Variation in noxiousness of different wheat species for celiac patients

Figures & data

Table 1 The main wheat (Triticum and Aegilops) species and their genome compositions.

Diploid species (genome)	Tetraploid species (genome AABB)	Hexaploid species (genome AABBDD)
T. urartu (AA)	T. turgidum ssp. dicoccoides	T. aestivum*
T. monococcum (AA)	T. turgidum ssp. dicoccum*	T. spelta*
Ae. speltoides (BB)	T. turgidum ssp. durum*	T. compactum
Ae. longissima (BB)	T. turgidum ssp. polonicum*	T. sphaerococcum
Ae. searsii (BB)	T. piramidale	T. vavilovii
Ae. tauschii (DD)	T. persicum	T. macha
*Cultivated wheats.

Figure 1.  (A) Fractionation of (A) gliadins by A-PAGE and (B) total proteins by SDS-PAGE in dicoccum landraces. (1–3) Prometeo and (4–12) Ersa 6. Gliadin biotypes c, d and e (A, lanes 1–3), and b (A, lanes 4–6) are shown.

Table 2 Agglutination of K562(S) cells by PT-digested prolamins from different wheat species.

PT-digested prolamins from	MAC (mg/l)*
Triticum monococcum
Cv. Monlis	n.a.
T. turgidum ssp. dicoccum
Cv. Prometeo	n.a.
L5563	n.a..
L5540	n.a.
L5558	n.a.
Ersa 6	73
Ersa 8	73
T. aestivum ssp. spelta	73
T. aestivum ssp. spelta+PP**	n.a.
T. aestivum L.
Cv. S. Pastore	73
n.a., no agglutination; *MAC, Minimal concentration of PT-digest required to agglutinate 100% of cells; **PP, protective peptide QQPQDAVQPF at 1 mg/ml concentration.

Table 3 Neutral red (NR) uptake, active caspases (AC) production, nitric oxide (NO) release and transepithelial electric resistance (TER) in Caco-2/TC7 cells treated with PT-digested prolamins from different wheat species.

	NR uptake (O.D. at 540 nm)
Origin of prolamins	48 h	72 h	96 h	AC production (RFU)	NO release (µM/mg prot.)	TER1 /TER0 ratio
No prolamin (control)	0.29	0.60	0.95	7.5	3.3	1.04
T. monococcum
Cv. Monlis	0.28	0.58	0.92	8.2	2.9	1.03
T. turgidum ssp. dicoccum
Prometeo	0.27	0.56	0.90	9.0	3.8	0.95
L5540	0.28	0.58	0.92	7.7	3.5	0.99
L5558	0.29	0.60	0.94	8.2	4.0	0.97
L5563	0.30	0.58	0.93	7.6	4.1	0.92
Ersa 6	0.17**	0.30**	0.65**	24.0**	5.8**	0.59**
Ersa 8	0.18**	0.32**	0.67**	23.2**	6.5**	0.62**
T. aestivum ssp. spelta	0.19**	0.47**	0.83**	22.4**	6.0**	0.47**
T. aestivum ssp. spelta + PP	0.23	0.57	0.91	12.1*	4.7*	0.76*
T. aestivum L. cv. S. Pastore	0.17**	0.44**	0.68**	23.7**	6.1**	0.42**
Data are the means of three experiments performed in duplicate; PP, protective peptide QQPQDAVQPF at 1 mg/ml concentration; Significant difference with respect to untreated (control) cells at *p≤0.05 or **p≤0.01.

Figure 2.  Microscopic immunofluorescence images of Caco-2/TC7 cells incubated with ZO-1 antibody. (1) Untreated cells and cells exposed to prolamins from (2) spelt wheat, (3) dicoccum wheat Ersa 6 and (4) dicoccum wheat L5540.

Figure 3.  (A) SDS-PAGE patterns of (1–5) native and (6–10) PT-digested endosperm proteins, and (B) their reactions with anti-FPGQQQPFPPQQP (P31–43) antiserum. The dicoccum wheat landraces analysed are (1, 6) Leonessa 1; (2, 7) Leonessa 4; (3, 8) Leonessa 5; (4, 9) Ersa 6 and (5, 10) Ersa 8.

Figure 4.  (A) SDS-PAGE patterns of (1, 2) PT-digested and (3, 4) native endosperm proteins and (B) their reactions with anti-QQPQDAVQPF antiserum. (1, 3) common wheat cv. FG; (2, 4) durum wheat cv Adamello.

Figure 5.  (A) Immunoblotting of anti-PP antiserum against (B) PT-digested prolamins fractionated by SDS-PAGE. The dicoccum wheat landraces are (1) L5563, (2) L5558, (3) L5540, (4) Filosini, (5) Prometeo, (6) Ersa 8, (7) Ersa 6, (8) Leonessa 5 and (9) Leonessa 4. Arrow indicates a band protein showing a weak immunoreaction.

Figure 6.  Immunoblotting of anti-PP antiserum against kernel proteins fractionated by SDS-PAGE. (1) T. monococcum; (2) T. aestivum line FG; (3) T. aestivum ssp. spelta and (4) Phaseolus vulgaris.