Catalogo Articoli (Spogli Riviste)

OPAC HELP

Titolo:
Purification of natural antikeratin autoantibodies from normal human serumand their effect on human keratinocytes cultured in vitro
Autore:
Li, CX; Wan, YH; Chi, SM; Wang, G; Sun, LC; Zhang, YG; Zhao, XD; Gao, TW; Liu, YF;
Indirizzi:
Xijing Hosp, Dept Dermatol, Xian, Peoples R China Xijing Hosp Xian Peoples R China , Dept Dermatol, Xian, Peoples R China Fourth Mil Med Univ, Dept Physiol, Xian 710033, Peoples R China Fourth MilMed Univ Xian Peoples R China 710033 710033, Peoples R China
Titolo Testata:
BRITISH JOURNAL OF DERMATOLOGY
fascicolo: 5, volume: 145, anno: 2001,
pagine: 737 - 748
SICI:
0007-0963(200111)145:5<737:PONAAF>2.0.ZU;2-F
Fonte:
ISI
Lingua:
ENG
Soggetto:
B-CELLS; FILAMENT AUTOANTIBODIES; RHEUMATOID-ARTHRITIS; ANTIBODIES; BINDING; INVITRO; PSORIASIS; ANTISERA; ANTIGENS; IGM;
Keywords:
affinity chromatography; antikeratin autoantibodies; human keratinocyte; natural autoantibodies;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
39
Recensione:
Indirizzi per estratti:
Indirizzo: Li, CX Xijing Hosp, Dept Dermatol, Xian, Peoples R China Xijing Hosp Xian Peoples R China Dermatol, Xian, Peoples R China
Citazione:
C.X. Li et al., "Purification of natural antikeratin autoantibodies from normal human serumand their effect on human keratinocytes cultured in vitro", BR J DERM, 145(5), 2001, pp. 737-748

Abstract

Background Antikeratin (AK) autoantibodies, circulating antibodies againstepidermal keratins, have been detected in all normal human sera. However, direct evidence on the biological significance of AK autoantibodies is still lacking. Objectives To purify AK autoantibodies from human serum and to make a preliminary study of their biological effects on human keratinocytes. Methods We first extracted keratin polypeptides from human stratum corneumand analysed their purity using sodium dodecyl sulphate-polyacrylamide gelelectrophoresis (SDS-PAGE). Next, a keratin affinity column was prepared with the extracted keratins, and AK autoantibodies were purified from poolednormal human serum. Antibodies obtained were identified with SDS-PAGE, enzyme-linked immunosorbent assay, immunoperoxidase staining, immunoelectron microscopy and Western blotting. The biological effect of AK autoantibodies on cultured human keratinocytes was studied using a DNA synthesis assay, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) colorimetric determination and cell cycle analysis. Results On average, 1.83 +/-0.24 mg of antibodies could be purified from 10 mL of pooled human serum. High-titre IgG (about 1:70) and low-titre IgM (about 1:30) AX autoantibodies were obtained. The DNA synthesis assay and MTT colorimetric determination demonstrated that AK autoantibodies have a significant dose-dependent inhibitory effect on cultured keratinocytes. Correlation coefficients in the two experiments were -0.583 and -0.797, respectively. Cell cycle analysis indicated that a small dose of AK autoantibodies leads to inhibition of proliferation of cultured keratinocytes, whereas a large dose of AK autoantibodies causes a visible hypodiploid peak, suggestingapoptosis of keratinocytes. Conclusions The present research lays a solid foundation for further investigation into the biological significance of natural AX autoantibodies.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 21/01/20 alle ore 06:54:39