Page 8 - Clinical Connections - Spring 2022

ÐÂÔÂÖ±²¥

 

 

 

 

 

Page 8 - Clinical Connections - Spring 2022
P. 8

RVC RESEARCH    STUDY    VETERINARY SERVICES       RVC.AC.UK
         Cancer Research

        USING FLOW CYTOMETRY FOR COMPANION
        ANIMAL CANCER RESEARCH


        Dirk Werling, Professor of Molecular Immunology

              ver the last two years, RVCÐÂÔÂÖ±²¥™s   labelled cells derived from bone marrow,   thousands  of  cells  per  second),  means
              research facilities have been   lymph nodes, or blood) is suspended in a   clinical oncology patients and RVC clinicians
        O revamped to stay at the cutting   fluid  and  injected  into  the  flow  cytometer   have  their  test results within hours.  This
        edge of changes in veterinary medicine.   instrument. A system of tubing, pipes, and   is  a  definite  game-changer,  particularly  in
        One of these changes is the greater impact   valves forces the cells to pass through the   cases where prompt treatment is needed to
        of basic research and research equipment   instrument in a single stream.  ensure better outcomes, and it can be easily
        to closer link research and clinical   When cells reach the interrogation point   used to run aside classical methods, such
        diagnostics, whether this is on a large scale   (or  laser  intercept),  they  pass  through  a   as  cytological  assessment.  Today,  cancer
        for farmed animals, or on a small scale   focused laser beam or a similar light source,   researchers commonly  use a  combination
        for  a  ÐÂÔÂÖ±²¥˜personalised  medicineÐÂÔÂÖ±²¥™  approach  in   causing  fluorescence  emission  and  light   of  fluorescent  monoclonal  antibodies  with
        companion animals.                  scattering. Fluorescence intensity indicates   flow cytometry.
          As part of the current investment of the   the quantity of cellular constituents, while
        Hawkshead Campus, with support from   light scatter determines the size (forward   Flow Cytometry and Cancer Research
        Hertfordshire  Local  Enterprise  Partnership   angle scatter) and structure (right angle   In  human  medicine,  flow  cytometry  has
        and the Animal Care Trust, we were able to   scatter) of individual cells. The data is then   become an essential component of the
        purchase two flow cytometers, one of which   collected and processed accordingly while   assessment,  diagnosis,  and  staging
        is a fluorescence-activated cell sorter, and   the cell sample is disposed of in the waste   of cancer. Here, it is used primarily for
        a new microscope, to enable researchers   container. In a cell sorter, however, the cell   immunophenotyping, which is the use of a
        to perform live-cell imaging, as well as the   sample is passed on to a collection tube for   panel of antibodies to identify cell lineage,
        multicolour ELISpot reader that allows   post-experimental use.         for  example  B  or  T  cell  lymphoma.  The
        precise enumeration of active immune cells.   Flow cytometry allows researchers to   range of immunophenotyping panels used
        The Flow Cytology Core Facility is available   simultaneously measure various parameters   to assess neoplasia in relevant veterinary
        to internal and external users.     within these samples, which can include the   species, such as cats and dogs, although
          For  over  30  years,  flow  cytometry  has   quantification of various cell populations and   increasingly used in veterinary diagnostics
        been  used  to  advance  the  understanding   the specific expression of tagged markers   is still constrained because of associated
        and diagnosis of cancer. As this technique   present on the cellÐÂÔÂÖ±²¥™s surface. However,   expense and a limited availability of species-
        evolves, researchers have been able   it  is  not  only  the  specificity  that  makes   specific antibodies.
        to  utilise  flow  cytometry  to  study  the   flow  cytometry  such  a  powerful  tool  for   In  veterinary  oncology,  flow  cytometry
        development and progression of cancer to   cancer diagnostics, but also its sensitivity.   is  most  often  applied  to  hemolymphoid
        improve patient care and ultimately develop   Researchers can detect the presence of a   neoplasia, osteosarcoma and transitional
        treatment options.                  small number of cancer cells even if other   cell carcinoma. Immunophenotyping can
                                            testing methods provide no evidence of the   also play a pivotal role in the assessment
        How does flow cytometry work?       disease, for example it is possible to detect   of  canine  leukaemia  by  defining  the  cell
        Flow cytometry is a laser technology that   as low as 1 in 10,000 (or even 70,000) cells.   lineage, stage of maturation and prognostic
        enables the user to quantitatively measure   Thus,  flow  cytometry  can  be  particularly   information.  This information is extremely
        the physical and chemical properties within   powerful in monitoring remission and   relevant when it comes to fast decision
        samples such as bone marrow, lymphoid   predicting disease recurrence.  making, resulting hopefully in a faster way
        tissue or blood.                     This  sensitivity,  in  connection  with  the   to treatment and potentially the increase in
          In this method, the sample (fluorescent-  speed of analysis (flow cytometry analyses   quality of life.


         Images to right: Live-dead cell staining
         by flow cytometry in a healthy individual
         (bottom row) and a cancer patient (top row).
         The most important image is the one on the
         far top right, showing more dead cells in a
         sample from a cancer patient compared to
         that of a healthy patient








                                                                                      For small animal referrals, please call:
                                                                                     01707 666399
                                                                                      Email:
                                                                                     qmhreception@rvc.ac.uk
        8   Spring 2022
   3   4   5   6   7   8   9   10   11   12