of a rapidly growing neck cancer in a four-month-old colt, resulting in a fatal outcome. Gross and immunohistological findings were most consistent with a poorly differentiated thymic carcinoma with sarcomatoid features and multifo...
In vitro cultivation of Trypanosoma equiperdum: Comparison of in vitro and in vivo antigens for an ethical and sustainable serological diagnosis of Dourine.
Veterinary parasitology    April 30, 2026   Volume 345 110790 doi: 10.1016/j.vetpar.2026.110790
Armillotta G, Capista S, Rodomonti D, Traini S, Testa L, Serroni A, Laguardia C, Palucci C, Di Pancrazio C, Di Febo T, Puglielli O, Ronchi GF....The protozoan Trypanosoma equiperdum is the causative agent of Dourine, a contagious sexually transmitted disease of equines that can manifest itself in chronic or acute form. Dourine has serious veterinary and economic implications, as there are no vaccines available for prevention and the management of the disease requires the culling of infected animals, together with rigorous sanitary control measures. This study compares the traditional production of T. equiperdum antigen grown in vivo, with an in vitro cultivation methodology developed to eliminate the use of animals. The in vitro and in...
Molecular and serological survey on Theileria equi and Babesia caballi infecting Arabian horses in the northern provinces of Syria.
Open veterinary journal    April 30, 2026   Volume 16, Issue 4 2203-2211 doi: 10.5455/OVJ.2026.v16.i4.22
Abdaljwad M, Al-Sabi MNS, Shawaf T, Mokahwi AA, Alfares W, Abdalkader A.Equine babesiosis is one of the prevalent diseases in horses worldwide, but its presence in Syria generally and in the northern region specifically is not known. In Syria, the civil war has severely disrupted veterinary services, hence disease control measures, creating a fertile ground for the spread of tick-borne diseases. Unassigned: This study aimed to conduct a comprehensive molecular and serological survey of and . in Arabian horses from the northern provinces of Syria, regions heavily impacted by the war. Unassigned: A total of 110 Arabian horses, aged between 2 and 10 years, were inc...
Allogeneic Platelet-Rich Gel Supernatant Reprograms the Cytokine and Growth Factor Microenvironment in an Equine In Vitro Inflammatory Tendon System.
International journal of molecular sciences    April 29, 2026   Volume 27, Issue 9 4006 doi: 10.3390/ijms27094006
Carmona JU, López C.Tendinopathy involves a dysregulated inflammatory microenvironment in which cytokines, growth factors (GF) and extracellular matrix components interact dynamically. Platelet-rich plasma (PRP) is widely used as a regenerative therapy, but its mechanisms of action in inflamed tendon remain unclear. This study evaluated whether platelet-rich gel supernatant (PRGS) reprograms the inflammatory-anabolic mediator network in an equine in vitro tendon explant system stimulated with lipopolysaccharide (LPS). Tendon explants were cultured under six experimental conditions, including non-stimulated contro...
Editorial: Biomarker identification and novel therapies in equine medicine.
Frontiers in veterinary science    April 28, 2026   Volume 13 1843968 doi: 10.3389/fvets.2026.1843968
Żak-Bochenek A, Siwińska N.No abstract available
Validation of a stall-side immunoglobulin assay for use in equine reproductive management.
Equine veterinary journal    April 28, 2026   doi: 10.1002/evj.70172
Moore L, McLain A, McDowell SH, Ledgerwood CJ, Bailie E, Nesbit MA, Moore T.Equine foals receive IgG from mare colostrum through passive transfer. Failure of passive transfer (FPT) is a significant risk to the foal's life, leaving them vulnerable to infection and sepsis. Radial Immunodiffusion (RID) and immunoturbidimetric assays quantify IgG present in a foal sample but require a laboratory to complete. Accurate, reproducible, stall-side testing to rapidly quantify IgG would allow for expedited clinical decisions, with potential to improve equine foal care and survival. Objective: To evaluate the analytical and clinical performance of a stall-side IgG lateral-flow te...
A scoping review of neuroendocrinological biomarkers with potential for use in equine behavioral medicine practice.
BMC veterinary research    April 27, 2026   doi: 10.1186/s12917-026-05509-5
Mendonça T, Ribeiro G, Casqueiro T, Prazeres J, Mills DS.BACKGROUND: Equine emotion and welfare are of increasing concern to the general public, equestrians and researchers. There is growing demand for more objective indicators of specific emotional states, as well as the well-being of horses in a variety of contexts. Such markers may also assist in both the diagnosis and selection of biological therapies within equine behavioral medicine. RESULTS: This scoping review summarizes and appraises the published findings relating to relevant biological measurements from the last two decades. Particular attention is given to methodological, subject-related...
Glanders: Past, Present and Future of a Neglected Zoonosis as a Threat in Veterinary and Human Health.
Veterinary medicine and science    April 21, 2026   Volume 12, Issue 3 e70950 doi: 10.1002/vms3.70950
Kanani Y, Oryan A, Derakhshandeh A.Burkholderia mallei is the causative agent of glanders, a serious zoonotic disease of major concern for both animal and human health. This highly contagious and potentially fatal bacterium is always classified as a biothreat agent. In this review, we covered a comprehensive overview of glanders, including the biology of the pathogen, epidemiology, pathogenesis, diagnosis, control strategies and ongoing eradication programs in Eurasia. We also summarized human case reports, differential diagnosis, treatment and organism's antimicrobial properties. The section on vaccine development is among the...
Multiple strategies, one mission: mesenchymal stromal cell-based mechanisms of action in osteoarthritis.
Frontiers in cell and developmental biology    April 20, 2026   Volume 14 1763344 doi: 10.3389/fcell.2026.1763344
Junuzović M, Troillet A, Burk J.Osteoarthritis (OA) is a cross-species, multifactorial joint disease characterized by the progressive degeneration of articular cartilage, morphological remodeling of the subchondral bone, and inflammatory and fibrotic changes of the joint capsule. These alterations arise from chronic, often subclinical, inflammatory processes and dysregulated cellular homeostasis, leading to profound shifts in the cellular and extracellular composition of the joint organ. Although the mechanisms driving persistent inflammation are only partially understood, their impact on all joint-associated tissues is well...
Multivalent Disabled Infectious Single Animal (DISA)-DIVA vaccine for all nine serotypes of African horse sickness virus (AHSV) is broadly protective in IFNAR (-/-) mice.
Veterinary research    April 16, 2026   doi: 10.1186/s13567-026-01753-7
Utrilla-Trigo S, Jiménez-Cabello L, van Gennip RGP, van Rijn PA, Ortego J, Calvo-Pinilla E.African horse sickness (AHS) is a highly severe, midge-borne disease of equids showing a mortality rate of > 90% in naive domestic horses. Outbreaks are caused by AHS virus (AHSV) and result in devastating economic losses in developing countries as equids play a crucial role in the agricultural industry and many households. The AHSV species encompasses nine serotypes showing no or limited cross protection, which is challenging regarding AHS vaccine development. Recently, and like bluetongue virus (BTV), the AHS Disabled Infectious Single Animal (DISA)-DIVA vaccine platform was developed...
Interleukins decrease epithelial protein and gene expression by K14 + CD105+ equine hoof progenitor cells.
Stem cell research & therapy    April 15, 2026   doi: 10.1186/s13287-026-05013-5
Yang Q, Lopez MJ.Background: Progenitor cell epithelial to mesenchymal transition (EMT) could impact healing in the epidermal-dermal junction of the equine hoof stratum internum (SI). The hypothesis of this study was that K14 + CD105+ equine hoof SI progenitor cells assume a mesenchymal phenotype in the presence of inflammatory interleukins in vitro.Methods: K14 + CD105+ progenitor cell percentages isolated from proliferative scarred (fibrous) and healthy SI were quantified. Decellularized matrix was prepared from and K14 + CD105+ cells localized in healthy SI. Gene expression (CD44, CD105, E-cadhe...
Terpenic compounds possess anthelmintic and immunomodulatory properties with potential for controlling equine cyathostomin infections.
International journal for parasitology. Drugs and drug resistance    April 12, 2026   Volume 31 100642 doi: 10.1016/j.ijpddr.2026.100642
Malsa J, Chereau A, Guégnard F, Serreau D, Gesbert A, Reigner F, Chamoin L, Guillot J, Vernudachi A, Mach N, Fleurance G, Lacroix-Lamandé S....The emergence of anthelmintic-resistant parasite isolates necessitates alternative control strategies. The use of dietary additives with anti-parasitic or immunomodulatory activities has been proposed as a solution, but whether such additives can simultaneously exert dual bioactivities within this context has rarely been explored. Here, we evaluated whether selected terpenes could exert dual bioactivities (anti-parasitic and immunomodulatory) with a view to their use as food additives that can limit strongyle infections in horses. In vitro tests with cyathostomin larval development, larval mig...
Learning in the arena: healthcare students’ lived experience of interprofessional education in equine-assisted services.
Journal of interprofessional care    April 9, 2026   1-12 doi: 10.1080/13561820.2026.2651764
Lungren L, Mazzarella J, Mumbauer-Pisano J.This qualitative study explored healthcare students' lived experiences during an interprofessional education (IPE) event incorporating equine-assisted services (EAS). Traditional IPE events often rely on classroom settings that may limit interprofessional engagement, collaboration, and application. Utilizing interpretive phenomenological analysis, researchers examined how the equine environment influenced interprofessional learning among fourteen graduate students from diverse healthcare disciplines at a western United States public university. Participants engaged in experiential demonstratio...
Mucosal T cell activation pathways are upregulated by equine herpesvirus type 1 infection.
Veterinary research    April 6, 2026   doi: 10.1186/s13567-026-01741-x
Holmes CM, Babasyan S, Wagner B.Equine herpesvirus type 1 (EHV-1) is a high morbidity and mortality virus that impacts horse populations worldwide. As a respiratory virus, it enters through the upper respiratory tract (URT), where mucosal immunity plays a crucial role in preventing severe disease. In this study, flow cytometry was used to characterize the nasal leukocyte population during EHV-1 infection, and RNA sequencing of nasal secretions was employed to assess transcriptional markers of the mucosal immune response. Horses with distinct immune statuses were compared at four stages: pre-infection, early (day 1 and 3 post...
Competitive enzyme-linked immunosorbent assay to quantify hepcidin in horse serum.
Journal of equine veterinary science    April 6, 2026   105887 doi: 10.1016/j.jevs.2026.105887
Gluck-Flynn CR, Han H, Valenzuela M, Routh P, Ostland VE, Pratt-Phillips SE.Hepcidin (Hepc) is a peptide hormone primarily produced by hepatocytes and considered the master regulator of iron homeostasis, with Hepc controlling the release of iron within the circulation. Despite the renewed interest in iron-related research due to Hepc discovery, there is limited data on serum hepcidin in horses. The objective of the study was to develop and partially validate a competitive enzyme-linked immunosorbent assay (ELISA) to quantify serum hepcidin in horses. Precision was evaluated using horse serum samples representing low, medium and high Hepc concentrations, yielding intra...
Seroprevalence of Sarcocystis neurona and Neospora hughesi in healthy horses from the province of Alberta, Canada.
Journal of veterinary internal medicine    April 5, 2026   Volume 40, Issue 2 aalag058 doi: 10.1093/jvimsj/aalag058
Sjolin E, Zakia LS, Galezowski A, Whitehead AE.Literature on the seroprevalence of antibodies to the causative agents of equine protozoal myeloencephalitis (EPM) in Canada is limited. Objective: To determine the prevalence of serum antibodies against Sarcocystis neurona and Neospora hughesi in apparently healthy horses without signs of neurologic disease which have always resided in Alberta, Canada, using commercially available immunodiagnostic tests. Methods: Samples from 116 healthy horses were obtained from a biobank containing horse serum collected across Alberta in 2021 and 2022. Samples were selected based on owner survey responses, ...
Evaluation of episcleral silicone matrix cyclosporine implants for presumed equine immune-mediated keratitis: a retrospective study.
Journal of equine veterinary science    April 3, 2026   105880 doi: 10.1016/j.jevs.2026.105880
Bruneel E, Hermange T, Casamatta JM, Pereira A, Pimenta J, Caseiro AR.Immune-mediated keratitis (IMMK) is a chronic inflammatory corneal disease in horses. Long-term topical therapy with cyclosporine is often impractical, highlighting the need for sustained-release alternatives. Objective: To assess clinical response, tolerability, and duration of effect of episcleral silicone matrix cyclosporine implants (ESMC) in horses with presumed IMMK. Methods: Medical records of 12 horses (14 eyes) treated between 2019 and 2023 were retrospectively reviewed. IMMK subtypes were classified as epithelial (n = 3 eyes), anterior stromal (n = 9), or mid-stromal (n =â€...
Development and evaluation of an enzyme-linked immunosorbent assay for measuring antibodies against VP8∗ receptor binding domain of equine rotavirus A.
Virology    April 2, 2026   Volume 619 110898 doi: 10.1016/j.virol.2026.110898
Naveed A, Eertink LG, Soni S, Kennedy MA, Adam E, Wang D, Li F.Equine rotavirus A (ERVA) poses a significant threat to foal health, necessitating reliable serological tools for assessing protective immune responses. This study focused on the development of an enzyme-linked immunosorbent assay (ELISA) for the measurement of antibodies against the VP8∗ receptor binding core domain of ERVA. Following the expression and purification of VP8∗ receptor binding domain from the bacterial expression system, the ELISA for detecting anti-ERVA VP8∗ antibodies with VP8∗ as a coating antigen was developed and its performance was validated by the comparative anal...
Comparative phenotypic and transcriptomic analysis of equine ovarian follicular aspirates-derived mesenchymal stromal cells and dermal fibroblasts.
Animal reproduction science    April 2, 2026   Volume 290 108179 doi: 10.1016/j.anireprosci.2026.108179
Del Prado Soriano-Campos M, Muñoz-García CC, Luis-Calero M, Gallardo-Soler A, González-Fernández L, Macías-García B.Mesenchymal stromal cells (MSCs) are widely used in human cell-based therapies and recent research is focused on the use of MSCs in equine regenerative medicine. Recently, MSCs have been isolated from equine follicular aspirates; however, a major concern during isolation is the potential contamination with fibroblasts, the predominant stromal cell type in many tissues. Furthermore, both cell types share the minimal criteria established by the International Society for Cellular Therapy (ISCT) for MSCs identification. Hence, our study aimed to compare the morphological characteristics, expansion...
In Vitro Characterization of an Equinized Anti-PD-L1 Antibody for Cancer Immunotherapy in Horses.
Veterinary sciences    April 1, 2026   Volume 13, Issue 4 343 doi: 10.3390/vetsci13040343
Horikawa T, Maekawa N, Okagawa T, Tiyamanee W, Ganbaatar O, Nakamura H, Ikehata M, Inoue M, Nakanishi T, Tachibana T, Kato Y, Yamamoto K, Suzuki Y....Horses develop spontaneous tumors, typically in old age. Although local tumor control can be achieved using conventional therapies, systemic therapies are required to treat recurrent and/or metastatic tumors. Immune checkpoint inhibitors, such as anti-PD-L1 antibodies, have been approved for the treatment of various tumor types in humans; however, little is known about the immunosuppressive roles of the PD-1/PD-L1 pathway in horses, and the therapeutic potential of these inhibitors remains to be elucidated. Previously, we reported that the rat monoclonal anti-PD-L1 antibody 6C11-3A11 cross-rea...
Cross-neutralization and antigenic characterization of simian and equine group A rotaviruses.
Journal of virology    March 31, 2026   e0019926 doi: 10.1128/jvi.00199-26
Soni S, Eertink LG, Shuisong N, Loynachan A, Barnum SM, Adam EN, Kennedy MA, Wang D, Li F.Rotaviral diarrhea in foals is caused by two dominant G3P[12] and G14P[12] genotypes of equine rotavirus A (ERVA). Protection of foals against ERVA infection and disease is achievable by passive antibody transfer through colostrum from mares receiving inactivated monovalent G3P[12] vaccine. Simian rotavirus SA11 G3P[2] is also included in ERVA vaccines to protect foals. Here, we characterized cross-neutralization and protection among three distinctive rotaviruses carrying the same or different G (defined by VP7) or P (defined by VP4) genotype by using a rabbit and equine monospecific antisera ...
Comparative analysis of uterine inflammation in jennies and mares after insemination with cryopreserved donkey semen.
Theriogenology    March 30, 2026   Volume 260 117917 doi: 10.1016/j.theriogenology.2026.117917
Sancler-Silva YFR, Costa LML, Freitas MS, Teixeira CS, Albino MV, García HDM, Fiala-Rechsteiner SMDE, Schultz ÉB, da Fonseca LA, Catalán J.The limited fertility of jennies inseminated with frozen donkey semen remains a major challenge for donkey reproduction and conservation, contrasting with the more favorable outcomes observed in mares. Species-specific differences in post-breeding uterine inflammation have been proposed as a key factor underlying this disparity. This study compared the uterine inflammatory response and pregnancy outcomes of jennies and mares inseminated with frozen donkey semen. Twelve jennies and eleven mares were monitored over 48 and 44 estrous cycles, respectively. Artificial insemination (AI) was performe...
Nucleolar proteins nucleolin and nucleophosmin 1 differentially relocalize during equid alphaherpesvirus 1 (EHV1) infection.
Veterinary research communications    March 30, 2026   Volume 50, Issue 3 239 doi: 10.1007/s11259-026-11187-3
Kellerer L, Conn KL.We investigated equine nucleolar proteins nucleophosmin 1 (NPM1) and nucleolin (NCL) during EHV1 infection to develop understanding of EHV1-nucleolar interactions. Equine NPM1 primarily localized throughout nucleoli whereas NCL accumulated in a peri-nucleolar ring and localized throughout the nucleoplasm. EHV1 did not notably affect NPM1 or NCL protein levels but did alter their subnuclear localization. NPM1 accumulated in EHV1 replication compartments and remained nucleolus-associated until very late infection stages, characterized by a kidney bean-shaped nucleus and round cell morphology. NC...
Synthetic progestin treatment alters cytokine expression and impairs vaccine response in horses.
Veterinary immunology and immunopathology    March 28, 2026   Volume 296 111111 doi: 10.1016/j.vetimm.2026.111111
Kapadia CL, Crook RA, Coleman S, Unger G, Douglas RH, Fedorka CE.Altrenogest is commonly used in equine veterinary medicine for the control of estrus, behavioral modifications, and maintenance of pregnancy. Recent studies have found that altrenogest may impact aspects of the immune system, including altering cytokine production in circulation and within the reproductive tract. Therefore, we hypothesize that this alteration may lead to a deviated immune response to vaccination. Sixteen mares were divided into two groups, with ten mares (n = 10) receiving long-acting injectable altrenogest (100 mg/mL; IM) administered every 10 days, and six mares (n =...
Serum-free medium modulates the immunomodulatory and anabolic function of equine bone marrow-derived mesenchymal stromal cells.
Experimental cell research    March 25, 2026   Volume 459, Issue 1 115000 doi: 10.1016/j.yexcr.2026.115000
Tehrani AH, Toth K, Osinchuk N, Dufour A, Krawetz R, Sen A, Sparks H.Adult bone marrow-derived mesenchymal stromal cells (BM-MSCs) hold significant therapeutic potential in human and veterinary medicine. Traditionally, BM-MSC cultures use fetal bovine serum (FBS) to promote growth, but this practice introduces batch variability and xenogeneic contamination that complicate clinical translation. While serum-free medium (SFM) alternatives have been optimized for human MSCs, limited options have been explored for alternative or veterinary species. Here, we compared a chemically defined, customizable SFM formulation with conventional serum-containing medium (SCM) fo...
Equine genital and ocular squamous cell carcinomas: clinical, histopathological, molecular and viral characterization with proposed histopathological classification system.
The veterinary quarterly    March 24, 2026   Volume 46, Issue 1 2648939 doi: 10.1080/01652176.2026.2648939
O'Brien K, Mair T, Mudhar HS, Pesavento P, Miller H, Priestnall SL, Suárez-Bonnet A.Equine squamous cell carcinomas (eSCCs) are common, and a proportion are likely induced by papillomavirus 2 (EcPV-2). Accurate prediction of clinical outcomes is challenging with no recognized prognostic criteria or consistent histopathological classification scheme for eSCC. The aims of this study were to histopathologically subtype a large case series of eSCCs (genital and ocular) and correlate them with p16 and HER-2 expression, equine papillomavirus infection status, and various clinical and histopathological parameters to predict tumour behavior and prognosis. One hundred and eighty-five...
Small Genome-wide association study of insect bite hypersensitivity in Hucul horses.
Journal of equine veterinary science    March 20, 2026   105857 doi: 10.1016/j.jevs.2026.105857
Błaszczak A, Olczak K, Stefaniuk-Szmukier M, Tomczyk-Wrona I, Musiał AD, Grzegorczyk J, Długosz B, Szmatoła T, Ropka-Molik K.Insect bite hypersensitivity (IBH) is a prevalent allergic skin disease in horses that significantly affects animal welfare and shows evidence of genetic predisposition. Objective: Understanding the genetic basis of IBH can improve management and breeding strategies. The Equine80K BeadChip microarray was used to identify genomic regions associated with the occurrence of IBH in Hucul horses. Methods: Hair samples were collected from 127 Hucul horses, including both affected individuals and controls. DNA was extracted and genotyped. A genome-wide association study (GWAS) was performed to identif...
Differential Expression of MITF, WNT3A, SLC7A11, and EDN3 in the Shoulder ‘Bider Marking’ of Dun Mongolian Horses.
Animals : an open access journal from MDPI    March 19, 2026   Volume 16, Issue 6 967 doi: 10.3390/ani16060967
An T, Dugarjaviin M.(1) Background: The "Bider marking" on the shoulder of the Dun Mongolian horse represents a unique pigmentation pattern, the molecular formation mechanism of which remains incompletely understood. This study investigates the differential expression and protein localization of pigment-related genes-specifically the core transcription factor , as well as , , and -in the skin. The analysis focuses on three distinct regions: the dark-colored area of the 'Bider marking' shoulder (BIDC), the light-colored area of the 'Bider marking' shoulder (BILC), and the non-Bider-marked shoulder area (NBIS). The...
Equine models in translational medicine: A comparative approach to human health.
Animal models and experimental medicine    March 16, 2026   doi: 10.1002/ame2.70180
Boozarjomehri Amnieh S, Ropka-Molik K.The horse is a distinctive translational model for bridging mechanistic discovery and clinically relevant investigation because of its physiological complexity, long lifespan, athletic phenotype, and broad spectrum of naturally occurring conditions that parallel aspects of human health and disease. Its utility extends from musculoskeletal and joint research to immunology, metabolic disorders, and exercise physiology, particularly where naturally developed disease, clinically applicable imaging, and longitudinal sampling are required. Additionally, it offers opportunities to examine both chroni...