ABSTRACTS – ORAL PRESENTATIONS - AMCA, spol. s r.o.

Acknowledgements
This work was supported by Slovak Cancer Research Foundation, VEGA grants 2/0171/13,
2/0130/13, 2/0088/11 and by Slovak Research and Development Agency under the
contract No. APVV-0230-11.
References
Januchowski, R, Wojtowicz, K, and Zabel, M (2013): The role of aldehyde dehydrogenase
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P76. CYTOKINE DYNAMICS AND IMMUNE CELLS PROPORTION DURING TUMOUR
GROWTH AND SUBSEQUENT EXCISION IN LEWIS RAT SARCOMA MODEL
Mishra R. 1,2 , Horák V. 1 , Kovalská J. 1,2 and Rajmon R. 2
1
Institute of Animal Physiology and Genetics, AV CR v.v.i., 277 21 Liběchov, Czech
Republic; mishra@iapg.cas.cz
2
Czech University of Life Sciences Prague, Faculty of Agrobiology, Food and Natural
Resources, 160 00 Prague, Czech Republic
The slow rate of systemic treatment of sarcoma is still a big challenge for researchers.
Among other methods, immunotherapy is one of the effective methods for the treatment
of patient with metastatic sarcoma (R. G.Maki; 2001, 2006). Rat sarcoma model has been
established in our laboratory in which tumour progression and regression occur and are
associated with distinct immunological changes (Holubova et al; 2012).
Moreover, changes in proportion of immune cell subpopulations in peripheral blood can
bring important information about tumour growth, its immunogenicity and efficiency
of therapy. In this study, our aim was to study changes in immune cell populations
and cytokine profile during tumour growth, after tumour excision and after second
inoculation by initially used cell line.
Thirty one Lewis rats (15 males and 16 females) were inoculated subcutaneously with the
R5-28-2 rat sarcoma cells (D6 clone). Four males and four females without cell inoculation
were used as controls. Tumours appeared in all rats 2 weeks after cell inoculation. They
were excised at the 5 th week reaching the average size around 35 mm. At the 11 th week
of experiment, second inoculation of sarcoma cells was done and rats were monitored
for next 4 weeks. Peripheral blood was taken once a week to determine haematological
parameters, proportion of lymphocyte subpopulation and to isolate serum for cytokine
screening and quantification.
Number of leukocytes increased with growing tumour size due to expanded population
of granulocytes. Four kind of immune cells (CD4 + , CD8 + , CD161 + and RT1B + ) were
analysed by flow cytometry. Proportion of CD4 + and CD8 + cells decreased with tumour
growth. Interestingly, these changes were more prominent in females than in males.
Increased serum levels of TNF-α, ICAM-1, MCP-1, IL-6 and L-Selectin were also observed
in tumour-bearing animals. Excision of tumour normalized all the parameters. The
second inoculation of sarcoma cells did not result tumour formation and changes in any
of the studied parameters.
Analytical Cytometry VII 179