Hyaluronan (HA) effects on immune response are suggested to be dependent on HA molecular weight (MW), as low MW HA should activate immune cells in contrast to high MW HA. However, some current studies do not support this conception and emphasize the importance of the form of preparation of HA, particularly with respect to its purity and origin. We compared the activation of mouse immune cells by HA samples (100kDa, 500kDa, and 997kDa) prepared from HA originating from rooster comb, and HA samples (71kDa, 500kDa, and 1000kDa) prepared from pharmacological grade HA originating from Streptococcus equi. Interestingly, in contrast to established theory, only middle and high MW HA originating from rooster comb induced the production of tumor necrosis factor-α by macrophages and in whole blood. Further, all tested preparations of HA failed to induce the expression of inducible nitric oxide synthase, the production of nitric oxide, or the expression of cyclooxygenase 2 in macrophages and splenocytes. Importantly, all HA samples originating from rooster comb were found to be contaminated by endotoxin (up to 1.23EU/ml). Hence, low MW HA did not reveal itself to have significantly higher immunostimulatory activity compared to HA of higher MW.

Contipro a s 401 561 02 Dolni Dobrouc Czech Republic

Institute of Biophysics Academy of Sciences of the Czech Republic 612 65 Brno Czech Republic

Institute of Experimental Biology Faculty of Science Masaryk University Brno Czech Republic

International Clinical Research Center Center of Biomolecular and Cellular Engineering St Anne's University Hospital Brno Czech Republic

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$a Šafránková, Barbora $u Contipro a.s. 401, 561 02 Dolni Dobrouc, Czech Republic; Institute of Biophysics, Academy of Sciences of the Czech Republic, 612 65 Brno, Czech Republic.

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$a Absence of differences among low, middle, and high molecular weight hyaluronan in activating murine immune cells in vitro / $c B. Šafránková, M. Hermannová, K. Nešporová, V. Velebný, L. Kubala,

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$a Hyaluronan (HA) effects on immune response are suggested to be dependent on HA molecular weight (MW), as low MW HA should activate immune cells in contrast to high MW HA. However, some current studies do not support this conception and emphasize the importance of the form of preparation of HA, particularly with respect to its purity and origin. We compared the activation of mouse immune cells by HA samples (100kDa, 500kDa, and 997kDa) prepared from HA originating from rooster comb, and HA samples (71kDa, 500kDa, and 1000kDa) prepared from pharmacological grade HA originating from Streptococcus equi. Interestingly, in contrast to established theory, only middle and high MW HA originating from rooster comb induced the production of tumor necrosis factor-α by macrophages and in whole blood. Further, all tested preparations of HA failed to induce the expression of inducible nitric oxide synthase, the production of nitric oxide, or the expression of cyclooxygenase 2 in macrophages and splenocytes. Importantly, all HA samples originating from rooster comb were found to be contaminated by endotoxin (up to 1.23EU/ml). Hence, low MW HA did not reveal itself to have significantly higher immunostimulatory activity compared to HA of higher MW.

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$a Hermannová, Martina $u Contipro a.s. 401, 561 02 Dolni Dobrouc, Czech Republic.

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$a Nešporová, Kristina $u Contipro a.s. 401, 561 02 Dolni Dobrouc, Czech Republic; Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic.

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$a Velebný, Vladimír $u Contipro a.s. 401, 561 02 Dolni Dobrouc, Czech Republic.

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$a Kubala, Lukáš $u Institute of Biophysics, Academy of Sciences of the Czech Republic, 612 65 Brno, Czech Republic; Institute of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic; International Clinical Research Center - Center of Biomolecular and Cellular Engineering, St. Anne's University Hospital, Brno, Czech Republic. Electronic address: kubalal@ibp.cz.

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