Evidence map›Paper›PMID 23937268›Full record

ArticleBMC cell biology2013

A simple methodology to assess endolysosomal protease activity involved in antigen processing in human primary cells.

Archana Vaithilingam, Nicole Y Lai, Ellen Duong, Julie Boucau, Yang Xu, Mariko Shimada, Malini Gandhi, Sylvie Le Gall

Open access · goldAbstract readEvaluation Study
In one paragraph

Article in BMC cell biology, 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.

0numbers the graph read from it
0cells of the map it votes in
12citing papers in PubMed
1.6field-weighted citation impact, top 19% of its field
1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

12 citing papers in PubMed, 19 citations in OpenAlex.

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4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

8 authors at 2 institutions in 1 country.

Archana VaithilingamRagon Institute of MGH, MIT and Harvard, 400 Technology Square, Cambridge, MA 02139, USA.
Nicole Y Lai
Ellen Duong
Julie Boucau
Yang Xu
Mariko Shimada
Malini Gandhi
Sylvie Le Gall
Ragon Institute of MGH, MIT and Harvard · USHarvard University · US

Funding

Mechanisms and optimization of epitope presentation by HIV-infectable cell subsetR01AI084753 · NIAID · MASSACHUSETTS GENERAL HOSPITAL · PI LE GALL, SYLVIE · 2009 to 2013
$2.3M
Immune responses against HIV-induced cell-derived neoepitopes and HIV controlR01AI084106 · NIAID · MASSACHUSETTS GENERAL HOSPITAL · PI LE GALL, SYLVIE · 2009 to 2012
$2.1M
NIAID NIH HHS R01 AI084106NIAID NIH HHS R01 AI084753PHS HHS R01 A1084753
6 · The paper itself

Abstract

backgroundEndolysosomes play a key role in maintaining the homeostasis of the cell. They are made of a complex set of proteins that degrade lipids, proteins and sugars. Studies involving endolysosome contribution to cellular functions such as MHC class I and II epitope production have used recombinant endolysosomal proteins, knockout mice that lack one of the enzymes or purified organelles from human tissue. Each of these approaches has some caveats in analyzing endolysosomal enzyme functions.

resultsIn this study, we have developed a simple methodology to assess endolysosomal protease activity. By varying the pH in crude lysate from human peripheral blood mononuclear cells (PBMCs), we documented increased endolysosomal cathepsin activity in acidic conditions. Using this new method, we showed that the degradation of HIV peptides in low pH extracts analyzed by mass spectrometry followed similar kinetics and degradation patterns as those performed with purified endolysosomes.

conclusionBy using crude lysate in the place of purified organelles this method will be a quick and useful tool to assess endolysosomal protease activities in primary cells of limited availability. This quick method will especially be useful to screen peptide susceptibility to degradation in endolysosomal compartments for antigen processing studies, following which detailed analysis using purified organelles may be used to study specific peptides.

Indexed as

Amino Acid SequenceAntigen PresentationAntigensCathepsinsCells, CulturedHumansHydrogen-Ion ConcentrationLeukocytes, MononuclearLysosomesMass SpectrometryMolecular Sequence DataPeptide HydrolasesProteinsAntigensCathepsinslysosomal proteinsPeptide HydrolasesProteins

Identifiers

PMID23937268
PMCPMC3751085
OpenAlexW2128122468

What OpenQuestion holds

Textmetadata
LicenceCC BY
Read underepoch 390

Registered trials

None linked

Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.