Publications (19)26.99 Total impact
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Article: Stem cells isolated from the human stromal limbus possess immunosuppressant properties.
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ABSTRACT: Mesenchymal stromal stem cells (MSC) are non-hemopoietic cells with the capacity to self-renewal and to differentiate into various cell lineages of mesenchymal origin. More recently, the immune regulatory potential of MSC has been focused on. Furthermore, mesenchymal stem cells obtained from diverse tissues possess immunomodulatory properties and inhibit proinflammatory immune reactions. The aim of this study was to determine the immunosuppressive characteristics of mesenchymal stem cells isolated from human limbal (L-MSC) tissue. L-MSC were enzymatically obtained from cadaveric sclero-corneal rims and expanded in vitro. The cells were characterized by flow cytometry using specific antibodies to mesenchymal stem cells markers. Clonogenic and tissue transdifferentiation in vitro assays were performed. The effect of L-MSC soluble factors on T cell proliferation was determined by flow cytometry. Cytokines such as transforming growth factor-b1 (TGF-β1) and Interleukin-10 (IL-10) on supernatants from L-MSC were identified by enzyme-linked immunosorbent assay (ELISA). Herein, we described that L-MSC cells in vitro-expanded were positive for the expression of vimentin, CD29, CD34, CD39, CD73 and CD105 mesenchymal stem cells markers; meanwhile, this cell population was negative to CD45 and HLA-DR hemopoietic markers as well as to cytokeratin expression. Clonogenic assays showed that these cells were able to form colonies. In addition, this L-MSC population had the ability to transdifferentiate into neurons and chondrocytes and to form tubular networks on matrigel in the presence of vascular endothelial growth factor (VEGF). These results indicated that these cells were stem cells. Additionally, soluble factors secreted by L-MSC were capable of mediating the suppression of T-cell receptor (TCR)-engagement lymphocyte proliferation. In an attempt to identify the possible immunosuppressive factors secreted by L-MSC, TGFβ1 and IL-10 cytokines were determined in the L-MSC supernatants by ELISA; interestingly, TGFβ1 was constitutively secreted by this cell population; in contrast, IL-10 was not detectable. Moreover, TGFβRII neutralizing antibodies were able to revert the TCR-engagement lymphocyte proliferation inhibition mediated by L-MSC. Thus, TGFβ1 secreted by L-MSC was able to suppress T cell proliferation. Taken together these results, explain in part the immunosuppressive features of this cell population obtained from the human limbus. All these characteristics make this cell population an excellent source to be used in the regenerative medicine.Molecular vision 01/2012; 18:2087-95. · 2.20 Impact Factor -
Article: Amniotic membrane is an immunosuppressor of peripheral blood mononuclear cells.
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ABSTRACT: Amniotic membrane (AM) is the inner layer of the placenta, which is in contact with the fetus; it has been used for transplantation in ocular surface diseases. It has been reported that amniotic membrane promotes epithelialization, inhibits angiogenesis and diminishes ocular inflammation. A persistent epithelial defect is the delay in epithelial wound healing caused by infiltrating inflammatory cells into the cornea and amniotic membrane transplantation has been successfully used in its treatment, however the mechanism of action in inhibiting inflammation it is not well understood. This study was aimed at determining whether denuded amniotic membrane (dAM) induces anti-inflammatory effects on peripheral blood mononuclear cells. Methods: Human peripheral blood mononuclear cells (PBMC) were cultured on dAM. Proliferation and apoptosis assays were performed on PBMC; and synthesis and secretion of pro-inflammatory cytokines by these cells was analyzed. Results: dAM induced apoptosis and inhibited cell proliferation of PBMC; and abolished the synthesis and the secretion of pro-inflammatory cytokines even when they were LPS stimulated. In contrast, when PBMC were cultured on hydrophilic membrane cell culture inserts, apoptosis was not significantly induced, cell proliferation was conserved, and synthesis and secretion of pro-inflammatory cytokines were not affected. Conclusions: Taken together, these results could explain the anti-inflammatory in vivo effects observed when the amniotic membrane is used as a transplant.Immunological Investigations 11/2010; 40(2):183-96. · 1.47 Impact Factor -
Article: [Down regulation of IL-8 and IL-6 in human limbal epithelial cells cultured with human dialyzable leukocyte extracts].
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ABSTRACT: Introduction: Human Limbal Epithelial Cells (hLEC) are stem cells that give rise to corneal epithelium. After corneal damage, hLEC produce large amounts of IL-8 and IL-6, inducing inflammation in cornea and conjunctiva. Despite inflammation is necessary to repair the ocular surface since this process may be potentially harmful and could lead to corneal opacity. Ophthalmic infectious diseases have been treated with human dialyzable leukocyte extracts (hDLE). Clinical observations in hDLE-treated patients, have suggested an apparent control of ocular inflammatory injuries, without changes in the re-epithelialization process. Objective: To determine the inflammatory cytokine profile in supernatants (SN) of hLEC cultured with hDLE. Methods: hLEC were obtained from cadaver donors. hDLE were added to the hLEC cultures, and SN were collected at different times (1h, 3h, 6h, and 24h). IL-1?, IL 6, IL-8, IL-12p70 and TNF-? were measured in SN with cytometric bead arrays. Results: The majority of isolated cells were CK19+/vimentin+/p63+, indicating that cultured-cells were limbal epithelial stem cells. Limbal cells responded to hDLE by diminishing the secretion of IL-8 and IL-6. Secretion of IL-8 and IL-6 was down-regulated significantly at 24h of culture with hDLE. Interestingly, hDLE did not induce secretion of IL-1 ?, TNF-?, and IL-12p70 in hLEC at any evaluated times. Conclusions: hDLE down-regulates secretion of IL-8 and IL-6 without induction of IL-1 ?, TNF-a, and IL-12p70 in hLEC. Our results provide a basis to understand some clinical effects, related to control ocular inflammation, that have been observed in patients treated with hDLE.Alergia 10/2010; 58(3):147-54. -
Article: Ocular involvement and blindness secondary to linear IgA dermatosis.
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ABSTRACT: A 43-year-old man with linear immunoglobulin A (IgA) dermatosis associated with gluten intolerance presented with progressive vision loss, pain and photosensitivity in both eyes. His visual acuity was light perception (LP) in both eyes. A physical examination revealed bullous, papular lesions with erythematous borders in periocular tissues, limbs, and thorax. Slit-lamp examination showed conjunctival hyperemia, fibrosis, corneal opacification, and vascularization with epithelial defects. Immunofluorescent skin and corneal surface biopsy studies showed linear IgA deposits. The patient was treated with keratolimbal allogenic transplantation and cryopreserved amniotic membrane in the right eye. Regardless of the treatment he persisted with torpid evolution developing retinal and choroidal detachments. After these events he was started on intravenous immune globulin (IVIG) and showed very slight improvement in ocular surface. These types of blistering diseases are rare in the eye. Even when adequate local treatment is given, systemic treatment is mandatory and ocular prognosis can be unsatisfactory.Journal of Ophthalmology 01/2010; 2010:280396. -
Article: The Amaranthus leucocarpus lectin enhances the anti-CD3 antibody-mediated activation of human peripheral blood CD4+ T cells.
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ABSTRACT: Activation of CD4(+) T cells plays a main role in adaptive immune response by regulating cellular and humoral immunity via processes associated with changes in cell surface oligosaccharide receptors. Lectins are glycoproteins that specifically recognize oligosaccharides and have been used to characterize changes in oligosaccharides present on T cell surface and their effects on activation. A lectin from Amaranthus leucocarpus seeds (ALL) is specific for glycoprotein structures containing galactose-N-acetylgalactosamine and is able to bind to human and murine CD4(+) T cells, however, its effect on activation remains unclear. We examined the effect of ALL on the activation of peripheral blood human CD4(+) T cells and analyzed cell proliferation, expression of the activation-associated molecule CD25, secretion of the activation-dependent cytokine interleukin (IL)-2 and intracellular calcium influx changes using flow cytometry. CD4(+) T cells were stimulated with anti-CD3 antibodies that provided the first activation signal in the presence or absence of ALL. ALL alone did not induce CD4(+) T cell activation but when also stimulated with anti-CD3 antibodies, ALL up-regulated CD25 expression, cell proliferation, IL-2 secretion and an intracellular calcium influx in a dose-dependent manner. In addition, ALL recognized CD4(+) T cells expressing the CD69 and Ki67 molecules expressed only by activated T cells and induced production of the TH1-type cytokine interferon-gamma. Our findings indicate that ALL binds to human activated CD4(+) T cells and enhances the degree of activation of CD4(+) T cells that are stimulated with anti-CD3 antibodies. ALL provides a new tool for analyzing T cell activation mechanisms.The Tohoku Journal of Experimental Medicine 01/2010; 221(4):271-9. · 1.24 Impact Factor -
Article: Effect of Bifidobacterium bifidum DSM 20082 cytoplasmic fraction on human immune cells.
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ABSTRACT: The objective of the present study was to determine the effect of the soluble cytoplasmic fraction from Bifidobacterium bifidum DSM 20082 (Bb) lysate on peripheral blood T cells. In peripheral blood mononuclear cells of healthy subjects, cytotoxic activity, proliferation, apoptosis, and up-regulation of CD8 or CD4 molecules in T cells were examined. When peripheral blood mononuclear cells were stimulated with Bb lysate, the main effect was observed in CD8+ cells as a significant increase of CD8 molecules in a dose-dependent manner, and this behavior was observed at 24, 48, and 72 h after stimulation; in contrast, stimulation with Bb lysate showed no effect on the up-regulation of CD4 molecules in T helper cells. Further Bb lysate did not induce proliferation activity in either CD8+ or CD4+ cells. Bb lysate induced activation of CD8+ cytotoxic activity against autologous monocytes. Around 80% of the cells stimulated with Bb lysate were positive to peanut agglutinin (PNA), suggesting that the stimulated CD8+ cells corresponded to activated/effector cellular populations. When apoptosis was determined, there were no differences between stimulated and non-stimulated cells. Our results indicate that Bb lysate is able to increase cytotoxic activity of peripheral CD8+ cells, without affecting lymphocyte survival.Immunological Investigations 02/2009; 38(1):104-15. · 1.47 Impact Factor -
Article: Study of the expression of CD30 in pterygia compared to healthy conjunctivas.
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ABSTRACT: The pterygium is characterized by a fibrovascular neoformation from the bulbar conjunctiva into the cornea. The recent discovery that abnormal markers associated with tumor diseases are identified in the pterygium strengthens the theory that the pterygium is a tumor-like disease rather than a degenerative disease. The CD30 molecule has been identified in neoplastic and normal epithelial proliferating cells. The aim of this study was to determine the expression of the CD30 molecule in the pterygium. Immunohistochemical staining using an antibody to CD30 and to the Ki-67 nuclear antigen was performed on 25 pterygial specimens and 10 healthy conjunctivas. Strong immunostaining against CD30 was observed in all pterygium specimens, in contrast to the healthy conjunctivas that showed weak immono-positivity in only three cases. The staining was diffused, predominantly to the basal epithelium. The Ki-67 antigen was observed in the nucleus of the basal epithelium of the pterygial specimens, and no staining was observed in the healthy conjunctiva. When serial sections were stained with CD30 and Ki-67, the cells that expressed CD30 also expressed Ki-67. The present study identified for the first time the existence of CD30 molecules in the basal epithelium of a pterygium. The fact that the positivity to Ki-67 in the basal epithelium of the pterygium correlated with the CD30 reactivity suggested that this protein could be associated with the uncontrolled cell proliferation of the epithelium in this pathology. The CD30 molecule could therefore be a suitable target to be inhibited using chimerical antibodies in pterygium diseases.Molecular vision 01/2009; 15:2068-73. · 2.20 Impact Factor -
Article: Comparative expression analysis of aquaporin-5 (AQP5) in keratoconic and healthy corneas.
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ABSTRACT: Keratoconus (KC) is a common progressive corneal disease characterized by excessive stromal thinning, central or paracentral conical protrusion, and disruptions in Bowman's layer. The etiology of KC is largely unknown, and a combination of genetic and environmental factors is believed to play a role in the origin of the disease. Recently, the absence of transcripts of the water channel, aquaporin-5 (AQP5), was demonstrated by reverse-transcription polymerase chain reaction (RT-PCR) in KC tissues and was proposed as a possible marker for KC. In this study, we sought to evaluate AQP5 mRNA and protein expression in KC and non-KC corneal tissues using a combination of techniques. A total of 69 samples of corneal tissue were analyzed including 39 corneal buttons from patients with advanced KC, 16 samples of non-KC corneal epithelium belonging to patients who underwent surface refractive surgery, 12 sclerocorneal rims obtained from healthy donor subjects, and two healthy corneal buttons. Determination of AQP5 transcript and protein expression patterns was performed by means of real time RT-PCR, immunohistochemistry, immunocytochemistry, and flow cytometry methods. Cell culture was performed to identify AQP5 protein expression in KC epithelial cells. AQP5 mRNA was expressed with no significant differences between KC and non-KC tissues. Moreover, AQP5 protein expression analysis did not reveal differences in protein levels and/or cell location among KC and non-KC tissues. Interestingly, AQP5 expression continues for up to 21 days in the isolated KC corneal epithelial cells. Our results do not support a role for AQP5 in KC etiopathogeny or as a disease marker. Genetic background differences or a distinct pathogenetic KC cascade specific to the analyzed population could account for the dissimilarities observed in KC-related AQP5 expression.Molecular vision 02/2008; 14:756-61. · 2.20 Impact Factor -
Article: Pars planitis is associated with an increased frequency of effector-memory CD57+ T cells.
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ABSTRACT: To evaluate the frequency, phenotype and the potential function of CD57+ T cell subsets in patients with pars planitis. CD4+CD57+ and CD8+CD57+ T cells were quantitated in peripheral blood from 15 patients with pars planitis and 15 healthy controls. To evaluate the phenotype and potential function of CD57+ T cell subsets CCR7, CD27, CD28, CD45RA, CD45RO, intracellular IFN-gamma, IL-4, perforin and granzyme-A expression were assessed by flow cytometry. CD57+ T cells subsets were increased in patients with pars planitis (p = 0.002). The majority of CD4+CD57+ T cells were CCR7-CD27-CD28-CD45RO+, while the most CD8+CD57+ T cells were CCR7-CD27-CD28-CD45RA+. The number of cells positive for intracellular IFN-gamma and IL-4 was higher in the CD57+ T cell populations. A greater number of CD8+CD57+ T cells than CD8+CD57- T cells were positive to perforin (p = 0.006) and granzyme-A (p = 0.01). CD57+ T cells had a phenotype associated with peripheral memory (CCR7-CD27-CD28-). Cytokine production by CD57+ T cells suggests that these cells may play a role in helper cell regulation. High expression of intracellular proteins involved in cytotoxicity suggests that CD8+CD57+ T cells may play an effector role. Taken together, this study proposes that CD57+ T cells function as memory-effector T cell subsets during pars planitis pathogenesis.British Journal of Ophthalmology 11/2007; 91(10):1393-8. · 2.90 Impact Factor -
Article: Increased expression of CD30 and CD57 molecules on CD4(+) T cells from children with atopic asthma: a preliminary report.
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ABSTRACT: T helper type 2 (Th2) cells play an important role in the onset and persistence of allergic airway inflammation. Consequently, many authors have attempted to identify cell surface markers associated with a Th2 phenotype. This work was aimed at correlating CD30 and CD57 expression on CD4(+) T cells with interleukin (IL)-4 production in peripheral blood mononuclear cells (PBMCs) from allergic patients. PBMCs from 17 children with atopic asthma and 12 nonatopic healthy control children were analyzed. The CD28, CD30, CD40L, CD57, CD62L, CD69, IL-4, and IFN-gamma expressions on CD4(+) T cells were determined by double immunofluorescence and flow cytometry in PBMCs ex vivo and after phorbol-12-myristate-13-acetate plus ionomycin (PMA/I) stimulation. An increased percentage of peripheral CD4(+)CD30(+) T cells was observed in asthmatic patients (p < 0.001). In addition, the percentage of CD4(+) T cells expressing IL-4, IFN-gamma, CD30, CD40L, CD57, or CD69 significantly increased (p < 0.01) after PMA/I stimulation, in asthmatic patients. The CD30 expression on CD4(+) T cells from asthmatic patients, after stimulation, correlated with both IL-4 and IFN-gamma production, whereas CD57 expression only correlated with IL-4 production. These data suggest that the expression of CD30 and CD57 cell markers on T cells could reflect circulating effector T cell early activation in the allergic airway disease.Allergy and Asthma Proceedings 10/2007; 28(6):659-66. · 2.17 Impact Factor -
Article: Lectin from Phaseolus acutifolius var. escumite: chemical characterization, sugar specificity, and effect on human T-lymphocytes.
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ABSTRACT: Purification of the lectin from Phaseolus acutifolius var. escumite was achieved by affinity chromatography on a column containing glutaraldehyzed membranes from blood group O erythrocytes. The lectin is a tetrameric glycoprotein of 121 kDa with 10% of sugar by weight composed by four subunits of 30 kDa as determined by SDS-PAGE. The lectin is composed of four isolectins as determined by ion-exchange chromatography on a mono-S column. The lectin and its isolectins showed identical NH2 terminal residues (ANDLSFNFQR FNETN) with homology to the PHA leucoagglutinin-precursor. Peptide mass fingerprint from each lectin isoform determined from tryptic peptides by MALDI-TOF (matrix assisted laser desorption ionization-time-of-flight) showed differences among subunits, thus suggesting microheterogeneity in their amino acid sequences or different glycosylation patterns. The lectin and its four isolectins agglutinated erythrocytes without serological specificity and showed mitogenic activity on human leukocytes; moreover, the main effect was rather toward CD8+ than to CD4+ human peripheral lymphocytes. The lectin from escumite was not inhibitable by simple sugars; however, the specificity of the lectin and its isoforms was mainly addressed toward galactose residues present in bi- or triantennary N-acetyllactosamine-type glycans.Journal of Agricultural and Food Chemistry 08/2007; 55(14):5781-7. · 2.82 Impact Factor -
Article: [In vitro phenotypic characterization of human limbal epithelial cells].
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ABSTRACT: Human limbal epithelial cells (huLEC) have been used for clinical purposes in ocular surface diseases to promote rapid re-epithelisation and restore corneal epithelium integrity. However, in Mexico this technique has not been fully developed. This study was conducted to characterize the huLEC phenotype expanded in vitro using a cell culture technique. Cells were obtained from limbal tissue, cultured in KSFM medium and analyzed for the expression of vimentin, K, K19, p63, K12, by flow cytometry and immuno-fluorescence. The phenotype of cultured cells was vimentin+K+K19+ p63+K12-. Our results suggest that under these culture conditions huLEC maintained their stem cell phenotype. This culture technique could be used for clinical purposes in Mexico.Gaceta medica de Mexico 01/2007; 143(3):183-7. · 0.22 Impact Factor -
Article: The N-acetyl-D-glucosamine specific adhesin from Mannheimia haemolytica activates bovine neutrophils oxidative burst.
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ABSTRACT: In this work we identified specific bovine leukocytes that were bound by the Mannheimia haemolytica adhesin molecule (MhA) and the biological effect on the leukocytes. Histochemical staining and flow cytometry showed that MhA bind neutrophils (90%) and monocytes (5%). MhA induced an oxidative response in purified neutrophils; this effect was 1.5-fold higher than the effect observed with control cells activated with Zymosan. Cellular binding by MhA was inhibited with GlcNAc and its oligomers, as well as by glycoproteins containing tri- and tetra-antennary N-glycosydically linked glycans. MhA-induced oxidative burst was significantly inhibited by GlcNAc, iodoacetamide, superoxide dismutase, and piroxicam (p<0.05). Our findings suggest that among bovine leukocytes, neutrophils are the main target for MhA, inducing production of oxidative radicals by non-opsonic mechanism that seem to play an important role in tissue damage during mannheimiosis.Veterinary Immunology and Immunopathology 09/2006; 113(1-2):148-56. · 2.08 Impact Factor -
Article: Comparative analysis of mononuclear cell surface markers in atopic processes--a preliminary study.
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ABSTRACT: Atopic disorders are driven by the Th2 cell subset. We have determined the expression of costimulatory molecules and cell surface markers on peripheral CD4+ T cells and antigen presenting cells, in different atopic diseases, and we have also tried to correlate the expression of these markers with the severity of the disease. Cells from patients with atopic and contact dermatitis, mild or severe asthma, and symptomatic and non-symptomatic atopic rhinitis were analyzed by flow cytometry. Our results showed that CD30, CD124, and CD152 expression on CD4+ T cells was significantly higher in atopic dermatitis than in contact dermatitis patients (p < 0.05). It was interesting to observe that the cell surface expression of CD80 in T and B cells from atopic dermatitis patients was not enhanced as opposed to the other atopic diseases we analyzed. Our results suggest that there are differences in the immune mechanisms involved in the different atopic diseases, and that expression of CD30 in CD4+ T cells might be a marker of disease activity in atopic dermatitis.Immunological investigations 03/2003; 32(1-2):95-104. · 1.16 Impact Factor -
Article: Peanut and Amaranthus leucocarpus lectins discriminate between memory and naive/quiescent porcine lymphocytes.
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ABSTRACT: Lectins are relevant tools to isolate and characterize different cellular sub-populations. In this work, we used the lectins Arachis hypogaea (Peanut agglutinin, PNA) and Amaranthus leucocarpus (ALL), specific for Galss1, 3GalNAc, to characterize naive and memory lymphocytes from pigs, experimentally infected with the porcine rubulavirus (RvP). Our results showed that both lectins recognized preferentially lymphocytes with the CD4(+)CD8(+) phenotype (P<0.05). The phenotypic analysis of the cells recognized by these lectins indicated that PNA(+) lymphocytes showed higher rate of the CD29 antigen (PNA(+)CD29(high)) than ALL(+) (ALL(+)CD29(low)). The number of PNA(+)CD29(high) lymphocytes increased after 8 weeks of experimental infection with RvP, and most of the ALL(+)CD29(low) cells became CD29(middle). PNA(+) lymphocytes isolated from infected pigs proliferated after stimulation with the RvP, whereas ALL(+) cells did not. In vitro assays indicated that the ALL(+) cells from previously infected pigs diminished from 7.5 +/- 2 to 0.5 +/- 0.3% after RvP stimulation; whereas PNA(+) cells increased from 4 +/- 1 to 42 +/- 2%, whereas no modification in ALL(+) or PNA(+) cellular population was identified in lymphocytes from naive animals after RvP stimulation. Our results suggest that the cellular distribution/organization of the O-glycosydically linked glycans on lymphocytes may correlate with biological functions, and that PNA could be a tool to isolate specifically porcine memory T cell subsets, whereas ALL could be useful to isolate naive/quiescent T lymphocytes.Veterinary Immunology and Immunopathology 01/2002; 84(1-2):71-82. · 2.08 Impact Factor -
Article: [Uveitis immunopathology: currents knowledge, clinical correlation and research perspectives in the immue-ocular area].
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ABSTRACT: The eye has multiple mechanisms of immune regulation implicated in the maintenance of ocular immune privilege. However, sometimes diseases or disorders appear and can cause clinical manifestations of intraocular inflammation; usually those diseases are collectively named "uveitis". Despite the uveitis is the main cause of eye morbidity and lost of visual function, the vast majority of the immune mechanisms involved in its generation remains unknown. In this article, the authors reviews the process of immune regulation inside the eye, the immunological mechanisms of damage implicated in the generation of uveitis, as well as the clinical aspects associated with the immune pathogenesis; in the last part of this paper we present the most recent trends in ocular research related to immunological damage in uveitis.Alergia 53(6):226-35. -
Article: Duration of botulinum toxin effect in the treatment of crocodile tears.
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ABSTRACT: To provide clinical evidence of the duration of botulinum toxin type A (BTX-A) effect when applied in the palpebral lobe of the lacrimal gland in patients with gustatory epiphora. Prospective, nonrandomized, nonblinded study. Patients with history of gustatory epiphora were included. A Schirmer test was performed to quantify tearing induced by chewing. Clinical examination included visual acuity, tear-duct syringing, slit lamp examination, corneal staining, and eyelid malpositions. A questionnaire was completed by each patient to asses the severity of hyperlacrimation. A single dose of 2.5 units of BTX-A was injected directly into the lacrimal gland palpebral lobe. Patients were evaluated before and at 1, 4, 12, and 24 weeks after injection. The same person performed the examination and the BTX-A injection. Descriptive statistics, using repeated measures and a paired t test, were used for statistical analysis. Fifteen patients were included. Mean age was 63 years. Before BTX-A injection, mean Schirmer test values were 5.47 mm in the unaffected eyes (NAE) and 12.07 mm in the affected eyes (AE). When comparing Schirmer test values in the AE before and after BTX-A injection, there were statistically significant differences (p < 0.05). Only 2 patients developed mild transitory ptosis. No other complications were noted. The effect of 2.5 units of BTX-A injected into the lacrimal gland lasted 6 months, a duration similar to that reported for other application sites.Ophthalmic Plastic and Reconstructive Surgery 22(6):453-6. · 0.69 Impact Factor -
Article: Peanut and Amaranthus leucocarpus lectins discriminate between memory and naive/quiescent porcine lymphocytes
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ABSTRACT: Lectins are relevant tools to isolate and characterize different cellular sub-populations. In this work, we used the lectins Arachis hypogaea (Peanut agglutinin, PNA) and Amaranthus leucocarpus (ALL), specific for Galß1, 3GalNAc, to characterize naive and memory lymphocytes from pigs, experimentally infected with the porcine rubulavirus (RvP). Our results showed that both lectins recognized preferentially lymphocytes with the CD4+CD8+ phenotype (P<0.05). The phenotypic analysis of the cells recognized by these lectins indicated that PNA+ lymphocytes showed higher rate of the CD29 antigen (PNA+CD29high) than ALL+ (ALL+CD29low). The number of PNA+CD29high lymphocytes increased after 8 weeks of experimental infection with RvP, and most of the ALL+CD29low cells became CD29middle. PNA+ lymphocytes isolated from infected pigs proliferated after stimulation with the RvP, whereas ALL+ cells did not. In vitro assays indicated that the ALL+ cells from previously infected pigs diminished from 7.5±2 to 0.5±0.3% after RvP stimulation; whereas PNA+ cells increased from 4±1 to 42±2%, whereas no modification in ALL+ or PNA+ cellular population was identified in lymphocytes from naive animals after RvP stimulation. Our results suggest that the cellular distribution/organization of the O-glycosydically linked glycans on lymphocytes may correlate with biological functions, and that PNA could be a tool to isolate specifically porcine memory T cell subsets, whereas ALL could be useful to isolate naive/quiescent T lymphocytes.Veterinary Immunology and Immunopathology 84:71-82. · 2.08 Impact Factor -
Article: Comparative evaluation of the CD4+CD8+ and CD4+CD8− lymphocytes in the immune response to porcine rubulavirus
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ABSTRACT: The porcine immune system is unique in the expression of CD4+CD8+ (double-positive, DP) lymphocytes. These cells have been associated with immunological memory due to their gradual increase with age, the expression of memory phenotype and their ability to respond to recall viral antigen. This work analyzes the biological function of CD4+CD8− and CD4+CD8+ lymphocytes in the immune response to porcine rubulavirus (PRv). CD4+CD8− cells isolated from pigs 3 weeks after infection with porcine rubulavirus proliferated in response to homologous virus and generated lymphoblasts which were predominantly of the CD4+CD8+ phenotype, whereas stimulation with mitogen induced proliferation but did not switch the phenotype. CD4+CD8− lymphocytes isolated after 10 weeks of infection proliferated in response to phytohemagglutinin (PHA) but did not proliferate in response to homologous virus and did not change their phenotype, whereas CD4+CD8+ lymphocytes proliferated in response to PHA and to viral antigen. The cytokine profile of both lymphocyte populations showed the presence of IL-2 and IL-10 transcripts, quantitation demonstrated that CD4+CD8+ cells expressed mainly IL-10, whereas CD4+CD8− lymphocytes expressed primarily IL-2. Our results show that CD4+CD8− lymphocytes in the early phase of porcine rubulavirus infection can be converted to double-positive cells expressing IL-10 in an antigen-dependent manner, and that CD4+CD8− T-cells late in infection do not acquire CD8.Veterinary Immunology and Immunopathology.
Top co-authors
Top Journals
Institutions
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2008–2012
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Instituto de Oftalmología Fundación Conde de Valenciana
Mexico City, The Federal District, Mexico
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2002–2010
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Instituto Nacional de Enfermedades Respiratorias
Mexico City, The Federal District, Mexico
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2009
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Université des Sciences et Technologies de Lille 1
Lille, Nord-Pas-de-Calais, France
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2007
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Instituto Nacional de Pediatría
Mexico City, The Federal District, Mexico
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