scholarly journals Generation of functional conjunctival epithelium, including goblet cells, from human iPSCs

Cell Reports ◽  
2021 ◽  
Vol 34 (5) ◽  
pp. 108715
Author(s):  
Kimihito Nomi ◽  
Ryuhei Hayashi ◽  
Yuki Ishikawa ◽  
Yuki Kobayashi ◽  
Tomohiko Katayama ◽  
...  
Keyword(s):  
Goblet Cells ◽  
Conjunctival Epithelium ◽  
Human Ipscs

scholarly journals Conjunctival Goblet Cell Responses to TLR5 Engagement Promote Activation of Local Antigen-Presenting Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Abiramy Logeswaran ◽  
Laura Contreras-Ruiz ◽  
Sharmila Masli
Keyword(s):  
Dendritic Cells ◽  
Goblet Cell ◽  
Ocular Surface ◽  
Primary Cultures ◽  
Goblet Cells ◽  
Microbial Colonization ◽  
Mucosal Inflammation ◽  
Conjunctival Epithelium ◽  
Cell Responses ◽  
Mucosal Homeostasis

Conjunctival epithelium forms a barrier between the ocular surface microbial flora and the ocular mucosa. In addition to secreting gel-forming mucins, goblet cells, located in the conjunctival epithelium, help maintain local immune homeostasis by secreting active TGFβ2 and promoting tolerogenic phenotype of dendritic cells in the vicinity. Although dendritic cell subsets, characteristic of mucosal tissues, are found in the conjunctiva, previous studies provided limited information about their location within the tissue. In this study, we examine immunostained conjunctiva explants to determine the location of CD11c-positive dendritic cells in the context of MUC5AC-positive goblet cells. Considering that conjunctival goblet cells are responsive to signaling induced by pathogen recognition receptors, we also assess if their responses to microbial product, flagellin, can contribute to the disruption of ocular mucosal homeostasis that promotes activation of dendritic cells and results in chronic ocular surface inflammation. We find that dendritic cells in the conjunctiva with an increased microbial colonization are located adjacent to goblet cells. While their cell bodies in the stromal layer are immediately below the epithelial layer, several extensions of dendritic cells are projected across the epithelium towards the ocular surface. Such trans-epithelial dendrites are not detectable in healthy ocular mucosa. In response to topically applied flagellin, increased proportion of CD11c-positive cells in the conjunctiva strongly express MHC class II relative to the untreated conjunctiva. This change is accompanied by reduced immunoreactivity to TGFβ-activating Thrombospondin-1 in the conjunctival epithelium. These findings are supported by in vitro observations in primary cultures of goblet cells that respond to the TLR5 stimulation with an increased expression of IL-6 and reduced level of active TGFβ. The observed changes in the conjunctiva after flagellin application correspond with the development of clinical signs of chronic ocular mucosal inflammation including corneal epitheliopathy. Collectively, these findings demonstrate the ability of ocular mucosal dendritic cells to extend trans-epithelial dendrites in response to increased microbial colonization at the ocular surface. Moreover, this study provides key insight into how goblet cell responses to microbial stimuli may contribute to the disruption of ocular mucosal homeostasis and chronic ocular mucosal inflammation.

scholarly journals Location and Clonal Analysis of Stem Cells and Their Differentiated Progeny in the Human Ocular Surface

1999 ◽  
Vol 145 (4) ◽  
pp. 769-782 ◽  
Author(s):  
Graziella Pellegrini ◽  
Osvaldo Golisano ◽  
Patrizia Paterna ◽  
Alessandro Lambiase ◽  
Stefano Bonini ◽  
...  
Keyword(s):  
Stem Cells ◽  
Life History ◽  
Ocular Surface ◽  
Goblet Cells ◽  
Clonal Analysis ◽  
Proliferative Potential ◽  
Conjunctival Epithelium ◽  
Proliferative Capacity ◽  
Differentiation Potential ◽  
Goblet Cell Differentiation

We have analyzed the proliferative and differentiation potential of human ocular keratinocytes. Holoclones, meroclones, and paraclones, previously identified in skin, constitute also the proliferative compartment of the ocular epithelium. Ocular holoclones have the expected properties of stem cells, while transient amplifying cells have variable proliferative potential. Corneal stem cells are segregated in the limbus, while conjunctival stem cells are uniformly distributed in bulbar and forniceal conjunctiva. Conjunctival keratinocytes and goblet cells derive from a common bipotent progenitor. Goblet cells were found in cultures of transient amplifying cells, suggesting that commitment for goblet cell differentiation can occur late in the life of a single conjunctival clone. We found that conjunctival keratinocytes with high proliferative capacity give rise to goblet cells at least twice in their life and, more importantly, at rather precise times of their life history, namely at 45–50 cell doublings and at ∼15 cell doublings before senescence. Thus, the decision of conjunctival keratinocytes to differentiate into goblet cells appears to be dependent upon an intrinsic “cell doubling clock.” These data open new perspectives in the surgical treatment of severe defects of the anterior ocular surface with autologous cultured conjunctival epithelium.

scholarly journals PADRONIZAÇÃO DA CITOLOGIA DE IMPRESSÃO DA SUPERFÍCIE OCULAR CANINA

2005 ◽  
Vol 10 (1) ◽  
Author(s):  
C.A.L. GODOY-ESTEVES ◽  
J.N. BARROS ◽  
L.S. CUNHA ◽  
V.L.D. MASCARO ◽  
A.L. HOFLING-LIMA ◽  
...  
Keyword(s):  
Filter Paper ◽  
Ocular Surface ◽  
Goblet Cells ◽  
Eye Disease ◽  
Conjunctival Epithelium ◽  
Human Beings ◽  
Impression Cytology ◽  
Feasible Method ◽  
Veterinary Hospital ◽  
Goblet Cell Density

Técnica de exame de citologia de impressão foi padronizada em olhos de cães sem alterações oculares. Foram realizados exames de citologia de impressão do epitélio corneano, conjuntival e tarsal em 30 olhos de 21 animais de raças e idades variadas. As amostras foram colhidas de cães atendidos no Hospital Veterinário da FMVZ-USP entre fevereiro e julho de 2003, sendo coradas e avaliadas no Laboratório de Doenças Externas Oculares da UNIFESP. A colheita foi bem tolerada pelos cães e o papel filtro utilizado removeu células em quantidade e morfologia adequadas para estudo citológico. Foi observado em 100% dos casos que o epitélio da conjuntiva bulbar canina apresenta aspecto “metaplasia-like”, com ausência de células caliciformes. Estas só foram encontradas na conjuntiva tarsal em 21,4% das amostras avaliadas dessa região. A citologia de impressão é um método factível para avaliação da superfície ocular em cães. Entretanto, a celularidade das amostras obtidas do tarso mostrou-se inadequada. Além disso, a pesquisa da densidade de células caliciformes em áreas bulbares, embora usada em seres humanos, pode não servir como indicador de alteração da superfície ocular para a espécie canina. Standardization of canine ocular surface impression cytology Abstract Impression cytology technique in dog eyes without ocular disease was standardized. Impression cytology was performed in corneal, conjunctival and tarsal epithelium in 30 eyes of 21 animals with different races and ages. Samples were obtained from dogs attended in FMVZ-USP Veterinary Hospital between February to July 2003, being stained and evaluated at UNIFESP´s External Eye Disease Laboratory. Sampling was well tolerated by dogs and the filter paper used removed cells with adequate morphology and quantity for cytologyc evaluation. In all cases canine bulbar conjunctival epithelium showed metaplasia-like features without goblet cells. Impression cytology is a feasible method for ocular surface evaluation in dogs. However, celularity was considered inadequated in samples obtained from tarsal conjunctiva. Furthermore, seeking goblet cell density in bulbar areas, although used in human beings, may not be used as an ocular surface disease indicator in canine species.

Histological examination of rat small intestine after surgical removal of obturation small bowel obstruction and peptide stimulation of lymph flow

2018 ◽  
pp. 157-162
Author(s):  
А.А. Коваленко ◽  
Г.П. Титова ◽  
В.К. Хугаева
Keyword(s):  
Small Intestine ◽  
Surgical Treatment ◽  
Survival Rate ◽  
Small Bowel ◽  
Goblet Cells ◽  
Intestinal Wall ◽  
Structure And Function ◽  
Lymph Flow ◽  
Intestinal Lumen ◽  
And Function

Оперативное лечение различных заболеваний кишечника сопровождается осложнениями в виде нарушений микроциркуляции в области анастомоза кишки. Ранее нами показана способность лимфостимуляторов пептидной природы восстанавливать нарушенную микроциркуляцию, что послужило основой для настоящего исследования. Цель работы - оценка влияния стимуляции лимфотока в стенке кишки на процессы восстановления микроциркуляции, структуры и функции тонкой кишки в области оперативного вмешательства. Методика. В экспериментах на наркотизированных крысах (хлоралгидрат в дозе 0,6 г/кг в 0,9% растворе NaCl) моделировали различные поражения тонкой кишки (наложение лигатуры, перевязка 1-3 брыжеечных артерий, перекрут петли кишки вокруг оси брыжейки, сочетание нескольких видов повреждений). Резекция поврежденного участка через 1 сут. с последующим созданием тонкокишечного анастомоза завершалась орошением операционного поля раствором пептида-стимулятора лимфотока (40 мкг/кг массы животного в 1 мл 0,9% раствора NaCl). На 7-е сут. после операции проводили гистологическое исследование фрагмента кишки в области анастомоза. Результаты. На 7-е сут. после резекции у выживших животных (летальность вследствие кишечной непроходимости составляла 30%) имеют место морфологические признаки острых сосудистых нарушений стенки кишки, изменений кровеносных и лимфатических микрососудов, интерстициальный отек всех слоев стенки кишки, дилатация просвета кишки, повреждение всасывающего эпителия ворсин с истончением щеточной каемки клеток, морфологические признаки гиперфункции бокаловидных клеток. Использование лимфостимулятора пептидной природы после операции увеличивало выживаемость животных на 24%. У части животных отмечалось уменьшение расширения просвета кишки, у других практически полная его нормализация. Восстанавливалась форма кишечных ворсин и распределение бокаловидных клеток. Отсутствовали признаки внутриклеточного и межмышечного отека. Отмечено умеренное полнокровие венул. Заключение. Использование лимфостимулятора при хирургическом лечении кишечной непроходимости увеличивает выживаемость животных на 24% по сравнению с контролем, способствует более раннему восстановлению структуры и функции тонкой кишки. Полученные результаты свидетельствуют о перспективности использования стимуляции лимфотока при операциях на кишечнике. Surgical treatment of bowel diseases is associated with complications that cause microcirculatory disturbances in the anastomosis area and may lead to a fatal outcome. This study was based on our previous finding that peptide-type lymphatic stimulators are able to restore impaired microcirculation. The aim of this work was stimulating the lymph flow in the intestinal wall to facilitate recovery of microcirculation, structure and function of the small intestine in the area of surgical intervention. Methods. In experiments on anesthetized rats (0.6 g/kg chloral hydrate in 0.9% NaCl), various small bowel lesions were modeled (bowel ligation, ligation of 1-3 mesenteric arteries, gut torsion, combination of several lesion types). In 24 h, the damaged area was resected, and a small intestine anastomosis was creased. The surgery was completed with irrigation of the operative field with a solution of lymph flow stimulating peptide (40 мg/kg body weight in 1 ml of 0.9% NaCl). A gut fragment from the anastomosis area was examined histologically on day 7 after the surgery. Results. On the 7th day after removing the intestinal obstruction, the surviving animals (lethality 30%) had morphological signs of acute vascular disorders in the intestinal wall; changes in blood and lymphatic microvessels; interstitial edema of all intestinal wall layers; dilatation of the intestinal lumen; damage to the absorptive epithelium of villi with thinning of the brush border, and hyperfunction of mucous (goblet) cells. The use of the peptide after surgery increased the survival rate of animals by 24% and provided a smaller dilatation of the intestinal lumen in some animals. In other animals, the lumen recovered. The shape of intestinal villi and distribution of goblet cells were restored. Signs of intracellular and intermuscular edema were absent. Moderate venular congestion was noticed. Conclusion. Using the lymphatic stimulator in surgical treatment of intestinal obstruction increases the survival rate of animals by 24% compared to the control, facilitates earlier restoration of the small intestine structure and function. The obtained results indicated the effectiveness of lymphatic stimulation in intestinal surgery.

scholarly journals Interleukin-17 and Th17 Lymphocytes Directly Impair Motoneuron Survival of Wildtype and FUS-ALS Mutant Human iPSCs

2021 ◽  
Vol 22 (15) ◽  
pp. 8042
Author(s):  
Mengmeng Jin ◽  
Katja Akgün ◽  
Tjalf Ziemssen ◽  
Markus Kipp ◽  
Rene Günther ◽  
...  
Keyword(s):  
Motor Neurons ◽  
Th17 Cells ◽  
Immune Cell ◽  
Cell Types ◽  
Positive Control ◽  
Interleukin 17 ◽  
Fused In Sarcoma ◽  
Th17 Lymphocytes ◽  
Human Ipscs ◽  
Als Patients

Amyotrophic lateral sclerosis (ALS) is a progressive disease leading to the degeneration of motor neurons (MNs). Neuroinflammation is involved in the pathogenesis of ALS; however, interactions of specific immune cell types and MNs are not well studied. We recently found a shift toward T helper (Th)1/Th17 cell-mediated, pro-inflammatory immune responses in the peripheral immune system of ALS patients, which positively correlated with disease severity and progression. Whether Th17 cells or their central mediator, Interleukin-17 (IL-17), directly affects human motor neuron survival is currently unknown. Here, we evaluated the contribution of Th17 cells and IL-17 on MN degeneration using the co-culture of iPSC-derived MNs of fused in sarcoma (FUS)-ALS patients and isogenic controls with Th17 lymphocytes derived from ALS patients, healthy controls, and multiple sclerosis (MS) patients (positive control). Only Th17 cells from MS patients induced severe MN degeneration in FUS-ALS as well as in wildtype MNs. Their main effector, IL-17A, yielded in a dose-dependent decline of the viability and neurite length of MNs. Surprisingly, IL-17F did not influence MNs. Importantly, neutralizing IL-17A and anti-IL-17 receptor A treatment reverted all effects of IL-17A. Our results offer compelling evidence that Th17 cells and IL-17A do directly contribute to MN degeneration.

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