TABLE 5.

GAG-binding preference and differential binding to glial and endothelial cells

Straina	Vero cell GAG recognizedb	C6 glioma			EA-Hy926
		% Boundc	% Inhibitiond		% Boundc	% Inhibitione
			Heparinase	Chondroitinase ABC		Heparinase	Chondroitinase ABC
N40	Hep-SO4	18.2 ± 1.9	30.1*	47.2*	13.6 ± 1.6	80.7*	18.1
PBi	Hep-SO4	40.4 ± 2.7	37.7*	42.1*	16.3 ± 3.2	58.8*	0.0
PBo	Derm-SO4, Hep-SO4	18.8 ± 2.4	39.3*	49.3*	4.0 ± 0.2	51.9*	4.1
VS461	Derm-SO4 > Hep-SO4	16.0 ± 2.5	42.1*	50.2*	2.3 ± 1.7	NA	NA
HB19 clone 1	None	14.9 ± 1.3	22.7*	46.5*	0.4 ± 0.4	NA	NA

^aSimilar results for N40 and HB19 were presented in Fig. 5; these results are included in this table for ease of comparison with other strains. 

^bThe putative class(es) of GAG that is an important mediator of bacterial attachment to Vero cells was inferred by the effect (or lack of effect) of heparinase or chondroitinase ABC digestion on bacterial binding (see Fig. 1 and 2). (For simplicity of presentation, the putative GAG that mediates bacterial attachment rather than the inhibitory lyase is given). Hep-SO₄, binding was significantly (P < 0.05) inhibited by digestion of the monolayer with heparinase or heparitinase; Derm-SO₄, binding was significantly inhibited by chondroitinase ABC; Derm-SO₄ > Hep-SO₄, both classes of lyase significantly inhibited binding, and chondroitinase ABC had a greater inhibitory effect than heparinase; None, binding was not significantly higher than to empty wells. 

^cBinding of radiolabeled bacteria to confluent monolayers was quantitated as described in Materials and Methods. Each datum point represents the mean ± the SD of four replicate samples. Less than 2% of bacteria bound to identically treated wells without mammalian cells (not shown). 

^dBinding to cells was measured after treatment of monolayers with the indicated lyase. For all strains, binding was inhibited by both chondroitinase ABC and heparinase digestion. Significant (P < 0.05) differences in binding to mock- versus lyase-treated monolayers were determined by t-test analysis and are indicated by asterisks. 

^eBinding to cells was measured after treatment of monolayers with the indicated lyase. For the three strains that bound significantly above the background levels (N40, PBi, and PBo), binding to EA-Hy926 cells was significantly inhibited by heparinase digestion but not by chondroitinase ABC digestion. Significant (P < 0.05) differences in binding to mock- versus lyase-treated monolayers were determined by t-test analysis and are indicated by asterisks.