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We have used the Galβ(l→3)GalNAc-specific Amaranthus leucocarpus lectin to isolate a thymus cell subpopulation which is different from that sorted with Arachis hypogaea lectin. The cells recognized by A. leucocarpus lectin were predominantly CD4+, whereas a minor proportion of CD8+ cells (approximately 11%) were also identified. The A. leucocarpus-positive cells were located in the thymus medulla and the cortico-medullary junction. The cortex was negative for A. leucocarpus cells. [ABSTRACT FROM AUTHOR]

A high morbidity and mortality of goats has been observed in the central area of Veracruz, Mexico, where goat production is practiced. The clinical signs showed by these goats may be related to mycoplasmosis, a highly contagious disease that causes serious economic losses. Thus, our objective was to evaluate the seroprevalence of Mycoplasma mycoides subspecies mycoides (Mmm) using a competitive ELISA (c-ELISA) and to assess risk factors associated with seropositivity. The seroprevalence of 556 goats sampled in random was 11.2%, the frequency by local municipality was 64.3%, and 25.3% per animal small production farm (SPF). The identified risk factors were associated with breeding in the municipalities of Coacoatzintla (OR=14.2; 95%CI: 6.9-29.5), Villa Aldama (OR=4.9; 95%CI:2.5-9.5) and Chiconquiaco (OR=7.71; 95%CI:3.6-16.47); the crossbreeding phenotype (OR=1.78; 95%CI:1.05-3.04), stage of suckling goat kids (OR=2.87; 95%CI:1.64-5.03), mobilization of animals (OR=4; 95%CI:2.33-6.88), semi intensive production system (OR=2.51; 95%CI:1.24-5.07) and coexistence with cattle (OR=3.28; 95%CI:1.53-7.02). Possible protective factors were associated with drink water from stream (OR=0.54; 95%CI: 0.29-0.98), well water (OR=0.11; 95% CI:0.01-0.82), Saanen breed (OR=0.39; 95%CI:0.19-0.8), purchased animals (OR=0.47; 95%CI:0.24-0.93) and the extensive system of production (OR=0.13; 95%CI:0.03-0.55). In conclusion, this study showed that the overall seroprevalence of Mmm in goats has a low general distribution, moderated by animal small production farms and high by municipality, in the central region of Veracruz, Mexico. Considering these results is required to perform further studies by use of molecular techniques for the precise identification of the etiological agent. [ABSTRACT FROM AUTHOR]

The COVID-19 disease continues to cause complications and deaths worldwide. Identifying effective immune protection strategies remains crucial to address this ongoing challenge. The Bacillus Calmette-Guérin (BCG) vaccine, developed initially to prevent pulmonary tuberculosis, has gained relevance due to its ability to induce cross-protection against other pathogens of the airways. This review summarizes research on the immunological protection provided by BCG, along with its primary clinical and therapeutic uses. It also explores the immunological features of COVID-19, the mechanisms implicated in host cell death, and its association with chronic pulmonary illnesses such as tuberculosis, which has led to complications in diagnosis and management. While vaccines against COVID-19 have been administered globally, uncertainty still exists about its effectiveness. Additionally, it is uncertain whether the utilization of BCG can regulate the immune response to pathogens such as SARS-CoV-2., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 Published by Elsevier Ltd.)

The global burden of cancer is on the rise, with varying national patterns. To gain a better understanding and control of cancer, it is essential to provide national estimates. Therefore, we present a comparative description of cancer incidence and mortality rates in Mexico from 1990 to 2019, by age and sex for 29 different cancer groups. Based on public data from the Global Burden of Disease Study 2019, we evaluated the national burden of cancer by analyzing counts and crude and age-standardized rates per 100,000 people with 95% uncertainty intervals for 2019 and trends using the annual percentage change from 1990 to 2019. In 2019, cancer resulted in 222,060 incident cases and 105,591 deaths. In 2019, the highest incidence of cancer was observed in non-melanoma skin cancer, prostate cancer, and breast cancer. Additionally, 53% of deaths were attributed to six cancer groups (lung, colorectal, stomach, prostate, breast, and pancreatic). From 1990 to 2019, there was an increasing trend in incidence and mortality rates, which varied by 10-436% among cancer groups. Furthermore, there were cancer-specific sex differences in crude and age-standardized rates. The results show an increase in the national cancer burden with sex-specific patterns of change. These findings can guide national efforts to reduce health loss due to cancer.

Background: Major depressive disorders (MDDs) occurs frequently in patients with tuberculosis (TB). Elevated serum pro-inflammatory cytokine levels in MDD patients is a well-established fact. Therefore, an integrated clinical practice should be considered. However, the inflammatory status of MDD-TB patients is unknown. In this study, we analyze cytokines in activated-cells and sera from MDD-TB, TB, MDD patients, and healthy controls., Methods: Flow cytometry was used to evaluate the intracellular production of interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-12, and IL-10 by peripheral blood mononuclear cells after a polyclonal stimulation. A Bio-Plex Luminex system was used to measure serum cytokine and chemokine levels in the study groups., Results: We observed a 40.6% prevalence of MDD in TB patients. The proportion of IFN-gamma-producing cells was higher in MDD-TB patients than other pathological groups. Nevertheless, the percentage of TNF-alpha- and IL-12-producing cells was similar between MDD-TB and TB patients. Likewise, MDD-TB and TB patients showed similar serum pro-inflammatory cytokine and chemokine levels, which were significantly lower than those in MDD patients. By multiple correspondence analyses, we observed that low levels of serum IL-4, IL-10, and IL-13 were powerfully associated with TB comorbidities with MDD., Conclusions: A high frequency of IFN-γ-producing cells is associated with low levels of serum anti-inflammatory cytokines in MDD-TB patients.

Background: Cystic fibrosis (CF) is a genetic disease in which thick, sticky mucus is produced in the lungs (and other organs) that impairs ciliary clearance, leading to respiratory problems, increased chronic bacterial infections, and decreased lung function. Staphylococcus aureus is one of the primary bacterial pathogens colonizing the lungs of CF patients. This study aimed to characterize the genetic relatedness of S. aureus, its presence in children with CF, and its cytotoxic activity in THP1 cell-derived macrophages (THP1m)., Methods: Genetic relatedness of S. aureus isolates from a cohort of 50 children with CF was determined by pulsed-field gel electrophoresis (PFGE). The VITEK 2 automated system was used to determine antimicrobial susceptibility, and methicillin-resistance S. aureus (MRSA) was determined by diffusion testing using cefoxitin disk. The presence of mecA and lukPV genes was determined by the polymerase chain reaction and cytotoxic activity of S.aureus on THP1m by CytoTox 96® assay., Results: From 51 S. aureus isolates from 50 children with CF, we identified 34pulsotypes by PFGE. Of the 50 children, 12 (24%) were colonized by more than one pulsotype, and 5/34 identified pulsotypes(14.7%) were shared between unrelated children. In addition, 3/34 pulsotypes (8.8%) were multidrug-resistant (MDR), and2/34 (5.9%) were MRSA. Notably, 30/34 pulsotypes (88.2%) exhibited cytotoxicity on THP1m cells and 14/34 (41.2%) alteredTHP1m monolayers. No isolate carried the lukPV gene., Conclusions: Although a low frequency of MRSA and MDR wasfound among clinical isolates, most of the S. aureus pulsotypes identified were cytotoxic on THP1m.

T lymphocyte activation begins with antigen/MHC recognition by the TCR/CD3 complex followed by a costimulatory signal provided by CD28. The search for novel costimulatory molecules has been extensive due to their potential use as immunotherapeutic targets. Although some molecules have been identified, they are unable to provide sustainable signaling to allow for proper T cell activation and proliferation. It has been shown that the Amaranthus leucocarpus lectin (ALL) can be used as an in vitro costimulator of CD4 + lymphocytes in the presence of anti-CD3 mAb; this lectin specifically recognizes O -glycans of the Galβ1-3GalNAc-O-Ser/Thr type, including a 70-kDa moesin-like protein that has been suggested as the costimulatory molecule. However, the identity of this molecule has not been confirmed and such costimulation has not been analyzed in CD8 + lymphocytes. We show herein that the expression kinetics of the glycoproteins recognized by ALL (gpALL) is different in CD4 + and CD8 + T cells, unlike moesin expression. Results from IP experiments demonstrate that the previously described 70-kDa moesin-like protein is an O -glycosylated form of moesin ( O -moesin) and that in vitro stimulation with anti-CD3 and anti-moesin mAb induces expression of the activation molecules CD69 and CD25, proliferation and IL-2 production as efficiently as cells costimulated with ALL or anti-CD28. Overall, our results demonstrate that O -moesin is expressed in CD4 + and CD8 + T lymphocytes and that moesin provides a new costimulatory activation signal in both T cell subsets., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Gómez-Henao, Saavedra, Chávez-Sánchez, Lascurain, Zenteno and Tenorio.)

Cystic fibrosis (CF) is a genetic disease affecting more than 70,000 people worldwide. It is caused by a mutation in the cftr gene, a chloride ion transporter localized in the plasma membrane of lung epithelial cells and other organs. The loss of CFTR function alters chloride, bicarbonate, and water transport through the plasma membrane, promoting the production of a thick and sticky mucus in which bacteria including Pseudomonas aeruginosa and Burkholderia cenocepacia can produce chronic infections that eventually decrease the lung function and increase the risk of mortality. Autophagy is a well-conserved lysosomal degradation pathway that mediates pathogen clearance and plays an important role in the control of bacterial infections. In this mini-review, we describe the principal strategies used by P. aeruginosa and B. cenocepacia to survive and avoid microbicidal mechanisms within the autophagic pathway leading to the establishment of chronic inflammatory immune responses that gradually compromise the lung function and the life of CF patients., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Flores-Vega, Vargas-Roldán, Lezana-Fernández, Lascurain, Santos-Preciado and Rosales-Reyes.)

The emergence of high-risk clones of priority pathogens exhibiting convergence of antimicrobial resistance and virulence is a critical issue worldwide. In a previous study, an extensively drug-resistant Pseudomonas aeruginosa was isolated from a chronically colonized pediatric patient with cystic fibrosis (CF). In this study, we analyzed genomic data of this strain (CF023-Psa42), extracting clinically and epidemiologically relevant information (i.e., the antimicrobial resistome, virulome, and sequence type). In this regard, we report the emergence of GES-19 (extended-spectrum β-lactamase)-producing P. aeruginosa with genotype exoU+. The CF023-Psa42 strain exhibited a broad resistome, belonging to the international high-risk clone sequence type ST235. The bla GES-19 gene was located on a class 1 integron, along to aac(6')-33, aac(6')-Ib-cr, bla OXA-2 , aadA1, sul1, and qacEΔ1 resistance genes. Relevant virulence genes such as lasA (proteolysis and elastolysis), toxA (exotoxin A), alg (alginate biosynthesis operon), and exoU (toxin of type III secretion systems) were predicted. Our findings reveal the convergence of broad resistome and virulome in P. aeruginosa ST235. Genomic surveillance is essential to monitor the emergence and dissemination of priority pathogens with epidemiological success., Competing Interests: Declaration of competing interest No potential conflict of interest was reported by the authors., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background: Long-term persistence of Pseudomonas aeruginosa in the lung of individuals with cystic fibrosis (CF) is associated with progressive selection of diverse genotypes and phenotypes. This bacterial adaptation leads to chronic infection and increased morbidity and mortality. The aim of this study was to establish the prevalence, clonal relatedness, antimicrobial susceptibility and virulence-associated phenotypes of P. aeruginosa isolates in a cohort of 50 Mexican children with CF-associated chronic lung infection., Methods: Clonal relatedness of P. aeruginosa isolates was verified by pulsed-field gel electrophoresis. The antimicrobial susceptibility was determined by an automated system that performs bacterial identificación and antibiotic susceptibility testing (VITEK 2) and/or broth microdilution method. Biofilm formation was quantified with the crystal violet method; swarming motility was measured on soft agar, and susceptibility to normal human serum determined by reduction of colony formed units (CFUs)., Results: High prevalence of P. aeruginosa colonization among Mexican children with CF was confirmed; 20% (10/49) of clones identified showed a multidrug-resistant phenotype and 8.2% (4/49) an extensive drug resistance phenotype; 26.5% (13/49) of the isolates were resistant to colistin, 42.9% (21/49) presented a phenotype of adaptation associated with chronic infection and 79.6% (39/49) showed increased ability to survive in normal human serum., Conclusions: This cohort of children with CF reveals that colonizing P. aeruginosa strains predominantly display resistance to several first-line antibiotics, although most isolates were susceptible to meropenem and tobramycin; 42.9% of isolates showed a phenotype consistent with adaptation to chronic lung infection.

Here, we present the draft genome sequence of a Pseudomonas aeruginosa isolate (strain CF16053) belonging to a novel sequence type (ST), ST3351, isolated from a pediatric patient with cystic fibrosis (CF). CF16053 shows high-level resistance to polymyxins associated with mutations in the pmrB gene. Biofilm, pyoverdine, exotoxin A, and type III secretion system (T3SS) genes were identified., (Copyright © 2020 Rosales-Reyes et al.)

Macrophages are essential cells of the innate immune response against microbial infections, and they have the ability to adapt under both pro- and anti-inflammatory conditions and develop different functions. A growing body of evidence regarding a novel macrophage subpopulation that expresses CD3 has recently emerged. Here, we explain that human circulating monocytes can be differentiated into CD3 + TCRαβ + and CD3 + TCRαβ - macrophages. Both cell subpopulations express on their cell surface HLA family molecules, but only the CD3 + TCRαβ + macrophage subpopulation co-express CD1 family molecules and transmembrane TNF (tmTNF). CD3 + TCRαβ + macrophages secrete IL-1β, IL-6 IP-10, and MCP-1 by both tmTNF- and CD3-dependent pathways, while CD3 + TCRαβ - macrophages specifically produce IFN-γ, TNF, and MIP-1β by a CD3-dependent pathway. In this study, we also used a mouse model of BCG-induced pleurisy and demonstrated that CD3 + myeloid cells (TCRαβ + and TCRαβ - cells) are increased at the infection sites during the acute phase (2 weeks post-infection). Interestingly, cell increment was mediated by tmTNF, and the soluble form of TNF was dispensable. BCG-infection also induced the expression of TNF receptor 2 on CD3 + myeloid cells, which increased after BCG-infection, suggesting that the tmTNF/TNFRs axis plays an important role in the presence or function of these cells in tuberculosis., (Copyright © 2019 Rodriguez-Cruz, Vesin, Ramon-Luing, Zuñiga, Quesniaux, Ryffel, Lascurain, Garcia and Chávez-Galán.)

Tuberculosis (TB) is an infectious disease caused by Mycobacterium tuberculosis (Mtb) and remains a major cause of morbidity and mortality worldwide. In the host's immune response system, T cells play a critical role in mediating protection against Mtb infection, but the role of CD8 + T cells is still controversial. We evaluated the phenotypical characterization and cytotoxic ability of CD8 + T cells by flow cytometry-based assay. Cytokine levels in serum were measured by multiplex cytokine assay. Our data show that cells from TB patients have an increased percentage of peripheral blood CD8 + αβ + T (p = 0.02) and CD56 + CD8 + T (p = 0.02) and a decreased frequency of NKG2D + CD8 + T (p = 0.02) compared with healthy donors. Unlike CD8 + T cells from healthy donors, CD8 + T cells from TB patients exhibit greater cytotoxicity, mediated by HLA class I molecules, on autologous monocytes in the presence of mycobacterial antigens (p = 0.005). Finally, TB patients have a proinflammatory profile characterized by serum high level of TNF-α (p = 0.02) and IL-8 (p = 0.0001), but, interestingly, IL-4 (p = 0.002) was also increased compared with healthy donors. Our data show evidence regarding the highly cytotoxic status of CD8 + T cells in Mtb infection. These cytotoxic cells restricted to HLA-A, B, and C could be used to optimize strategies for designing new TB vaccines or for identifying markers of disease progression., (© 2019 The Authors. Microbiology and Immunology published by The Societies and John Wiley & Sons Australia, Ltd.)

Background: Light at night creates a conflicting signal to the biological clock and disrupts circadian physiology. In rodents, light at night increases the risk to develop mood disorders, overweight, disrupted energy metabolism, immune dysfunction and cancer. We hypothesized that constant light (LL) in rats may facilitate tumor growth via disrupted metabolism and increased inflammatory response in the host, inducing a propitious microenvironment for tumor cells., Methods: Male Wistar rats were exposed to LL or a regular light-dark cycle (LD) for 5 weeks. Body weight gain, food consumption, triglycerides and glucose blood levels were evaluated; a glucose tolerance test was also performed. Inflammation and sickness behavior were evaluated after the administration of intravenous lipopolysaccharide. Tumors were induced by subcutaneous inoculation of glioma cells (C6). In tumor-bearing rats, the metabolic state and immune cells infiltration to the tumor was investigated by using immunohistochemistry and flow cytometry. The mRNA expression of genes involved metabolic, growth, angiogenes and inflammatory pathways was measured in the tumor microenvironment by qPCR. Tumor growth was also evaluated in animals fed with a high sugar diet., Results: We found that LL induced overweight, high plasma triglycerides and glucose levels as well as reduced glucose clearance. In response to an LPS challenge, LL rats responded with higher pro-inflammatory cytokines and exacerbated sickness behavior. Tumor cell inoculation resulted in increased tumor volume in LL as compared with LD rats, associated with high blood glucose levels and decreased triglycerides levels in the host. More macrophages were recruited in the LL tumor and the microenvironment was characterized by upregulation of genes involved in lipogenesis (Acaca, Fasn, and Pparγ), glucose uptake (Glut-1), and tumor growth (Vegfα, Myc, Ir) suggesting that LL tumors rely on these processes in order to support their enhanced growth. Genes related with the inflammatory state in the tumor microenvironment were not different between LL and LD conditions. In rats fed a high caloric diet tumor growth was similar to LL conditions., Conclusions: Data indicates that circadian disruption by LL provides a favorable condition for tumor growth by promoting an anabolic metabolism in the host.

The extracellular domains of some membrane proteins can be shed from the cell. A similar phenomenon occurs with β1 integrins (α1β1 and α2β1) in guinea pig. The putative role of β1 integrin subunit alterations due to shedding in airway smooth muscle (ASM) in an allergic asthma model was evaluated. Guinea pigs were sensitized and challenged with antigen. Antigenic challenges induced bronchoobstruction and hyperresponsiveness at the third antigenic challenge. Immunohistochemistry and immunoelectronmicroscopy studies showed that the cytosolic and extracellular domains of the β1 integrin subunit shared the same distribution in airway structures in both groups. Various polypeptides with similar molecular weights were detected with both the cytosolic and extracellular β1 integrin subunit antibodies in isolated airway myocytes and the connective tissue that surrounds the ASM bundle. Flow cytometry and Western blot studies showed that the expression of cytosolic and extracellular β1 integrin subunit domains in ASM was similar between groups. An increment of ITGB1 mRNA in ASM was observed in the asthma model group. RACE-PCR of ITGB1 in ASM did not show splicing variants. The expression levels of integrin-linked kinase (ILK) and paxillin diminished in the asthma model, but not talin. The levels of phosphorylation of myosin phosphatase target subunit 1 (MYPT1) at Thr(696) increased in asthma model. Our work suggests that β1 integrin is secreted in guinea pig airway wall. This secretion is not altered in asthma model; nevertheless, β1 integrin cytodomain assembly proteins in focal cell adhesions in which ILK and paxillin are involved are altered in asthma model. J. Cell. Biochem. 117: 2385-2396, 2016. © 2016 Wiley Periodicals, Inc., (© 2016 Wiley Periodicals, Inc.)

T cells from patients with systemic lupus erythematosus (SLE) show a decreased activation threshold and increased apoptosis. These processes seem to be regulated by glycosylated molecules on the T cell surface. Here, we determined through flow cytometry the expression of mucin-type O-glycans on T helper cells in peripheral blood mononuclear cells (PBMC) from 23 SLE patients and its relation with disease activity. We used lectins specific for the disaccharide Gal-GalNAc, such as Amaranthus leucocarpus lectin (ALL), Artocarpus integrifolia lectin (jacalin) and Arachis hypogaea lectin (peanut agglutinin, PNA), as well as lectins for sialic acid such as Sambucus nigra agglutinin (SNA) and Maakia amurensis agglutinin (MAA). The results showed that ALL, but not jacalin or PNA, identified significant differences in O-glycan expression on T helper cells from active SLE patients (n = 10). Moreover, an inverse correlation was found between the frequency of T helper cells recognized by ALL and SLE Disease Activity Index (SLEDAI) score in SLE patients. In contrast, SNA and MAA lectins did not identify any differences between CD4(+) T cells from SLE patients. There was no difference in the recognition by ALL on activated T helper cells and T regulatory (Treg) cells. Our findings point out that activation of SLE disease diminishes the expression of O-glycans in T helper cells; ALL could be considered as a marker to determine activity of the disease.

Adoptive immunotherapy requires the isolation of CD8+ T cells specific for tumor-associated antigens, their expansion in vitro and their transfusion to the patient to mediate a therapeutic effect. MUC1 is an important adenocarcinoma antigen immunogenic for T cells. The MUC1-derived SAPDTRPA (MUC1-8-mer) peptide is a potent epitope recognized by CD8+ T cells in murine models. Likewise, the T2 cell line has been used as an antigen-presenting cell to activate CD8+ T cells, but so far MUC1 has not been assessed in this context. We evaluated whether the MUC1-8-mer peptide can be presented by T2 cells to expand CD25+CD8+ T cells isolated from HLA-A2+ lung adenocarcinoma patients with stage III or IV tumors. The results showed that MUC1-8-mer peptide-loaded T2 cells activated CD8+ T cells from cancer HLA-A2+ patients when anti-CD2, anti-CD28 antibodies and IL-2 were added. The percentage of CD25+CD8+ T cells was 3-fold higher than those in the non-stimulated cells (P=0.018). HLA-A2+ patient cells showed a significant difference (2.3-fold higher) in activation status than HLA-A2+ healthy control cells (P=0.04). Moreover, 77.6% of MUC1-8-mer peptide-specific CD8+ T cells proliferated following a second stimulation with MUC1-8-mer peptide-loaded T2 cells after 10 days of cell culture. There were significant differences in the percentage of basal CD25+CD8+ T cells in relation to the cancer stage; this difference disappeared after MUC1-8-mer peptide stimulation. In conclusion, expansion of CD25+CD8+ T cells by MUC1-8 peptide-loaded T2 cells plus costimulatory signals via CD2, CD28 and IL-2 can be useful in adoptive immunotherapy.

The Galβ1,3GalNAcα1,O-Ser/Thr specific lectin from Amaranthus leucocarpus (ALL) binds a ∼70 kDa glycoprotein on murine T cell surface. We show that in the absence of antigen presenting cells, murine CD4(+) T cells activated by an anti-CD3 antibody plus ALL enhanced cell proliferation similar to those cells activated via CD3/CD28 at 48 h of culture. Moreover, ALL induced the production of IL-4, IL-10, TNF-alpha, and TGF-beta in CD3-activated cells. Proteomic assay using two-dimensional electrophoresis and far-Western blotting, ALL recognized two prominent proteins associated to the lipid raft microdomains in CD3/CD28-activated CD4(+) T cells. By mass spectrometry, the peptide fragments from ALL-recognized proteins showed sequences with 33% homology to matricin (gi|347839 NCBInr) and 41% identity to an unnamed protein related to moesin (gi|74186081 NCBInr). Confocal microscopy analysis of CD3/CD28-activated CD4(+) T cells confirmed that staining by ALL colocalized with anti-moesin FERM domain antibody along the plasma membrane and in the intercellular contact sites. Our findings suggest that a moesin-like O-glycoprotein is the ALL-recognized molecule in lipid rats, which induces costimulatory signals on CD4(+) T cells.

Background: Vernal keratoconjunctivitis (VKC) is a severe form of allergic conjunctivitis, in which inflammatory infiltrates of the conjunctiva are characterized by CD3+ and CD30+ cells. Until today, the functional involvement of CD30+ T cells in VKC was unclear. Our aim was to evaluate the functional characteristics of CD30+ T cells after allergen stimulation in peripheral blood mononuclear cells obtained from patients with VKC., Methods: Seventeen consecutive patients at the Institute of Ophthalmology with active forms of VKC were included., Results: After allergen stimulation, we observed the frequency of CD30+ T cells increased compared with non-stimulated cells (p<0.0001). The CD30+ T cells responded to the specific allergen-inducing expression of intracellular interleukin-4 (IL-4), IL-5, and interferon-gamma (IFN-γ) compared with the CD30- T cells (p<0.0001). Increased early secretion of soluble CD30 was observed in the supernatant of the cultured cells from patients with keratoconjunctivitis, compared with healthy controls (p=0.03). Blockage with IL-4 significantly diminished CD30 frequency in the allergen-stimulated cells., Conclusions: Our results suggest that after allergenic stimulation, CD4+CD30+ cells are the most important source of IL-4, IL-5, and IFN-γ. IL-4 acts as an activation loop that increases CD30 expression on T cells after specific stimulation. These findings suggest that CD4+CD30+ T cells are effector cells and play a significant role in the immune pathogenic response in patients with vernal keratoconjunctivitis.

Background: Caveolin-1 is a fundamental signalling scaffold protein involved in contraction; however, the role of caveolin-1 in airway responsiveness remains unclear. We evaluated the relationship between caveolin-1 expression in airway smooth muscle (ASM) and antigen-induced airway responsiveness and obstruction in a guinea pig asthma model., Methods: Airway obstruction in sensitised guinea pigs, induced by antigenic (ovalbumin) challenges administered every 10 days, was measured. Antigen-induced responsiveness to histamine and the expression of caveolin-1 and cavin 1, 2 and 3 were evaluated at the third ovalbumin challenge. The control group received saline solution instead of ovalbumin., Results: After the first challenge, antigen exposure induced a transient airway obstruction and airway hyperresponsiveness, high levels of IL-4 and IL-5 in lung and airway globet cells proliferation at the third antigenic challenge. Caveolin-1 mRNA levels in total lung decreased in the experimental group compared with controls. Flow cytometric analysis of ASM from the experimental group showed a high number of cells expressing caveolin-1 compared with controls. This increase was confirmed by western blot. Airway obstruction and hyperresponsiveness correlated with the degree of increased caveolin-1 expression in ASM cells (P < 0.05; r = 0.69 and -0.52, respectively). The expression of cavins 1, 2 and 3 in ASM also increased in the experimental group compared to controls. Immunohistochemical findings reveal that differences in ASM caveolin-1 were not evident between groups. Nevertheless, a marked decrease in caveolin-1 and caspase 3 was observed in the pulmonary vascular smooth muscle of asthma model compared with controls. Histological analysis did not reveal differences in smooth muscles mass or subepithelial fibrosis levels in airways between groups. However, an enlargement of smooth muscle mass was observed in the pulmonary microvessels of experimental animals. This enlargement did not induce changes in pulmonary or systemic arterial pressures., Conclusions: Our data suggest that caveolin-1 expression in ASM has a crucial role in the development of antigen-induced airway obstruction and hyperresponsiveness in a guinea pig asthma model. In addition, the asthma model in guinea pigs appears to induce a contractile smooth muscle phenotype in the airways and a proliferative smooth muscle phenotype in pulmonary vessels.

Bovine tuberculosis (bTB) is a major economic problem in animal husbandry and is a public health risk in nonindustrialized countries. It is generally accepted that protection against TB is generated through cell-mediated immunity. Previous investigations have shown that WC1(+) γδ, CD4(+) and CD8(+) T-cell subpopulations are important in the immune response to bTB. It is known that changes in the immune balance from a dominant T helper 1 (Th1)-type response toward a more prominent Th2 response may be observed during disease progression. In this study, we aimed to investigate immune peripheral blood cells in tuberculin reactor cattle that are seropositive or seronegative for Mycobacterium bovis antigens, using flow cytometry and hematological analysis. The evaluation of the T cell subpopulations revealed a decrease in CD8(+) T cells of the seropositive and seronegative animals compared with the control animals (p=0.0001). Moreover, the seropositive group exhibited a lower percentage of CD8(+) T cells than the seronegative group. The percentage of B cells was significantly increased in the seropositive group compared with the seronegative group and the control group (p=0.0009). No difference was observed in the percentage of WC1(+) γδ and CD4(+) T cells among the groups. Furthermore, following 24h of peripheral blood culture with bovine purified protein derivative (PPD), both apparently infected groups showed an increase in the levels of cellular activation compared with the control group (p<0.0001). The seropositive group displayed a higher level of cellular activation than the seronegative group. In both apparently infected groups, the hematological analysis showed an increase in total leukocyte (p=0.0012), lymphocyte (p=0.0057), monocyte (p=0.0010) and neutrophil (p=0.0320) counts in comparison with the healthy animals. Our results demonstrated differences in immune peripheral blood cells of tuberculin reactor cattle that are seropositive or seronegative for M. bovis antigens, probably due to different stages of bTB among the groups. The percentages of CD8(+) T cells, B cells and the T cell activation levels may represent biomarkers for the progression of the disease. However, general characteristics shared by both apparently infected groups as lymphocytosis and monocytosis may also be indicative of the disease. Further experiments are required to understand the variations between cellular and humoral immunities throughout the course of bTB infection. A detailed knowledge of the peripheral blood cells involved in all stages of the bTB immune response of naturally infected cattle is essential for the optimal exploitation of diagnosis and vaccination models., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Exposition to tobacco smoke has been established as the main risk factor to develop chronic obstructive pulmonary disease (COPD), by inducing inflammation of the airways. Several cell populations participate in this inflammatory process. It has been accepted that a maladaptive modulation of inflammatory responses plays a critical role in the development of the disease. Regulatory T cells (Treg) are a subset of T CD4(+) lymphocytes that modulate the immune response through secretion of cytokines. The role of the Treg cells in chronic obstructive pulmonary disease is not clearly known, that is why it is important to focus in understanding their participation in the pathogenesis of the disease. To elaborate a systematic review of original articles in which we could describe Treg cells (their ontogeny, mechanisms of action) and their role in COPD, we made a systematic literature search in some data bases (MEDLINE, AMED, PubMed and Scielo) looking through the next keywords: "COPD and Regulatory T cells/EPOC y células T reguladoras", «Inflammation and COPD/Inflamación y EPOC», «Regulatory T cells/Células T reguladoras». We included basic science articles, controlled and non-controlled clinical trials, meta-analysis and guides. From this search we conclude that Treg cells are a subpopulation of T CD4(+) lymphocytes and their major functions are the suppression of immune responses and the maintenance of tolerance to self-antigens. A disruption in the regulatory mechanisms of the Treg cells leads to the development and perpetuation of inflammation in COPD., (Copyright © 2012 Instituto Nacional de Cardiología Ignacio Chávez. Published by Masson Doyma México S.A. All rights reserved.)

O-glycosidically-linked glycans have been involved in development, maturation, homing, and immune regulation in T cells. Previous reports indicate that Amaranthus leucocarpus lectin (ALL), specific for glycans containing galactose-N-acetylgalactosamine and N-acetylgalactosamine, recognizes human naïve CD27(+)CD25(+)CD4(+) T cells. Our aim was to evaluate the phenotype of CD4(+) T cells recognized by ALL in peripheral blood mononuclear cells obtained from healthy volunteers. CD4(+) T cells were isolated by negative selection using magnetic beads-labeled monoclonal antibodies; the expression of T regulatory cell phenotypic markers was assessed on ALL-recognized cells. In addition, IL-4, IL-10, IFN-γ, and TGF-β intracellular production in ALL (+) cells was also evaluated. The analyses of phenotypic markers and intracellular cytokines were performed through flow cytometry. ALL-recognized CD4(+) T cells were mainly CD45RA(+), CCR7(+) cells. Although 52 ± 10% CD25(+)Foxp3(+) cells were positive to ALL, only 34 ± 4% of ALL (+) cells corresponded to CD25(+)Foxp3(-) cells. Intracellular cytokines in freshly obtained ALL (+)CD4(+) T cells exhibited 8% of IL-4, 15% of IL-10, 2% of IFN-γ, and 15% of TGF-β, whereas ALL (-)CD4(+) T cells depicted 1% of IL-4, 2% of IL-10, <1% of IFN-γ, and 6% of TGF-β. Our results show that galactose-N-acetylgalactosamine and N-galactosamine-bearing CD4(+) T cells expressed phenotypic markers of NnTreg cells.

Experimental models have shown that lipoproteins from Mycobacterium tuberculosis induce apoptosis via Toll-like receptor 2 (TLR2) in the THP-1 cell line and in monocyte-derived macrophages from healthy volunteers. We found an increased percentage of circulating monocytes in patients with tuberculosis (TB) in comparison to healthy controls. Patients with TB showed a higher TLR2 and TLR4 expression density on monocytes, and a higher proportion of TLR2(+) monocytes, as well as increased serum tumour necrosis factor-α level. In culture, monocytes from TB patients were more susceptible to death than monocytes from healthy controls. Moreover, death-susceptible monocytes were positive to both TLR2 and TLR4 at the start of culture. Freshly obtained monocytes from TB patients exhibited cleaved caspase 9 and denaturalized cytochrome c. For levels of caspase 8, apoptosis-regulating signal kinase 1, and phospho-p38 mitogen-activated protein kinase there was no difference between samples from TB patients and from healthy controls. The culture filtrate antigen extract from M. tuberculosis H37Rv strain induced the death of monocytes from patient with TB after a 4-hr incubation, which was abrogated by neutralizing antibodies for TLR2 but not TLR4. Similarly, Pam3CSK4, a synthetic agonist triacylated ligand to TLR2, also induced the death of monocytes, although it did not increase levels of cleaved caspase 9. Our findings suggest that monocytes from TB patients are more susceptible to death, probably through mitochondrial damage, and that cell death increases in the presence of mycobacterial antigen by a TLR2-dependent pathway., (© 2011 The Authors. Immunology © 2011 Blackwell Publishing Ltd.)

The Galβ1,3GalNAc-specific lectin from Amaranthus leucocarpus (ALL) shows a differential binding pattern on murine thymocytes, peripheral and activated CD4(+) and CD8(+) T cells. Although ALL detects activation-related changes in T cell surface carbohydrate moieties, no study has been performed to examine the effect of ALL on T cell activation. In this study, we analyzed the anti-CD3-dependent activation of murine T cells in the presence of ALL by measuring proliferation, surface activation marker expression, and IL-2 secretion using total cells from the lymph node. The results showed that ALL did not significantly induce T cell activation but did enhance anti-CD3-dependent activation of both CD4(+) and CD8(+) T cells. In addition, ALL protected T cells from spontaneous apoptosis and increased cell survival in serum-free culture conditions. Our findings indicate that ALL alone does not affect T cell activation, but do suggest that ALL has an anti-CD3-dependent co-stimulatory-like effect on T cell activation. Moreover, ALL promotes cell survival in regular and serum-free culture conditions. This study is the first report of a non-mitogenic T cell-binding lectin that can induce a possible costimulatory-like effect and provides a new tool for understanding how glycosylation impacts the T cell response.

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.

Pulmonary tuberculosis is currently considered a serious health problem worldwide. To understand tuberculosis infection we need to know the interaction between the host's immune response and the microorganism causing tuberculosis. Ample research has been carried out to identify new molecules and genes involved in the evasion mechanism of the host's immune response. This knowledge has generated the possibility that future therapeutic schemes will be designed to fight these particular molecules or genes. We reviewed recent experimental evidence on the mechanisms that will successfully fight against mycobacteria.

Histoplasma capsulatum is an intracellular fungal pathogen that causes respiratory and systemic disease by proliferating within phagocytic cells. The binding of H. capsulatum to phagocytes may be mediated by the pathogen's cell wall carbohydrates, glucans, which consist of glucose homo and hetero-polymers and whose glycosydic linkage types differ between the yeast and mycelial phases. The alpha-1,3-glucan is considered relevant for H. capsulatum virulence, whereas the beta-1,3-glucan is antigenic and participates in the modulation of the host immune response. H. capsulatum cell wall components with lectin-like activity seem to interact with the host cell surface, while host membrane lectin-like receptors can recognize a particular fungal carbohydrate ligand. This review emphasizes the relevance of the main H. capsulatum and host carbohydrate-driven interactions that allow for binding and internalization of the fungal cell into phagocytes and its subsequent avoidance of intracellular elimination.

Background and Objective: Specific CD8+ T-cell cytotoxicity has been recognized as being involved in the elimination of drug-susceptible tuberculosis (DS-TB). Given that there is currently no information on the cytotoxic effector functions of CD8+ T cells in multi-drug-resistant tuberculosis (MDR-TB), our objective was to analyse the cytotoxic activity, both basal and stimulated, of CD8+ T cells from MDR-TB patients and compare it with that of DS-TB patients, as well as purified protein derivative (PPD)+ and PPD- subjects., Methods: Cytotoxic activity of CD8+ T cells from MDR-TB patients, DS-TB patients, PPD+ and PPD- subjects was measured by a colorimetric assay, using H37Rv culture filtrate protein as the antigenic stimulus., Results: Twenty-eight subjects were studied (7 MDR-TB patients, 7 DS-TB patients, 7 PPD+ subjects and 7 PPD- subjects). In the presence of the antigenic stimulus, the cytotoxic activity of CD8+ T cells from MDR-TB patients (% lysis) increased from 6.7% to 59.6% (P < 0.001). In DS-TB patients lysis increased from 3.2% to 22.5% (P < 0.001), whereas in PPD+ subjects it increased from 2.7% to 12.0% (P < 0.001) and in PPD- subjects from 1.3% to 3.2% (P < 0.001). Basal cytotoxic activity was significantly higher for MDR-TB patients than PPD+ and PPD- subjects (P = 0.003), but not compared with that for DS-TB patients (P = 0.05). Stimulated cytotoxic activity was highest for MDR-TB patients., Conclusions: CD8+ T cells from MDR-TB patients showed an exaggerated cytotoxic activity after antigenic stimulation. Further studies are required to elucidate the role of this response in the immunopathogenesis of MDR-TB.

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.

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.

Neutrophils participate in host protection and central to this process is the regulation of oxidative mechanisms. We purified by affinity chromatography the receptor for the GlcNAc-specific WGA from CD14+ CD16+ cell lysates (WGAr). The receptor is a 141 kDa glycoprotein constituted by two subunits of 78 and 63 kDa. It is mainly composed of Ser, Asx, and Gly, and, in a minor proportion, His, Cys, and Pro. Its glycan portion contains GlcNAc, Gal, and Man; NeuAc and GalNAc were identified in a minor proportion. The amino acid sequence of the WGA receptor was predicted from tryptic peptides by MALDI-TOF, both subunits showed homology with cytokeratin type II (26 and 29% for the 78 and 63 kDa subunits, respectively); the 78 kDa subunit showed also homology with the human transferrin receptor (24%). Antibodies against WGAr induce higher oxidative burst than WGA, determined by NBT reduction; however, this effect was inhibited (p < 0.05) with GlcNAc suggesting that WGAr participates as mediator in signal transduction in neutrophils.