Guidelines for the use of flow cytometry and cell sorting in immunological studies

Introduction: Guidelines for the use of flow cytometry in immunology

The marriage between immunology and cytometry is one of the most stable and productive in the recent history of science. A rapid search in PubMed shows that, as of July 2017, using “flow cytometry immunology” as a search term yields more than 68 000 articles, the first of which, interestingly, is not about lymphocytes.

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Leonard Herzenberg (1931-2013)

Leonard Herzenberg died on October 27, 2013 aged 81. He had been hospitalized since he suffered a severe stroke on October 8. In cooperation with his wife and scientific partner, Lee Herzenberg, he brought the power of flow cytometry to the field of immunology, and developed fluorescence activated cell sorting, the basis to purify stem cells today. Without the Herzenbergs, tens of thousands of people now alive would not be. I'm glad I had the privilege to meet him and his wife. I will never forget you, Len.


Intercalar Workshop 2012

9 -10 July 2012. Facultad de Medicina, Valencia
Coordinators: Alberto Álvarez Barrientos and José Enrique O’Connor

Through keynote conferences, round tables and practical activities, international experts will discuss new tools and advanced cytometric approaches that can be integrated into the diagnosis, prognosis and clinical research.
In collaboration with the Industry, the Workshop will show the progress in reagents, analysis software and instrumentation, highlighting the novel flow cytometers, small but powerful, and the image-in-flow cytometry systems.

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Identified key mechanisms in the biology of cancer stem cells

Although many ABC transporters have been identified as drug resistance proteins, they may play an additional role regulating the traffic of endogenous or other physiological substrates. Particularly, ABCB1 and ABCG2 transporters are highly conserved in normal stem cells, and may shield the stem compartment from the action of a series of ligands involved in signal transduction. Now, we have identified how the highly conserved Sonic Hedgehog (Hh) pathway can be modulated through the activity of ABCG2 in SP cells with stem-like properties. This upstream regulation of Hh signaling intermediate levels might preserve the stemness of the SP compartment. Cytometry Part A, 2011


Cutting Edge Cytometry at CYTO 2011

Dear Cytometry Colleague,
As we look forward to CYTO 2011 we would very much like to recruit your help on spreading information about this exciting ISAC congress to your co-workers, collaborators etc.
Each attached pdf has a different themed attraction. You may wish to select different ones to send to individuals in your own networks.
The CYTO 2011 flyers highlight:
Our main purpose is to attract interest and encourage abstract submission for poster sessions.
Please use the following link to submit an abstract
We hope that you can help us at this critical point in our congress calendar.
Best wishes,
Paul Smith,
ISAC President
Andreas Radbruch
CYTO 2011 Chair

XMRV Workshop


A new XMRV Workshop has taken place in Vall Hebron Hospital in Barcelona. We have had five lectures, all of them full of the history of all the papers that have been published since Dr. Judy Mikovits’s paper came out in September 2009. But each speaker has also given a summary of what are they working on.

Dr. José Alegre opened the session and was the first to speak. He is one of the most well known specialists in CFS, and as soon as the Science paper was published, he immediately tried to involve a group of Biologists from his Hospital in this research.

Then, Dr. Cecilia Cabrera from IrsiCaixa talked for an hour. She mentioned the great importance of Dr. Ila Singh’s study with animals, and all they have learned about XMRV. Then she said that they have tested 11 patients and done 4 controls, that they have identified sequences of XMRV (not polytrophic virus) in B cells of CFS patients and controls, and that they are developing models of infection in human tissues in vitro, and in this way was able to study the viral pathogenity.

Dr. José Montoya of Stanford University was next. He started explaining that six years ago he met his first CFS patient, and since then he has visited more than 450. He is sure that you can develop CFS after a viral infection, and he believes that 11 % of patients that suffer EBV, Q fever, Ross River virus, etc, can end up with a CFS. About Dr. Judy Mikovit’s paper he remarked that since it was published everybody is turning to CFS with the intention to find out all they can because they have completely changed their opinion.

In a random, double-blind, placebo-controlled study they have found that there was a clinical improvement in patients who were taking valganciclovir for a period of more than six months, compared with placebo. CFS patients met Foukuda’s international criteria and had high antibody data against HHV-6 and EBV. Several immunological markers changed significantly in the treated patients that were not seen in patients taking placebo. This study will be submitted for publication this month, and provides evidence that CFS is a real illness that can be caused by an infectious agent that can be treated with prolonged antiviral intervention. They are also working on the hypothesis that besides the antiviral mechanism there could also be an immonumodulator component in the good results.

And the last to talk was Dr. Jordi Petriz, of the Vall Hebron Hospital. He told us that the aim of his research with Dr. Alegre has to do with the development of functional tests that help to find a better classification of subtypes of CFS. They will use cytometric techniques (a very sophisticated technology) on the whole blood of people with CFS infected with XMRV, and compare the results with appropriate controls, in the hope of finding a pattern and goal differential characteristic of CFS. These techniques will also permit the analysis of the lymphocytes in the study of CFS patient’s immune system. He has also widely described the study that is focused on neutrophil oxidative metabolism and how this may affect their function.

In conclusion, all of them have mentioned the importance of an animal model to study the pathogenesis of XMRV. It’s also very important to have accepted tools to work with, and this means the urgent need of a universal kit. They are all working in the same direction, they are sharing lots of data, samples, etc, and they all agreed with the great complicity between the different groups, which they had not observed so far, for the other diseases they have worked on. Good long term perspectives.