scholarly journals A quantitative methodology to detect surface antigens on red blood cells using optical cell-detachment technique

2020 ◽  
Author(s):  
CHIH-LANG LIN ◽  
SHYANG-GUANG WANG ◽  
MENG-TSUNG TIEN ◽  
CHUNG-HAN CHIANG ◽  
YI-CHIEH LEE ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Antigen Expression ◽  
Surface Antigens ◽  
Cell Detachment ◽  
Quantitative Methodology ◽  
Optical Cell ◽  
Surface Antigen Expression ◽  
Alternative Methodology ◽  
Coated Surface

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.

scholarly journals A simple and intuitive methodology for estimating red blood cell surface antigen expression variation using optical cell-detachment technique

2021 ◽  
Author(s):  
CHIH-LANG LIN ◽  
SHYANG-GUANG WANG ◽  
MENG-TSUNG TIEN ◽  
CHUNG-HAN CHIANG ◽  
YI-CHIEH LEE ◽  
...  
Keyword(s):  
Optical Tweezers ◽  
Surface Antigen ◽  
Antigenic Variation ◽  
Antigen Expression ◽  
Cell Detachment ◽  
Optical Cell ◽  
Expression Variation ◽  
Surface Antigen Expression ◽  
Alternative Methodology ◽  
Coated Surfaces

Abstract The analysis of surface antigens on cells, especially red blood cells (RBCs), has attracted increasing attention due to the recognition of antigenic variation that can facilitate early diagnoses. This paper presents an alternative methodology to estimate the variation of surface antigen expressions using an optical cell-detachment technique to validate the binding of individual RBCs stuck on corresponding antibody-coated surfaces. The detachment tests were implemented by an optical tweezers with gradually decreasing laser powers associated with serial antibody dilutions. Then, the antigen expression variation was estimated based on the known antibody dilution folds. The B- and B3-types of RBCs were selected for the demonstration subjects. With the semi-quantitative analysis, the proposed methodology was successfully verified for evaluating the variation of the RBC surface antigen expressions. The analysis result shows good consistency with the literature’s findings.

scholarly journals Cross-reactive lysis of trinitrophenyl (TNP)-derivatized H-2 incompatible target cells by cytolytic T lymphocytes generated against syngeneic TNP spleen cells.

1976 ◽  
Vol 144 (6) ◽  
pp. 1609-1620 ◽  
Author(s):  
S J Burakoff ◽  
R N Germain ◽  
B Benacerraf
Keyword(s):  
T Lymphocytes ◽  
Red Blood Cells ◽  
Target Cell ◽  
Blood Cells ◽  
Surface Antigens ◽  
Lymphoid Cells ◽  
Target Cells ◽  
Cytolytic T Lymphocytes ◽  
Spleen Cells ◽  
Normal Spleen

Normal spleen cells, when cultured with irradiated trinitrophenyl (TNP)-derivatized syngeneic spleen cells, develop cytotoxic effectors that lyse most effectiviely a TNP-derivatized target that is H-2 compatible with the effector. However, these effectors also lyse to a lesser extent TNP tumor and TNP spleen targets that are H-2 incompatible. This cross-reactive lysis correlates with the degree of cytolysis seen on the TNP-derivatized syngeneic target; it appears to be medicated by Thy 1.2-bearing cells and is inhibited by antisera to the K and/or D loci of the target cell and not by antisera to non-K or non-D surface antigens. Nonradiolabeled TNP-derivatized lymphoid cells syngeneic to either the stimulator or the target are able to competitively inhibit cross-reactive lysis, while TNP chicken red blood cells are unable to specifically inhibit lysis. These data on cross-reactive lysis of TNP-conjugated targets are most consistent with the altered-self hypothesis.

scholarly journals Murine Red Blood Cells Lack Ligands for B Cell Siglecs, Allowing Strong Activation by Erythrocyte Surface Antigens

2017 ◽  
Vol 200 (3) ◽  
pp. 949-956 ◽  
Author(s):  
Fernando Spiller ◽  
Corwin M. Nycholat ◽  
Chika Kikuchi ◽  
James C. Paulson ◽  
Matthew S. Macauley
Keyword(s):  
Red Blood Cells ◽  
B Cell ◽  
Blood Cells ◽  
Surface Antigens ◽  
Erythrocyte Surface

scholarly journals The Vsa Proteins Modulate Susceptibility of Mycoplasma pulmonis to Complement Killing, Hemadsorption, and Adherence to Polystyrene

2003 ◽  
Vol 71 (10) ◽  
pp. 5733-5738 ◽  
Author(s):  
Warren L. Simmons ◽  
Kevin Dybvig
Keyword(s):  
Red Blood Cells ◽  
Blood Cells ◽  
Tandem Repeats ◽  
Surface Antigens ◽  
Terminal Region ◽  
Respiratory Pathogen ◽  
Mycoplasma Pulmonis ◽  
Variable Regions ◽  
Variable Surface ◽  
Nonspecific Interactions

ABSTRACT The variable surface antigens (Vsa) of the murine respiratory pathogen Mycoplasma pulmonis are associated with the virulence of the microorganism in the lung. In strain UAB CT, the antigens consist of an N-terminal region that is combined with one of seven different C-terminal variable regions comprised of tandem repeats. M. pulmonis producing a VsaA protein with about 40 tandem repeats (R40) does not adhere to red blood cells or polystyrene. Strains that produce VsaH contain a short C-terminal region that lacks tandem repeats and adhere to red blood cells and plastic. We isolated and analyzed M. pulmonis strain CT variants (CT182 and derivatives) that produced a VsaA protein with only three tandem repeats (R3). These variants adhered to plastic and red blood cells similarly to the VsaH-producing strain. When the R3-producing CT182 strain or the VsaH-producing strains were incubated with normal guinea pig serum, they were efficiently killed. Killing was abolished when the serum was heat inactivated. In contrast, the M. pulmonis strains that produced VsaA R40 were highly resistant to complement killing. CT182R3 variants that survived the complement killing reactions all produced the R40 form of VsaA and were resistant to complement killing. VsaA R40 is the first mycoplasmal protein shown to be associated with resistance to complement. As both VsaH and VsaA can mediate adherence to plastic, cytadherence, and susceptibility to complement, we propose that Vsa modulates these phenotypes by nonspecific interactions.

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

1988 ◽  
Vol 30 (2) ◽  
pp. 63-67 ◽  
Author(s):  
Beatriz J. Celeste ◽  
M. Carolina S. Guimarães
Keyword(s):  
Culture Medium ◽  
Growth Curves ◽  
Antigen Expression ◽  
Surface Antigens ◽  
Leishmania Braziliensis ◽  
Mucocutaneous Leishmaniasis ◽  
Before And After ◽  
Human Sera ◽  
Surface Antigen Expression ◽  
Biphasic Medium

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.

scholarly journals Differences in pulmonary innate lymphoid cells are dependent on mouse age, sex and strain

2020 ◽  
Author(s):  
Svenja Loering ◽  
Guy J. M Cameron ◽  
Nirmal P Bhatt ◽  
Gabrielle T Belz ◽  
Paul S Foster ◽  
...  
Keyword(s):  
Cell Surface ◽  
Antigen Expression ◽  
Surface Antigens ◽  
Innate Lymphoid Cells ◽  
Lymphoid Cells ◽  
Cell Surface Antigens ◽  
Adult Mice ◽  
Reporter Mice ◽  
Surface Antigen Expression ◽  
The Impact

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.

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