Functional Characteristics Of Cells Research Articles

Recent Advancements in High-Frequency Ultrasound Applications from Imaging to Microbeam Stimulation.

Ultrasound is a versatile and well-established technique using sound waves with frequencies higher than the upper limit of human hearing. Typically, therapeutic and diagnosis ultrasound operate in the frequency range of 500 kHz to 15 MHz with greater depth of penetration into the body. However, to achieve improved spatial resolution, high-frequency ultrasound (>15 MHz) was recently introduced and has shown promise in various fields such as high-resolution imaging for the morphological features of the eye and skin as well as small animal imaging for drug and gene therapy. In addition, high-frequency ultrasound microbeam stimulation has been demonstrated to manipulate single cells or microparticles for the elucidation of physical and functional characteristics of cells with minimal effect on normal cell physiology and activity. Furthermore, integrating machine learning with high-frequency ultrasound enhances diagnostics, including cell classification, cell deformability estimation, and the diagnosis of diabetes and dysnatremia using convolutional neural networks (CNNs). In this paper, current efforts in the use of high-frequency ultrasound from imaging to stimulation as well as the integration of deep learning are reviewed, and potential biomedical and cellular applications are discussed.

Influence of sugar concentration on the vesicle compactness, deformation and membrane poration induced by anionic nanoparticles.

Sugar plays a vital role in the structural and functional characteristics of cells. Hence, the interaction of NPs with cell membranes in the presence of sugar concentrations is important for medicinal and pharmacological innovations. This study integrated three tools: giant unilamellar vesicles (GUVs), anionic magnetite nanoparticles (NPs), and sugar concentrations, to understand a simplified mechanism for interactions between the vesicle membranes and NPs under various sugar concentrations. We focused on changing the sugar concentration in aqueous solution; more precisely, sucrose inside the GUVs and glucose outside with equal osmolarity. 1,2-dioleoyl-sn-glycero-3-phospho-(1′-rac-glycerol) (sodium salt) (DOPG) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) were used to prepare the charged membranes of 40mole%DOPG/60mole%DOPC-GUVs, whereas only DOPC was used to prepare the neutral membranes. Phase contrast fluorescence microscopy shows that the adherence of 18 nm magnetite NPs with anionic charge depends on the sugar concentration. The alterations of GUVs induced by the NPs are characterized in terms of i) vesicle compactness, ii) deformation, and iii) membrane poration. The presence of sugar provides additional structural stability to the GUVs and reduces the effects of the NPs with respect to these parameters; more precisely, the higher the sugar concentration, the smaller the alteration induced by the NPs. The differences in NPs effects are explained by the change in the type of interaction between sugar molecules and lipid membranes, namely enthalpy and entropy-driven interaction, respectively. In addition, such alterations are influenced by the surface charge density of the lipid bilayer. The surface pressure of membranes due to the adsorption of NPs is responsible for inducing the poration in membranes. The differences in deformation and poration in charged and neutral GUVs under various sugar concentrations are discussed based on the structure of the head of lipid molecules.

Lyophilized Human Cord Blood Leukoconcentrate to Treat Brain Ischemia in Rats

The paper outlines the experimental data on the impact of lyophilization on preservation of structural and functional characteristics of cells from human cord blood leukoconcentrate (HCBL). Lyophilization of HCBL was shown to ensure the preservation of cells with immune modulating potential in a heterogeneous population of HCBL. Lyophilized HCBL (lHCBL), to the same extent as cryopreserved one (cHCBL), revealed an equal immune correcting effect during ischemic brain injury in vivo in the experimental model of ischemic stroke (IS). The inclusion of lHCBL in therapy of brain ischemia demonstrated the improved indices of IFN-ү+ and IL-10+ spleen cells, adhesive and phagocytic activity of peritoneal cavity cells in rats with IS. The efficiency of using both lHCBL and cHCBL during IS may be associated with the implementation of therapeutic effect by presented among them cells and mediators, including multi-vector regulatory systems that maintain homeostatic stability of the body (immune, endocrine, nervous ones).

Morphological and functional characteristics of satellite glial cells in the peripheral nervous system.

Satellite glial cells are specialised cells that form a functional perineuronal sheath around sensory ganglion neurons. There are a large number of studies that reveal the morphological and functional characteristics of these cells. Satellite glial cells have been studied both in intact ganglions and in tissue cultures, using light and transmission electron microscopy, immunohistochemical and other methods. Satellite glial cells have polygonal form; they are mononuclear and have developed synthetic organelles, numerous receptors, adhesion molecules and ion channels, which enable them to interact with adjacent neurons, as well as transmit signals in the ganglions of the peripheral nervous system. Based on the literature data, satellite glial cells thanks to their characteristics can receive signals from other cells and react to changes in their surroundings. Previous studies have investigated the potential role of satellite glial cells in the formation of the blood-nervous tissue barrier of the peripheral nervous system, as well as in the neuropathic pain genesis. Some recent discoveries support the fact that satellite glial cells can participate in controlling of local viral infections and protecting pseudounipolar neurons from mentioned infections.

Non-genetic inheritance restraint of cell-to-cell variation

Heterogeneity in physical and functional characteristics of cells (e.g. size, cycle time, growth rate, protein concentration) proliferates within an isogenic population due to stochasticity in intracellular biochemical processes and in the distribution of resources during divisions. Conversely, it is limited in part by the inheritance of cellular components between consecutive generations. Here we introduce a new experimental method for measuring proliferation of heterogeneity in bacterial cell characteristics, based on measuring how two sister cells become different from each other over time. Our measurements provide the inheritance dynamics of different cellular properties, and the 'inertia' of cells to maintain these properties along time. We find that inheritance dynamics are property specific and can exhibit long-term memory (∼10 generations) that works to restrain variation among cells. Our results can reveal mechanisms of non-genetic inheritance in bacteria and help understand how cells control their properties and heterogeneity within isogenic cell populations.

Vitamin E Derivative with Modified Side Chain Induced Apoptosis by Modulating the Cellular Lipids and Membrane Dynamics in MCF7 Cells.

The vitamin E derivative with side chain modification (TC6OAc) has been shown to possess anticancer activity in our earlier in vivo studies. It was hypothesized that, as Vitamin E (VE) and VE derivative are fat soluble lipophilic molecules, they exert their function by modulating the lipid metabolism and related pathways. This study aimed to evaluate the cellular impact of this VE derivative (2,5,7,8-Tetramethyl-2-(4'-Methyl-3'-Pentenyl)-6-Acetoxy Chromane-TC6OH), using α-tocopherol as a reference compound throughout the experiments. Their effects on the cellular metabolism, the biophysical properties of cellular lipids and the functional characteristics of cells were monitored in human estrogen receptor (ER) positive breast cancer cells. It has been documented that TC6OH treatment induces tumor cell apoptosis by dissipating the mitochondrial membrane potential, modulating the lipid, transportation and degradation as well as downregulating certain anti-apoptotic and growth factor related proteins. Due to resistance of ER positive cells to the established therapies, the findings of this study are of translational value.

Antiviral Activity of Exopolysaccharides Produced by Lactic Acid Bacteria of the Genera Pediococcus, Leuconostoc and Lactobacillus against Human Adenovirus Type 5

Background and objectives: The use of antagonistic probiotic microorganisms and their byproducts represents a promising approach for the treatment of viral diseases. In the current work, the effect of exopolysaccharides (EPSs) produced by lactic acid bacteria from different genera on the structural and functional characteristics of cells and the development of adenoviral infection in vitro was studied. Materials and Methods: Cytotoxicity of six EPSs of lactic acid bacteria of the genera Lactobacillus, Leuconostoc and Pediococcus was determined by MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay. The influence of the EPSs on the infectivity of human adenovirus type 5 (HAdV-5) and on the cell cycle under a condition of adenovirus infection was studied using plaque reduction assay and flow cytometric analysis, respectively. Results: It was shown that exopolysaccharides were non-toxic to Madin-Darby bovine kidney cells (MDBK) as they reduced their viability by 3–17%. A change in the distribution of the cell cycle phases in the non-infected cell population treated with EPSs was observed. The analysis demonstrated an increase in the number of cells in the S phase by 47% when using EPSs 15a and a decrease in the number of cells in the G1 phase by 20–27% when treated with the EPSs 15a, 33a, and 19s. The use of EPSs did not led to the normalization of the life cycle of HAdV-5 infected cells to the level of non-infected cells. The EPSs showed low virucidal activity and reduced the HAdV-5 infectivity to 85%. Among the studied exopolysaccharides, anti-adenovirus activity was found for EPS 26a that is produced by Lactobacillus spp. strain. The treatment of cells with the EPS following virus adsorption completely (100%) suppressed the formation and release of HAdV-5 infectious. Conclusions: EPS 26a possessed distinct anti-HAdV-5 activity and the obtained data demonstrate the potential of using exopolysaccharides as anti-adenoviral agents.

Label-free analysis of the characteristics of a single cell trapped by acoustic tweezers

Single-cell analysis is essential to understand the physical and functional characteristics of cells. The basic knowledge of these characteristics is important to elucidate the unique features of various cells and causative factors of diseases and determine the most effective treatments for diseases. Recently, acoustic tweezers based on tightly focused ultrasound microbeam have attracted considerable attention owing to their capability to grab and separate a single cell from a heterogeneous cell sample and to measure its physical cell properties. However, the measurement cannot be performed while trapping the target cell, because the current method uses long ultrasound pulses for grabbing one cell and short pulses for interrogating the target cell. In this paper, we demonstrate that short ultrasound pulses can be used for generating acoustic trapping force comparable to that with long pulses by adjusting the pulse repetition frequency (PRF). This enables us to capture a single cell and measure its physical properties simultaneously. Furthermore, it is shown that short ultrasound pulses at a PRF of 167 kHz can trap and separate either one red blood cell or one prostate cancer cell and facilitate the simultaneous measurement of its integrated backscattering coefficient related to the cell size and mechanical properties.

Anatomical and functional implications of corticotrophin-releasing hormone neurones in a septal nucleus of the avian brain: an emphasis on glial-neuronal interaction via V1a receptors in vitro.

Previously, we showed that corticotrophin-releasing hormone immunoreactive (CRH-IR) neurones in a septal structure are associated with stress and the hypothalamic-pituitary-adrenal axis in birds. In the present study, we focused upon CRH-IR neurones located within the septal structure called the nucleus of the hippocampal commissure (NHpC). Immunocytochemical and gene expression analyses were used to identify the anatomical and functional characteristics of cells within the NHpC. A comparative morphometry analysis showed that CRH-IR neurones in the NHpC were significantly larger than CRH-IR parvocellular neurones in the paraventricular nucleus of the hypothalamus (PVN) and lateral bed nucleus of the stria terminalis. Furthermore, these large neurones in the NHpC usually have more than two processes, showing characteristics of multipolar neurones. Utilisation of an organotypic slice culture method enabled testing of how CRH-IR neurones could be regulated within the NHpC. Similar to the PVN, CRH mRNA levels in the NHpC were increased following forskolin treatment. However, dexamethasone decreased forskolin-induced CRH gene expression only in the PVN and not in the NHpC, indicating differential inhibitory mechanisms in the PVN and the NHpC of the avian brain. Moreover, immunocytochemical evidence also showed that CRH-IR neurones reside in the NHpC along with the vasotocinergic system, comprising arginine vasotocin (AVT) nerve terminals and immunoreactive vasotocin V1a receptors (V1aR) in glia. Hence, we hypothesised that AVT acts as a neuromodulator within the NHpC to modulate activity of CRH neurones via glial V1aR. Gene expression analysis of cultured slices revealed that AVT treatment increased CRH mRNA levels, whereas a combination of AVT and a V1aR antagonist treatment decreased CRH mRNA expression. Furthermore, an attempt to identify an intercellular mechanism in glial-neuronal communication in the NHpC revealed that brain-derived neurotrophic factor (BDNF) and its receptor (TrkB) could be involved in the signalling mechanism. Immunocytochemical results further showed that both BDNF and TrkB receptors were found in glia of the NHpC. Interestingly, in cultured brain slices containing the NHpC, the use of a selective TrkB antagonist decreased the AVT-induced increase in CRH gene expression levels. The results from the present study collectively suggest that CRH neuronal activity is modulated by AVT via V1aR involving BDNF and TrkB glia in the NHpC.

Isolation of Live Fibroblasts by Fluorescence-Activated Cell Sorting.

Flow cytometry is a powerful tool in cell biology in that it allows real-time characterization of cellular phenotypes. Additionally, through the process of fluorescence-activated cell sorting (FACS), living cells can be isolated for future in vitro experiments, including single cell analysis. Here we describe the isolation of live fibroblasts from the dermis of the skin in mice using FACS. This method circumvents the need for ex vivo expansion in cell culture, which can alter phenotypic and functional characteristics of cells.

A step further toward glyphosate-induced epidermal cell death: Involvement of mitochondrial and oxidative mechanisms

A deregulation of programmed cell death mechanisms in human epidermis leads to skin pathologies. We previously showed that glyphosate, an extensively used herbicide, provoked cytotoxic effects on cultured human keratinocytes, affecting their antioxidant capacities and impairing morphological and functional cell characteristics. The aim of the present study, carried out on the human epidermal cell line HaCaT, was to examine the part of apoptosis plays in the cytotoxic effects of glyphosate and the intracellular mechanisms involved in the apoptotic events. We have conducted different incubation periods to reveal the specific events in glyphosate-induced cell death. We observed an increase in the number of early apoptotic cells at a low cytotoxicity level (15%), and then, a decrease, in favor of late apoptotic and necrotic cell rates for more severe cytotoxicity conditions. At the same time, we showed that the glyphosate-induced mitochondrial membrane potential disruption could be a cause of apoptosis in keratinocyte cultures.

Human hepatocytes and endothelial cells in organotypic membrane systems

The realization of organotypic liver model that exhibits stable phenotype is a major challenge in the field of liver tissue engineering. In this study we developed liver organotypic co-culture systems by using synthetic and biodegradable membranes with primary human hepatocytes and human umbilical vein endothelial cells (HUVEC). Synthetic membranes prepared by a polymeric blend constituted of modified polyetheretherketone (PEEK-WC) and polyurethane (PU) and biodegradable chitosan membranes were developed by phase inversion technique and used in homotypic and organotypic culture systems. The morphological and functional characteristics of cells in the organotypic co-culture membrane systems were evaluated in comparison with homotypic cultures and traditional systems. Hepatocytes in the organotypic co-culture systems exhibit compact polyhedral cells with round nuclei and well demarcated cell–cell borders like in vivo, as a result of heterotypic interaction with HUVECs. In addition HUVECs formed tube-like structures directly through the interactions with the membranes and hepatocytes and indirectly through the secretion of ECM proteins which secretion improved in the organotypic co-culture membrane systems. The heterotypic cell–cell contacts have beneficial effect on the hepatocyte albumin production, urea synthesis and drug biotransformation. The developed organotypic co-culture membrane systems elicit liver specific functions in vitro and could be applied for the realization of engineered liver tissues to be used in tissue engineering, drug metabolism studies and bioartificial liver devices.

319 VALIDATION OF THE CARBOXYFLUORESCEIN DIACETATE ASSOCIATED WITH PROPIDIUM IODIDE FOR MURINE SPERM ACROSOME AND PLASMA MEMBRANE INTEGRITY

The spermatozoa is an ideal vehicle for genetic modification, production of transgenic animals, as well as a biotechnological tool for sperm-mediated gene transfer. However, in order to achieve successful sperm fertilization and exogenous DNA integration, it is necessary for viable cells to remain intact, allowing the sperm to penetrate the oocyte. Fluorescent probes allow evaluation of morphological and functional characteristics of cells, which can be evaluated separately or simultaneously. Therefore, the aim of this study was to validate the simultaneous evaluation of the integrity of plasma and acrosomal membranes of murine sperm using the probes carboxyfluorescein diacetate (CF) and propidium iodide (PI). In order to validate, a standard curve was performed. Sperm were obtained from epididymis and vas deferens from CD-1 mice (8 to 16 weeks of age). Recovered samples were diluted in PBS and then divided into 2 aliquots: one prepared with fresh semen (FS) and the other submitted to Percoll gradient (45%/90%) followed by flash-freezing in liquid nitrogen and thawing (FTP) to induce acrosome damage. Samples were prepared with the following average of FS:FTP: 100:0 (T100), 50 : 50 (T50), and 0 : 100 (T0). Samples were stained using 2 μL Hoescht 33342 (40 μLmL-1 in Dulbecco’s phosphate buffered saline), 3 μL of PI (0.5 mg mL-1 in PBS), 3 μL of CF (0.46 mg mL-1 in DMSO), and were incubated for 8 min at room temperature. After staining, the samples were placed on a slide, coverslipped, and evaluated immediately by epifluorescent microscopy. The Hoescht, PI, and CF fluorescence was detected using a filter with excitation at 352, 538, and 495 nm and emission at 455, 617, and 517 nm, respectively. Approximately 200 sperm cells per slide were examined and classified based on the fluorescence emitted from each probe. Spermatozoa CF+/IP- were considered as intact membranes, CF+/PI+ as acrosome membrane intact and plasma damaged, CF-/PI+ as damaged membranes, and CF-/PI- as acrosome membrane damaged and plasma intact. Hoeschst was used as positive dye. This experiment was replicated 6 times per group, and for statistical analyses, the data of plasma and acrosomal membrane integrity (dependent variables) in the treatments T0, T50, and T100 (independent variables) were submitted to simple linear regression analysis by STATVIEW software (SAS Institute Inc., Cary, NC, USA). The CFDA/PI probes were suitable for the analysis of acrosomal and membrane status of murine sperm and showed a high determination coefficient to plasma membrane integrity (R2 = 0.81; Y = 0.5412x + 6.375) and acrosome integrity (R2 = 0.85; Y = 0.5653x + 11.653). The described protocol was efficient for the simultaneous evaluation of plasma and acrosomal membrane integrity of murine spermatozoa, proving that CFDA can be employed to access acrosomal integrity as an alternative to FITC-PSA. Financial support: FAPESP.

Functional analysis of cells obtained from bronchoalveolar lavage fluid (BALF) of lung cancer patients

BALF from tumor segments provides access to immune system cells in contact with lung tumors. We analyzed BALF cells as to their production of H 2O 2 and NO, comparing tumor-affected to non-affected lung segments. Twelve patients were studied (4 NSCLC, 3 SCC, 5 Adenocarcinoma). The cell numbers recovered from BALF varied, and, in adenocarcinoma patients, smaller numbers were recovered from tumor-affected segments. H 2O 2 production (up to 6.3 nmoles/2 × 10 5cells) was obtained in 7/12 patients and, in these, it was more frequent in non-affected segments (7/7) than in affected segments (2/7). After culture, NO production was observed in three patients (6 to 314 μM) that also produced H 2O 2. These functional characteristics of cells in contact with neoplasia may have a role in determining the fate of the interactions between the immune system and lung cancer.

Method — A nonviral gene transfer method for transfecting multipotent adult progenitor cells (MAPC)

Multipotent adult progenitor cells (MAPC) are stem cells isolated primarily from adult bone marrow that have the ability to differentiate in vitro into cells with phenotypic and functional characteristics of cells from the three germ layers, namely endoderm, mesoderm and neuroectoderm. In addition, MAPCs proliferate for extended periods of time without obvious senescence. Hence, MAPCs may be ideal cells for therapy of genetic disorders, provided that the main impediment to gene therapy, namely efficient gene transfer and persistent gene expression, can be overcome. Most commercially available lipid-based methods that very highly efficiently transfect cell lines and primary cells, fail to transfect MAPC. However, 10–15% transfection of MAPC can be achieved using Superfect. However, this approach requires high density culture of MAPCs which subsequently differentiate. Between 12 and 15% transfection is achieved in MAPCs using electroporation, but this is highly toxic to MAPCs. Here, using transient expression of an enhanced green fluorescent protein (EGFP) gene, we report that nucleofection results in a transfection efficiency of over 25% with mouse MAPC without perturbing the conditions suitable for MAPC growth and maintenance, and with significantly less toxicity than electroporation. Similar results were seen for rat and human MAPC. Efficient transfection of MAPC by nucleofection offers an appealing non-viral mode of gene delivery and can be used to overexpress genes of interest in these cells. In addition, it should easily accomodate emerging site-specific integration technology, thereby culminating eventually in MAPC-mediated long-term gene therapy for genetic disorders.

Gene-expression analysis of single cells-nested polymerase chain reaction after laser microdissection.

The structural and functional characteristics of cells are dependent on the specific gene expression profile. The ability to study and compare gene expression at the cellular level will therefore provide valuable insights into cell physiology and pathophysiology. Individual cells were isolated from frozen colon tissue sections using laser microdissection. DNA as well as RNA were extracted, and total RNA was reversely transcribed to complementary DNA (cDNA). Both DNA and cDNA were analyzed by nested polymerase chain reaction (PCR). The quality of isolated DNA and RNA was satisfactory. Single cells were successfully microdissected using an ultraviolet laser micromanipulator. Nested PCR amplification products of DNA and cDNA of single cells could clearly be visualized by agarose gel electrophoresis. The combined use of laser microdissection and nested-PCR provides an opportunity to analyze gene expression in single cells. This method allows the analysis and identification of specific genes which are involved in physiological and pathophysiological processes in a complex of variable cell phenotypes.

Paclitaxel vs cyclophosphamide in peripheral blood stem cell mobilization: Comparative studies in a murine model

Paclitaxel is a promising drug for the treatment of breast and ovarian cancer. It also may play a role in mobilization of peripheral blood stem cells (PBSC), as an alternative to cyclophosphamide (Cy). We investigated the PBSC-mobilizing potential of paclitaxel compared to Cy in a murine model. C57Bl/6 mice were primed with intraperitoneal injections of Cy (200 mg/kg) or paclitaxel (60 mg/kg) and were sacrificed 4, 6, 8, or 10 days later. Spleens were harvested and processed to obtain low-density mononuclear cells that were used as PBSC. The number of hematopoietic progenitors (CFU-C) on day 4 was significantly higher in the paclitaxel group when compared to mice receiving Cy (72.0 ± 1.8 vs 9.8 ± 2.8, p < 0.001). By day 6, CFU-C became significantly higher in the Cy-treated group compared to the paclitaxel-treated group (195.6 ± 31.9 vs 95.8 ± 20.7, p < 0.05) and this trend was maintained. However, the total number of CFU-C recovered per spleen was greater in the paclitaxel-treated group (1.27 × 10 5 ± 0.53 × 10 5 vs 1.06 × 10 5 ± 0.36 × 10 5, NS). In contrast to paclitaxel, mobilization with Cy was associated with marked perturbation in the proportion of lymphoid cell subsets in the PBSC population along with functional impairment of lymphocytes. After 24 hours of in vitro IL-2 activation, the cytotoxic effector cell function of the Cy-mobilized PBSC population was lower than that of paclitaxel-mobilized cells when tested against three tumor cell lines (B16, melanoma; C1498, AML; and Yak-1, lymphoma). These results indicate that paclitaxel is an efficient mobilizer of PBSC, leading to early (day 4 to 6) mobilization of PBSC when compared to Cy (day 6 to 8). In addition, paclitaxel was associated with less perturbation of phenotypic and functional characteristics of cells contained within the mobilized PBSC population.

Placental cell release of transferrin: Analysis by reverse haemolytic plaque assay

Recent demonstrations of transferrin (TF) mRNA in placental tissue raised the possibility that the placenta may serve as an extra-hepatic source of this iron-binding protein during development. In this study, we first confirmed these findings using cRNA probes for TF, and then adapted the TF reverse haemolytic plaque assay for use with rat placental cells to identify the location and functional characteristics of cells secreting this product. We found TF releasing cells in placentae (day 19-21) with greater proportions present in cultures from basal than labyrinth zone regions. These cells appeared quite stable as evidenced by observations that fresh populations and 24, 48 and 72 h cultures from the same placental regions all contained similar percentages of secretors. The rate of TF plaque formation was greatly enhanced in the presence of tumour necrosis factor (0.1 ng/ml) for basal zone and yolk sac cells, but not for labyrinth zone cells, suggesting a potent but regionally specific modulation of TF release. When taken together, these findings demonstrate clearly that TF is released from cells of the placenta. Moreover, the regional differences in frequency and modulation of these cells suggests that the release of placental TF is a dynamic and carefully controlled process.

Culture of human renal cortex epithelial cells

A procedure is described for the establishment and propagation of epithelial cell rich cultures derived from normal human kidney cortex (NHK-C cells). Cells are harvested from tissue fragments of donor human kidney by progressive enzymatic dissociation. NHK-C cultures are morphologically heterogeneous but exhibit, predominantly, the functional characteristics of cells of the kidney proximal tubule.

Isolation and characterization of lymphocytes from bovine intestinal epithelium and lamina propria

The lymphocyte populations of the bovine gut lamina proprial (LP) and epithelial tissues were isolated and characterized with respect to cells bearing surface and cytoplasmic immunoglobulin (Ig). Functional characteristics of cells from the two tissues, including responsiveness to Concanavalin A (Con A), anti-bovine immunoglobulin (anti-Ig), Con A supernatants of bovine peripheral blood lymphocytes (bConA sup) and recombinant human IL-2 (rhIL-2), were also assessed. Less than 1% of the mononuclear cells in the epithelial tissue (IEL) stained for cytoplasmic Ig, and 9% stained positively for surface Ig. IEL did not proliferate in response to anti-Ig, although cells of this population did respond to Con A, bConA sup, and rhIL-2. Twenty-seven percent of bovine gut LP lymphocytes stained for surface Ig, while 39% of these cells were positive for cytoplasmic Ig. LP lymphocytes proliferated in response to all four stimulants used, Con A, anti-Ig, bConA sup and rhIL-2.