differences in blood collection

Marty Bigos mbigos at gladstone.ucsf.edu
Mon Mar 11 12:04:34 EST 2002


Information from Jeff Harris related to this topic. Please contact
him if you need further information.

Marty



>X-Webmail-User: jharris at itsa.ucsf.edu
>
>I don't know the references off the top of my head, but it is definitely
>described that the sodium heparin green top tubes can activate
>lymphocytes-  that's why many labs use lavender/purple tops (using EDTA
>as anticoagulant) or yellow top ACD (acid citrate/dextrose - citrate is
>also an anticoagulant) tubes for culture/stimulation methods and even
>phenotyping when you are worried about a labile marker - I must say
>however, that for robust assays I don't think the heparin is a huge
>effert, because we use them for the CFCs in the core lab and our
>"non-stimulated" controls for cytokine secretion are quite negative, but
>there is some small background activation by markers like CD69; similarly
>we sort samples based on CD62L, which is also quite labile and sensitive
>to stimulation/temperature/etc. but we try to use the blood freshly and
>move quickly to separate the lymphocytes by Ficoll and keep them cold on
>ice to avoid surface marker changes - whereas the core and clinical labs
>use only EDTa-containing tubes for phenotyping for this reason.
>
>
>
>On Sat, 9 Mar 2002 19:55:35 -0800 Marty Bigos wrote:
>
>>  Have you ever looked at this or anything similar?  Marty
>>
>>  >Date: Thu, 07 Mar 2002 17:03:50 -0400 (EDT)
>>  >From: MIGUEL-ANGEL PERALES <mperales at OPAL.TUFTS.EDU>
>>  >Subject: differences in blood collection
>>  >To: Cytometry Mailing List <cytometry at flowcyt.cyto.purdue.edu>
>>  >X-PMFLAGS: 34078848 0 1 18543.cnm
>>  >
>>  >
>>  >Not exactly a flow question, but somewhat relevant: We recently did a
>>  >study looking at T cell responses to a vaccine by Tetramer. Some
>>  >patients had green tops collected and others underwent leukapheresis.  It
>>  >seemed that T cell responses were higher in green top samples.  We didn't
>>  >compare the 2 methods in the same patient however.  Has anyone had a
>>  >similar experience or tested this?  Also, has anyone compared T cell
>>  >reponses in samples collected in green top vs, CPT tubes.
>>  >
>>  >Thanks,
>>  >
>>  >Miguel Perales
>>
>>
>
>
>
>Jeffrey M. Harris, M.D., Ph.D.
>Asst. Adj. Professor, Pediatrics
>University of California, San Francisco
>Visiting Investigator
>Gladstone Institute of Virology and Immunology

--
Marty Bigos
Director, Flow Core
Gladstone Institute of Virology and Immunology
Building 3 SFGH Rm 509
415-695-3832



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