ABCMdb

PMID: 21811577

Sorio C, Buffelli M, Angiari C, Ettorre M, Johansson J, Vezzalini M, Viviani L, Ricciardi M, Verze G, Assael BM, Melotti P
Defective CFTR expression and function are detectable in blood monocytes: development of a new blood test for cystic fibrosis.
PLoS One. 2011;6(7):e22212. Epub 2011 Jul 21., [PubMed]

Sentences

No. Mutations Sentence Comment

38 ABCC7 p.Trp1282* Cell lines Cell lines utilized were the following: CFBE41o2 , (kind gift from DC Gruenert, California Pacific Medical Center Research Institute, San Francisco, CA, USA) [10], 16HBE14o- (kindly provided by P. Davis, Case Western Reserve University School of Medicine, Cleveland, OH, USA) [11], the bronchial epithelial cell line IB3-1 derived from a CF patient with genotype W1282X/ F508del (kind gift of Pamela Zeitlin, Johns Hopkins Hospital, Baltimore, USA) [12], pancreatic cell lines Suit-2 derived from a liver metastasis of a human pancreatic adenocarcinoma [13] and CFPAC-1 [14]. Login to comment 105 ABCC7 p.Arg1162* ABCC7 p.Arg1162* ABCC7 p.Glu585* ABCC7 p.Arg1158* The specificity of the signal detected with the antibodies is further strengthened by the observation of the loss of the C-terminal epitope in monocytes derived from patients carrying two nonsense mutations (R1158X/E585X; R1162X/R1162X). Login to comment 110 ABCC7 p.Arg1162* ABCC7 p.Arg1162* The membrane localization of CFTR expressed by non-CF monocytes is confirmed by confocal microscopy analysis (Figure 2, bottom right panel), where non-CF monocytes expressing membrane-localized CFTR are shown in comparison with the monocytes isolated from patients carrying a R1162X/R1162X, predicted to produce a truncated form of the protein as confirmed by western blotting data (Figure 1, panel C) that appears to loose membrane localization, as detected by confocal microscopy analysis (Figure 2, bottom left panel). Login to comment 130 ABCC7 p.Trp1282* CFBE41o2 (CFBE) (F508del/F508del) cells express a single band corresponding to a core-glycosylated CFTR (band B, ACL-006 and 24-1 antibodies) while CFTR expression level in IB3-1 (W1282X/F508del) is below the detection limit of the assay for ACL-006 antibody but display a band using the 24-1 antibody. Login to comment 134 ABCC7 p.Gln39* Note that the bands corresponding to a full length CFTR present in wild-type (WT) or obligate heterozygotes (WT/Q39X) are missing in monocytes derived from patients homozygous for nonsense mutations (13-1 antibody). Login to comment 157 ABCC7 p.Arg1162* ABCC7 p.Arg1162* Lower Panel: Confocal microscopy analysis performed using the 13-1 antibody on monocytes derived from subjects carrying a truncated (R1162X/R1162X) and wild type forms of CFTR. Login to comment 158 ABCC7 p.Arg1162* ABCC7 p.Arg1162* Membrane localization is detected in WT monocytes while R1162X/R1162X ones display a disorganized distribution of the signal. Login to comment 202 ABCC7 p.Trp1282* ABCC7 p.Arg553* ABCC7 p.Arg553* ABCC7 p.Asn1303Lys ABCC7 p.Gly542* ABCC7 p.Gly542* ABCC7 p.Gly542* ABCC7 p.Gly542* ABCC7 p.Gly542* ABCC7 p.Gly542* ABCC7 p.Gly542* ABCC7 p.Arg1162* ABCC7 p.Arg1162* ABCC7 p.Arg1162* ABCC7 p.Arg1162* ABCC7 p.Arg1162* ABCC7 p.Arg1162* ABCC7 p.Arg1162* ABCC7 p.Arg1162* ABCC7 p.Arg1162* ABCC7 p.Arg1162* ABCC7 p.Arg1162* ABCC7 p.Arg1162* ABCC7 p.Gly85Glu ABCC7 p.Arg1158* ABCC7 p.Gln39* ABCC7 p.Gln552* ABCC7 p.Gln552* ABCC7 p.Asp1152His ABCC7 p.Trp57* ABCC7 p.Gly1249Arg Case Gender Age at diagnosis (years) CFTR genotype* Age (years) Sweat Cl- mEq/L** FEV1 % mean values 2009 Pa PI NPD results*** CF-index 1 F 0 3132delTG 1497delGG 34 129 75 yes yes nd 222,10 2 F 0 R1162X R1162X 43 144 52 yes yes nd 229,65 3 M 0 R1162X R1162X 10 102 59 no yes 1,02 210,18 4 M 0 R1162X R1162X 25 115 81 no yes 1,07 267,11 5 M 7 G542X 711+5 G.A 24 105 59 yes yes nd 25,84 6 M 1 CFTRdele1 G542X 36 107 22 yes yes nd 2113,92 7 M 0 G542X G542X 16 110 71 yes yes 0,97 280,20 8 F 1 Q552X CFTRdele17a-18 35 99 72 yes yes 2,08 2219,81 9 M 16 R1162X 3849+10 Kb C.T 42 74 43 yes no 1,02 271,47 10 M 0 R1162X R1162X 32 105 45 yes yes 1,43 2114,67 11 M 1 F508del F508del 16 86 71 no yes nd 260,04 12 F 0 F508del F508del 16 88 118 no yes nd 248,20 13 M 0 F508del F508del 33 118 51 yes yes nd 265,49 14 M 7 F508del F508del 37 89 37 yes yes nd 2359,82 15 F 0 F508del F508del 27 118 71 yes yes nd 267,26 16 F 8 1717-1 G.A F508del 38 140 74 yes yes nd 2136,80 17 F 0 R1158X F508del 32 95 60 yes yes 1,77 228,31 18 M 7 G542X F508del 39 110 46 yes yes nd 247,52 19 M 0 Q39X F508del 17 101 79 no yes 1,11 264,20 20 F 1 R1162X F508del 41 188 60 no yes 0,94 296,73 21 M 13 3849+10 Kb C.T F508del 24 76 78 yes no 4,67 26,33 22 M 0 W1282X 621+1G.T 33 119 77 yes yes 1,27 242,74 23 F 4 R553X 2789+5 G.A 31 92 44 yes no 7,4 260,94 24 F 11 F508del R553X 39 116 55 yes yes nd 2113,67 25 M 12 F508del 3849+10 Kb C.T 27 51 71 yes no 1,12 298,84 26 F 0 F508del G542X 19 109 109 yes yes nd 2173,24 27 F 0 F508del R1162X 32 94 86 yes yes 1,34 270,16 28 F 0 F508del W57X (TAG) 27 99 78 yes yes 1,21 269,33 29 M 0 F508del Q552X 24 94 41 yes yes 1,50 272,75 30 M 20 F508del 3849+10 Kb C.T 43 58 60 no no 1,13 2112,56 31 M 0 F508del R1162X 12 99 65 no yes 2,14 280,92 32 M 4 F508del 3849+10 Kb C.T 17 60 100 no no nd 2121,31 33 F 1 F508del 1717-1 G.A 26 105 73 yes yes 2,05 255,66 34 F 11 F508del 3849+10 Kb C.T 40 85 59 yes no nd 2152,23 35 F 4 F508del 1717-1 G.A 44 130 97 yes yes nd 2116,56 36 M 13 F508del 3849+10 Kb C.T 43 70 65 yes no CF 265,10 37 F 19 F508del unknown 29 95 100 no no nd 240,53 38 M 6 F508del unknown 15 92 87 yes no nd 270,17 39 F 0 G542X N1303K 34 108 97 yes yes nd 296,14 40 M 50 G1249R IVS8 T5TG12 50 61 74 no no nd 2199,15 41 F 10 2183 AA.G IVS8 T5TG15/T7TG10 45 79 29 yes no 1,9 286,27 42 F 1 G85E unknown 43 120 107 yes no nd 249,21 43 F 0 3272-26 A.G I507del 21 113 88 no no nd 236,79 44 M 8 F508del D1152H 10 77 107 no no nd 210,85 *Cystic Fibrosis mutation database reference: http://www3.genet.sickkids.on.ca/cftr/app. Login to comment