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Topic:Immunophenotyping

Immunophenotyping is a laboratory technique used to analyze the expression of specific cell surface and intracellular markers in horses. This method involves using flow cytometry or similar technologies to identify and quantify various immune cell populations based on their phenotypic characteristics. Immunophenotyping provides insights into the composition and functional status of the equine immune system, aiding in the understanding of immune responses during health and disease. It can be applied to study conditions such as infections, allergies, and autoimmune disorders. This page compiles peer-reviewed research studies and scholarly articles that explore the methodologies, applications, and findings related to immunophenotyping in equine immunology.
Single-cell resolution landscape of equine peripheral blood mononuclear cells reveals diverse cell types including T-bet+ B cells.
BMC biology    January 22, 2021   Volume 19, Issue 1 13 doi: 10.1186/s12915-020-00947-5
Patel RS, Tomlinson JE, Divers TJ, Van de Walle GR, Rosenberg BR.Traditional laboratory model organisms represent a small fraction of the diversity of multicellular life, and findings in any given experimental model often do not translate to other species. Immunology research in non-traditional model organisms can be advantageous or even necessary, such as when studying host-pathogen interactions. However, such research presents multiple challenges, many stemming from an incomplete understanding of potentially species-specific immune cell types, frequencies, and phenotypes. Identifying and characterizing immune cells in such organisms is frequently limited ...
Equine Intestinal Lymphoma: Clinical-Pathological Features, Immunophenotype, and Survival.
Veterinary pathology    March 23, 2020   Volume 57, Issue 3 369-376 doi: 10.1177/0300985820906889
Bacci B, Stent AW, Walmsley EA.Lymphoma is the most common intestinal neoplasm in horses, but its clinical-pathological features are poorly characterized. Primary intestinal lymphoma was diagnosed in 20 horses on biopsy samples and further confirmed by postmortem examination in 16 cases. Lymphoma was found in the small intestine in 12 of 20 (60%), in the colon in 5 of 20 (25%), and in both small and large intestines in 3 of 20 (15%) cases. Gross findings included thickening of the intestinal wall (45%), mural nodules or masses (30%), and both thickening and nodules (10%). Cases were classified according to the human World H...
Autologous Platelet Lysate Does Not Enhance Chondrogenic Differentiation of Equine Bone Marrow-Derived Mesenchymal Stromal Cells Despite Increased TGF-β1 Concentration.
Stem cells and development    January 6, 2020   Volume 29, Issue 3 144-155 doi: 10.1089/scd.2019.0239
Chapman HS, Gale AL, Dodson ME, Linardi RL, Ortved KF.Bone marrow-derived mesenchymal stromal cells (BM-MSCs) are being investigated for their potential in the treatment of musculoskeletal injuries, including tendon and ligament lesions, and cartilage lesions. Culture expansion of cells has traditionally been performed in medium supplemented with fetal bovine serum (FBS), however, concerns regarding the antigenicity and potential viral or prion contamination of FBS have prompted interest in alternative medium supplements. Platelet lysate (PL) contains elevated concentrations of growth factors, including transforming growth factor-β (TGF-β), pla...
Comparison of the Chondrogenic Differentiation Potential of Equine Synovial Membrane-Derived and Bone Marrow-Derived Mesenchymal Stem Cells.
Frontiers in veterinary science    June 6, 2019   Volume 6 178 doi: 10.3389/fvets.2019.00178
Gale AL, Linardi RL, McClung G, Mammone RM, Ortved KF.Focal cartilage injury occurs commonly and often precipitates OA. Mesenchymal stem cells (MSCs) may be useful for repairing cartilage lesions, thereby preventing joint degeneration. Although MSCs isolated from bone marrow have been shown to have chondrogenic potential, synovial membrane-derived MSCs (SM-MSCs) may have superior chondrogenic abilities due to a common progenitor cell between synovium and cartilage. The objective of this study was to directly compare the immunophenotype, proliferative capabilities, and chondrogenic potential of equine SM-MSCs and bone marrow-derived MSCs (BM-MSCs)...
Persistence of fluorescent nanoparticle-labelled bone marrow mesenchymal stem cells in vitro and after intra-articular injection.
Journal of tissue engineering and regenerative medicine    January 23, 2019   Volume 13, Issue 2 191-202 doi: 10.1002/term.2781
Grady ST, Britton L, Hinrichs K, Nixon AJ, Watts AE.Mesenchymal stem cells (MSCs) improve the osteoarthritis condition, but the fate of MSCs after intra-articular injection is unclear. We used fluorescent nanoparticles (quantum dots [QDs]) to track equine MSCs (QD-labelled MSCs [QD-MSCs]) in vivo after intra-articular injection into normal and osteoarthritic joints. One week after injection of QD-MSCs, unlabelled MSCs, or vehicle, we determined the presence of QD-MSCs in synovium and articular cartilage histologically. In vitro, we evaluated the persistence of QDs in MSCs and whether QDs affected proliferation, immunophenotype, or differentiati...
Clinical and immunophenotypic findings in 4 forms of equine lymphoma.
The Canadian veterinary journal = La revue veterinaire canadienne    January 18, 2019   Volume 60, Issue 1 33-40 doi: 10.1111/eve.12850
Miglio A, Morelli C, Gialletti R, Lauteri E, Sforna M, Marenzoni ML, Antognoni MT.The clinical, histological, and immunophenotypic findings are presented for 4 horses affected by different types of lymphoma. Diagnoses of a monomorphic epitheliotropic intestinal T-cell lymphoma, a diffuse splenic large B-cell lymphoma, a peripheral T-cell lymphoma, and a T-cell rich large B-cell lymphoma of the third eyelid were made. Constatations cliniques et immunophénotypiques pour quatre formes de lymphomes équins. Les constatations cliniques, histologiques et immunophénotypiques sont présentées pour quatre chevaux affectés par différents types de lymphome. Des diagnostics d’un...
Mapping of equine mesenchymal stromal cell surface proteomes for identification of specific markers using proteomics and gene expression analysis: an in vitro cross-sectional study.
Stem cell research & therapy    October 25, 2018   Volume 9, Issue 1 288 doi: 10.1186/s13287-018-1041-8
Bundgaard L, Stensballe A, Elbæk KJ, Berg LC.Stem cells have great potential for tissue regeneration, but before stem cell populations can be used in the clinic, it is crucial that the stem cells have been definitely characterized by a set of specific markers. Although there have been attempts to identify a set of immunophenotypic markers to characterize equine mesenchymal stromal cells (MSCs), immunophenotyping of equine MSCs is still challenging due to the limited availability of suitable antibodies of high quality and consistent performance across different laboratories. The aim of this study was to evaluate a strategy for mapping the...
Serum-free human MSC medium supports consistency in human but not in equine adipose-derived multipotent mesenchymal stromal cell culture.
Cytometry. Part A : the journal of the International Society for Analytical Cytology    September 19, 2017   Volume 93, Issue 1 60-72 doi: 10.1002/cyto.a.23240
Schubert S, Brehm W, Hillmann A, Burk J.For clinical applications of multipotent mesenchymal stromal cells (MSCs), serum-free culture is preferable to standardize cell products and prevent contamination with pathogens. In contrast to human MSCs, knowledge on serum-free culture of large animal MSCs is limited, despite its relevance for preclinical studies and development of veterinary cellular therapeutics. This study aimed to evaluate the suitability of a commercially available serum-free human MSC medium for culturing equine adipose-derived MSCs in comparison with human adipose MSCs. Enzyme-free isolation by explant technique and e...
Acute leukemia in six horses (1990-2012). Barrell EA, Asakawa MG, Felippe MJB, Divers TJ, Stokol T.Acute leukemia is rare in horses. Herein we describe historical, clinicopathologic, and postmortem findings in 6 horses with acute leukemia. Medical records of horses with >20% bone marrow blasts and cytochemical or immunophenotyping results were reviewed. Affected horses were 2-8 y of age and of different breeds and sex. Horses were presented acutely with nonspecific signs (e.g., fever, lethargy). Characteristic hemogram findings were bi- or pancytopenia with low blast numbers. Histologic examination revealed extramedullary infiltrates, especially in lymph nodes, spleen, kidney, liver, and lu...
Phenotypical and functional characteristics of mesenchymal stem cells derived from equine umbilical cord blood.
Cytotechnology    December 9, 2014   Volume 68, Issue 4 795-807 doi: 10.1007/s10616-014-9831-z
Mohanty N, Gulati BR, Kumar R, Gera S, Kumar S, Kumar P, Yadav PS.Mesenchymal stem cells (MSCs) offer promise as therapeutic aid in the repair of tendon and ligament injuries in race horses. Fetal adnexa is considered as an ideal source of MSCs due to many advantages, including non-invasive nature of isolation procedures and availability of large tissue mass for harvesting the cells. However, MSCs isolated from equine fetal adnexa have not been fully characterized due to lack of species-specific markers. Therefore, this study was carried out to isolate MSCs from equine umbilical cord blood (UCB) and characterize them using cross-reactive markers. The plastic...
Comparative immunophenotyping of equine multipotent mesenchymal stromal cells: an approach toward a standardized definition.
Cytometry. Part A : the journal of the International Society for Analytical Cytology    June 3, 2014   Volume 85, Issue 8 678-687 doi: 10.1002/cyto.a.22491
Paebst F, Piehler D, Brehm W, Heller S, Schroeck C, Tárnok A, Burk J.Horses are an approved large animal model for therapies of the musculoskeletal system. Especially for tendon disease where cell-based therapy is commonly used in equine patients, the translation of achieved results to human medicine would be a great accomplishment. Immunophenotyping of equine mesenchymal stromal cells (MSCs) remains the last obstacle to meet the criteria of the International Society for Cellular Therapy (ISCT) definition of human MSCs. Therefore, the surface antigen expression of CD 29, CD 44, CD 73, CD 90, CD 105, CD 14, CD 34, CD 45, CD 79α, and MHC II in equine MSCs from a...
Equine mesenchymal stem cells from bone marrow, adipose tissue and umbilical cord: immunophenotypic characterization and differentiation potential.
Stem cell research & therapy    February 21, 2014   Volume 5, Issue 1 25 doi: 10.1186/scrt414
Barberini DJ, Freitas NP, Magnoni MS, Maia L, Listoni AJ, Heckler MC, Sudano MJ, Golim MA, da Cruz Landim-Alvarenga F, Amorim RM.Studies with mesenchymal stem cells (MSCs) are increasing due to their immunomodulatory, anti-inflammatory and tissue regenerative properties. However, there is still no agreement about the best source of equine MSCs for a bank for allogeneic therapy. The aim of this study was to evaluate the cell culture and immunophenotypic characteristics and differentiation potential of equine MSCs from bone marrow (BM-MSCs), adipose tissue (AT-MSCs) and umbilical cord (UC-MSCs) under identical in vitro conditions, to compare these sources for research or an allogeneic therapy cell bank. Methods: The BM-MS...
Characterization and profiling of immunomodulatory genes of equine mesenchymal stromal cells from non-invasive sources.
Stem cell research & therapy    January 13, 2014   Volume 5, Issue 1 6 doi: 10.1186/scrt395
De Schauwer C, Goossens K, Piepers S, Hoogewijs MK, Govaere JL, Smits K, Meyer E, Van Soom A, Van de Walle GR.Mesenchymal stromal cells (MSCs) have been extensively studied for their promising capabilities in regenerative medicine. Although bone marrow is the best-known source for isolating equine MSCs, non-invasive alternative sources such as umbilical cord blood (UCB), umbilical cord matrix (UCM), and peripheral blood (PB) have also been reported. Methods: Equine MSCs from three non-invasive alternative sources were isolated from six individual mares (PB) and their foals (UCB and UCM) at parturition. To minimize inter-horse variability, the samples from the three sources were matched within the same...
Successful isolation of equine mesenchymal stromal cells from cryopreserved umbilical cord blood-derived mononuclear cell fractions.
Equine veterinary journal    December 4, 2012   Volume 45, Issue 4 518-522 doi: 10.1111/evj.12003
De Schauwer C, van de Walle GR, Piepers S, Hoogewijs MK, Govaere JL, Meyer E, van Soom A.The therapeutic potential of mesenchymal stromal cells for cellular therapy has generated increasing interest in human as well as veterinary medicine. Considerable research has been performed on the cryopreservation of expanded mesenchymal stromal cells, but little information is available on the cryopreservation of the original mononuclear cell fraction. Objective: The present study describes a protocol to expand equine mesenchymal stromal cells after cryopreserving the mononuclear cells of umbilical cord blood. Methods: To this end, mononuclear cells were isolated from 7 umbilical cord blood...
Culture and characterisation of equine peripheral blood mesenchymal stromal cells.
Veterinary journal (London, England : 1997)    June 18, 2012   Volume 195, Issue 1 107-113 doi: 10.1016/j.tvjl.2012.05.006
Spaas JH, De Schauwer C, Cornillie P, Meyer E, Van Soom A, Van de Walle GR.Although the use of mesenchymal stromal cells (MSCs) for the treatment of orthopaedic injuries in horses has been reported, no official guidelines exist that classify a particular cell as an equine MSC. Given the limited characterisation of peripheral blood (PB)-derived equine MSCs in particular, this study aimed to provide more detailed information in relation to this cell type. Mesenchymal stromal cells were isolated from equine PB samples and colony forming unit (CFU) assays as well as population doubling times (PDTs) (from P(0) to P(10)) were performed. Two types of colonies, 'fingerprint'...
In search for cross-reactivity to immunophenotype equine mesenchymal stromal cells by multicolor flow cytometry.
Cytometry. Part A : the journal of the International Society for Analytical Cytology    March 12, 2012   Volume 81, Issue 4 312-323 doi: 10.1002/cyto.a.22026
De Schauwer C, Piepers S, Van de Walle GR, Demeyere K, Hoogewijs MK, Govaere JL, Braeckmans K, Van Soom A, Meyer E.During recent years, cell-based therapies using mesenchymal stem cells (MSC) are reported in equine veterinary medicine with increasing frequency. In most cases, the isolation and in vitro differentiation of equine MSC are described, but their proper immunophenotypic characterization is rarely performed. The lack of a single marker specific for MSC and the limited availability of monoclonal antibodies (mAbs) for equine MSC in particular, strongly hamper this research. In this study, 30 commercial mAbs were screened with flow cytometry for recognizing equine epitopes using the appropriate posit...
Immunophenotype and gene expression profiles of cell surface markers of mesenchymal stem cells derived from equine bone marrow and adipose tissue.
Veterinary immunology and immunopathology    July 2, 2011   Volume 144, Issue 1-2 147-154 doi: 10.1016/j.vetimm.2011.06.033
Ranera B, Lyahyai J, Romero A, Vázquez FJ, Remacha AR, Bernal ML, Zaragoza P, Rodellar C, Martín-Burriel I.Bone marrow and adipose tissue are the two main sources of mesenchymal stem cell (MSC). The aim of this work was to analyse the immunophenotype of 7 surface markers and the expression of a panel of 13 genes coding for cell surface markers in equine bone marrow and adipose tissue-derived MSCs obtained from 9 horses at third passage. The tri-lineage differentiation was confirmed by specific staining. Equine MSCs from both sources were positive for the MSC markers CD29 and CD90, while were negative for CD44, CD73, CD105, CD45 and CD34. The gene expression of these molecules was also evaluated by ...
Markers of stemness in equine mesenchymal stem cells: a plea for uniformity.
Theriogenology    December 31, 2010   Volume 75, Issue 8 1431-1443 doi: 10.1016/j.theriogenology.2010.11.008
De Schauwer C, Meyer E, Van de Walle GR, Van Soom A.Mesenchymal stromal cells (MSC) are a very promising subpopulation of adult stem cells for cell-based regenerative therapies in veterinary medicine. Despite major progress in the knowledge on adult stem cells during recent years, a proper identification of MSC remains a challenge. In human medicine, the Mesenchymal and Tissue Stem Cell Committee of the International Society for Cellular Therapy (ISCT) recently proposed three criteria to define MSC. Firstly, cells must be plastic-adherent when maintained under standard culture conditions. Secondly, MSC must express CD73, CD90 and CD105, and lac...
Evaluation of the osteogenic and chondrogenic differentiation capacities of equine adipose tissue-derived mesenchymal stem cells.
American journal of veterinary research    October 6, 2010   Volume 71, Issue 10 1228-1236 doi: 10.2460/ajvr.71.10.1228
Braun J, Hack A, Weis-Klemm M, Conrad S, Treml S, Kohler K, Walliser U, Skutella T, Aicher WK.To evaluate the proliferative behavior, telomere length, immunophenotype, and differentiation capacity of equine adipose tissue-derived mesenchymal stem cells (AT-MSCs). Methods: 6 adult racing horses treated for articular Injury but otherwise healthy. Methods: AT-MSCs were Isolated from horses and expanded In Dulbecco modified Eagle medium enriched with fetal bovine serum and antimicrobials. Expression of cell surface antigens and telomere length were Investigated via flow cytometry Differentiation of MSCs Into chondrocytes, osteoblasts, and adipocytes was Induced In vitro by specific stimuli...
Isolation and immunophenotypic characterization of mesenchymal stem cells derived from equine species adipose tissue.
Veterinary immunology and immunopathology    June 30, 2009   Volume 132, Issue 2-4 303-306 doi: 10.1016/j.vetimm.2009.06.014
de Mattos Carvalho A, Alves AL, Golim MA, Moroz A, Hussni CA, de Oliveira PG, Deffune E.The purpose of this work was to isolate and cultivate mesenchymal stem cells (MSC) derived from equine adipose tissue and conduct cellular characterization with the following markers: CD90, CD44 and CD13. Adipose tissue collection was performed at the base of the horses' tails, followed by immediate isolation and cultivation of the MSC and posterior characterization by flow cytometry for the interspecies reaction test using mouse anti-rat CD90 monoclonal antibody (mAb), fluorescein isothiocyanate (FITC), and tests with specific mAb mouse anti-horse CD13 and mouse anti-horse CD44. The technique...
Characterization of an equine macrophage cell line: application to studies of EIAV infection.
Veterinary microbiology    November 1, 2008   Volume 136, Issue 1-2 8-19 doi: 10.1016/j.vetmic.2008.10.010
Fidalgo-Carvalho I, Craigo JK, Barnes S, Costa-Ramos C, Montelaro RC.EIAV is a monocyte/macrophage tropic virus. To date, even though EIAV has been under investigation for numerous years, very few details have been elucidated about EIAV/macrophage interactions. This is largely due to the absence of an equine macrophage cell line that would support viral replication. Herein we describe the spontaneous immortalization and generation of a clonal equine macrophage-like (EML) cell line with the functional and immunophenotype characteristics of differentiated equine monocyte derived macrophage(s) (eMDM(s)). These cells possess strong non-specific esterase (NSE) activ...
Bone marrow and lymph node evaluation.
The Veterinary clinics of North America. Equine practice    July 26, 2008   Volume 24, Issue 2 261-v doi: 10.1016/j.cveq.2008.04.001
Tornquist SJ.Evaluation of equine bone marrow and lymph node samples can provide the definitive diagnosis in some cases, and may provide useful information in other cases. Some newer techniques, including immunophenotyping of cells and clonality assays, provide the capability to more precisely identify cells, both as to origin and malignancy. Use of these techniques on equine bone marrow and lymph node samples, and compiling of the data, will eventually provide invaluable information about equine neoplasia that will greatly improve the ability to predict tumor behavior and response to therapy.
Clinical, immunophenotypic and functional characterisation of T-cell leukaemia in six horses.
Equine veterinary journal    December 11, 2007   Volume 39, Issue 6 522-528 doi: 10.2746/042516407X2163271
Rendle DI, Durham AE, Thompson JC, Archer J, Mitchell M, Saunders K, Millere J, Paillot R, Smith KC, Kydd JH.Lymphoid leukaemia (LL) is rare in equids. In man, immunophenotypic classification identifies distinct leukaemic types with different treatment strategies. Improved understanding and classification of equine LL may allow similar advances. Objective: To document the clinical, immunophenotypic and functional characteristics in 6 cases of equine LL of T-cell origin. Methods: The clinical records and pathological findings from 6 cases of equine LL were analysed. Immunohistochemistry to identify T or B lymphocytes was performed on paraffin embedded tissues in 4 cases. Peripheral blood mononuclear c...
Lymphocyte responses and immunophenotypes in horses with Sarcocystis neurona infection.
Equine veterinary journal    January 5, 2002   Volume 33, Issue 7 726-729 doi: 10.2746/042516401776249255
Tornquist SJ, Boeder LJ, Mattson DE, Cebra CK, Bildfell RJ, Hamir AN.No abstract available
Immunophenotypic classification of leukemia in 3 horses.
Journal of veterinary internal medicine    April 13, 2001   Volume 15, Issue 2 144-152 doi: 10.1892/0891-6640(2001)015<0144:icolih>2.3.co;2
McClure JT, Young KM, Fiste M, Sharkey LC, Lunn DP.No abstract available
Equine malignant lymphomas: morphologic and immunohistochemical classification.
Veterinary pathology    July 31, 1998   Volume 35, Issue 4 241-252 doi: 10.1177/030098589803500402
Kelley LC, Mahaffey EA.Gross lesions, microscopic appearance, and immunophenotyping are reported in a retrospective study of 31 cases of equine malignant lymphoma. Immunohistochemical studies were performed on archived formalin-fixed, paraffin-embedded tissues. Monoclonal antibodies to surface glycoprotein BLA.36 and intracytoplasmic domains of mb-1 and B29 were used to document the presence of B lymphocytes in the equine tumors. Polyclonal antibody to CD3 and monoclonal antibodies to T-lymphocyte markers CD3 and CD5 revealed the presence of variable numbers of T cells within the equine lymphomas. The neoplastic com...
Regression of subcutaneous lymphoma following removal of an ovarian granulosatheca cell tumor in a horse.
Journal of the American Veterinary Medical Association    May 20, 1998   Volume 212, Issue 9 1419-1422 
Henson KL, Alleman AR, Cutler TJ, Ginn PE, Kelley LC.A 9-year-old Arabian mare was admitted for evaluation of multiple subcutaneous nodules and infertility. Fine-needle aspiration of one of the subcutaneous nodules resulted in a cytologic diagnosis of histiolymphocytic lymphoma. Palpation per rectum and transrectal ultrasonography revealed a mass associated with the left ovary. Excision of the ovarian tumor was performed, and a histopathologic diagnosis of granulosa-theca cell tumor was made. After removal of the granulosa-theca cell tumor, subcutaneous nodules regressed. The referring veterinarian reported that the nodules had also disappeared ...