From: jschmitz@bidmc.harvard.edu
Date: Sun Mar 02 2003 - 16:40:36 EST
I totally agree with James Crowe on the fact that "Unfixed human specimens fall under the category of Universal Precautions, ...). That is the reason as to why we treat ALL non-fixed specimens exactly in the same way: Use the protection suit. I am sure that many members, who subscribe to this mailing list, have a longer experience in flow cytometry than I. However, in the last 13 year I have seen a lot: i.e. mouse cells turned up to be human specimens from HIV-infected patients, an HIV-negative cell line transfected with some construct and expressing GFP was highly infected with HIV, ... . The kind of cell sorting facility we have built is not cheap. Many institutions may not have the funds to build such a facility. Even more, some principle investigators, who supervise a flow cytometry facility, even don't see the needs for rigorous protection. Interestingly enough many of those PIs never have sorted any cells in their life. Finally, here is another piece of advice: "Be careful out there!" Joern -----Original Message----- From: Crowe, James To: cyto-inbox Mailing List" Sent: 3/1/03 11:09 AM Subject: RE: Sorting live human lymphocytes <<<<If you can trust your principle investigators that they know of and tell you exactly what potential pathogens the cell subsets contain>>> Unfixed human specimens fall under the category of Universal Precautions, which has been in place since 1987 ( http://www.cdc.gov/ncidod/hip/Blood/UNIVERSA.HTM <http://www.cdc.gov/ncidod/hip/Blood/UNIVERSA.HTM> ), which considers that "blood and certain body fluids of all patients are considered potentially infectious for HIV, HBV and other bloodborne pathogens." Therefore, every specimen should be considered potentially infectious unless that particular specimen has been tested for those agents. One way to address this we have found to address this is to use lymphocytes from Red Cross donations (Weitkamp JH, Crowe JE Jr. Blood donor leukocyte reduction filters as a source of human B lymphocytes. Biotechniques. 2001 Sep;31(3):464, 466. ). These specimens are tested such that the red cells are actually reinfused into patients from 450 ml donations, so that they are as safe as is currently available for lymphocyte studies. In terms of the trust level between operators and investigators, maybe we should refocus the issue away from the operator/investigator interface. Likely investigators should be registering protocols with potentially infectious materials with the Institutional Biosafety Committee (IBC). These committees then become the responsible for the review and enforcement of compliance of the procedures suggested by the PI's protocols, rather than the operator. Therefore, the facility could ask each investigator if their flow cytometry procedures and cell sources are reviewed and approved by the IBC. If not, then the institution can't support it. Our IBC reviews the use not only of infectious material, but also retroviral vector use, recombinant DNA, etc (old functions of RAC). There still is some level of trust required, but the discussion of whether the basic procedure is safe or not is an instituional one with federal oversight. jc
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