Journal of Immunological Methods
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Adjuvant effect of diphtheria toxin after mucosal administration in both wild type and diphtheria toxin receptor engineered mouse strains.
We used Foxp3(DTR) mice to attempt local depletion of Foxp3+ cells in the lung in a model of tolerance breakdown. Intratracheal administration of DTx resulted in robust depletion of lung Foxp3+ cells. However, DTx administration was accompanied by significant local inflammation, even in control C57B...
Development of a diphtheria toxin-based recombinant porcine IL-2 fusion toxin for depleting porcine CD25(+) cells.
Regulatory T cells (Tregs) have been widely recognized as crucial players in controlling immune responses. Because their major role is to ensure that the immune system is not over reactive, Tregs have been the focus of multiple research studies including those investigating transplantation tolerance...
The two hit hypothesis: an improved method for siRNA-mediated gene silencing in stimulated primary human T cells.
We found that the recommended programs for nucleofection of stimulated primary human T cells with siRNAs inhibited cellular proliferation and were associated with a significant loss of cell viability. Furthermore, viable cells that survived the nucleofection procedure were perturbed in their ability...
Design and characterization of a recombinant colorimetric SAG1-alkaline phosphatase conjugate to detect specific antibody responses against ...
We demonstrate that the recombinant SAG1-AP can detect specific T. gondii antibodies in one-step procedure and can successfully discriminate between T. gondii immune and non-immune patients, in agreement with the standard gold test. In conclusion, the present study shows that the genetic fusion prot...
Peripheral blood natural killer (NK) cell function in healthy adults assessed using the target-induced NK loss (TINKL) assay.
We provide data useful for the clinical application of the target-induced Natural Killer (NK) loss (TINKL) assay. The TINKL assay is a sensitive flow cytometry-based assay for measuring NK cell function. Loss of NK cells from the lymphocyte gate occurs following culture with K562 (the prototypic tar...
Development of a highly-sensitive multi-plex assay using monoclonal antibodies for the simultaneous measurement of kappa and lambda immunogl...
We describe an improved method using specific mouse anti-human FLC monoclonal antibodies (mAbs). Anti-κ and anti-λ FLC mAbs were, separately, covalently coupled to polystyrene Xmap® beads and assayed, simultaneously, in a multi-plex format by Luminex® (mAb assay). The mAbs displayed no cross-reactiv...
Performance evaluation of the MBio Diagnostics point-of-care CD4 counter.
We evaluate the performance of a prototype point-of-care device (POC) to quantify CD4 T cells from MBio Diagnostics, Inc. Whole blood samples, both venous and capillary (finger stick), were collected from known HIV-infected participants at the University of California, San Diego Antiviral Research C...
TRAIL protein localization in human primary T cells by 3D microscopy using 3D interactive surface plot: A new method to visualize plasma mem...
We investigated whether primary CD4(+) T cells expressed TRAIL in their intracellular compartment and whether TRAIL is relocalized on the plasma membrane under HIV activation. We found that TRAIL protein was stocked in intracellular compartment in non activated CD4(+) T cells and that the total leve...
Purification of functional eosinophils from human bone marrow.
We describe the development of a novel, cost effective protocol for the purification of eosinophils from human bone marrow that allows us to obtain bone marrow eosinophils of near 100% purity after an 8-day culture system. Furthermore, we demonstrate that bone marrow eosinophils have characteristics...
Designing of novel antigenic peptide cocktail for the detection of antibodies to HIV-1/2 by ELISA.
HIV (human immunodeficiency virus) infection has now become endemic worldwide and AIDS ranks fourth among the world's top killers of mankind. A rapid and accurate HIV testing assay is a pre-requisite for practical applicability of diagnostic tests. The aim of this present study was to design peptide...