scholarly journals Growth curves of Leishmania braziliensis braziliensis Promastigotes and surface antigen expression before and after adaptation to Schneider's drosophila medium as assessed by anti-Leishmania human sera

Author(s):  
Beatriz J. Celeste ◽  
M. Carolina S. Guimarães

Leishmania braziliensis braziliensis(MHOM/BR/75/M2903) was grown in Schneider's Drosophila medium. In one set of experiments promastigotes were already adapted to the medium by means of serial passages whereas in the second cells were grown in a biphasic medium and transfered to the liquid. Growth was more abundant for culture medium adapted cells; degenerate cells in small numbers as well as dead ones were present from day 5 for promastigotes adapted to liquid medium and from day 3 for newly adapted cells. Synthesis of surface antigens differed according to length of cell culture as assessed by the titer of five mucocutaneous leishmaniasis sera on subsequent days. Five days of culture for cells already adapted to the culture medium and 3 days for newly adapted ones were judged to be the best for the preparation of immunofluorescence antigens.

2020 ◽  
Author(s):  
CHIH-LANG LIN ◽  
SHYANG-GUANG WANG ◽  
MENG-TSUNG TIEN ◽  
CHUNG-HAN CHIANG ◽  
YI-CHIEH LEE ◽  
...  

Abstract The quantitative analysis of surface antigens on cells, especially red blood cells (RBCs), has attracted increasing attention due to the recognition of antigenic changes that can facilitate early diagnoses. This paper presents an alternative methodology developed using the optical cell-detachment technique to evaluate antibody-antigen interactions and quantitatively analyze the RBC surface antigen expression. RBC subtyping was used to verify the proposed detection principle based on a comparison of the bonding strengths between individual RBCs and antibody coatings. The bonding strengths were measured with serial antibody dilutions with gradually decreasing laser powers, for which a single cell was optically detached from the corresponding antibody-coated surface. With the quantitatively analysis, the proposed alternative methodology was verified as a highly sensitive technique for detecting antigen expression on the RBC surface.


2020 ◽  
Author(s):  
Svenja Loering ◽  
Guy J. M Cameron ◽  
Nirmal P Bhatt ◽  
Gabrielle T Belz ◽  
Paul S Foster ◽  
...  

AbstractInnate lymphoid cells (ILC) are resident in the lung and are involved in both the maintenance of homeostasis and the pathogenesis of respiratory diseases. In this study, murine lung ILC were characterised using flow cytometry and the impact of mouse age, sex and strain were assessed. Lung ILC were found as early as postnatal day 4 and numbers peaked at 2 weeks, and then decreased as the lung matured. During postnatal lung development, ILC expressed differential amounts of ILC2-associated cell surface antigens including ST2, CD90.2 and ICOS. Using Il5venusIl13td-tomato dual reporter mice, neonates were found to have increased constitutive IL-13 expression compared to adult mice. Neonates and adults had similar ratios of IL-5+CD45+ leukocytes, however, these cells were mostly composed of ILC in neonates and T cells in adults. Sex-specific differences in ILC numbers were also observed, with females having greater numbers of lung ILC than males in both neonatal and adult mice. Female lung ILC also expressed higher levels of ICOS and decreased KLRG1. Mouse strain also impacted on lung ILC with BALB/c mice having more ILC in the lung and increased expression of ST2 and ICOS compared with C57BL/6J mice. Collectively, these data show that lung ILC numbers, cell surface antigen expression, IL-5 and IL-13 levels differed between neonatal and adult lung ILC. Additionally, cell surface antigens commonly used for ILC2 quantification, such as ST2, CD90.2, and ICOS, differ depending on age, sex and strain and these are important considerations for consistent universal identification of lung ILC2.


2017 ◽  
Vol 35 (15_suppl) ◽  
pp. 7042-7042
Author(s):  
Madlen Jentzsch ◽  
Marius Bill ◽  
Luba Schuhmann ◽  
Juliane Grimm ◽  
Julia Schulz ◽  
...  

7042 Background: Surface antigen expression evaluation is part of the standard work-up at acute myeloid leukemia (AML) diagnosis. The biological & prognostic implications of surface antigen expression patterns in normal karyotype (NK) AML patients (pts) remain unknown. Methods: The diagnostic antigen expression patterns of mononuclear cells in bone marrow (BM) of 111 NK-AML pts were assessed using a standard flow cytometric panel. At diagnosis common AML gene mutations (mut) & expression levels were analyzed. Pts received stem cell transplantation (SCT, 98% allogeneic, 2% autologous; median age 63 years [y, range 26-74y]) after induction therapy at our institution. Median follow up was 3.3y. With R’s gplot package unsupervised hierarchical clustering of surface antigens was performed & revealed 4 distinct clusters. Results: Pts in cluster 1 (n = 36) had higher expression of immature, in cluster 2 (n = 31) of thrombocytic/T-cell/erythroid, in cluster 3 (n = 24) of monocytic & in cluster 4 (n = 20) of myeloid surface antigens. All 4 clusters associated with distinct clinical & molecular features. At diagnosis, compared to all others, pts in cluster 1 had a higher CD34+/CD38- cell burden ( P< .001), higher blood blasts ( P< .03) & BM blasts ( P< .06) by trend. They had less NPM1 mut ( P< .001) & DNMT3A mut ( P= .02), were more likely to be EVI1 positive ( P= .03) & had higher EZH2 ( P= .02), RUNX1 ( P= .009), BAALC ( P< .001), ERG ( P= .02) & MN1 ( P< .001) expression. Compared to all others, pts in cluster 1 had a higher cumulative incidence of relapse (CIR, P= .002, at 1y 41% vs 15%) & shorter event-free survival (EFS, P= .02, at 1y 50% vs 69%). In multivariate analysis, cluster 1 pts had a significantly higher CIR (Hazard Ratio [HR] 5.4, P= .01) after adjustment for FLT3-ITD & shorter EFS (HR 2.1, P= .02) after adjustment for FLT3-ITD, age & disease status at SCT. Conclusions: Pts in cluster 1 had high expression of immature surface antigens (eg CD34, CD117, CD13), genes involved in stem cell renewal & worse outcome. Our data indicate a relationship between easily accessible surface antigen expression patterns at diagnosis, molecular disease features & aggressiveness of the NK-AML phenotype.


Thorax ◽  
2001 ◽  
Vol 56 (6) ◽  
pp. 450-455
Author(s):  
E Mund ◽  
B Christensson ◽  
K Larsson ◽  
R Grönneberg

BACKGROUNDAge related changes in the immune system have been studied frequently but a possible relation to sex has not, to our knowledge, previously been examined. The effect of age and sex on the composition of lymphocyte subsets in bronchoalveolar lavage (BAL) fluid and peripheral blood was therefore examined.METHODSBronchoscopy with lavage was performed in 32 healthy non-atopic, non-smoking volunteers (16 women aged 26–63 years (mean 44) and 16 men aged 23–63 years (mean 39)). Cytospin preparations for differential counts of BAL fluid cells and surface antigen expression of lymphocytes from BAL fluid and blood were analysed by flow cytometry.RESULTSMost parameters in the BAL fluid changed with age in women. The percentage of CD4+ lymphocytes increased with age from a mean of 48 (SD10)% in women aged ⩽40 years to 69 (11)% in women aged >43 years (p=0.001). The percentage of CD8+ lymphocytes tended to decrease with age and the CD4/CD8 ratio was 5.8 (1.2) in women aged >43 years compared with 2.1 (0.7) in those aged ⩽40 years (p<0.0001). Women aged >43 years differed from men aged >43 years as well as from younger subjects of both sexes with respect to CD4+ cells and CD4/CD8 ratio, and from younger women with respect to CD8+ cells. There was no age related change in the CD4/CD8 ratio in blood. No sex related differences were seen in the blood or BAL fluid of adults below the age of 40 years.CONCLUSIONSThe composition of lymphocytes with different phenotypes in the lower respiratory tract changes with age in women but not in men. This may have implications for some clinical conditions such as chronic dry cough which are observed predominantly in women.


1997 ◽  
Vol 107 (3) ◽  
pp. 593-600 ◽  
Author(s):  
F. BARRAT ◽  
B. M. LESOURD ◽  
A. LOUISE ◽  
H.-J. BOULOUIS ◽  
S. VINCENT-NAULLEAU ◽  
...  

Lung Cancer ◽  
2000 ◽  
Vol 30 (3) ◽  
pp. 169-174 ◽  
Author(s):  
Tokujiro Yano ◽  
Kenji Sugio ◽  
Koji Yamazaki ◽  
Shinichiro Kase ◽  
Masafumi Yamaguchi ◽  
...  

2002 ◽  
Vol 45 (1) ◽  
pp. 155-167 ◽  
Author(s):  
Aleida Vazquez-Macias ◽  
Perla Martinez-Cruz ◽  
Maria Cristina Castaneda-Patlan ◽  
Christine Scheidig ◽  
Jurg Gysin ◽  
...  

2007 ◽  
Vol 75 (9) ◽  
pp. 4272-4281 ◽  
Author(s):  
Qilong Xu ◽  
Sunita V. Seemanaplli ◽  
Kristy McShan ◽  
Fang Ting Liang

ABSTRACT Tight regulation of surface antigenic expression is crucial for the pathogenic strategy of the Lyme disease spirochete, Borrelia burgdorferi. Here, we report the influence of increasing expression of decorin-binding protein A (DbpA), one of the most investigated spirochetal surface adhesins, on the 50% infectious dose (ID50), dissemination, tissue colonization, pathogenicity, and persistence of B. burgdorferi in the murine host. Our in vitro assays showed that increasing DbpA expression dramatically increased the interaction of B. burgdorferi with decorin and sensitivity to growth inhibition/killing by anti-DbpA antibodies; however, this increased interaction did not affect spirochetal growth and replication in the presence of decorin. Increasing DbpA expression significantly reduced ID50 values and severely impaired dissemination in severe combined immunodeficiency (SCID) and immunocompetent mice. During infection of SCID mice, B. burgdorferi with increased DbpA expression was able to effectively colonize heart and skin tissues, but not joint tissues, completely abrogating arthritis virulence. Although increasing DbpA expression did not affect spirochetal persistence in the skin, it diminished the ability of B. burgdorferi to persist in the heart and joint tissues during chronic infection of immunocompetent mice. Taken together, the study highlights the importance of controlling surface antigen expression in the infectivity, dissemination, tissue colonization, pathogenicity, and persistence of B. burgdorferi during mammalian infection.


Sign in / Sign up

Export Citation Format

Share Document