A Non-invasive Way to Isolate and Phenotype Cells from the Conjunctiva

Bose, Tanima; Hou, Aihua; Lee, Ryan; Tong, Louis; Chandy, K. George

doi:10.3791/55591

Cited by 5 publications

(4 citation statements)

References 22 publications

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“…The superficial layer of conjunctiva cells, containing IELs, has often been studied with techniques such as brush technology and impression cytology. 62 , 63 And the ocular surface cytology has been used to evaluate many ocular surface diseases, including keratoconjunctivitis sicca, vernal keratoconjunctivitis, keratoconus, and so on. 64 – 66 We use brush technology to obtain the ocular surface cytology sample and analyze the IELs by flow cytometry.…”

Section: Discussionmentioning

confidence: 99%

“…The total number of CD45 + cells per individual varied from 800 to 5000 cells, similar to what has been previously reported. 62 Owing to the limited cell numbers, we have selected the surface marker to analyze the proportion of γδ T cells. Future efforts are, therefore, warranted to analyze the production of IL-17A and IFN-γ in a more comprehensive manner.…”

Section: Discussionmentioning

confidence: 99%

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Conjunctiva Resident γδ T Cells Expressed High Level of IL-17A and Promoted the Severity of Dry Eye

Zhu

et al. 2022

Invest. Ophthalmol. Vis. Sci.

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Purpose Conjunctival inflammation promotes ocular surface disorders in dry eye disease (DED). Here we identified γδ T cells as the predominant source of IL-17A in the murine conjunctiva and assessed their contribution to the pathogenesis of DED. Methods We enrolled 22 patients with DED, and analyzed the proportion of γδ T cells in the conjunctival epithelial samples by flow cytometry. Adult C57Bl/6 wild-type and TCRδ − / − mice were used to induce DED models to investigate the role of γδ T cells. The characteristics of immune cell infiltration and the expression of immune-related cytokines or markers in mouse conjunctiva were analyzed by flow cytometry, Western blot, and quantitative polymerase chain reaction. Results The proportion of γδ T cells in the human DED conjunctiva is significantly higher in patients with severe corneal epithelial defects than in mild ones, which is consistently observed in the murine DED model. Further, a high level of IL-17A but not IFN-γ is detected in the conjunctiva of mice. The increased murine IL-17A–producing cells on the conjunctiva are identified as γδ T cells predominantly and Th17 cells to a lesser extent. Ablation of γδ T cells by antibody depletion or genetic deletion of TCRδ alleviates ocular surface damage in the murine DED model. Conclusions Our studies evaluate human and experimental murine DED for evidence of γδ T-cell–mediated inflammation and highlight a potential therapeutic synergy by targeting IL-17 and γδ T cells in DED treatment.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Conjunctiva Resident γδ T Cells Expressed High Level of IL-17A and Promoted the Severity of Dry Eye

Zhu

et al. 2022

Invest. Ophthalmol. Vis. Sci.

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“…[ 32 ] Another advantage of impression cytology is the ability to analyze immune cells with flow cytometry. [ 33 ] Flow cytometry on tear fluid washings detects neutrophils and natural killer cells, and a study reported a higher proportion of the former and a lower proportion of the latter in ADDE with corneal staining compared to those without staining and to controls. [ 34 ] Similarly, the quantification of tear inflammatory cytokines may have interesting applications in DED, but one of the practical issues to overcome is the wide range of tear cytokine levels in the general population and the effect of increasing age on the normal range.…”

Section: The Dry Eye Patientmentioning

confidence: 99%

Evaluation of ocular surface and tear function - A review of current approaches for dry eye

Rao

Koh

Srinivas

et al. 2022

Indian J Ophthalmol

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An increasing prevalence of dry eye disease in the past decade has resulted in a greater focus on diagnostic methods for this condition. There has been a proliferation of technologies that attempt to quantify various aspects of tear function and ocular surface health. However, a cost-effective, simple, and efficient method remains elusive. In the Indian context, the majority of these patients present to the general ophthalmologist, and a clinical approach that is quick and easy to perform would allow widespread usage for accurate diagnosis. This article reviews currently available methods and their relevance to the general ophthalmologist.

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“…The procedure of using Eyeprim for IC with subsequent flow cytometry is shown in a video publication. [54] The proportions of tissue resident memory cells (TRM) and recirculating T cell subsets at the ocular surface in both healthy controls and patients with DED were determined using this procedure. It was found that the normal human ocular surface was protected by two subsets of TRM cells and four subsets of recirculating T cells.…”

Section: Recent Studies On the Use Of Impression Cytologymentioning

confidence: 99%

Update on the role of impression cytology in ocular surface disease

Thia

Tong

2019

Taiwan J Ophthalmol

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Understanding of the molecular pathology of ocular surface disease (OSD) is poor, and treatment is highly unsatisfactory. To facilitate treatment of OSD, a relatively noninvasive procedure, i.e. impression cytology (IC) has been shown to be useful. Recently, the technologies employed in research studies using IC in OSD have vastly improved, and standardized IC has even been used in clinical trials of dry eye. Here, this review aims to describe the advances of IC in the last 10 years, which serves as an update on the progress in this field since the last major review of IC. OSD that has been recently evaluated include meibomian gland dysfunction, Sjogren's syndrome, Steven–Johnson syndrome, and postmenopausal dry eye. The recent studies (4 longitudinal, 18 cross-sectional analyses) which utilized IC analyzed DNA, RNA, proteins, and ocular surface cells, including memory T-lymphocytes, dendritic cells (DCs), neutrophils, conjunctival epithelial cells, and goblet cells. These studies employed quantification of transcripts associated with inflammation, proteins involved in oxidative stress, enzymes such as matrix metalloproteinases, and cell surface proteins by flow cytometry, such as HLA-DR, cytokine and chemokine receptors, markers for T cell differentiation, and DC activation, in addition to the more traditional morphological evaluation of squamous metaplasia and staining for goblet cells. Some challenges in the clinical use of IC have also been described, including issues related to storage and normalization of data. In summary, advances in IC have permitted a more robust evaluation of the ocular surface and will facilitate progress in the understanding and treatment of OSD.

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A Non-invasive Way to Isolate and Phenotype Cells from the Conjunctiva

Cited by 5 publications

References 22 publications

Conjunctiva Resident γδ T Cells Expressed High Level of IL-17A and Promoted the Severity of Dry Eye

Conjunctiva Resident γδ T Cells Expressed High Level of IL-17A and Promoted the Severity of Dry Eye

Evaluation of ocular surface and tear function - A review of current approaches for dry eye

Update on the role of impression cytology in ocular surface disease

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